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Sample records for black-crowned night-heron embryos

  1. Swimming black-crowned night-herons (Nycticorax nycticorax) Kleptoparasitize American coots (Fulica americana)

    DEFF Research Database (Denmark)

    Graves, Gary R.

    2015-01-01

    I observed black-crowned night-herons (Nycticorax nycticorax) swimming and kleptoparasitizing American coots (Fulica americana) at an artificial lake in Pinal County, Arizona. This appears to be the first record of interspecific kleptoparasitism by a swimming ardeid.......I observed black-crowned night-herons (Nycticorax nycticorax) swimming and kleptoparasitizing American coots (Fulica americana) at an artificial lake in Pinal County, Arizona. This appears to be the first record of interspecific kleptoparasitism by a swimming ardeid....

  2. Observations of indirect filial cannibalism in response to nest failure of Black-crowned Night-Herons (Nycticorax nycticorax)

    Science.gov (United States)

    Brussee, Brianne E.; Coates, Peter S.; Dwight, Ian; Young, Laura G.

    2017-01-01

    During 2011, four separate instances of indirect filial cannibalism, whereby adults consumed their young that died from unknown causes, were observed using video-monitoring techniques in a nesting colony of Black-crowned Night-Herons (Nycticorax nycticorax) on Alcatraz Island. Though they were not observed actively killing their young, in all four observations adult Black-crowned Night-Herons consumed their young following death (i.e., indirect filial cannibalism). We could not determine cause of chick mortality, but parental neglect was likely a contributing factor in at least two instances. Indirect filial cannibalism is not commonly documented among birds, and understanding how cannibalism contributes to nest failure can help researchers better understand factors that limit nesting populations.

  3. Lead accumulation in feathers of nestling black-crowned night-herons (Nycticorax nycticorax) experimentally treated in the field

    Science.gov (United States)

    Golden, N.H.; Rattner, B.A.; Cohen, J.B.; Hoffman, D.J.; Russek-Cohen, E.; Ottinger, M.A.

    2003-01-01

    Although lead can attain high concentrations in feathers, interpretation of the biological significance of this phenomenon is difficult. As part of an effort to develop and validate non-invasive methods to monitor contaminant exposure in free-ranging birds, lead uptake by feathers of nestling black-crowned night-herons (Nycticorax nycticorax) was evaluated in a controlled exposure study. Four to six day-old heron nestlings (one/nest) at Chincoteague Bay, Virginia, received a single intraperitoneal injection of dosing vehicle (control; n=7) or a dose of lead nitrate in water (0.01, 0.05, or 0.25 mg Pb/g body weight of nestling; n=6 or 7/dose) chosen to yield feather lead concentrations found at low to moderately polluted sites. Nestlings were euthanized at 15 days of age. Lead accumulation in feathers was associated with concentrations in bone, kidney, and liver (r = 0.32 - 0.74, p < 0.02), but exhibited only modest dose-dependence. Blood delta-aminolevulinic acid dehydratase activity was inhibited by lead, although effects on other biochemical endpoints were marginal. Tarsus growth rate was inversely related to feather lead concentration. Culmen growth rate was depressed in nestlings treated with the highest dose of lead, but not correlated with feather lead concentration. These findings provide evidence that feathers of nestling herons are a sensitive indicator of lead exposure and have potential application for the extrapolation of lead concentrations in other tissues and the estimation of environmental lead exposure in birds.

  4. Mercury and drought along the lower Carson River, Nevada: II. Snowy egret and black-crowned night-heron reproduction on Lahontan Reservoir, 1997-2006

    Science.gov (United States)

    Hill, E.F.; Henny, C.J.; Grove, R.A.

    2008-01-01

    Mercury concentrations in the floodplain of the Carson River Basin in northwestern Nevada are some of the highest ever reported in a natural system. Thus, a portion of the basin including Lahontan Reservoir was placed on the U.S. Environmental Protection Agency's Natural Priorities List for research and cleanup. Preliminary studies indicated that reproduction in piscivorous birds may be at risk. Therefore, a 10-year study (1997-2006) was conducted to evaluate reproduction of snowy egrets (Egretta thula) and black-crowned night-herons (Nycticorax nycticorax) nesting on Gull Island in Lahontan Reservoir. Special attention was given to the annual flow of the Carson River, the resultant fluctuation of this irrigation reservoir, and the annual exposure of snowy egrets and night-herons to methylmercury (MeHg). The dynamic character of the river due to flooding and drought (drought effect) influenced snowy egret and night-heron reproduction more so than did MeHg contamination of eggs. During an extended drought (2000-2004) in the middle of the study, snowy egret nests containing eggs with concentrations of MeHg (measured as total mercury [THg] ??? 100% MeHg) ???0.80 ??g THg/g, ww, all failed, but in 1997 and 2006 (wet years with general flooding), substantial numbers of young were produced (but fewer than at nests where eggs contained Science+Business Media, LLC.

  5. Mercury and drought along the lower Carson River, Nevada: I. Snowy egret and black-crowned night-heron annual exposure to mercury, 1997-2006

    Science.gov (United States)

    Henny, C.J.; Hill, E.F.; Grove, R.A.; Kaiser, J.L.

    2007-01-01

    The dynamic nature of the annual volume of water discharged down the Carson River over a 10-year period, which included a century flood and drought, was examined in order to gain a better understanding of mercury movement, biological availability, and exposure to waterbirds nesting at Lahontan Reservoir. Total annual water discharge directly influenced total mercury (THg) in unfiltered water above the reservoir and downstream of a mining area, whereas methyl mercury (MeHg) at the same site was negatively related to annual discharge. Annual water storage at Lahontan Reservoir in the spring and early summer, as expected, was directly related to annual Carson River discharge. In contrast to the findings from above the reservoir, annual MeHg concentrations in water sampled below the reservoir were positively correlated with the total discharge and the amount of water stored in the reservoir on 1 July; that is, the reservoir is an important location for mercury methylation, which agrees with earlier findings. However, unfiltered water MeHg concentrations were about 10-fold higher above than below the reservoir, which indicated that much MeHg that entered as well as that produced in the reservoir settled out in the reservoir. Avian exposure to mercury at Lahontan Reservoir was evaluated in both eggs and blood of young snowy egrets (Egretta thula) and black-crowned night-herons (Nycticorax nycticorax). Annual MeHg concentrations in unfiltered water below the reservoir, during the time period (Julian Days 90-190) when birds were present, correlated significantly with mercury concentrations in night-heron blood (r 2= 0.461, p = 0.027), snowy egret blood (r 2= 0.474, p = 0.024), and night-heron eggs (r 2 = 0.447, p = 0.029), but not snowy egret eggs. A possible reason for lack of an MeHg water correlation with snowy egret eggs is discussed and relates to potential exposure differences associated with the food habits of both species. THg concentrations in water collected

  6. Absorption and biotransformation of polybrominated diphenyl ethers DE-71 and DE-79 in chicken (Gallus gallus), mallard (Anas platyrhynchos), American kestrel (Falco sparverius) and black-crowned night-heron (Nycticorax nycticorax) eggs

    Science.gov (United States)

    McKernan, Moira A.; Rattner, Barnett A.; Hatfield, Jeff S.; Hale, Robert C.; Ottinger, Mary Ann

    2010-01-01

    We recently reported that air cell administration of penta-brominated diphenyl ether (penta-BDE; DE-71) evokes biochemical and immunologic effects in chicken (Gallus gallus) embryos at very low doses, and impairs pipping (i.e., stage immediately prior to hatching) and hatching success at 1.8 ug g-1 egg (actual dose absorbed) in American kestrels (Falco sparverius). I n the present study, absorption of polybrominated diphenyl ether (PBDE) congeners was measured following air cell administration of a penta-BDE mixture (11.1 ug DE-71 g-1 egg) or an octa-brominated diphenyl ether mixture (octa-BDE; DE-79; 15.4 ug DE-79 g-1 egg). Uptake of PBDE congeners was measured at 24 h post-injection, midway through incubation, and at pipping in chicken, mallard (Anas platyrhynchos), and American kestrel egg contents, and at the end of incubation in black-crowned night-heron (Nycticorax nycticorax) egg contents. Absorption of penta-BDE and octa-BDE from the air cell into egg contents occurred throughout incubation; at pipping, up to 29.6% of penta-BDE was absorbed, but only 1.40-6.48% of octa-BDE was absorbed. Higher brominated congeners appeared to be absorbed more slowly than lower brominated congeners, and uptake rate was inversely proportional to the log Kow of predominant BDE congeners. Six congeners or co-eluting pairs of congeners were detected in penta-BDE-treated eggs that were not found in the dosing solution suggesting debromination in the developing embryo, extraembryonic membranes, and possibly even in the air cell membrane. This study demonstrates the importance of determining the fraction of xenobiotic absorbed into the egg following air cell administration for estimation of the lowest-observed-effect level.

  7. Mercury and drought along the Lower Carson River, Nevada: III. effects on blood and organ biochemistry and histopathology of snowy egrets and black-crowned night-herons on Lahontan Reservoir, 2002-2006

    Science.gov (United States)

    Hoffman, David J.; Henny, Charles J.; Hill, Elwood F.; Grove, Robert A.; Kaiser, James L.; Stebbins, Katherine R.

    2009-01-01

    A 10-year study (1997-2006) was conducted to evaluate reproduction and health of aquatic birds in the Carson River Basin of northwestern Nevada (on the U.S. Environmental Protection Agency Natural Priorities List) due to high mercury (Hg) concentrations from past mining activities. This part of the study evaluated physiological associations with blood Hg in young snowy egrets (Egretta thula) and black-crowned night-herons (Nycticorax nycticorax), and organ biochemistry and histopathological effects in snowy egrets on Lahontan Reservoir (LR) from the period 2002-2006. LR snowy egret geometric mean total Hg concentrations (μg/g ww) ranged from 1.5 to 4.8 for blood, 2.4 to 3.1 liver, 1.8 to 2.5 kidneys, 1.7 to 2.4 brain, and 20.5 to 36.4 feathers over these years. For night-herons, mean Hg for blood ranged from 1.6 to 7.4. Significant positive correlations were found between total Hg in blood and five plasma enzyme activities of snowy egrets suggesting hepatic stress. Histopathological findings revealed vacuolar changes in hepatocytes in LR snowy egrets as well as correlation of increased liver inflammation with increasing blood and tissue Hg. Hepatic oxidative effects were manifested by decreased hepatic total thiol concentration and glutathione reductase activity and elevated hepatic thiobarbituric acid-reactive substances (TBARS), a measure of lipid peroxidation. However, other hepatic changes indicated compensatory mechanisms in response to oxidative stress, including decreased oxidized glutathione (GSSG) concentration and decreased ratio of GSSG to reduced glutathione. In young black-crowned night-herons, fewer correlations were apparent. In both species, positive correlations between blood total Hg and plasma uric acid and inorganic phosphorus were suggestive of renal stress, which was supported by histopathological findings. Both oxidative effects and adaptive responses to oxidative stress were apparent in kidneys and brain. Vacuolar change and inflammation in

  8. Colonial nesting Yellow-crowned Night Herons on the San Antonio River Walk

    Science.gov (United States)

    Boal, Clint W.

    2010-01-01

    Yellow-crowned Night Herons (Nyctinassa violacea) typically nest as single pairs or in small colonies of about four pairs with high internest distances. They are also reported as susceptible to disturbance and to avoid habitat with high human use. However, some Yellowcrowned Night Herons habituate to human-dominated landscapes and nest in residential areas. I located a colony of nesting Yellow-crowned Night Herons in San Antonio, Texas on the River Walk, a popular tourist destination with an estimated 2.5 million visitors annually. I located 68 and 71 active nests in 2008 and 2009, respectively. This suggests the breeding population of the colony was 142 adult birds (77 adult herons/linear km of River Walk) in 2009. Herons occurred in a colony with three nesting aggregations situated 241 (±14 SD) m apart. Aggregations averaged 23.7 (±8.7 SD) nests each with one–nine nests per tree; nest trees within each aggregation were usually adjacent. Nests averaged 16.7 m (±4.1 SD) above ground, with 56% of nests over the river, 23% over sidewalks, 17% over dining areas, and 3% over landscaping. Only bald cypress (Taxodium distichum) was used for nest trees, and these were significantly taller and larger in diameter than random bald cypress trees. The herons were habituated to pedestrian activities, often perching only a few meters over sidewalks or dining areas, and foraging along the water’s edge as pedestrians passed within 4–5 m. Nests located over dining areas and sidewalks do impose some management issues. It is apparent the species is capable of habituating to human activities to exploit suitable urban settings for nesting and foraging habitat.

  9. The impact of capture, domestication and trade on Black Crowned ...

    African Journals Online (AJOL)

    The inland delta of the Niger, a vast floodplain, is the only refuge of Black Crowned Cranes in Mali. This study, centred on the regions of Mopti, Tenenkou and Youwarou, aimed to: (1) census the number of cranes in the delta and in captivity in the towns of Mopti and Bamako; (2) examine exploitation at different levels (local ...

  10. Avian species differences in the intestinal absorption of xenobiotics (PCB, dieldrin, Hg2+)

    Science.gov (United States)

    Serafin, J.A.

    1984-01-01

    1. Intestinal absorption of a polychlorinated biphenyl, dieldrin, and mercury (from HgCl2) was measured in adult Northern bobwhites, Eastern screech owls, American kestrels, black-crowned night-herons and mallards in vivo by an in situ luminal perfusion technique.2. Bobwhites, screech owls and kestrels absorbed much more of each xenobiotic than black-crowned night-herons and mallards.3. Mallards absorbed less dieldrin and mercury than black-crowned night-herons.4. Mercury absorption by kestrels was more than twice that in screech owls and eight times that observed in mallards.5. Pronounced differences in xenobiotic absorption rates between bobwhites, screech owls and kestrels on the one hand, and black-crowned night-herons and mallards on the other, raise the possibility that absorptive ability may be associated with the phylogenetic classification of birds.

  11. Effects of contaminants on reproductive success of aquatic birds nesting at Edwards Air Force Base, California

    Science.gov (United States)

    Hothem, R.L.; Crayon, J.J.; Law, M.A.

    2006-01-01

    Contamination by organochlorine pesticides (OCs), polychlorinated biphenyls, metals, and trace elements at Edwards Air Force Base (EAFB), located in the Mojave Desert, could adversely affect nesting aquatic birds, especially at the sewage lagoons that comprise Piute Ponds. Estimates of avian reproduction, in conjunction with analyses of eggs and avian foods for contaminant residues, may indicate the potential for negative effects on avian populations. From 1996 to 1999, we conducted studies at the Piute Ponds area of EAFB to evaluate the impacts of contaminants on nesting birds. Avian reproduction was evaluated in 1999. Eggs were collected for chemical analyses in 1996 and 1999, and African clawed frogs (Xenopus laevis), a likely food source, were collected for chemical analyses in 1998. Avian species occupying the higher trophic levels-black-crowned night-heron (Nycticorax nycticorax), white-faced ibis (Plegadis chihi), and American avocet (Recurvirostra americana)-generally bioaccumulated higher concentrations of contaminants in their eggs. Reproductive success and egg hatchability of night-herons and white-faced ibises in the Piute Ponds were similar to results observed at other western colonies. Deformities were observed in only one embryo in this study, but concentrations of contaminants evaluated in this ibis embryo were considered insufficient to have caused the deformities. Because clawed frogs, a primary prey item for night-herons at Piute Ponds, had no detectable levels of any OCs, it is likely that OCs found in night-heron eggs were acquired from the wintering grounds rather than from EAFB. The presence of isomers of dichlorodiphenyltrichloroethane (DDT) in ibis eggs indicated recent exposure, but invertebrates used for food by ibises were not sampled at Piute Ponds, and conclusions about the source of OCs in ibis eggs could not be made. Concentrations of contaminants in random and failed eggs of individual species were not different, and we concluded

  12. Embryo splitting

    OpenAIRE

    Karl Illmensee; Mike Levanduski

    2010-01-01

    Mammalian embryo splitting has successfully been established in farm animals. Embryo splitting is safely and efficiently used for assisted reproduction in several livestock species. In the mouse, efficient embryo splitting as well as single blastomere cloning have been developed in this animal system. In nonhuman primates embryo splitting has resulted in several pregnancies. Human embryo splitting has been reported recently. Microsurgical embryo splitting under Institutional Review Board appr...

  13. Embryo splitting

    Directory of Open Access Journals (Sweden)

    Karl Illmensee

    2010-04-01

    Full Text Available Mammalian embryo splitting has successfully been established in farm animals. Embryo splitting is safely and efficiently used for assisted reproduction in several livestock species. In the mouse, efficient embryo splitting as well as single blastomere cloning have been developed in this animal system. In nonhuman primates embryo splitting has resulted in several pregnancies. Human embryo splitting has been reported recently. Microsurgical embryo splitting under Institutional Review Board approval has been carried out to determine its efficiency for blastocyst development. Embryo splitting at the 6–8 cell stage provided a much higher developmental efficiency compared to splitting at the 2–5 cell stage. Embryo splitting may be advantageous for providing additional embryos to be cryopreserved and for patients with low response to hormonal stimulation in assisted reproduction programs. Social and ethical issues concerning embryo splitting are included regarding ethics committee guidelines. Prognostic perspectives are presented for human embryo splitting in reproductive medicine.

  14. Selenium and metal concentrations in waterbird eggs and chicks at Agassiz National Wildlife Refuge, Minnesota

    Science.gov (United States)

    Custer, T.W.; Custer, Christine M.; Eichhorst, B.A.; Warburton, D.

    2007-01-01

    Exceptionally high cadmium (Cd) and chromium (Cr) concentrations were reported in eggs, feathers, or livers of selected waterbird species nesting at Agassiz National Wildlife Refuge (Agassiz) in 1994. Ten- to 15-day-old Franklin's gull (Larus pipixcan), black-crowned night-heron (Nycticorax nycticorax), and eared grebe (Podiceps nigricollis) chicks were collected in 1998, 1999, and 2001 at Agassiz and analyzed for selenium (Se) and metals including Cd and Cr. Freshly laid eggs were collected in 2001 from Franklin's gull, black-crowned night-heron, eared grebe, and pied-billed grebe (Podilymbus podiceps) nests at Agassiz. Based on a multivariate analysis, the pattern of Se and metal concentrations differed among species for eggs, chick feathers, and chick livers. Low Cd and Cr concentrations were measured in eggs, chick livers, and chick feathers of all four species. Mercury concentrations in black-crowned night-heron and eared grebe eggs collected from Agassiz in 2001 were lower than concentrations reported in 1994. Se and metal concentrations, including Cd and Cr, in waterbird eggs and chicks collected at Agassiz in 1998, 1999, and 2001 were not at toxic levels. ?? 2007 Springer Science+Business Media, LLC.

  15. Embryos, microscopes, and society.

    Science.gov (United States)

    Maienschein, Jane

    2016-06-01

    Embryos have different meanings for different people and in different contexts. Seen under the microscope, the biological embryo starts out as one cell and then becomes a bunch of cells. Gradually these divide and differentiate to make up the embryo, which in humans becomes a fetus at eight weeks, and then eventually a baby. At least, that happens in those cases that carry through normally and successfully. Yet a popular public perception imagines the embryo as already a little person in the very earliest stages of development, as if it were predictably to become an adult. In actuality, cells can combine, pull apart, and recombine in a variety of ways and still produce embryos, whereas most embryos never develop into adults at all. Biological embryos and popular imaginations of embryos diverge. This paper looks at some of the historical reasons for and social implications of that divergence. Copyright © 2016 The Author. Published by Elsevier Ltd.. All rights reserved.

  16. Major Rehabilitation Effort, Mississippi River, Locks and Dams 2-22: Illinois Waterway from LA Grange to Lockport Locks and Dams: Iowa, Illinois, Missouri, Minnesota and Wisconsin.

    Science.gov (United States)

    1989-03-01

    Demographics Services Center. Madison, Wt. Unpublished Source. - IS-36 (I0 00 ) 1*1 L (Baox) mammozqY&0 KTMES8TnfeTastigLcs edo EIS-7 NaIS~ton t L/D10 (986... redhead . Numerous bird species also utilize this reach. A rookery consisting of great blue, little blue, and black-crowned night herons is located at...lesser scaup, ringneck, goldeneye, canvasback, and redhead . Breeding ducks include wood duck, mallard, and blue-winged teal. 3.146 Marsh and shorebirds

  17. Embryo-maternal communication

    DEFF Research Database (Denmark)

    Østrup, Esben; Hyttel, Poul; Østrup, Olga

    2011-01-01

    Communication during early pregnancy is essential for successful reproduction. In this review we address the beginning of the communication between mother and developing embryo; including morphological and transcriptional changes in the endometrium as well as epigenetic regulation mechanisms dire...... directing the placentation. An increasing knowledge of the embryo-maternal communication might not only help to improve the fertility of our farm animals but also our understanding of human health and reproduction.......Communication during early pregnancy is essential for successful reproduction. In this review we address the beginning of the communication between mother and developing embryo; including morphological and transcriptional changes in the endometrium as well as epigenetic regulation mechanisms...

  18. Manipulating early pig embryos.

    Science.gov (United States)

    Niemann, H; Reichelt, B

    1993-01-01

    On the basis of established surgical procedures for embryo recovery and transfer, the early pig embryo can be subjected to various manipulations aimed at a long-term preservation of genetic material, the generation of identical multiplets, the early determination of sex or the alteration of the genetic make-up. Most of these procedures are still at an experimental stage and despite recent considerable progress are far from practical application. Normal piglets have been obtained after cryopreservation of pig blastocysts hatched in vitro, whereas all attempts to freeze embryos with intact zona pellucida have been unsuccessful. Pig embryos at the morula and blastocyst stage can be bisected microsurgically and the resulting demi-embryos possess a high developmental potential in vitro, whereas their development in vivo is impaired. Pregnancy rates are similar (80%) but litter size is reduced compared with intact embryos and twinning rate is approximately 2%. Pig blastomeres isolated from embryos up to the 16-cell stage can be grown in culture and result in normal blastocysts. Normal piglets have been born upon transfer of blastocysts derived from isolated eight-cell blastomeres, clearly underlining the totipotency of this developmental stage. Upon nuclear transfer the developmental capacity of reconstituted pig embryos is low and culture. Sex determination can be achieved either by separation of X and Y chromosome bearing spermatozoa by flow cytometry or by analysing the expression of the HY antigen in pig embryos from the eight-cell to morula stage. Microinjection of foreign DNA has been successfully used to alter growth and development of transgenic pigs, and to produce foreign proteins in the mammary gland or in the bloodstream, indicating that pigs can be used as donors for valuable human pharmaceutical proteins. Another promising area of gene transfer is the increase of disease resistance in transgenic lines of pigs. Approximately 30% of pig spermatozoa bind

  19. Ethical Challenges of Embryo Donation in Embryo Donors and Recipients

    OpenAIRE

    Taebi, Mahboubeh; Bahrami, Reyhane; Bagheri-Lankarani, Narges; Shahriari, Mohsen

    2018-01-01

    Background: Embryo donation, as one of the novel assisted reproductive technologies (ART), has remained a controversial issue. This is due to this methods' need for individuals from outside the family circle. Their presence can cause many ethical issues and complicate the designing and planning of the embryo donation process. The present study was conducted with the aim to assess the ethical challenges of embryo donation from the view point of embryo donors and recipients. Material and Method...

  20. Rabbit whole embryo culture.

    Science.gov (United States)

    Marshall, Valerie A; Carney, Edward W

    2012-01-01

    Although the rabbit is used extensively in developmental toxicity testing, relatively little is known about the fundamental developmental biology of this species let alone mechanisms underlying developmental toxicity. This paucity of information about the rabbit is partly due to the historic lack of whole embryo culture (WEC) methods for the rabbit, which have only been made available fairly recently. In rabbit WEC, early somite stage embryos (gestation day 9) enclosed within an intact amnion and attached to the visceral yolk sac are dissected from maternal tissues and placed in culture for up to 48 h at approximately 37°C and are continuously exposed to an humidified gas atmosphere mixture in a rotating culture system. During this 48 h culture period, major phases of organogenesis can be studied including cardiac looping and segmentation, neural tube closure, and development of anlagen of the otic system, eyes and craniofacial structures, somites and early phases of limb development (up to bud stage), as well as expansion and closure of the visceral yolk sac around the embryo. Following completion of the culture period, embryos are evaluated based on several growth and development parameters and also are assessed for morphological abnormalities. The ability to sustain embryo development independent of the maternal system allows for exposure at precise development stages providing the opportunity study the direct action of a teratogen or one of its metabolites on the developing embryo. Rabbit WEC is perhaps most useful when used in conjunction with rodent WEC methods to investigate species-specific mechanisms of developmental toxicity.

  1. The First Human Cloned Embryo.

    Science.gov (United States)

    Cibelli, Jose B.; Lanza, Robert P.; West, Michael D.; Ezzell, Carol

    2002-01-01

    Describes a process known as parthenogenesis which produces cloned, early-stage embryos and human embryos generated only from eggs. Speculates that this technology puts therapeutic cloning within reach. (DDR)

  2. Ovarian stimulation and embryo quality

    NARCIS (Netherlands)

    Baart, Esther; Macklon, Nick S.; Fauser, Bart J. C. M.

    To Study the effects of different ovarian stimulation approaches on oocyte and embryo quality, it is imperative to assess embryo quality with a reliable and objective method. Embryos rated as high quality by standardized morphological assessment are associated with higher implantation and pregnancy

  3. impact on embryo quality

    Directory of Open Access Journals (Sweden)

    Marijan Tandara

    2013-05-01

    Conclusions: In men with poorer semen quality, evaluated by standard semen parameters, a higher proportion of sperm with damaged DNA can also be expected. Higher sperm DNA damage, established by Halosperm test, also had an impact on embryo quality in this group of patients.

  4. Embryo Aggregation in Pig Improves Cloning Efficiency and Embryo Quality.

    Science.gov (United States)

    Buemo, Carla Paola; Gambini, Andrés; Moro, Lucia Natalia; Hiriart, María Inés; Fernández-Martín, Rafael; Collas, Philippe; Salamone, Daniel Felipe

    2016-01-01

    In this study, we analyzed the effects of the cloned embryo aggregation on in vitro embryo development and embryo quality by measuring blastocyst diameter and cell number, DNA fragmentation levels and the expression of genes associated with pluripotency, apoptosis, trophoblast and DNA methylation in the porcine. Zona-free reconstructed cloned embryos were cultured in the well of the well system, placing one (1x non aggregated group) or three (3x group) embryos per microwell. Our results showed that aggregation of three embryos increased blastocyst formation rate and blastocyst diameter of cloned pig embryos. DNA fragmentation levels in 3x aggregated cloned blastocysts were significantly decreased compared to 1x blastocysts. Levels of Oct4, Klf4, Igf2, Bax and Dnmt 1 transcripts were significantly higher in aggregated embryos, whereas Nanog levels were not affected. Transcripts of Cdx2 and Bcl-xl were essentially non-detectable. Our study suggests that embryo aggregation in the porcine may be beneficial for cloned embryo development and embryo quality, through a reduction in apoptotic levels and an improvement in cell reprogramming.

  5. Ethical Challenges of Embryo Donation in Embryo Donors and Recipients.

    Science.gov (United States)

    Taebi, Mahboubeh; Bahrami, Reyhane; Bagheri-Lankarani, Narges; Shahriari, Mohsen

    2018-01-01

    Embryo donation, as one of the novel assisted reproductive technologies (ART), has remained a controversial issue. This is due to this methods' need for individuals from outside the family circle. Their presence can cause many ethical issues and complicate the designing and planning of the embryo donation process. The present study was conducted with the aim to assess the ethical challenges of embryo donation from the view point of embryo donors and recipients. This descriptive, cross-sectional study was conducted on 192 couples (96 embryo donators and 96 embryo recipients) referring to Isfahan Fertility and Infertility Center and Royan Institute, Iran. The subjects were selected through convenience sampling. The data collection tool was the researcher-made Ethical Challenges Questionnaire. Data were analyzed in SPSS software. Embryo donors and recipients expresses the most important ethical challenges of embryo donation in the principle of justice (70.20%) and respect for autonomy (42.57%), respectively. The four ethical principles are important in the view of embryo donors and recipients; however, they highlighted the importance of the principle of respect for autonomy considering the existing barriers in the services of infertility centers. Legislators and relevant authorities must take measures toward the development of guidelines for this treatment method in the framework of ethics principles and incorporate all four principles independently.

  6. Ethical challenges of embryo donation in embryo donors and recipients

    Directory of Open Access Journals (Sweden)

    Mahboubeh Taebi

    2018-01-01

    Full Text Available Background: Embryo donation, as one of the novel assisted reproductive technologies (ART, has remained a controversial issue. This is due to this methods' need for individuals from outside the family circle. Their presence can cause many ethical issues and complicate the designing and planning of the embryo donation process. The present study was conducted with the aim to assess the ethical challenges of embryo donation from the view point of embryo donors and recipients. Material and Methods: This descriptive, cross-sectional study was conducted on 192 couples (96 embryo donators and 96 embryo recipients referring to Isfahan Fertility and Infertility Center and Royan Institute, Iran. The subjects were selected through convenience sampling. The data collection tool was the researcher-made Ethical Challenges Questionnaire. Data were analyzed in SPSS software. Results: Embryo donors and recipients expresses the most important ethical challenges of embryo donation in the principle of justice (70.20% and respect for autonomy (42.57%, respectively. Conclusions: The four ethical principles are important in the view of embryo donors and recipients; however, they highlighted the importance of the principle of respect for autonomy considering the existing barriers in the services of infertility centers. Legislators and relevant authorities must take measures toward the development of guidelines for this treatment method in the framework of ethics principles and incorporate all four principles independently.

  7. Gender determination of avian embryo

    Energy Technology Data Exchange (ETDEWEB)

    Daum, Keith A. (Idaho Falls, ID); Atkinson, David A. (Idaho Falls, ID)

    2002-01-01

    Disclosed is a method for gender determination of avian embryos. During the embryo incubation process, the outer hard shells of eggs are drilled and samples of allantoic fluid are removed. The allantoic fluids are directly introduced into an ion mobility spectrometer (IMS) for analysis. The resulting spectra contain the relevant marker peaks in the positive or negative mode which correlate with unique mobilities which are sex-specific. This way, the gender of the embryo can be determined.

  8. Who abandons embryos after IVF?

    LENUS (Irish Health Repository)

    Walsh, A P H

    2010-04-01

    This investigation describes features of in vitro fertilisation (IVF) patients who never returned to claim their embryos following cryopreservation. Frozen embryo data were reviewed to establish communication patterns between patient and clinic; embryos were considered abandoned when 1) an IVF patient with frozen embryo\\/s stored at our facility failed to make contact with our clinic for > 2 yrs and 2) the patient could not be located after a multi-modal outreach effort was undertaken. For these patients, telephone numbers had been disconnected and no forwarding address was available. Patient, spouse and emergency family contact\\/s all escaped detection efforts despite an exhaustive public database search including death records and Internet directory portals. From 3244 IVF cycles completed from 2000 to 2008, > or = 1 embryo was frozen in 1159 cases (35.7%). Those without correspondence for > 2 yrs accounted for 292 (25.2%) patients with frozen embryos; 281 were contacted by methods including registered (signature involving abandoned embryos did not differ substantially from other patients. The goal of having a baby was achieved by 10\\/11 patients either by spontaneous conception, adoption or IVF. One patient moved away with conception status unconfirmed. The overall rate of embryo abandonment was 11\\/1159 (< 1%) in this IVF population. Pre-IVF counselling minimises, but does not totally eliminate, the problem of abandoned embryos. As the number of abandoned embryos from IVF accumulates, their fate urgently requires clarification. We propose that clinicians develop a policy consistent with relevant Irish Constitutional provisions to address this medical dilemma.

  9. Embryos, genes, and birth defects

    National Research Council Canada - National Science Library

    Ferretti, Patrizia

    2006-01-01

    ... Structural anomalies The genesis of chromosome abnormalities Embryo survival The cause of high levels of chromosome abnormality in human embryos Relative parental risks - age, translocations, inversions, gonadal and germinal mosaics 33 33 34 35 36 44 44 45 4 Identification and Analysis of Genes Involved in Congenital Malformation Syndromes Peter J. Scambler Ge...

  10. In Vitro Embryo as a Person

    Directory of Open Access Journals (Sweden)

    محمد حسن صادقی مقدم

    2016-03-01

    Full Text Available Based on moral principles and the natural laws, an in vitro embryo is considered as a natural embryo. The difference in the course of natural growing of an in vitro embryo and natural embryo, does not result in a difference of the governing rules, since both are embryos. It is argued that an in vitro embryo has the same rights as the natural embryo. Considering this claim, despite the position of some scholars who believe that such an embryo is subject to ownership, this article aims to critique this idea and argue that an in vitro embryo has the same personhood as a natural embryo from the moment of conception. Therefore, the embryo cannot be transferred as a gift or otherwise traded in the form of a stipulation in a contract.

  11. Oxygen diffusion in fish embryos

    NARCIS (Netherlands)

    Kranenbarg, S.

    2002-01-01

    All vertebrate embryos pass through a developmental period of remarkably low morphological variability. This period has been called phylotypic period. During the phylotypic period, organogenesis takes place, including blood vessel development. Before the phylotypic

  12. Proteomics of early zebrafish embryos

    Directory of Open Access Journals (Sweden)

    Heisenberg Carl-Philipp

    2006-01-01

    Full Text Available Abstract Background Zebrafish (D. rerio has become a powerful and widely used model system for the analysis of vertebrate embryogenesis and organ development. While genetic methods are readily available in zebrafish, protocols for two dimensional (2D gel electrophoresis and proteomics have yet to be developed. Results As a prerequisite to carry out proteomic experiments with early zebrafish embryos, we developed a method to efficiently remove the yolk from large batches of embryos. This method enabled high resolution 2D gel electrophoresis and improved Western blotting considerably. Here, we provide detailed protocols for proteomics in zebrafish from sample preparation to mass spectrometry (MS, including a comparison of databases for MS identification of zebrafish proteins. Conclusion The provided protocols for proteomic analysis of early embryos enable research to be taken in novel directions in embryogenesis.

  13. Chromatin remodeling in mammalian embryos.

    Science.gov (United States)

    Cabot, Birgit; Cabot, Ryan A

    2018-03-01

    The mammalian embryo undergoes a dramatic amount of epigenetic remodeling during the first week of development. In this review, we discuss several epigenetic changes that happen over the course of cleavage development, focusing on covalent marks (e.g., histone methylation and acetylation) and non-covalent remodeling (chromatin remodeling via remodeling complexes; e.g., SWI/SNF-mediated chromatin remodeling). Comparisons are also drawn between remodeling events that occur in embryos from a variety of mammalian species. © 2018 Society for Reproduction and Fertility.

  14. In vitro tagging of embryos with nanoparticles

    OpenAIRE

    Fynewever, Tricia L.; Agcaoili, Evelyn S.; Jacobson, John D.; Patton, William C.; Chan, Philip J.

    2006-01-01

    Purpose: To develop an in vitro method for tagging embryos and to compare the development of the embryos after nanoparticles injection versus externally-applied nanoparticles derived from either polystyrene or polyacrylonitrile.

  15. Cryopreservation of biopsied cleavage stage human embryos.

    Science.gov (United States)

    Stachecki, James J; Cohen, Jacques; Munné, Santiago

    2005-12-01

    The aim was to develop a method to optimize cryopreservation of biopsied multi-celled human embryos. Human day 3 embryos that were donated to research, along with those found to be chromosomally abnormal after blastomere biopsy and fluorescence in-situ hyridization (FISH), were cryopreserved using a slow-freezing protocol in either standard embryo cryopreservation solution [embryo transfer freezing medium (ETFM), a conventional sodium-based medium] or CJ3 (a choline-based, sodium-free medium). After thawing, the number of intact cells was recorded and the previously biopsied embryos were re-analysed using FISH. Biopsied embryos had a lower proportion of intact blastomeres after cryopreservation as compared with intact embryos. However, a significantly (P < 0.05) higher proportion of blastomeres from intact and biopsied embryos cryopreserved in CJ3 (84.1 and 80.1% respectively) survived after thaw than those in ETFM (73.6 and 50.5% respectively). The proportion of aneuploid and mosaic embryos was not statistically different between the two groups. In addition, the frequency of lost cells by aneuploid and mosaic embryos was similar. This study describes a new method that improves the survival of cryopreserved biopsied embryos, and shows that it may also be beneficial for the storage of intact human multi-celled embryos.

  16. Improving embryo quality in assisted reproduction

    NARCIS (Netherlands)

    Mantikou, E.

    2013-01-01

    The goal of this thesis was to improve embryo quality in assisted reproductive technologies by gaining more insight into human preimplantation embryo development and by improving in vitro culture conditions. To do so, we investigated an intriguing feature of the human preimplantation embryo, i.e.

  17. Mechanistic dissection of plant embryo initiation

    NARCIS (Netherlands)

    Radoeva, T.M.

    2016-01-01

    Land plants can reproduce sexually by developing an embryo from a fertilized egg cell, the zygote. After fertilization, the zygote undergoes several rounds of controlled cell divisions to generate a mature embryo. However, embryo formation can also be induced in a variety of other cell types in

  18. The development of ovary in quail's embryo

    African Journals Online (AJOL)

    user

    2011-01-24

    Jan 24, 2011 ... The experiment was conducted to study the development of ovary in quails' embryos which were incubated for 4 to 17 days and incubated out for 1 day. The quails' embryos or gonads were cut out and. HE staining was carried out. The results showed that when embryo was hatched for 4 days, lots of.

  19. Mechanistic dissection of plant embryo initiation

    NARCIS (Netherlands)

    Radoeva, T.M.

    2016-01-01

    Land plants can reproduce sexually by developing an embryo from a fertilized egg cell, the zygote. After fertilization, the zygote undergoes several rounds of controlled cell divisions to generate a mature embryo. However, embryo formation can also be induced in a variety of other cell types in many

  20. Embryo adoption: Some further considerations

    Science.gov (United States)

    Patterson, Colin

    2015-01-01

    Recent discussions of embryo adoption have sought to make sense of the teaching of the Congregation for the Doctrine of the Faith (CDF) document Dignitas personae which appeared to provide a negative judgment on such a practice. This article aims to provide a personalist account of the process of fertilization and implantation that might serve as the basis for the negative judgment of the CDF document. In doing so, it relies upon the idea that a person, including an embryo, is not to be considered in isolation, but always in relation to God and to others. This approach extends the substantialist conceptualizations commonly employed in discussions of this issue. More generally, the article seeks to highlight the value of a personalist re-framing for an understanding of the moral questions surrounding the beginning of life. Lay summary: This article seeks to make sense of what appears to be a clear-cut rejection, set out in the Congregation for the Doctrine of the Faith (CDF) document Dignitas personae, of the proposal for women to “adopt” surplus frozen embryos. It draws upon more recently developed modes of philosophical/theological reasoning to argue that, in human procreation, both fertilization and implantation represent constitutive dimensions of divine creative activity and so must be protected from manipulative technological intervention. Since embryo adoption requires this kind of technology, it makes sense for the Church document not to approve it. PMID:25698841

  1. In vitro culture of Arabidopsis embryos.

    Science.gov (United States)

    Sauer, Michael; Friml, Jirí

    2008-01-01

    Embryogenesis of Arabidopsis thaliana follows a nearly invariant cell division pattern and provides an ideal system for studies of early plant development. However, experimental manipulation with embryogenesis is difficult, as the embryo develops deeply inside maternal tissues. Here, we present a method to culture zygotic Arabidopsis embryos in vitro. It enables culturing for prolonged periods of time from the first developmental stages on. The technique omits excision of the embryo by culturing the entire ovule, which facilitates the manual procedure. It allows pharmacological manipulation of embryo development and does not interfere with standard techniques for localizing gene expression and protein localization in the cultivated embryos.

  2. Diagnosis of human preimplantation embryo viability.

    Science.gov (United States)

    Gardner, David K; Meseguer, Marcos; Rubio, Carmen; Treff, Nathan R

    2015-01-01

    Transfer of more than a single embryo in an IVF cycle comes with the finite possibility of a multiple gestation. Even a twin pregnancy confers significant risk to both mother and babies. The move to single-embryo transfer for all patients will be greatly facilitated by the ability to quantify embryo viability. Developments in time-lapse incubation systems have provided new insights into the developmental kinetics of the human preimplantation embryo. Advances in molecular methods of chromosomal analysis have created platforms for highly effective screening of biopsied embryos, while noninvasive analysis of embryo physiology reveals more about the embryo than can be determined by morphology alone. Recent developments in time-lapse microscopy, molecular karyotyping and in proteomics and metabolomics have been assessed and presented here in a descriptive review. New algorithms are being created for embryo selection based on their developmental kinetics in culture, and the impact of factors such as patient etiology and treatment are being clarified. Potential links between morphokinetic data and embryo karyotype are being elucidated. The introduction of new molecular methods of determining embryo chromosomal complement is proving to be accurate and reproducible, with the future trending toward CGH arrays or next generation sequencing as a rapid and reliable means of analysis, that should be suitable for each IVF clinic to adopt. A relationship between embryo metabolism and viability is established and is now being considered together with morphokinetic data to create more robust algorithms for embryo selection. Microfluidic devices have the capacity and potential to be used in human IVF clinics for the routine diagnosis of embryo biomarkers. © The Author 2015. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  3. Nucleolar ultrastructure in bovine nuclear transfer embryos

    DEFF Research Database (Denmark)

    Kaňka, Jiří; Smith, Steven Dale; Soloy, Eva

    1999-01-01

    all three cell cycles. In the eight-cell stage embryo, a primary vacuole appeared as an electron lucid area originating in the centre of the nucleolar precursor body. In nuclear transfer embryos reconstructed from nonactivated cytoplasts, the nuclear envelope was fragmented or completely broken down...... vacuoles. A nucleolar precursor body typical for the two-cell stage control embryos was never observed. None of the reconstructed embryos of this group reached the eight-cell stage. Nuclear transfer embryos reconstructed from activated cytoplasts, in contrast, exhibited a complete nuclear envelope at all...... time intervals after fusion. In the two-cell stage nuclear transfer embryo, the originally reticulated nucleolus of the donor blastomere had changed into a typical nucleolar precursor body consisting of a homogeneous fibrillar structure. A primary vacuole appeared in the four-cell stage nuclear...

  4. Nucleolar remodeling in nuclear transfer embryos

    DEFF Research Database (Denmark)

    Laurincik, Jozef; Maddox-Hyttel, Poul

    2007-01-01

    nucleoli are not apparent until the 5th cell cycle, whereas in somatic cell nuclear transfer embryos the functional nucleoli emerge already during the 3rd cell cycle. Intergeneric reconstructed embryos produced by the fusion of bovine differentiated somatic cell to a nonactivated ovine cytoplast fail...... the developmental potential of embryos originating from varied nuclear transfer protocols. In bovine in vivo developed embryos, functional ribosome-synthesizing nucleoli become structurally distinct toward the end of the 4th post-fertilization cell cycle. In embryonic cell nuclear transfer embryos, fully developed...... is completed toward the end of the 4th cell cycle. A substantial proportion of bovine embryos produced by nuclear transfer of embryonic or somatic cells to bovine ooplasts display aberrations in protein localization in one or more blastomers. This information is indicative of underlying aberrations in genomic...

  5. Somatic embryogenesis and plant regeneration from embryo rescue ...

    African Journals Online (AJOL)

    The application of tissue culture techniques, particularly in the area of embryo rescue, has had a major impact on the maintenance and development of hybrid embryo from wide crosses. Embryo rescue techniques are directed towards obtaining more efficient survival of embryos in situations where very immature embryos ...

  6. Methanol as a cryoprotectant for equine embryos.

    Science.gov (United States)

    Bass, L D; Denniston, D J; Maclellan, L J; McCue, P M; Seidel, G E; Squires, E L

    2004-09-15

    Equine embryos (n=43) were recovered nonsurgically 7-8 days after ovulation and randomly assigned to be cryopreserved in one of two cryoprotectants: 48% (15M) methanol (n=22) or 10% (136 M) glycerol (n=21). Embryos (300-1000 microm) were measured at five intervals after exposure to glycerol (0, 2, 5, 10 and 15 min) or methanol (0, 15, 35, 75 and 10 min) to determine changes (%) in diameter over time (+/-S.D.). Embryos were loaded into 0.25-ml plastic straws, sealed, placed in a programmable cell freezer and cooled from room temperature (22 degrees C) to -6 degrees C. Straws were then seeded, held at -6 degrees C for 10 min and then cooled to -33 degrees C before being plunged into liquid nitrogen. Two or three embryos within a treatment group were thawed and assigned to be either cultured for 12 h prior to transfer or immediately nonsurgically transferred to a single mare. Embryo diameter decreased in all embryos upon initial exposure to cryoprotectant. Embryos in methanol shrank and recovered slightly to 76+/-8 % of their original diameter; however, embryos in glycerol continued to shrink, reaching 57+/-6 % of their original diameter prior to cryopreservation. Survival rates of embryos through Day 16 of pregnancy were 38 and 23%, respectively (P>0.05) for embryos cryopreserved in the presence of glycerol or methanol. There was no difference in pregnancy rates of mares receiving embryos that were cultured prior to transfer or not cultured (P>0.05). Preliminary experiments indicated that 48% methanol was not toxic to fresh equine embryos but methanol provided no advantage over glycerol as a cryoprotectant for equine blastocysts.

  7. Proteome profiling of embryo chick retina

    OpenAIRE

    Mizukami, Mina; Kanamoto, Takashi; Souchelnytskyi, Nazariy; Kiuchi, Yoshiaki

    2008-01-01

    Abstract Background Little is known regarding the molecular pathways that underlie the process of retinal development. The purpose of this study was to identify proteins which may be involved in development of retina. We used a proteomics-based approach to identify proteins that are up- or down-regulated during the development of the embryo chick retina. Results Two-dimensional gel electrophoresis was performed with the retina of embryo chicken, which was obtained from embryos of day 7 (ED7) ...

  8. Die Behandlung menschliches Embryos und Menschenwurde

    OpenAIRE

    Matsui, Fumio

    2002-01-01

    We are confronted with an old and new problem, which has come up with the progress of modern biotechnologies: what is a life or when does a life begin? The expectation of order-made medicine has build up since the discovery of Embryo Stem cell called "a dream master cell", while there is any condemnation against the destruction of human embryo in order to gain it. It is a question whether a human embryo is a human being in the world. Human dignity(=HD) is a principle that keeps human embryos ...

  9. Theory about the Embryo Cryo-Treatment.

    Science.gov (United States)

    Vladimirov, Iavor K; Tacheva, Desislava; Diez, Antonio

    2017-04-01

    To create hypothesis, which can give a logical explanation related to the benefits of freezing/thawing embryos. Cryopreservation is not only a technology used for storing embryos, but also a method of embryo treatment that can potentially improve the success rate in infertile couples. From the analysis of multiple results in assisted reproductive technology, which have no satisfactory explanation to date, we found evidence to support a 'therapeutic' effect of the freezing/thawing of embryos on the process of recovery of the embryo and its subsequent implantation. Freezing/thawing is a way to activate the endogenous survival and repair responses in preimplantation embryos. Several molecular mechanisms can explain the higher success rate of ET using thawed embryos compared to fresh ET in women of advanced reproductive age, the higher miscarriage rate in cases of thawed blastocyst ET compared to thawed ET at early cleavage embryo, and the higher perinatal parameters of born children after thawed ET. Embryo thawing induces a stress. Controlled stress is not necessarily detrimental, because it generates a phenomenon that is counteracted by several known biological responses aimed to repair mitochondrial damage of membrane and protein misfolding. The term for favorable biological responses to low exposures to stress is called hormesis. This thesis will summarize the role of cryopreservation in the activation of a hormetic response, preserving the mitochondrial function, improving survival, and having an impact on the process of implantation, miscarriage, and the development of pregnancy.

  10. Nano-nutrition of chicken embryos

    DEFF Research Database (Denmark)

    Sawosz, Filip; Pineda, Lane Manalili; Hotowy, Anna

    2013-01-01

    It has been suggested that the quantity and quality of nutrients stored in the egg might not be optimal for the fast rate of chicken embryo development in modern broilers, and embryos could be supplemented with nutrients by in ovo injection. Recent experiments showed that in ovo feeding reduces...... broiler eggs was randomly divided into a Control group without injection and injected groups with hydrocolloids of Nano-Ag, ATP or a complex of Nano-Ag and ATP (Nano-Ag/ATP). The embryos were evaluated on day 20 of incubation. The results indicate that the application of ATP to chicken embryos increases...

  11. Adoption first? The disposition of human embryos.

    Science.gov (United States)

    Murphy, Timothy F

    2014-06-01

    Anja Karnein has suggested that because of the importance of respect for persons, law and policy should require some human embryos created in vitro to be available for adoption for a period of time. If no one comes forward to adopt the embryos during that time, they may be destroyed (in the case of embryos left over from fertility medicine) or used in research (in the case of embryos created for that purpose or left over from fertility medicine). This adoption option would increase the number of embryos available for couples looking for help in having children, but that effect is less important--Karnein argues--than the observance of respect for human persons. As possible persons, she holds that embryos ought to be treated, as if they will become children, if only for a while. If enacted as a matter of law and policy, an 'adoption option' would wrongly interfere with the dispositional rights women and men ought to have over embryos they create in the course of trying to have children. Karnein's proposal would also deprive researchers of certainty that the embryos they create for research would actually be available that way, leading to increased burdens of time and money and maybe even to more embryos than would otherwise be produced. Karnein's analysis does not show, moreover, that any duty of rescue applies to embryos. No woman is required to adopt any embryo, which significantly undercuts the justification for an obligatory adoption period. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.

  12. Effects of Fluoxetine on Human Embryo Development.

    Science.gov (United States)

    Kaihola, Helena; Yaldir, Fatma G; Hreinsson, Julius; Hörnaeus, Katarina; Bergquist, Jonas; Olivier, Jocelien D A; Åkerud, Helena; Sundström-Poromaa, Inger

    2016-01-01

    The use of antidepressant treatment during pregnancy is increasing, and selective serotonin reuptake inhibitors (SSRIs) are the most widely prescribed antidepressants in pregnant women. Serotonin plays a role in embryogenesis, and serotonin transporters are expressed in two-cell mouse embryos. Thus, the aim of the present study was to evaluate whether fluoxetine, one of the most prescribed SSRI antidepressant world-wide, exposure influences the timing of different embryo developmental stages, and furthermore, to analyze what protein, and protein networks, are affected by fluoxetine in the early embryo development. Human embryos (n = 48) were randomly assigned to treatment with 0.25 or 0.5 μM fluoxetine in culture medium. Embryo development was evaluated by time-lapse monitoring. The fluoxetine-induced human embryo proteome was analyzed by shotgun mass spectrometry. Protein secretion from fluoxetine-exposed human embryos was analyzed by use of high-multiplex immunoassay. The lower dose of fluoxetine had no influence on embryo development. A trend toward reduced time between thawing and start of cavitation was noted in embryos treated with 0.5 μM fluoxetine (p = 0.065). Protein analysis by shotgun mass spectrometry detected 45 proteins that were uniquely expressed in fluoxetine-treated embryos. These proteins are involved in cell growth, survival, proliferation, and inflammatory response. Culturing with 0.5 μM, but not 0.25 μM fluoxetine, caused a significant increase in urokinase-type plasminogen activator (uPA) in the culture medium. In conclusion, fluoxetine has marginal effects on the timing of developmental stages in embryos, but induces expression and secretion of several proteins in a manner that depends on dose. For these reasons, and in line with current guidelines, the lowest possible dose of SSRI should be used in pregnant women who need to continue treatment.

  13. 9 CFR 98.16 - The embryo collection unit.

    Science.gov (United States)

    2010-01-01

    ... breeding them (either natural breeding or artificial insemination). (b) Embryo collection area. The embryo... equipment used for artificial insemination or for collection, processing, or storage of embryos. The walls...

  14. Embryo forming cells in carrot suspension cultures

    NARCIS (Netherlands)

    Toonen, M.A.J.

    1997-01-01


    Somatic cells of many plant species can be cultured in vitro and induced to form embryos that are able to develop into mature plants. This process, termed somatic embryogenesis, was originally described in carrot (Daucus carota L.). Somatic embryos develop through the same characteristic

  15. Human stem cell ethics: beyond the embryo.

    Science.gov (United States)

    Sugarman, Jeremy

    2008-06-05

    Human embryonic stem cell research has elicited powerful debates about the morality of destroying human embryos. However, there are important ethical issues related to stem cell research that are unrelated to embryo destruction. These include particular issues involving different types of cells used, the procurement of such cells, in vivo use of stem cells, intellectual property, and conflicts of interest.

  16. Future aspects of micromanipualtion with embryos for

    African Journals Online (AJOL)

    Embryo micromanipulation techniques and their potential genetic impact in dairy cattle are discussed. In addition, some aspects of gene transfer are mentioned. Only the technique of splitting bovine embryos and the subsequent transfer of halfembryos has reached a stage which might make its application to cattle breeding ...

  17. Embryo transfer using cryopreserved Boer goat blastocysts ...

    African Journals Online (AJOL)

    The aim of this trial was to evaluate the effect of embryo cryopreservation techniques on the survivability of embryos and fertility following transfer to Boer goat does. The oestrous cycles of 27 mature recipients Boer goat does were synchronised using controlled internal drug release dispensers (CIDR's) for 16 days. At CIDR ...

  18. Neural network classification of sweet potato embryos

    Science.gov (United States)

    Molto, Enrique; Harrell, Roy C.

    1993-05-01

    Somatic embryogenesis is a process that allows for the in vitro propagation of thousands of plants in sub-liter size vessels and has been successfully applied to many significant species. The heterogeneity of maturity and quality of embryos produced with this technique requires sorting to obtain a uniform product. An automated harvester is being developed at the University of Florida to sort embryos in vitro at different stages of maturation in a suspension culture. The system utilizes machine vision to characterize embryo morphology and a fluidic based separation device to isolate embryos associated with a pre-defined, targeted morphology. Two different backpropagation neural networks (BNN) were used to classify embryos based on information extracted from the vision system. One network utilized geometric features such as embryo area, length, and symmetry as inputs. The alternative network utilized polar coordinates of an embryo's perimeter with respect to its centroid as inputs. The performances of both techniques were compared with each other and with an embryo classification method based on linear discriminant analysis (LDA). Similar results were obtained with all three techniques. Classification efficiency was improved by reducing the dimension of the feature vector trough a forward stepwise analysis by LDA. In order to enhance the purity of the sample selected as harvestable, a reject to classify option was introduced in the model and analyzed. The best classifier performances (76% overall correct classifications, 75% harvestable objects properly classified, homogeneity improvement ratio 1.5) were obtained using 8 features in a BNN.

  19. Noninvasive metabolomic profiling as an adjunct to morphology for noninvasive embryo assessment in women undergoing single embryo transfer

    NARCIS (Netherlands)

    Seli, E.; Vergouw, C.G.; Morita, H.; Botros, L.; Roos, P.; Lambalk, C.B.; Yamashita, N.; Kato, O.; Sakkas, D.

    2010-01-01

    Objective: To determine whether metabolomic profiling of spent embryo culture media correlates with reproductive potential of human embryos. Design: Retrospective study. Setting: Academic and a private assisted reproductive technology (ART) programs. Patient(s): Women undergoing single embryo

  20. Rape embryogenesis. III. Embryo development in time

    Directory of Open Access Journals (Sweden)

    Teresa Tykarska

    2014-01-01

    Full Text Available It was found that the growth curve of the rape embryo axis is of triple sigmoid type. Embryo growth occurs in 3 phases corresponding to 3 different periods of development. Phase I includes growth of the apical cell up to it's division into two layers of octants. Phase II comprises the increase of the spherical proembryo to the change of its symmetry from radial to bilateral. Phase III includes, growth of the embryo from the heart stage up to the end of embryogenesis. In each phase the relative growth rate increases drastically and then diminishes. The differences in growth intensity during the same phase are several-fold. The growth intensity maximum of the embryo axis occurs in phase II. The phasic growth intensity maxima occur: in phase I during apical cell elongation, :before its division, and in phases II and III in the periods of cell division ;growth in globular and torpedo-shaped -shaped embryos.

  1. Nucleolar remodeling in nuclear transfer embryos

    DEFF Research Database (Denmark)

    Laurincik, Jozef; Maddox-Hyttel, Poul

    2007-01-01

    the developmental potential of embryos originating from varied nuclear transfer protocols. In bovine in vivo developed embryos, functional ribosome-synthesizing nucleoli become structurally distinct toward the end of the 4th post-fertilization cell cycle. In embryonic cell nuclear transfer embryos, fully developed...... to develop fully functional nucleoli. In bovine in vivo developed embryos, a range of important nucleolar proteins (e.g., topoisomerase I, upstream binding factor and RNA polymerase I, fibrillarin, nucleophosmin and nucleolin) become localized to the nucleolar anlage over several cell cycles. This relocation......Transcription of the ribosomal RNA (rRNA) genes occurs in the nucleolus and results in ribosome biogenesis. The rRNA gene activation and the associated nucleolus formation may be used as a marker for the activation of the embryonic genome in mammalian embryos and, thus serve to evaluate...

  2. Embryo transfer in domestic South American camelids.

    Science.gov (United States)

    Sumar, Julio B

    2013-01-10

    Intraspecific and interspecific embryo transfer in domestic South American camelids is developing into a well-established technique. Reports reveal many benefits of using reproductive biotechnologies to allow rapid propagation of alpacas and llamas of high genetic merit (e.g., high fiber quality, preserve color variation). The objective of this review is to provide up-to-date information about embryo transfer in domestic South American camelids. Specific information is provided on criteria for male selection, donor and recipient synchronization, the practice of single- vs. super-ovulation protocols, embryo recovery and transfer techniques, advances in cryopreservation of embryos, results of intra- and inter-specific transfer, and the future of the embryo transfer in domestic South American camelids. Copyright © 2012. Published by Elsevier B.V.

  3. Potential of human twin embryos generated by embryo splitting in assisted reproduction and research.

    Science.gov (United States)

    Noli, Laila; Ogilvie, Caroline; Khalaf, Yacoub; Ilic, Dusko

    2017-03-01

    Embryo splitting or twinning has been widely used in veterinary medicine over 20 years to generate monozygotic twins with desirable genetic characteristics. The first human embryo splitting, reported in 1993, triggered fierce ethical debate on human embryo cloning. Since Dolly the sheep was born in 1997, the international community has acknowledged the complexity of the moral arguments related to this research and has expressed concerns about the potential for reproductive cloning in humans. A number of countries have formulated bans either through laws, decrees or official statements. However, in general, these laws specifically define cloning as an embryo that is generated via nuclear transfer (NT) and do not mention embryo splitting. Only the UK includes under cloning both embryo splitting and NT in the same legislation. On the contrary, the Ethics Committee of the American Society for Reproductive Medicine does not have a major ethical objection to transferring two or more artificially created embryos with the same genome with the aim of producing a single pregnancy, stating that 'since embryo splitting has the potential to improve the efficacy of IVF treatments for infertility, research to investigate the technique is ethically acceptable'. Embryo splitting has been introduced successfully to the veterinary medicine several decades ago and today is a part of standard practice. We present here an overview of embryo splitting experiments in humans and non-human primates and discuss the potential of this technology in assisted reproduction and research. A comprehensive literature search was carried out using PUBMED and Google Scholar databases to identify studies on embryo splitting in humans and non-human primates. 'Embryo splitting' and 'embryo twinning' were used as the keywords, alone or in combination with other search phrases relevant to the topics of biology of preimplantation embryos. A very limited number of studies have been conducted in humans and non

  4. Human rights of an embryo.

    Science.gov (United States)

    Wennergren, Bertil

    1991-01-01

    Spontaneous ethical views very often spring just from feelings. But an ethical standpoint of any lasting value cannot be based just upon feelings. Relevant facts and values are indispensable. The purpose of the article is to draw the attention to the set of values that is enshrined in the Universal Declaration of Human Rights, particularly the values of human life, human dignity, human liberty, human physical integrity and the brotherhood of man. More generally recognized values are difficult to find. The usefulness of them as criteria in ethical contexts is tested by applying them to a judging pro and con of research on human embryos. The testing is performed against the background of the report of the Warnock Committee.

  5. Embryo cryopreservation and preeclampsia risk.

    Science.gov (United States)

    Sites, Cynthia K; Wilson, Donna; Barsky, Maya; Bernson, Dana; Bernstein, Ira M; Boulet, Sheree; Zhang, Yujia

    2017-11-01

    To determine whether assisted reproductive technology (ART) cycles involving cryopreserved-warmed embryos are associated with the development of preeclampsia. Retrospective cohort study. IVF clinics and hospitals. A total of 15,937 births from ART: 9,417 singleton and 6,520 twin. We used linked ART surveillance, birth certificate, and maternal hospitalization discharge data, considering resident singleton and twin births from autologous or donor eggs from 2005-2010. We compared the frequency of preeclampsia diagnosis for cryopreserved-warmed versus fresh ET and used multivariable logistic regression to adjust for confounders. Among pregnancies conceived with autologous eggs resulting in singletons, preeclampsia was greater after cryopreserved-warmed versus fresh ET (7.51% vs. 4.29%, adjusted odds ratio = 2.17 [95% CI 1.67-2.82]). Preeclampsia without and with severe features, preeclampsia with preterm delivery, and chronic hypertension with superimposed preeclampsia were more frequent after cryopreserved-warmed versus fresh ET (3.99% vs. 2.55%; 2.95% vs. 1.41%; 2.76 vs. 1.48%; and 0.95% vs. 0.43%, respectively). Among pregnancies from autologous eggs resulting in twins, the frequency of preeclampsia with severe features (9.26% vs. 5.70%) and preeclampsia with preterm delivery (14.81% vs. 11.74%) was higher after cryopreserved versus fresh transfers. Among donor egg pregnancies, rates of preeclampsia did not differ significantly between cryopreserved-warmed and fresh ET (10.78% vs. 12.13% for singletons and 28.0% vs. 25.15% for twins). Among ART pregnancies conceived using autologous eggs resulting in live births, those involving transfer of cryopreserved-warmed embryos, as compared with fresh ETs, had increased risk for preeclampsia with severe features and preeclampsia with preterm delivery. Copyright © 2017 American Society for Reproductive Medicine. All rights reserved.

  6. [Scientific ethics and frozen embryos].

    Science.gov (United States)

    Valenzuela, C Y

    2001-05-01

    Scientific Ethics is the theory and praxis of decisions. Philosophical Ethics is presented as the theory and praxis of the good. As the good differs among cultures, Philosophical Ethics is dependent on the endo-cultural good conception. The decision (included that one of adhesion or not to a world vision) depends on neuro-psychic specific factors: i) cognitive factors that include mostly the knowledge of the alternatives and their consequences and the ideological or religious conception of good in relation to the alternatives; ii) affective factors that make alternatives pleasant, unpleasant or neutral, attractive, repulsive or neutral; iii) emotional factors that associate to alternatives anger, peace or neutrality, sadness, happiness or neutrality; iv) value factors that assign importance, triviality or neutrality to alternatives, or assign them significance, irrelevancy or neutrality. There are unspecific factors such as the psychic energy, desire or others. Mixed factors such as attitude, motivation, intention and others. Scientific Ethics deals with the mind as a materio-energetic process which is different from the soul, eggs and embryos of any species are full individuals of that species, because, they have initiated a copy of their genome that specify, give autonomy and define them as individuals. For Scientific Ethics to leave frozen embryos like that for ever, to defrost and get rid of them or to use their cells for science are synonymous of killing them. To defrost them to use their cells as stem cells for somatic cell therapy or to implant them into uteri to continue their development is to maintain alive their cells, but only the implantation allows their maintenance as individuals, thus, being the only compatible with the Christian ethics. The compatibility of these alternatives with other ethics is discussed.

  7. [Association of human chorionic gonadotropin level in embryo culture media with early embryo development].

    Science.gov (United States)

    Wang, Haiying; Zhang, Renli; Han, Dong; Liu, Caixia; Cai, Jiajie; Bi, Yanling; Wen, Anmin; Quan, Song

    2014-06-01

    To investigate the association of human chorionic gonadotropin (HCG) level on day 3 of embryo culture with embryo development. Spent culture media were collected from individually cultured embryos on day 3 of in vitro fertilization and embryo transfer (IVF-ET) cycles. HCG concentration in the culture media was measured using an ELISA kit and its association with embryo development was assessed. In the 163 samples of embryo culture media from 60 patients, HCG was positive in 153 sample (93.8%) with a mean level of 0.85 ± 0.43 mIU/ml. The concentration of hCG in the culture media increased gradually as the number of blastomeres increased (F=2.273, P=0.03), and decreased as the morphological grade of the embryo was lowered (F=3.900, P=0.02). ELISA is capable of detecting HCG levels in spent culture media of embryos on day 3 of in vitro culture. The concentration of HCG in spent culture media is positively correlated with the status of early embryo development and implantation rate and thus serves as a useful marker for embryo selection in IVF-ET procedure.

  8. Support for selling embryos among infertility patients.

    Science.gov (United States)

    Jain, Tarun; Missmer, Stacey A

    2008-09-01

    To determine the opinions of infertility patients regarding selling extra embryos, and to investigate the relation between patient choice and demographic and socioeconomic characteristics. Cross-sectional, self-administered survey. University hospital-based fertility center. 1350 consecutive women who presented for infertility care. None. Patient opinion regarding selling extra embryos to other couples, and correlations with their demographic and socioeconomic background. Of respondents with a definitive opinion, 56% felt that selling extra embryos to other couples should be allowed. After adjustment for observed predictors favoring selling extra embryos, we found statistically significantly lower support for selling embryos among patients who were Hispanic (relative to Caucasians) or had never been pregnant, whereas significantly greater support was observed among Hindu and secular women, patients being treated for male factor infertility, and those who in the past had or were currently undergoing intrauterine insemination. Age, education, marital status, and parity were not statistically significantly associated with the opinions about selling extra embryos to other couples. A large proportion of infertility patient participants approved of selling leftover embryos to other couples. However, some demographic and reproductive factors are significantly associated with patient opinion.

  9. In-vivo Centrifugation of Drosophila Embryos

    Science.gov (United States)

    Tran, Susan L.; Welte, Michael A.

    2010-01-01

    A major strategy for purifying and isolating different types of intracellular organelles is to separate them from each other based on differences in buoyant density. However, when cells are disrupted prior to centrifugation, proteins and organelles in this non-native environment often inappropriately stick to each other. Here we describe a method to separate organelles by density in intact, living Drosophila embryos. Early embryos before cellularization are harvested from population cages, and their outer egg shells are removed by treatment with 50% bleach. Embryos are then transferred to a small agar plate and inserted, posterior end first, into small vertical holes in the agar. The plates containing embedded embryos are centrifuged for 30 min at 3000g. The agar supports the embryos and keeps them in a defined orientation. Afterwards, the embryos are dug out of the agar with a blunt needle. Centrifugation separates major organelles into distinct layers, a stratification easily visible by bright-field microscopy. A number of fluorescent markers are available to confirm successful stratification in living embryos. Proteins associated with certain organelles will be enriched in a particular layer, demonstrating colocalization. Individual layers can be recovered for biochemical analysis or transplantation into donor eggs. This technique is applicable for organelle separation in other large cells, including the eggs and oocytes of diverse species. PMID:20613707

  10. [Chapter 10. The embryo, a particular thing].

    Science.gov (United States)

    Marais, Astrid

    2018-03-07

    By allowing a woman to terminate the life of the embryo, the law of 17 January 1975 leads us to ask the question about how to qualify the embryo. Is it a thing or a person ? Subordinate to the birth of a viable, living being, the embryo can have no acknowledged legal personality. Its particularity must nevertheless be noted : it is the only thing in law that is likely to become a person. So, conjugated in the present (The reality of the present : the embryo is a thing), the embryo is without doubt a thing.This qualification is implicit and results from an a contrario interpretation of article 16 CC. By not qualifying the human being from the start of his life as a person, the legislator treats him as a thing. That explains why the term homicide cannot be used against someone whose fault has provoked the termination of the pregnancy of a pregnant woman.The qualification as a thing does not prevent the protection of the embryo, attached to its quality as a human being. This protection is different from that applied to a person. In fact, it does not present an obstacle to the destruction of the embryo. It only consists in providing a framework for any violations that the embryo might suffer. The protection is variable according to whether it applies to the embryo in vitro or in utero and increases as the parental project progresses.Conjugated in the future, the embryo will become a person (The fiction taken from the future : the embryo is a person). This perspective allows us to qualify it fictitiously as a person, from conception, by means of an analysis, either retrospective or prospective.The retrospective analysis allows, once the child is born alive, a ?return to the future? by recognising that the embryo has rights, from conception, by applying the adage infans conceptus.The prospective analysis leads to anticipating the future and the quality as a person that will be acknowledged for the embryo at birth. Inspired by the law of goods and in particular the notion

  11. [Philosophical aspects. Why the embryo is a issue].

    Science.gov (United States)

    Grassin, Marc

    2018-03-07

    How raising the embryo's issue today ? Far from evidences and certainties, we have to face the confrontations to understand them. Embryo's issue crystalise all cultural and human tensions. Beyond ontological disagrement on the embryo being what can we expect for mankind today and tomorrow ? Through an anthropological thinking, embryo's issue challenges us in our common capability of decision.

  12. 9 CFR 98.9 - Embryos refused entry.

    Science.gov (United States)

    2010-01-01

    ... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Embryos refused entry. 98.9 Section 98.9 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE... Disease; and Embryos of Horses and Asses § 98.9 Embryos refused entry. Any embryo refused entry into the...

  13. Timing embryo biopsy for PGD - before or after cryopreservation?

    Science.gov (United States)

    Shinar, S; Kornecki, N; Schwartz, T; Mey-Raz, N; Amir, H; Almog, B; Shavit, T; Hasson, J

    2016-09-01

    Pre-implantation genetic diagnosis (PGD) is required in order to screen and diagnose embryos of patients at risk of having a genetically affected offspring. A biopsy to diagnose the genetic profile of the embryo may be performed either before or after cryopreservation. The aim of this study was to determine which biopsy timing yields higher embryo survival rates. Retrospective cohort study of all PGD patients in a public IVF unit between 2010 and 2013. Inclusion criteria were patients with good-quality embryos available for cryopreservation by the slow freezing method. Embryos were divided into two groups: biopsy before and biopsy after cryopreservation. The primary outcome was embryo survival rates post thawing. Sixty-five patients met inclusion criteria. 145 embryos were biopsied before cryopreservation and 228 embryos were cryopreserved and biopsied after thawing. Embryo survival was significantly greater in the latter group (77% vs. 68%, p Cryopreservation preceding biopsy results in better embryo survival compared to biopsy before cryopreservation.

  14. Multiple-embryo transfer for studying very early maternal-embryo interactions in cattle.

    Science.gov (United States)

    Gómez, E; Muñoz, M

    2015-08-01

    In the present paper, we highlight the need to study very early maternal-embryo interactions and discuss how these interactions can be addressed. Bovine species normally carry one or, less frequently, two embryos to term; there are very rare cases of triplets or higher-order multiple pregnancies in which all the offspring are born alive. Multiple-embryo transfer (MET) in cattle allows for the detection of endometrial responses in scenarios where single-embryo transfer would not. Although MET is non-physiological, the present study shows that at the very early embryonic stages, a uterus carrying zona-enclosed embryos does not exhibit non-physiological reactions. On the contrary, MET should be considered the sum of multiple individual effects triggered by developing embryos. We provide arguments to support our hypothesis that describe a rationale for current work with MET, and we discuss alternative hypotheses. Using cattle as a model, we describe how technical approaches to analyzing zona-enclosed early embryo-maternal interactions (i.e., transcriptomics, proteomics, and endometrial cell culture) can help identify molecular changes that may be difficult to observe when only a single embryo is present. We conclude that MET can be used for studying very early maternal-embryo interactions in vivo in monotocous species. Free Spanish abstract: A Spanish translation of this abstract is freely available at http://www.reproduction-online.org/content/150/2/R35/suppl/DC1. © 2015 Society for Reproduction and Fertility.

  15. Bovine in vitro embryo production : An overview

    Directory of Open Access Journals (Sweden)

    V. S. Suthar

    Full Text Available Dairy industry perfected the application of the first reproductive biotechnology, i.e. artificial insemination (AI - a great success story and also remains the user of embryo transfer technology (ETT. In addition, recently the researchers taking interest to embraced the field of Transvaginal OocyteRecovery (TVOR and in vitro production (IVEP of embryos. IVF provides the starting point for the generation of reproductive material for a number of advanced reproduction techniques including sperm microinjection and nuclear transfer (cloning. In several countries commercial IVF facilities are already being employed by cattle ET operators. Various research groups have reported on modification of TVOR technique to give greater efficiency. Much research is still needed in domestic animal (Especially Indian species on mechanisms controlling embryo development and on development of totally in vitro system for embryo culture. [Vet World 2009; 2(12.000: 478-479`

  16. Transcervical Embryo recovery by transcervical technique in ...

    African Journals Online (AJOL)

    Vicente Freitas

    1970s that similar technologies were adopted in goat breeding. Since then, embryos from thousands of goats have been collected and transferred. Three countries, Australia, New Zealand and South Africa, are particularly prominent in this field ...

  17. Predator recognition in rainbowfish, Melanotaenia duboulayi, embryos.

    Directory of Open Access Journals (Sweden)

    Lois Jane Oulton

    Full Text Available Exposure to olfactory cues during embryonic development can have long term impacts on birds and amphibians behaviour. Despite the vast literature on predator recognition and responses in fishes, few researchers have determined how fish embryos respond to predator cues. Here we exposed four-day-old rainbowfish (Melanotaenia duboulayi embryos to cues emanating from a novel predator, a native predator and injured conspecifics. Their response was assessed by monitoring heart rate and hatch time. Results showed that embryos have an innate capacity to differentiate between cues as illustrated by faster heart rates relative to controls. The greatest increase in heart rate occurred in response to native predator odour. While we found no significant change in the time taken for eggs to hatch, all treatments experienced slight delays as expected if embryos are attempting to reduce exposure to larval predators.

  18. Valuing embryos as both commodities and singularities.

    Science.gov (United States)

    Legge, Michael; Fitzgerald, Ruth

    2016-03-11

    An argument put forward against gamete and embryo donation, sale and research, is that to do so would treat the gametes or embryos as objects with no intrinsic value as human. Instead, gametes and embryos created and used for donation, sale or research, can be considered more like a commodity created and traded for economic exchange--something that is valuable only for the amount of money or other goods and services that others are willing to exchange. While Kant asserts that humans have dignity rather than object worth, the provision of human gametes and embryos are progressively becoming utilities for resolving childlessness and for certain research investigations. In this paper we discuss the commodity market and the relationship to human reproduction material.

  19. Role of melatonin in embryo fetal development

    OpenAIRE

    Voiculescu, SE; Zygouropoulos, N; Zahiu, CD; Zagrean, AM

    2014-01-01

    Melatonin is an indoleamine produced by the pineal gland and secreted in a circadian manner. In the past few decades, research over this topic has been enhanced. Melatonin has many important roles in the human physiology: regulator of the circadian rhythms, sleep inducer, antioxidant, anticarcinogenic. This paper reviews the involvement of melatonin in embryo fetal development. The pineal gland develops completely postpartum, so both the embryo and the fetus are dependent on the maternal mela...

  20. Embryo disposition and the new death scene

    Directory of Open Access Journals (Sweden)

    Ellison, David

    2011-01-01

    Full Text Available In the IVF clinic - a place designed principally for the production and implantation of embryos - scientists and IVF recipients are faced with decisions regarding the disposition of frozen embryos. At this time there are hundred of thousands of cryopreserved embryos awaiting such determinations. They may be thawed for transfer to the woman herself, they may be donated for research or for use by other infertile couples, they may remain in frozen storage, or they may variously be discarded by being allowed to 'succumb', or 'perish'. Where the choice is discard, some IVF clients have chosen to formalise the process through ceremony. A new language is emerging in response to the desires of the would-be-parents who might wish to characterise the discard experience as a ‘good death’. This article examines the procedure known as ‘compassionate transfer’ where the embryo to be discarded is placed in the woman’s vagina where it is clear that it will not develop further. An alternate method has the embryo transferred in the usual manner but without the benefit of fertility-enhancing hormones at a point in the cycle unreceptive to implantation. The embryo destined for disposal is thus removed from the realm of technological possibility and ‘returned’ to the female body for a homely death. While debates continue about whether or not embryos constitute life, new practices are developing in response to the emotional experience of embryo discard. We argue that compassionate transfer is a death scene taking shape. In this article, we take the measure of this new death scene’s fabrication, and consider the form, significance, and legal complexity of its ceremonies.

  1. [Assisted reproductive technologies and the embryo status].

    Science.gov (United States)

    Englert, Y

    The status of the human embryo has always be a subject of philosophical and theological thoughts with major social consequences, but, until the 19th century, it has been mainly an abstraction. The arrival of the human embryo in vitro, materialized by Louise Brown's birth in 1978 and above all by the supernumerary embryos produced by the Australian team of Trounson and Wood following the introduction of ovarian stimulation, will turn theoretical thoughts into a reality. Nobody may ignore the hidden intentions behind the debate, as to recognise a status to a few days old embryo will immediately have a major impact on the status of a few weeks old foetus and therefore on the abortion rights. We will see that the embryo status, essentially based as well on a vision on the good and evil as on social order, cannot be based on a scientific analysis of the reproduction process but comes from a society's choice, by essence " arbitrary " and always disputable. This does not preclude the collectivity right and legitimacy to give a precise status and it is remarkable to observe the law is careful not to specify which status to give to the human embryo. It is more thru handling procedures and functioning rules that the law designed the embryo position, neither with a status of a person, nor of a thing. It nevertheless remains true that there is a constant risk that the legislation gives the embryo a status that would call into question it's unique characteristic of early reproductive stage, jeopardizing at once the hard-won reproductive freedom (reproductive choice) as well as freedom of research on embryonic stem cells, one of the most promising field of medical research.

  2. Cryopreservation of Arachis pintoi (leguminosae) somatic embryos.

    Science.gov (United States)

    Rey, H Y; Faloci, M; Medina, R; Dolce, N; Engelmann, F; Mroginski, L

    2013-01-01

    In this study, we successfully cryopreserved cotyledonary somatic embryos of diploid and triploid Arachis pintoi cytotypes using the encapsulation-dehydration technique. The highest survival rates were obtained when somatic embryos were encapsulated in calcium alginate beads and precultured in agitated (80 rpm) liquid establishment medium (EM) with daily increasing sucrose concentration (0.50, 0.75, and 1.0 M). The encapsulated somatic embryos were then dehydrated with silica gel for 5 h to 20% moisture content (fresh weight basis) and cooled either rapidly (direct immersion in liquid nitrogen, LN) or slowly (1 degree C per min from 25 degree C to -30 degree C followed by immersion in LN). Beads were kept in LN for a minimum of 1 h and then were rapidly rewarmed in a 30 degree C water-bath for 2 min. Finally, encapsulated somatic embryos were post-cultured in agitated (80 rpm) liquid EM with daily decreasing sucrose concentration (0.75 and 0.5 M) and transferred to solidified EM. Using this protocol, we obtained 26% and 30% plant regeneration from cryopreserved somatic embryos of diploid and triploid cytotypes. No morphological abnormalities were observed in any of the plants regenerated from cryopreserved embryos and their genetic stability was confirmed with 10 isozyme systems and nine RAPD profiles.

  3. Manipulation and imaging of Kryptolebias marmoratus embryos.

    Science.gov (United States)

    Mourabit, Sulayman; Kudoh, Tetsuhiro

    2012-12-01

    The self-fertilizing mangrove killifish, Kryptolebias marmoratus, is an upcoming model species for a range of biological disciplines. To further establish this model in the field of developmental biology, we examined several techniques for embryonic manipulation and for imaging that can be used in an array of experimental designs. These methodological approaches can be divided into two categories: handling of embryos with and without their chorionic membrane. Embryos still enclosed in their chorion can be manipulated using an agarose bed or a methyl cellulose system, holding them in place and allowing their rotation to more specific angles and positions. Using these methods, we demonstrate microinjection of embryos and monitoring of fluorescent yolk syncytial nuclei (YSN) using both stereo and compound microscopes. For higher magnification imaging using compound microscopes as well as time-lapse analyses, embryos were dechorionated and embedded in low-melting-point agarose. To demonstrate this embedding technique, we further examined fluorescent YSN and also analyzed the yolk surface of K. marmoratus embryos. The latter was observed to provide an excellent imaging platform for study of the behavior and morphology of cells during embryonic development, for various types of cells. Our data demonstrate that K. marmoratus is an excellent model species for research in developmental biology, as methodological approaches for the manipulation and imaging of embryos are efficient and readily available.

  4. Proteome profiling of embryo chick retina

    Directory of Open Access Journals (Sweden)

    Souchelnytskyi Nazariy

    2008-01-01

    Full Text Available Abstract Background Little is known regarding the molecular pathways that underlie the process of retinal development. The purpose of this study was to identify proteins which may be involved in development of retina. We used a proteomics-based approach to identify proteins that are up- or down-regulated during the development of the embryo chick retina. Results Two-dimensional gel electrophoresis was performed with the retina of embryo chicken, which was obtained from embryos of day 7 (ED7 and of day 11 (ED11. The protein spots showing significant differences were selected for identification by MALDI mass spectrometry. Thirteen proteins were differentially expressed; seven proteins were up-regulated in embryo retina of chicken at ED 11 and six proteins were down-regulated. Significant proteins were also evaluated in embryo day 15 (ED15. Some of identified proteins were known to regulate cell proliferation, cell death, transport, metabolism, organization and extracellular matrix, and others also included novel proteins. Conclusion We identified thirteen proteins which differentially expressed in embryonal retina of chicken at day 7, as compared to the retina of embryo of day 11. They were various regulatory proteins for cellular signaling.

  5. Effects of alpha particles on zebrafish embryos

    International Nuclear Information System (INIS)

    Yum, E.H.W.; Choi, V.W.Y.; Yu, K.N.; Li, V.W.T.; Cheng, S.H.

    2008-01-01

    Full text: Ionizing radiation such as X-ray and alpha particles can damage cellular macromolecules, which can lead to DNA single- and double-strand breaks. In the present work, we studied the effects of alpha particles on dechorionated zebrafish embryos. Thin polyallyldiglycol carbonate (PADC) films with a thickness of 16 μm were prepared from commercially available PADC films (with thickness of 100 μm) by chemical etching and used as support substrates for holding zebrafish embryos for alpha-particle irradiation. These films recorded alpha-particle hit positions, quantified the number and energy of alpha particles actually incident on the embryo cells, and thus enabled the calculation of the dose absorbed by the embryo cells. Irradiation was made at 1.25 hours post fertilization (hpf) with various absorbed dose. TdT-mediated dUTP Nick-End Labeling (TUNEL) assay was performed on the embryos at different time stages after irradiation. Marked apoptosis was detected only in embryos at earlier time stages. The results showed that DNA double-strand break during zebrafish embryogenesis can be induced by alpha-particle irradiation, which suggests that zebrafish is a potential model for assessing the effects of alpha-particle radiation

  6. Der Embryo: Eine junge Erfindung? The Embryo: A recent Invention?

    Directory of Open Access Journals (Sweden)

    Ute Frietsch

    2003-07-01

    Full Text Available Die Veröffentlichung des Max-Planck-Institutes für Geschichte sammelt Beiträge, die im Rahmen einer Konferenz der internationalen und interdisziplinären Arbeitsgruppe zur Geschichte der Geburt 1999 in Göttingen entstanden sind. Schwangerschaftserfahrung von Frauen und wissenschaftliche Zugriffe von Theologen, Anatomen, Medizinern und Juristen auf Ungeborene, Frauenkörper und Gesellschaftskörper werden programmatisch miteinander konfrontiert. Gearbeitet wird mit unterschiedlichen Perspektiven, Zeitausschnitten und Größenverhältnissen. Sowohl Divergenzen als auch Zusammenhänge von Körpergeschichte und Bevölkerungs- oder Biowissenschaft werden schlaglichtartig in Fallstudien beleuchtet. Die Biologisierung der Schwangerschaft in der Neuzeit wird aus ihrer Verwissenschaftlichung erklärt.10 interdisciplinary contributions dealing with the conflicting fields between personal experiences and the biologisation of pregnancy in modern times. This anthology published by the Max Planck Institute for History features a variety of contributions which came into being during a conference of the international and interdisciplinary working group on the history of birth. The conference took place in Göttingen in 1999. The anthology features the experiences with pregnancy of different women as well as scientific perspectives on embryos, women’s bodies, and societal bodies by theologians, anatomists, physicians, and jurists. The different contributions employ different perspectives, time periods, and scales. Case studies introduce and highlight diverging and converging perspectives on the history of the body, population studies, and biology, and explain how the advancement of science as well as societal processes caused pregnancy to become biologised in modern times.

  7. In vitro bovine embryo production in a synthetic medium: embryo development, cryosurvival, and establishment of pregnancy.

    Science.gov (United States)

    Moreno, D; Neira, A; Dubreil, L; Liegeois, L; Destrumelle, S; Michaud, S; Thorin, C; Briand-Amirat, L; Bencharif, D; Tainturier, D

    2015-10-15

    The aim of this study was to develop an in vitro embryo culture medium without either fetal calf serum or BSA, using various growth factors and cytokines (GFs-CYKs; IGF-I, IGF-II, bFGF, LIF, GM-CSF, TGF-β1, and PDGF-BB), and other molecules with surfactant and embryotrophic properties, such as recombinant albumin (RA) and hyaluronan (HA). The first part of the study was dedicated to define the best combination of GFs-CYKs + RA + HA for optimal embryonic development. Next, we compared development rates and embryo quality (inner cell mass [ICM]-to-total cell number [TCN] ratio), and postthaw survival and hatching rates using this synthetic medium (T1) and a control medium: synthetic oviduct fluid + BSA + ITS (insulin, transferrin, and selenium). The blastocyst rates were significantly higher with T1 than those with the control at 7 and 8 days after fertilization. There was no significant difference in TCN or the ICM/TCN ratio between the two treatments. Survival and hatching rates 48 hours after thawing were similar for both treatments. Finally, nine embryo transfers were conducted using fresh and previously frozen Day-7 blastocysts to evaluate the in vivo viability of embryos produced in this synthetic medium; four gestations were obtained from six fresh embryos and one gestation from three frozen embryos. In conclusion, the fetal calf serum and BSA-free medium, supplemented with GFs-CYKs + RA + HA, improved embryo development and gave comparable ICM/TCN ratios and postthaw survival rates to the control with BSA. Fresh and frozen embryos produced in this medium are viable for embryo transfer. This fully synthetic method of embryo culture is a useful means of reducing the risk of disease transmission via embryo transfer. Copyright © 2015 Elsevier Inc. All rights reserved.

  8. Miniaturized embryo array for automated trapping, immobilization and microperfusion of zebrafish embryos.

    Directory of Open Access Journals (Sweden)

    Jin Akagi

    Full Text Available Zebrafish (Danio rerio has recently emerged as a powerful experimental model in drug discovery and environmental toxicology. Drug discovery screens performed on zebrafish embryos mirror with a high level of accuracy the tests usually performed on mammalian animal models, and fish embryo toxicity assay (FET is one of the most promising alternative approaches to acute ecotoxicity testing with adult fish. Notwithstanding this, automated in-situ analysis of zebrafish embryos is still deeply in its infancy. This is mostly due to the inherent limitations of conventional techniques and the fact that metazoan organisms are not easily susceptible to laboratory automation. In this work, we describe the development of an innovative miniaturized chip-based device for the in-situ analysis of zebrafish embryos. We present evidence that automatic, hydrodynamic positioning, trapping and long-term immobilization of single embryos inside the microfluidic chips can be combined with time-lapse imaging to provide real-time developmental analysis. Our platform, fabricated using biocompatible polymer molding technology, enables rapid trapping of embryos in low shear stress zones, uniform drug microperfusion and high-resolution imaging without the need of manual embryo handling at various developmental stages. The device provides a highly controllable fluidic microenvironment and post-analysis eleuthero-embryo stage recovery. Throughout the incubation, the position of individual embryos is registered. Importantly, we also for first time show that microfluidic embryo array technology can be effectively used for the analysis of anti-angiogenic compounds using transgenic zebrafish line (fli1a:EGFP. The work provides a new rationale for rapid and automated manipulation and analysis of developing zebrafish embryos at a large scale.

  9. Meanings of the embryo in Japan: narratives of IVF experience and embryo ownership

    NARCIS (Netherlands)

    Kato, M.; Sleeboom-Faulkner, M.

    2011-01-01

    This article explores the sociocultural meanings of the embryo implied in the narratives of 58 women who have undergone in vitro fertilisation in Japan over a period from 2006 to 2008. We argue that a lack of sufficient analysis of the sociocultural meanings of the embryo result in a situation where

  10. Development of interspecies cloned embryos in yak and dog.

    Science.gov (United States)

    Murakami, Masao; Otoi, Takeshige; Wongsrikeao, Pimprapar; Agung, Budiyanto; Sambuu, Rentsenkhand; Suzuki, Tatsuyuki

    2005-01-01

    Interspecies nuclear transfer (NT) could be an alternative to replicate animals when supply of recipient oocytes is limited or in vitro embryo production systems are incomplete. In the present study, embryonic development was assessed following interspecies NT of donor cumulus cells derived from yak and dog into the recipient ooplasm of domestic cow. The percentages of fusion and subsequent embryo development to the eight-cell stage of interspecies NT embryos were comparable to those of intraspecies NT embryos (cow-cow NT embryos). The percentage of development to blastocysts was significantly lower (p dog-cow NT embryos, only one embryo (0.4%) developed to the blastocyst stage. These results indicate that interspecies NT embryos possess equally developmental competence to the eight-cell stage as intraspecies NT embryos, but the development to blastocysts is very low when dog somatic cells are used as the donor nuclei.

  11. Cryopreservation of peach palm zygotic embryos.

    Science.gov (United States)

    Steinmacher, Douglas A; Saldanha, Cleber W; Clement, Charles R; Guerra, Miguel P

    2007-01-01

    Cryopreservation is a safe and cost-effective option for long-term germplasm conservation of non-orthodox seed species, such as peach palm (Bactris gasipaes). The objective of the present study was to establish a cryopreservation protocol for peach palm zygotic embryos based on the encapsulation-dehydration technique. After excision, zygotic embryos were encapsulated with 3 percent sodium alginate plus 2 M glycerol and 0.4 M sucrose, and pre-treated or not with 1 M sucrose during 24 h, followed by air-drying. Fresh weight water contents of beads decreased from 83 percent and 87 percent to 18 percent and 20 percent for pre-treated or non-pretreated beads, respectively, after 4 h of dehydration. Sucrose pre-treatment at 1 M caused lower zygotic embryo germination and plantlet height in contrast to non-treated beads. All the variables were statistically influenced by dehydration time. Optimal conditions for recovery of cryopreserved zygotic embryos include encapsulation and dehydration for 4 h in a forced air cabinet to 20 percent water content, followed by rapid freezing in liquid nitrogen (-196 degree C) and rapid thawing at 45 degree C. In these conditions 29 percent of the zygotic embryos germinated in vitro. However, plantlets obtained from dehydrated zygotic embryos had stunted haustoria and lower heights. Histological analysis showed that haustorium cells were large, vacuolated, with few protein bodies. In contrast, small cells with high nucleus:cytoplasm ratio formed the shoot apical meristem of the embryos, which were the cell types with favorable characteristics for survival after exposure to liquid nitrogen. Plantlets were successfully acclimatized and showed 41+/-9 percent and 88+/-4 percent survival levels after 12 weeks of acclimatization from cryopreserved and non-cryopreserved treatments, respectively.

  12. Patients' Attitudes towards the Surplus Frozen Embryos in China

    Directory of Open Access Journals (Sweden)

    Xuan Jin

    2013-01-01

    Full Text Available Background. Assisted reproductive techniques have been used in China for more than 20 years. This study investigates the attitudes of surplus embryo holders towards embryos storage and donation for medical research. Methods. A total of 363 couples who had completed in vitro fertilization (IVF treatment and had already had biological children but who still had frozen embryos in storage were invited to participate. Interviews were conducted by clinics in a narrative style. Results. Family size was the major reason for participants’ (discontinuation of embryo storage; moreover, the moral status of embryos was an important factor for couples choosing embryo storage, while the storage fee was an important factor for couples choosing embryo disposal. Most couples discontinued the storage of their embryos once their children were older than 3 years. In our study, 58.8% of the couples preferred to dispose of surplus embryos rather than donate them to research, citing a lack of information and distrust in science as significant reasons for their decision. Conclusions. Interviews regarding frozen embryos, including patients’ expectations for embryo storage and information to assist them with decisions regarding embryo disposal, are beneficial for policies addressing embryo disposition and embryo donation in China.

  13. Embryos, individuals, and persons: an argument against embryo creation and research.

    Science.gov (United States)

    Tollefsen, C

    2001-01-01

    One strategy for arguing that it should be legally permissible to create human embryos, or to use spare human embryos, for scientific research purposes involves the claim that such embryos cannot be persons because they are not human individuals while twinning may yet take place. Being a human individual is considered to be by most people a necessary condition for being a human person. I argue first that such an argument against the personhood of embryos must be rationally conclusive if their destruction in public places such as laboratories is to be countenanced. I base this argument on a popular understanding of the role that the notion of privacy plays in abortion laws. I then argue that such arguments against personhood are not rationally conclusive. The claim that the early embryos is not a human individual is not nearly as obvious as some assert.

  14. Metabolite profiling of somatic embryos of Cyclamen persicum in comparison to zygotic embryos, endosperm and testa

    Directory of Open Access Journals (Sweden)

    Traud eWinkelmann

    2015-08-01

    Full Text Available Somatic embryogenesis has been shown to be an efficient in vitro plant regeneration system for many crops such as the important ornamental plant Cyclamen persicum, for which this regeneration pathway of somatic embryogenesis is of interest for the vegetative propagation of parental lines as well as elite plants. However, somatic embryogenesis is not commercially used in many crops due to several unsolved problems, such as malformations, asynchronous development, deficiencies in maturation and germination of somatic embryos. In contrast, zygotic embryos in seeds develop and germinate without abnormalities in most cases. Instead of time-consuming and labor-intensive experiments involving tests of different in vitro culture conditions and plant growth regulator supplements, we follow a more directed approach. Zygotic embryos served as a reference and were compared to somatic embryos in metabolomic analyses allowing the future optimization of the in vitro system. The aims of this study were to detect differences in the metabolite profiles of torpedo stage somatic and zygotic embryos of C. persicum. Moreover, major metabolites in endosperm and testa were identified and quantified.Two sets of extracts of two to four biological replicates each were analyzed. In total 52 metabolites were identified and quantified in the different tissues. One of the most significant differences between somatic and zygotic embryos was that the proline concentration in the zygotic embryos was about 40 times higher than that found in somatic embryos. Epicatechin, a scavenger for reactive oxygen species, was found in highest abundance in the testa. Sucrose, the most abundant metabolite was detected in significantly higher concentrations in zygotic embryos. Also, a yet unknown trisaccharide, was significantly enriched in zygotic embryos.

  15. Formation of somatic embryos in Persea americana Mill var Catalina from immature zygotic embryos.

    Directory of Open Access Journals (Sweden)

    Lillien Fajardo Rosabal

    2005-04-01

    Full Text Available The establishment of embryogenic culture of avocado have been achieved in different genotypes, usually the immature zygotic embryos are the initial explants and the process has been described in several variety. In the present paper the induction of the somatic embryogenesis in avocado (Catalina variety from zygotic embryos is proposed. Zygotic embryos taken from unripe fruits were used as explants . The fruits were divided into five groups according to their size. The embryos were cultured in a medium containing 4-amino-3,5,6 trichlorpicolinic acid (Picloram in concentrations of 0.1, 0.4, and 0.6 uM. The culture medium used for the induction of the somatic embryogenesis consisted of: Macro B5, Micro MS, thiamine (0.8 mg.l-1, myo-inositol (100 mg.l-1, sucrose (30g.l-1 and pH 5.7. The number of zygotic embryos with opened cotyledonal leaves was evaluated starting from the third day of culture. It was also evaluated the number of fenolized zygotic embryos at the third week of culture and the presence of somatic embryos five weeks after the culture initiation. The formation of somatic embryos was achieved in all the treatments. The highest number of explants that formed somatic embryos was achieved when a concentration of 0.6 uM of Picloram was used and the second group of size (0.71 x 0.65 mm observing significant differences between the different groups of fruit size. Keywords: avocado, cotyledonal leafs, somatic embryo,

  16. [The dignity of the frozen human embryo].

    Science.gov (United States)

    Zurriaráin, R Germán

    2007-01-01

    The issue of the dignity of the human embryo is one of the key questions in bioethics debate today. The human being's dignity lies in the original and unique individuality that every embryo has. If there is no respect and defense of the human body from the very first moment of its appearance, it is impossible to assert the dignity of any human being. For this reason, it does not seem good for human dignity that human beings, in their incipient phase, should suffer from the halting of their biological functions. The practice of cryopreservation of human embryos shows a loss of the sense of the value of each individual human being. This paper is developed from a descriptive and interdisciplinary study of key ethical concepts in this subject. 1. Respect for the embryo's dignity must indeed exist from the beginning of its life. 2. For this reason, this respect does not depend on any operation, but on the eminence of its being. 3. Not only is freezing a habitat which contradicts the dignity of life, but it is also the expression of a will that determines and decides the human life of weaker beings. The embryo's dignity is thus reduced to the value of use where it is not out of date.

  17. Embryo aggregation does not improve the development of interspecies somatic cell nuclear transfer embryos in the horse.

    Science.gov (United States)

    Gambini, Andrés; De Stéfano, Adrián; Jarazo, Javier; Buemo, Carla; Karlanian, Florencia; Salamone, Daniel Felipe

    2016-09-01

    The low efficiency of interspecies somatic cell nuclear transfer (iSCNT) makes it necessary to investigate new strategies to improve embryonic developmental competence. Embryo aggregation has been successfully applied to improve cloning efficiency in mammals, but it remains unclear whether it could also be beneficial for iSCNT. In this study, we first compared the effect of embryo aggregation over in vitro development and blastocyst quality of porcine, bovine, and feline zona-free (ZF) parthenogenetic (PA) embryos to test the effects of embryo aggregation on species that were later used as enucleated oocytes donors in our iSCNT study. We then assessed whether embryo aggregation could improve the in vitro development of ZF equine iSCNT embryos after reconstruction with porcine, bovine, and feline ooplasm. Bovine- and porcine-aggregated PA blastocysts had significantly larger diameters compared with nonaggregated embryos. On the other hand, feline- and bovine-aggregated PA embryos had higher blastocyst cell number. Embryo aggregation of equine-equine SCNT was found to be beneficial for embryo development as we have previously reported, but the aggregation of three ZF reconstructed embryos did not improve embryo developmental rates on iSCNT. In vitro embryo development of nonaggregated iSCNT was predominantly arrested around the stage when transcriptional activation of the embryonic genome is reported to start on the embryo of the donor species. Nevertheless, independent of embryo aggregation, equine blastocyst-like structures could be obtained in our study using domestic feline-enucleated oocytes. Taken together, these results reported that embryo aggregation enhance in vitro PA embryo development and embryo quality but effects vary depending on the species. Embryo aggregation also improves, as expected, the in vitro embryo development of equine-equine SCNT embryos; however, we did not observe positive effects on equine iSCNT embryo development. Among oocytes

  18. [Chapter 9. The embryo in comparative law].

    Science.gov (United States)

    Mastor, Wanda

    2018-03-07

    On the boundaries of life and, as a result, almost a question of metaphysics, still dividing science and continually fuelling debates, one question does seem to be legally insoluble, ie the question of the status of the human embryo. A comparatist look allows us to put into perspective the various national postures with regard to the embryo in order to confront them, by putting forward the areas where they converge or diverge. Although a very global approach allows us to note certain similarities, a more precise study of the question of abortion in particular reflects the evidence of the contextualisation of the embryo. It is what it is, subject or object, enjoying absolute or very relative protection, a simply legislative or constitutional status, only with regard to legal systems, but also moral and religious systems in which it takes its place.

  19. Fixation of Ilyanassa snail embryos and larvae.

    Science.gov (United States)

    Gharbiah, Maey; Cooley, James; Leise, Esther M; Nakamoto, Ayaki; Rabinowitz, Jeremy S; Lambert, J David; Nagy, Lisa M

    2009-04-01

    The marine gastropod Ilyanassa obsoleta is a long-standing and very useful model for studies of embryonic development. It is an especially important model for spiralian development, and for studies of asymmetric cell division. The embryos are amenable to classic embryological manipulation techniques, as well as a growing number of molecular approaches. Ilyanassa is also an important model for studies of metamorphosis, the ecology of parasitism, the effects of environmental contaminants on morphology and sexual function, and comparative neurobiology. Ilyanassa embryos are particularly well suited for RNA and protein localization studies because of the relatively large cells and favorable properties for imaging. This protocol describes how to fix and store Ilyanassa embryos and larvae for use in whole-mount in situ hybridization and immunohistochemical studies.

  20. Learning to segment mouse embryo cells

    Science.gov (United States)

    León, Juan; Pardo, Alejandro; Arbeláez, Pablo

    2017-11-01

    Recent advances in microscopy enable the capture of temporal sequences during cell development stages. However, the study of such sequences is a complex task and time consuming task. In this paper we propose an automatic strategy to adders the problem of semantic and instance segmentation of mouse embryos using NYU's Mouse Embryo Tracking Database. We obtain our instance proposals as refined predictions from the generalized hough transform, using prior knowledge of the embryo's locations and their current cell stage. We use two main approaches to learn the priors: Hand crafted features and automatic learned features. Our strategy increases the baseline jaccard index from 0.12 up to 0.24 using hand crafted features and 0.28 by using automatic learned ones.

  1. Survival of embryo irradiated with gamma rays by embryo culture in Brassica pekinensis Rupr

    International Nuclear Information System (INIS)

    Moue, T.

    1984-01-01

    The effect of irradiation on the survival rates and embryonic development of Brassica pekinensis RUPR. (Varieties; Kashin, Kohai 65 nichi and kairyochitose) was investigated. The purpose of this study was to seek ways of increasing the survival rates of embryos such as B.oleracea obtained through embryo culture techniques after irradiation doses affecting seed fertility and germination, for the purpose of increasing mutation rates. Embryos at different developmental stages ranging from the globular to the early heart stages were irradiated with 20 KR of gamma rays at the daily rate 0L 20 KR or 10 KR (Fig.1 and Table 1). The embryos were excised from ovules 4 to 10 days after irradiation and cultured on White's medium. The shooting and rooting rates on the 34th day of culture were higher at the dose of 10 KR/day than 20 KR/day and were lower when the materials were irradiated at the young embryonic stage (Table 3). Varietal differences in the shooting and rooting rates were also observed. The irradiated embryos survived mainly in the state of callus. It was concluded that the embryo culture technique was successful when applied to irradiated embryos excised at the young embryonic stage and that the technique affected B.pekinensis less than B.oleracea

  2. RECYCLING OF CATHETER FOR EMBRYO RECOVERY: A TOOL FOR COSTS REDUCTION IN EQUINE EMBRYO TRANSFER

    Directory of Open Access Journals (Sweden)

    Alberto Lopes Gusmao

    2010-06-01

    Full Text Available The embryo transfer is becoming a widespread practice.Most embryos are collected from spontaneous single ovulatingmares and result in 50% of embryo recovery, increasing the costsof production. To illustrate, the price of a catheter for embryosrecovery range from US$ 194.00 to US$ 250.00 (R$ 350.00 to R$450.00. Therefore, the aim of this work was to verify if catheterwith damaged balloon can be recuperated and reused withoutaltering its efficiency. For this study, two groups were used: acontrol group (GI, n=10, on which the nonsurgical recovery of theembryos of mares was performed with the catheter with originalballoon; and another group (GII, n=20, in which a restored catheterwas utilized. The mares of GI had an embryo recovery rate of60%, and GII mares had an embryo recovery rate of 55%. Therewas not statistical difference between groups I and II (P>0.05.Considering that the material used to restore the catheter costsUS$16.66 (R$30.00, this data show that the recuperation of thecatheters for embryo recovery in mares may reduce costs withoutcompromising the rates of embryo recovery.

  3. Arabidopsis mitochondrial protein slow embryo development1 is essential for embryo development

    Energy Technology Data Exchange (ETDEWEB)

    Ju, Yan; Liu, Chunying; Lu, Wenwen; Zhang, Quan; Sodmergen, E-mail: sodmergn@pku.edu.cn

    2016-05-27

    The plant seeds formation are crucial parts in reproductive process in seed plants as well as food source for humans. Proper embryo development ensure viable seed formation. Here, we showed an Arabidopsis T-DNA insertion mutant slow embryo development1 (sed1) which exhibited retarded embryogenesis, led to aborted seeds. Embryo without SED1 developed slower compared to normal one and could be recognized at early globular stage by its white appearance. In later development stage, storage accumulated poorly with less protein and lipid body production. In vitro culture did not rescue albino embryo. SED1 encoded a protein targeted to mitochondria. Transmission electron microscopic analysis revealed that mitochondria developed abnormally, and more strikingly plastid failed to construct grana in time in sed1/sed1 embryo. These data indicated that SED1 is indispensable for embryogenesis in Arabidopsis, and the mitochondria may be involved in the regulation of many aspects of seed development. -- Highlights: •Arabidopsis SED1 is essential for embryo development. •The sed1 embryo accumulates less storage and has abnormal ultrastructure. •SED1 localizes to the mitochondrion.

  4. Arabidopsis mitochondrial protein slow embryo development1 is essential for embryo development

    International Nuclear Information System (INIS)

    Ju, Yan; Liu, Chunying; Lu, Wenwen; Zhang, Quan; Sodmergen

    2016-01-01

    The plant seeds formation are crucial parts in reproductive process in seed plants as well as food source for humans. Proper embryo development ensure viable seed formation. Here, we showed an Arabidopsis T-DNA insertion mutant slow embryo development1 (sed1) which exhibited retarded embryogenesis, led to aborted seeds. Embryo without SED1 developed slower compared to normal one and could be recognized at early globular stage by its white appearance. In later development stage, storage accumulated poorly with less protein and lipid body production. In vitro culture did not rescue albino embryo. SED1 encoded a protein targeted to mitochondria. Transmission electron microscopic analysis revealed that mitochondria developed abnormally, and more strikingly plastid failed to construct grana in time in sed1/sed1 embryo. These data indicated that SED1 is indispensable for embryogenesis in Arabidopsis, and the mitochondria may be involved in the regulation of many aspects of seed development. -- Highlights: •Arabidopsis SED1 is essential for embryo development. •The sed1 embryo accumulates less storage and has abnormal ultrastructure. •SED1 localizes to the mitochondrion.

  5. In vitro manipulation techniques of porcine embryos

    DEFF Research Database (Denmark)

    Liu, Ying; Li, Juan; Løvendahl, Peter

    2015-01-01

    to analyse data collected from all published articles with a focus on zygotes and embryos for transfer, pregnancy, full-term development and piglets born. It was generally concluded that an increasing level of in vitro manipulation of porcine embryos decreased the overall efficiency for production of piglets...... higher litter size. More complete information is needed in future scientific articles about these in vitro manipulation techniques to establish a more solid basis for the evaluation of their status and to reveal and further investigate any eventual problems...

  6. Incomplete methylation reprogramming in SCNT embryos.

    Science.gov (United States)

    Peat, Julian R; Reik, Wolf

    2012-09-01

    The cloning of Dolly the sheep was a remarkable demonstration of the oocyte's ability to reprogram a specialized nucleus. However, embryos derived from such somatic cell nuclear transfer (SCNT) very rarely result in live births-a fate that may be linked to observed epigenetic defects. A new genome-wide study shows that epigenetic reprogramming in SCNT embryos does not fully recapitulate the natural DNA demethylation events occurring at fertilization, resulting in aberrant methylation at some promoters and repetitive elements that may contribute to developmental failure.

  7. Using fertile couples as embryo donors: An ethical dilemma.

    Science.gov (United States)

    Alizadeh, Leila; Omani Samani, Reza

    2014-03-01

    The use of donated embryos has offered hope for infertile couples who have no other means to have children. In Iran, fertility centers use fertile couples as embryo donors. In this paper, the advantages and disadvantages of this procedure will be discussed. We conclude that embryo-donation should be performed with frozen embryos thus preventing healthy donors from being harmed by fertility drugs. There must be guidelines for choosing the appropriate donor families. In countries where commercial egg donation is acceptable, fertile couples can be procured as embryo donors thus fulfilling the possible shortage of good quality embryos. Using frozen embryos seems to have less ethical, religious and legal problems when compared to the use of fertile embryo donors.

  8. Cryopreservation of embryos and oocytes in human assisted reproduction.

    Science.gov (United States)

    Konc, János; Kanyó, Katalin; Kriston, Rita; Somoskői, Bence; Cseh, Sándor

    2014-01-01

    Both sperm and embryo cryopreservation have become routine procedures in human assisted reproduction and oocyte cryopreservation is being introduced into clinical practice and is getting more and more widely used. Embryo cryopreservation has decreased the number of fresh embryo transfers and maximized the effectiveness of the IVF cycle. The data shows that women who had transfers of fresh and frozen embryos obtained 8% additional births by using their cryopreserved embryos. Oocyte cryopreservation offers more advantages compared to embryo freezing, such as fertility preservation in women at risk of losing fertility due to oncological treatment or chronic disease, egg donation, and postponing childbirth, and eliminates religious and/or other ethical, legal, and moral concerns of embryo freezing. In this review, the basic principles, methodology, and practical experiences as well as safety and other aspects concerning slow cooling and ultrarapid cooling (vitrification) of human embryos and oocytes are summarized.

  9. Cryopreservation of Embryos and Oocytes in Human Assisted Reproduction

    Directory of Open Access Journals (Sweden)

    János Konc

    2014-01-01

    Full Text Available Both sperm and embryo cryopreservation have become routine procedures in human assisted reproduction and oocyte cryopreservation is being introduced into clinical practice and is getting more and more widely used. Embryo cryopreservation has decreased the number of fresh embryo transfers and maximized the effectiveness of the IVF cycle. The data shows that women who had transfers of fresh and frozen embryos obtained 8% additional births by using their cryopreserved embryos. Oocyte cryopreservation offers more advantages compared to embryo freezing, such as fertility preservation in women at risk of losing fertility due to oncological treatment or chronic disease, egg donation, and postponing childbirth, and eliminates religious and/or other ethical, legal, and moral concerns of embryo freezing. In this review, the basic principles, methodology, and practical experiences as well as safety and other aspects concerning slow cooling and ultrarapid cooling (vitrification of human embryos and oocytes are summarized.

  10. Omics as a window to view embryo viability.

    Science.gov (United States)

    Krisher, Rebecca L; Schoolcraft, William B; Katz-Jaffe, Mandy G

    2015-02-01

    The advent of advanced omics technologies and the application of these techniques to the analysis of extremely limited material have opened the door to the investigation of embryo physiology in a focused, in-depth approach never before possible. The application of noninvasive metabolomic and proteomic platforms to understanding embryo viability permits the characterization of individual embryos in culture. Initial clinical data have highlighted the promise of these technologies for the development of noninvasive embryo selection criteria. In this way, a complex view of embryo function can be compiled and related to embryo development, quality, and outcome. Application of knowledge gained from omics will transform both our understanding of embryo physiology as well as our ability to select viable embryos for transfer in assisted reproductive technology. Copyright © 2015 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  11. Developmental competence of porcine chimeric embryos produced by aggregation

    DEFF Research Database (Denmark)

    Li, Juan; Jakobsen, Jannik E.; Xiong, Qiang

    2015-01-01

    either by parthenogenetic activation (PA) or handmade cloning (HMC). Results showed that the developmental competence of chimeric embryos, evaluated based on their blastocyst rate and total cell number per blastocyst, was increased when two whole 2-cell stage embryos (PA or HMC) were aggregated....... In comparison, when two blastomeres were aggregated, the developmental competence of the chimeric embryos decreased if the blastomeres were either from PA or from HMC embryos, but not if they were from different sources, i.e. one PA and one HMC blastomere. To evaluate the cell contribution in embryo formation......, aggregation was made with HMC embryos cloned using EGFP transgenic cells; the cell contribution in the formation of the inner cell mass or trophectoderm was random in chimeric blastocysts. Finally, two blastomeres from 2-cell stage embryos were fused to construct tetraploid embryos, and when diploid...

  12. Sourcing human embryos for embryonic stem cell lines: Problems & perspectives

    Directory of Open Access Journals (Sweden)

    Rajvi H Mehta

    2014-01-01

    Full Text Available The ability to successfully derive human embryonic stem cells (hESC lines from human embryos following in vitro fertilization (IVF opened up a plethora of potential applications of this technique. These cell lines could have been successfully used to increase our understanding of human developmental biology, transplantation medicine and the emerging science of regenerative medicine. The main source for human embryos has been ′discarded′ or ′spare′ fresh or frozen human embryos following IVF. It is a common practice to stimulate the ovaries of women undergoing any of the assisted reproductive technologies (ART and retrieve multiple oocytes which subsequently lead to multiple embryos. Of these, only two or maximum of three embryos are transferred while the rest are cryopreserved as per the decision of the couple. In case a couple does not desire to ′cryopreserve′ their embryos then all the embryos remaining following embryo transfer can be considered ′spare′ or if a couple is no longer in need of the ′cryopreserved′ embryos then these also can be considered as ′spare′. But, the question raised by the ethicists is, "what about ′slightly′ over-stimulating a woman to get a few extra eggs and embryos? The decision becomes more difficult when it comes to ′discarded′ embryos. As of today, the quality of the embryos is primarily assessed based on morphology and the rate of development mainly judged by single point assessment. Despite many criteria described in the literature, the quality assessment is purely subjective. The question that arises is on the decision of ′discarding′ embryos. What would be the criteria for discarding embryos and the potential ′use′ of ESC derived from the ′abnormal appearing′ embryos? This paper discusses some of the newer methods to procure embryos for the derivation of embryonic stem cell lines which will respect the ethical concerns but still provide the source material.

  13. Cryopreservation of somatic embryos of paradise tree (Melia azedarach L.)

    OpenAIRE

    Scocchi, A.; Vila, S.; Mroginski, L.; Engelmann, Florent

    2007-01-01

    In paradise tree (Melia azedarach L.), immature zygotic embryos sampled from immature fruits are the starting material for the production of somatic embryos. These somatic embryos are employed for freezing experiments. Immature fruits could be stored at 25 degrees C for up to 80 days without impairing the embryogenic potential of zygotic embryos, which represents a four-fold increase in immature fruit storage duration, compared with previous studies. Among the three cryopreservation technique...

  14. Embryo transfer and related technologies in sheep reproduction

    OpenAIRE

    Loi, Pasqualino; Ptak, Grazyna; Dattena, Maria; Ledda, Sergio; Naitana, Salvatore; Cappai, Pietro

    1998-01-01

    This paper reviews the status of embryo transfer and the major technologies applied to preimplantation of embryos in sheep. Embryo production from superovulated ewes is hindered by an unpredictable response to hormonal treatment. Progress in this area should be expected by an appropriated control of follicular development with gonadotropin-releasing hormone (GnRH) agonist or antagonist prior to gonadotrophin administration. Simple protocols for the cryopreservation of sheep embryos by vitrifi...

  15. Sourcing human embryos for embryonic stem cell lines: problems & perspectives.

    Science.gov (United States)

    Mehta, Rajvi H

    2014-11-01

    The ability to successfully derive human embryonic stem cells (hESC) lines from human embryos following in vitro fertilization (IVF) opened up a plethora of potential applications of this technique. These cell lines could have been successfully used to increase our understanding of human developmental biology, transplantation medicine and the emerging science of regenerative medicine. The main source for human embryos has been 'discarded' or 'spare' fresh or frozen human embryos following IVF. It is a common practice to stimulate the ovaries of women undergoing any of the assisted reproductive technologies (ART) and retrieve multiple oocytes which subsequently lead to multiple embryos. Of these, only two or maximum of three embryos are transferred while the rest are cryopreserved as per the decision of the couple. in case a couple does not desire to 'cryopreserve' their embryos then all the embryos remaining following embryo transfer can be considered 'spare' or if a couple is no longer in need of the 'cryopreserved' embryos then these also can be considered as 'spare'. But, the question raised by the ethicists is, "what about 'slightly' over-stimulating a woman to get a few extra eggs and embryos? The decision becomes more difficult when it comes to 'discarded' embryos. As of today, the quality of the embryos is primarily assessed based on morphology and the rate of development mainly judged by single point assessment. Despite many criteria described in the literature, the quality assessment is purely subjective. The question that arises is on the decision of 'discarding' embryos. What would be the criteria for discarding embryos and the potential 'use' of ESC derived from the 'abnormal appearing' embryos? This paper discusses some of the newer methods to procure embryos for the derivation of embryonic stem cell lines which will respect the ethical concerns but still provide the source material.

  16. Precocious germination and its regulation in embryos of triticale caryopses

    OpenAIRE

    Stanisław Weidner

    2014-01-01

    Triticale var. Lasko embryos, isolated from grain gathered at milk ripeness, the beginning of wax ripeness and at full ripeness, were allowed to germinate for 48 h on agar with glucose. The highest incorporation of tritiated adenosine into polyribosomal RNA during germination was found in the ribosome fractions from embryos of grain gathered at full ripeness, lower incorporation was in preparations from embryos of milk ripe grain and the lowest in preparations from embryos of wax ripe grain. ...

  17. Sexing bovine pre-implantation embryos using the polymerase ...

    African Journals Online (AJOL)

    Yomi

    2012-03-06

    Mar 6, 2012 ... The paper aims to present a bovine model for human embryo sexing. Cows were super-ovulated, artificially inseminated and embryos were recovered 7 days later. Embryo biopsy was performed; DNA was extracted from blastomeres and amplified using bovine-specific and bovine-Y-chromosome-.

  18. Correlation between embryo morphology and development and chromosomal complement

    Directory of Open Access Journals (Sweden)

    Vy Phan

    2014-06-01

    Conclusions: Embryo development rate and morphological parameter such as degree, type of fragmentation and the symmetry of the blastomeres to a large extent reflect the cytogenetic status of the embryo and thus are important in the selection of embryos with the highest implantation potential.

  19. The development of ovary in quail's embryo | Rong | African Journal ...

    African Journals Online (AJOL)

    The experiment was conducted to study the development of ovary in quails' embryos which were incubated for 4 to 17 days and incubated out for 1 day. The quails' embryos or gonads were cut out and HE staining was carried out. The results showed that when embryo was hatched for 4 days, lots of primordial germ cells ...

  20. Sexing bovine pre-implantation embryos using the polymerase ...

    African Journals Online (AJOL)

    The paper aims to present a bovine model for human embryo sexing. Cows were super-ovulated, artificially inseminated and embryos were recovered 7 days later. Embryo biopsy was performed; DNA was extracted from blastomeres and amplified using bovine-specific and bovine-Y-chromosomespecific primers, followed ...

  1. Debating Elective Single Embryo Transfer after in vitro Fertilization ...

    African Journals Online (AJOL)

    transfer (eSET) to reduce the risk of multiple gestation and. Debating Elective Single Embryo Transfer ... the American Society for Reproductive Medicine, elective SET is defined as “the transfer of a single embryo at ... or should they transfer just one embryo, which reduces the risk of complications and is in line with current ...

  2. Bovine in-vitro embryo production and its contribution towards ...

    African Journals Online (AJOL)

    Bovine in-vitro embryo production and its contribution towards improved food security in Kenya. ... African Journal of Food, Agriculture, Nutrition and Development ... be low valued, it can be raised to higher reproductive potential using current reproductive technologies such as in-vitro embryo production and embryo transfer.

  3. Mouse embryos cultured in amniotic fluid | Oettle | South African ...

    African Journals Online (AJOL)

    The embryos were cultured for 72 hours at 37°C in 5% carbon dioxide in air. Osmolarity, pH, partial arterial carbon dioxide pressure and HCO; were checked before culture and again at the end of culture. Embryos were assessed according to the stage of development, namely degenerate embryos, morulae and blastocysts.

  4. What Drives Embryo Development? Chromosomal Normality or Mitochondria?

    Directory of Open Access Journals (Sweden)

    A. Bayram

    2017-01-01

    Full Text Available Objective. To report the arrest of euploid embryos with high mtDNA content. Design. A report of 2 cases. Setting. Private fertility clinic. Patients. 2 patients, 45 and 40 years old undergoing IVF treatment. Interventions. Mature oocytes were collected and vitrified from two ovarian stimulations. Postthaw, survived mature oocytes underwent fertilization by intracytoplasmic sperm injection (ICSI. Preimplantation genetic screening (PGS and mitochondrial DNA (mtDNA copy number were done using next generation sequencing (NGS. The only normal embryo among the all-biopsied embryos had the highest “Mitoscore” value and was the only arrested embryo in both cases. Therefore, the embryo transfer was cancelled. Main Outcome Measures. Postthaw survival and fertilization rate, embryo euploidy, mtDNA copy number, and embryo development. Results. In both patients, after PGS only 1 embryo was euploid. Both embryos had the highest mtDNA copy number from all tested embryos and both embryos were arrested on further development. Conclusions. These cases clearly demonstrate the lack of correlation between mtDNA value (Mitoscore and chromosomal status of embryo.

  5. Mouse embryos cultured In amnIotIc fluid

    African Journals Online (AJOL)

    Mouse embryos are used as a monitor of human in vicro fertilisation (IVF) culture conditions. Embryo culture media are often complicated solutions requiring careful monitoring to ensure uniformity for successful embryo culture.1-3 Much of the quality control is outside the scope of the IVF laboratory, for example the source of ...

  6. Ultrastructural changes in goat interspecies and intraspecies reconstructed early embryos

    DEFF Research Database (Denmark)

    Tao, Yong; Gheng, Lizi; Zhang, Meiling

    2008-01-01

    The low efficiency of somatic cell nuclear transfer may be related to the ultrastructural deviations of reconstructed embryos. The present study investigated ultrastructural differences between in vivo-produced and cloned goat embryos, including intra- and interspecies embryos. Goat ear fibroblas...

  7. Xenopus laevis embryos and tadpoles as models for testing for ...

    African Journals Online (AJOL)

    The toxicity of bio available Zn, Cu, Pb, and Cd on the life stages of Xenopus laevis embryos and tadpoles was investigated. Cu and Cd were found to affect the hatching success of the embryos, with a strong negative relationship existing between the increase in Cu concentrations and the hatching of the embryos.

  8. Superovulation and embryo recovery in Boer goats treated with ...

    African Journals Online (AJOL)

    Lehloenya

    These pregnancy rates obtained were also indicative of an acceptable procedure of embryo transfer used in this trial. The differences obtained could also be attributed to the stage of development of the embryos, as in this trial blastocysts were used. In most trials transferable embryos (morulae and blastocysts) were used.

  9. 9 CFR 98.20 - Embryos refused entry.

    Science.gov (United States)

    2010-01-01

    ... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Embryos refused entry. 98.20 Section...-and-Mouth Disease Exists § 98.20 Embryos refused entry. If any embryos are determined to be ineligible for importation into the United States upon arrival at the port of entry, the importer must remove the...

  10. Acephalous lamb from an in vitro-produced sheep embryo

    OpenAIRE

    Shirazi, Abolfazl; Ahmadi, Ebrahim; Jadidi, Majid; Shams-Esfandabadi, Naser; Heidari, Banafsheh

    2009-01-01

    This is the first report of an acephalous lamb from the transfer of an in vitro-produced sheep embryo. Twelve in vitro-fertilized embryos were transferred to 4 recipient ewes (3 embryos/ewe). Two ewes remained pregnant: one delivered a normal female lamb, the other a male acephalous lamb. Possible contributing factors are discussed.

  11. Plant regeneration in wheat mature embryo culture

    African Journals Online (AJOL)

    Kamil Haliloğlu

    2011-11-09

    Nov 9, 2011 ... medium and same amount (8 mg/l) of 2,4-D and dicamba, respectively except for non-endosperm sup- ported mature embryos and hormones, there was no plant regeneration in method #12 in which dicamba was used as auxine. Filippov et al. (2006) obtained the best plant regeneration in 10 mg/l doses of ...

  12. Mouse embryo manipulations with OCT guidance

    Science.gov (United States)

    Garcia, Monica D.; Syed, Saba H.; Coughlin, Andrew J.; Wang, Shang; West, Jennifer L.; Larin, Kirill V.; Larina, Irina V.

    2014-03-01

    Optical coherence tomography (OCT) is a three-dimensional, non-invasive optical imaging technique that relies on low-coherence interferometry. OCT has the capability of imaging 2 - 3 mm into tissue, which enables imaging of deeper structures within the embryo with a relatively high spatial resolution (2 - 15μm). Within the past decade, OCT has been increasingly used as a live imaging tool for embryonic cardiovascular research in several animal models. Research in our lab has recently shown that OCT can be used in combination with embryo culture for the visualization of early mammalian cardiovascular development (E7.5 - E10.0). Here, we demonstrate that OCT can be used for the guided microinjection of gold-silica nanoshell suspension into the cardiovascular system in live embryos without deleterious effect. This approach shows a promising application for the OCT guided delivery of contrast agents, viral vectors, therapeutic or pharmacological agents, signaling molecules or dyes to specific organ systems or tissues in live embryos and demonstrates a great potential for gold-silica nanoshells as a contrast agent in embryonic studies.

  13. Interspecies embryo reconstruction in Tibetan antelope Pantholops ...

    African Journals Online (AJOL)

    user

    2011-03-21

    Mar 21, 2011 ... improves the development of interspecies reconstructed giant panda. (Aluropoda melanoleuca) embryos. Chin. Sci. Bull. 47: 467-469. Li Y, Dai Y, Du W, Zhao C, Wang H, Wang L, Li R, Liu Y, Wan R, Li N. (2006). Cloned endangered species takin (Budorcas taxicolor) by inter-species nuclear transfer and ...

  14. [Chapter 3. Governing the research on embryos].

    Science.gov (United States)

    Bioy, Xavier; Rial-Sebbag, Emmanuelle

    2018-03-07

    The bioethics laws since their first adoption in 1994 until the latest proposals, have been widely commented and analyzed, regarding the provisions related to the legal qualification and the legal status of the embryo and of the embryonic stem cells. The legal issues raised by these hesitations imply instability for both researchers and health professionals about what they can and cannot do when it comes to the embryo and its cells. Thus, while the protection of the embryo was the subject of a consensus until the end of the 2000s, it now appears as the object of a political will, from researchers and state agencies, to support this research. The legal frameworks have been modified step by step leading to an administrative police that reconciles freedom of research and ethical issues whose legal enforcement remains weak and uncertain. It will therefore be important to highlight the legal and institutional milestones that led to the acceptance of embryo research and to the liberalization of the legal framework. Some doubts will then be expressed about the benefits of this liberalization notably because cases law on the patentability of inventions resulting from this research are a bit of a threat to encourage development perspectives and because embryonic stem cells could remain marginal because of the economic and scientific contexts.

  15. [Chapter 2. The embryo in abortion legislation].

    Science.gov (United States)

    Vigneau, Daniel

    2018-03-07

    For a long time, the legal condition of the child conceived, the embryo or the human fœtus, to use the language of biomedicine, was dominated by two main principles : one, having its roots in Roman law, made it possible to count it, by anticipation, among beings with a legal existence, so as to grant it its rights, as if it was already born (the infans conceptus maxim) : the other, protecting its life in utero, and so its chances of being born, by the penal incrimination of abortion. The legalisation of abortion by the ?Veil? law of 17 January 1975 upset this traditional approach of the law. The subsequent evolution of the legislation has accentuated the phenomenon. The promotion of the freedom of women to obtain an abortion has been accompanied by a correlated decline in the protection of the embryo. At the same time, abortion resting on medical techniques has become a means, precious for research, to obtain embryo or foetal cells or tissues. Eliminated on the one hand, the embryo has finally become on the other hand the object of medical and scientific stakes.

  16. Effects of fluoxetine on human embryo development

    NARCIS (Netherlands)

    Kaihola, Helena; Yaldir, Fatma G.; Hreinsson, Julius; Hornaeus, Katarina; Bergquist, Jonas; Olivier, Jocelien D. A.; Akerud, Helena; Sundstrom-Poromaa, Inger

    2016-01-01

    The use of antidepressant treatment during pregnancy is increasing, and selective serotonin reuptake inhibitors (SSRIs) are the most widely prescribed antidepressants in pregnant women. Serotonin plays a role in embryogenesis, and serotonin transporters are expressed in two-cell mouse embryos. Thus,

  17. Transcriptome analysis of Anopheles stephensi embryo using ...

    Indian Academy of Sciences (India)

    2013-04-18

    Apr 18, 2013 ... SignalP/) (Nielsen et al. 1997). All ESTs are submitted in NCBI GenBank database ..... 2001). Expression of these genes hint at conserved pathways of epigenetic regulation associated with. GBR stage embryos. Developmental timing of gene expres- sion is regulated by a set of evolutionarily conserved.

  18. Mapping selection within Drosophila melanogaster embryo's anatomy

    DEFF Research Database (Denmark)

    Salvador-Martínez, Irepan; Coronado-Zamora, Marta; Castellano, David

    2018-01-01

    We present a survey of selection across Drosophila melanogaster embryonic anatomy. Our approach integrates genomic variation, spatial gene expression patterns and development, with the aim of mapping adaptation over the entire embryo's anatomy. Our adaptation map is based on analyzing spatial gen...

  19. Genetic transformation of olive somatic embryos through ...

    African Journals Online (AJOL)

    Administrator

    2011-06-20

    Jun 20, 2011 ... 2Department of Biochemistry, National Center of Genetic Engineering and Biotechnology, Tehran, Iran. Accepted 9 March, 2011. Transformed olive plants were regenerated from inoculated somatic embryos with Agrobacterium tumefacience strain GV3101, which carries the plasmid pBI-P5CS containing ...

  20. The fate of the mosaic embryo: Chromosomal constitution and development of Day 4, 5 and 8 human embryos

    NARCIS (Netherlands)

    M.A. Santos; G. Teklenburg (Gijs); N.S. Macklon (Nick); D. van Opstal (Diane); G.H. Schuring-Blom (Heleen); P-J. Krijtenburg (Pieter-Jaap); J. de Vreeden-Elbertse (Johanna); B.C.J.M. Fauser (Bart); E.B. Baart (Esther)

    2010-01-01

    textabstractBackground: Post-zygotic chromosome segregation errors are very common in human embryos after in vitro fertilization, resulting in mosaic embryos. However, the significance of mosaicism for the developmental potential of early embryos is unknown. We assessed chromosomal constitution and

  1. Effect of embryo density on in vitro developmental characteristics of bovine preimplantative embryos with respect to micro and macroenvironments.

    Science.gov (United States)

    Hoelker, M; Rings, F; Lund, Q; Phatsara, C; Schellander, K; Tesfaye, D

    2010-10-01

    To overcome developmental problems as a consequence of single embryo culture, the Well of the Well (WOW) culture system has been developed. In this study, we aimed to examine the effect of embryo densities with respect to both microenvironment and macroenvironment on developmental rates and embryo quality to get a deeper insight into developmentally important mechanisms. WOW diameter and depth significantly affected developmental rates (p WOWs with diameter of 500 μm reached significantly higher blastocyst rates (32.5 vs 21.1% vs 20.3%) compared to embryos cultured in WOWs of 300 μm diameter or plain cultured controls. Embryos cultured in WOWs with 700 μm depth reached significant higher developmental rates compared with embryos cultured in WOWs of 300 μm depth and control embryos (30.6 vs 22.6% vs 20.3%). Correlation of the embryo per WOW volume with developmental rates was higher (r(2) = 0.92, p = 0.0004) than correlation of WOW diameter or WOW depth with developmental rates. However, the embryo per WOW volume did not affect differential cell counts. An embryo per culture dish volume of 1 : 30 μl was identified to be optimal when the embryo per WOW volume was 1 : 0.27 μl increasing developmental rates up to the level of mass embryo production. Giving the opportunity to track each embryo over the complete culture period while keeping high developmental rates with normal mitotic dynamics, the results of this work will provide benefit for the single culture of embryos in human assisted reproduction, mammalian embryos with high economic interest as well as for scientific purpose. © 2009 Blackwell Verlag GmbH.

  2. Somatic Embryos in Catharanthus roseus: A Scanning Electron Microscopic Study

    Directory of Open Access Journals (Sweden)

    Junaid ASLAM

    2014-06-01

    Full Text Available Catharanthus roseus (L. G. Don is an important medicinal plant as it contains several anti-cancerous compounds, like vinblastine and vincristine. Plant tissue culture technology (organogenesis and embryogenesis has currently been used in fast mass propagating raw materials for secondary metabolite synthesis. In this present communication, scanning electron microscopic (SEM study of somatic embryos was conducted and discussed. The embryogenic callus was first induced from hypocotyls of in vitro germinated seeds on which somatic embryos, differentiated in numbers, particularly on 2,4-D (1.0 mg/L Murashige and Skoog (MS was medium. To understand more about the regeneration method and in vitro formed embryos SEM was performed. The SEM study revealed normal somatic embryo origin and development from globular to heart-, torpedo- and then into cotyledonary-stage of embryos. At early stage, the embryos were clustered together in a callus mass and could not easily be detached from the parental tissue. The embryos were often long cylindrical structure with or without typical notch at the tip. Secondary embryos were also formed on primary embryo structure. The advanced cotyledonary embryos showed prominent roots and shoot axis, which germinated into plantlets. The morphology, structure and other details of somatic embryos at various stages were presented.

  3. Evaluation of somatic embryos of alfalfa for recombinant protein expression.

    Science.gov (United States)

    Fu, Guohua; Grbic, Vojislava; Ma, Shengwu; Tian, Lining

    2015-02-01

    Somatic embryos of alfalfa can accumulate higher levels of recombinant proteins comparing to vegetative organs. Somatic embryos may be explored as a new system for new protein production for plants. Plants have been explored via genetic engineering as an inexpensive system for recombinant protein production. However, protein expression levels in vegetative tissues have been low, which limits the commercial utilization of plant expression systems. Somatic embryos resemble zygotic embryos in many aspects and may accumulate higher levels of proteins as true seed. In this study, somatic embryo of alfalfa (Medicago sativa L.) was investigated for the expression of recombinant proteins. Three heterologous genes, including the standard scientific reporter uid that codes for β-glucuronidase and two genes of interest: ctb coding for cholera toxin B subunit (CTB), and hIL-13 coding for human interleukin 13, were independently introduced into alfalfa via Agrobacterium-mediated transformation. Somatic embryos were subsequently induced from transgenic plants carrying these genes. Somatic embryos accumulated approximately twofold more recombinant proteins than vegetative organs including roots, stems, and leaves. The recombinant proteins of CTB and hIL-13 accumulated up to 0.15 and 0.18 % of total soluble protein in alfalfa somatic embryos, respectively. The recombinant proteins expressed in somatic embryos also exhibited biological activities. As somatic embryos can be induced in many plant species and their production can be scaled up via different avenues, somatic embryos may be developed as an efficient expression system for recombinant protein production.

  4. Comprehensive embryo testing. Experts' opinions regarding future directions: an expert panel study on comprehensive embryo testing.

    Science.gov (United States)

    Hens, Kristien; Dondorp, Wybo J; Geraedts, Joep P M; de Wert, Guido M

    2013-05-01

    What do scientists in the field of preimplantation genetic diagnosis (PGD) and preimplantation genetic screening (PGS) consider to be the future direction of comprehensive embryo testing? Although there are many biological and technical limitations, as well as uncertainties regarding the meaning of genetic variation, comprehensive embryo testing will impact the IVF/PGD practice and a timely ethical reflection is needed. Comprehensive testing using microarrays is currently being introduced in the context of PGD and PGS, and it is to be expected that whole-genome sequencing will also follow. Current ethical and empirical sociological research on embryo testing focuses on PGD as it is practiced now. However, empirical research and systematic reflection regarding the impact of comprehensive techniques for embryo testing is missing. In order to understand the potential of this technology and to be able to adequately foresee its implications, we held an expert panel with seven pioneers in PGD. We conducted an expert panel in October 2011 with seven PGD pioneers from Belgium, The Netherlands, Germany and the UK. Participants expected the use of comprehensive techniques in the context of PGD. However, the introduction of these techniques in embryo testing requires timely ethical reflection as it involves a shift from choosing an embryo without a particular genetic disease (i.e. PGD) or most likely to result in a successful pregnancy (i.e. PGS) to choosing the best embryo based on a much wider set of criteria. Such ethical reflection should take account of current technical and biological limitations and also of current uncertainties with regard to the meaning of genetic variance. However, ethicists should also not be afraid to look into the future. There was a general agreement that embryo testing will be increasingly preceded by comprehensive preconception screening, thus enabling smart combinations of genetic testing. The group was composed of seven participants from

  5. Classification of embryo sacs in the Eragrostis curvula Complex

    Directory of Open Access Journals (Sweden)

    T. B. Vorster

    1984-12-01

    Full Text Available At each of 17 collecting points between Johannesburg and Brits in the Transvaal, three plants which belong to the  Eragrostis curvula Complex were collected and studied. A total o f 3 902 embryo sacs was examined in this sample. Of the embryo sacs examined, 3 306 were apomictic by means of diplospory, whereas 99 were sexual monosporic Polygonum-type embryo sacs. One hundred and nineteen embryo sacs were abnormal or divergent, and 378 were degenerated. There are indications that seasonal climatic fluctuations may be responsible for embryo sacs developing abnormally or degenerating. Simple and multiple correlations confirmed that sexual embryo sacs usually do not develop abnormally or degenerate during the later developmental stages. This finding lends credence to both the system of classification of individual embryo sacs and to the validity of the estimate of the proportion of sexuality of the plants sampled at each sampling point.

  6. Tissue densities in developing avian embryos. [under acceleration stresses

    Science.gov (United States)

    Smith, A. H.; Abbott, U. K.; Morzenti, A.

    1984-01-01

    The density changes in the components of the incubated egg, the embryo, and the embryo's body parts were measured in the course of 21 days of incubation. In the first two-thirds of the incubation period there is a sequence of increasing density among egg contents: amniotic fluid, embryo, yolk, and albumin. As a result, the embryo is located at the bottom of the amniotic fluid, but at the top of the albumin. This position provides the embryo with mechanical protection and a proximity to the egg's air cell. The observed density changes and the asymmetry of these changes among various body parts of the embryo suggest a functional relationship. The density distributions among the body parts are particularly important in gravitational investigations of embryogenesis since they will produce forces tending to dislocate parts of the embryo.

  7. Assessing embryo development using swept source optical coherence tomography

    Science.gov (United States)

    Caujolle, S.; Cernat, R.; Silvestri, G.; Marques, M. J.; Bradu, A.; Feuchter, T.; Robinson, G.; Griffin, D.; Podoleanu, A.

    2018-03-01

    A detailed assessment of embryo development would assist biologists with selecting the most suitable embryos for transfer leading to higher pregnancy rates. Currently, only low resolution microscopy is employed to perform this assessment. Although this method delivers some information on the embryo surface morphology, no specific details are shown related to its inner structure. Using a Master-Slave Swept-Source Optical Coherence Tomography (SS-OCT), images of bovine embryos from day 7 after fertilization were collected from different depths. The dynamic changes inside the embryos were examined, in detail and in real-time from several depths. To prove our ability to characterize the morphology, a single embryo was imaged over 26 hours. The embryo was deprived of its life support environment, leading to its death. Over this period, clear morphological changes were observed.

  8. Effect of supplementation of different growth factors in embryo culture medium with a small number of bovine embryos on in vitro embryo development and quality.

    Science.gov (United States)

    Ahumada, C J; Salvador, I; Cebrian-Serrano, A; Lopera, R; Silvestre, M A

    2013-03-01

    When embryos are cultured individually or in small groups, blastocyst yield efficiency and quality are usually reduced. The aim of this work was to investigate the effect of supplementation of the embryo culture medium (CM) with several growth factors (GFs) on embryo development and apoptosis rate when a reduced number of embryos were in vitro cultured. Two experimental studies (ES) were carried out. In ES 1, five treatments were tested to study the effect of GF on embryo development: Control (∼30 to 50 embryos cultured in 500 μl of CM); Control 5 (Five embryos cultured in 50 μl microdrops of CM), without addition of GF in either of the two control groups; epidermal GF (EGF); IGF-I; and transforming GF-α (TGF-α) (Five embryos were cultured in 50 μl microdrops of CM with 10 ng/ml EGF, 10 ng/ml IGF-I or 10 ng/ml TGF-α, respectively). In ES 2, following the results obtained in ES 1, four different treatments were tested to study their effect on embryo development and quality (number of cells per blastocyst and apoptotic rate): Control; Control 5; EGF, all three similar to ES 1; EGF + IGF-I group (five embryos cultured in 50 μl microdrops of CM with 10 ng/ml EGF and 10 ng/ml IGF-I). In both ESs, it was observed that a higher proportion of embryos cultured in larger groups achieved blastocyst stage than embryos cultured in reduced groups (22.6% v. 14.0%, 12.6% and 5.3% for Control v. Control 5, IGF-I, TGF-α groups in ES 1, and 24.9% v. 17.1% and 19.0% for Control v. Control 5 and EGF in ES 2, respectively; P embryos cultured in medium supplemented with EGF (18.5%) or with EGF + IGF-I (23.5%), in ES 1 and ES 2, respectively. With regard to blastocyst quality, embryos cultured in reduced groups and supplemented with EGF, alone or combined with IGF-I, presented lower apoptosis rates than embryos cultured in reduced groups without GF supplementation (11.6% and 10.5% v. 21.9% for EGF, EGF + IGF-I and Control 5 groups, respectively; P embryos, increasing blastocyst

  9. Microfluidic EmbryoSort technology: towards in flow analysis, sorting and dispensing of individual vertebrate embryos

    Science.gov (United States)

    Fuad, Nurul M.; Wlodkowic, Donald

    2013-12-01

    The demand to reduce the numbers of laboratory animals has facilitated the emergence of surrogate models such as tests performed on zebrafish (Danio rerio) or African clawed frog's (Xenopus levis) eggs, embryos and larvae. Those two model organisms are becoming increasingly popular replacements to current adult animal testing in toxicology, ecotoxicology and also in drug discovery. Zebrafish eggs and embryos are particularly attractive for toxicological analysis due their size (diameter 1.6 mm), optical transparency, large numbers generated per fish and very straightforward husbandry. The current bottleneck in using zebrafish embryos for screening purposes is, however, a tedious manual evaluation to confirm the fertilization status and subsequent dispensing of single developing embryos to multitier plates to perform toxicity analysis. Manual procedures associated with sorting hundreds of embryos are very monotonous and as such prone to significant analytical errors due to operator's fatigue. In this work, we present a proofof- concept design of a continuous flow embryo sorter capable of analyzing, sorting and dispensing objects ranging in size from 1.5 - 2.5 mm. The prototypes were fabricated in polymethyl methacrylate (PMMA) transparent thermoplastic using infrared laser micromachining. The application of additive manufacturing processes to prototype Lab-on-a-Chip sorters using both fused deposition manufacturing (FDM) and stereolithography (SLA) were also explored. The operation of the device was based on a revolving receptacle capable of receiving, holding and positioning single fish embryos for both interrogation and subsequent sorting. The actuation of the revolving receptacle was performed using a DC motor and/or microservo motor. The system was designed to separate between fertilized (LIVE) and non-fertilized (DEAD) eggs, based on optical transparency using infrared (IR) emitters and receivers.

  10. Cholinoreceptors of early (preneural) sea urchin embryos.

    Science.gov (United States)

    Buznikov, G A; Rakich, L

    2000-01-01

    Agonists of nicotinic cholinoreceptors (n-AChR) and 1-acetyl-4-methylpiperazine (100 microM) had no effect on early embryogenesis in sea urchins, while in the presence of phorbol-12-myristate-13-acetate (PMA) and various other protein kinase C activators, these agents induced rapid lysis of oocytes or early embryos, as a result of calcium shock. Many n-AChR ligands which do not penetrate into the cytoplasm (not being antagonists of muscarinic cholinoreceptors) protected against this cytotoxic effect. In the presence of PMA, acetylcholine and carbachol had actions which were much weaker than those of nicotine, while muscarine was completely inactive in these conditions. Thus, the surfaces of sea urchin oocytes and early embryos bear receptor structures, presumably n-AChR, which are functionally linked with second messengers which are endogenous protein kinase C activators.

  11. Cells, embryos and development in space

    Science.gov (United States)

    Krikorian, A. D.

    1984-01-01

    Work continues to focus on the demonstrable totipotency of cultured somatic cells of various higher plants and has examined the conditions which regulate this propensity to be controllably released. This was done with special reference to cells obtained from cultured explants of daylily and carrot. For purposes of identifying the variables in question, work was carried out almost exclusively in liquid media. The events that intervene between the aseptic isolation of tissue explants, the culture of small derived units and free cells and the propagation in large numbers of adventive or somatic embryos to plantlets were traced and certain definitive stages at which control is exercised were identified. In daylily, morphologically competent units are now propagated with a high degree of precision in rotated liquid cultures in bulk, and under the conditions of continuous neutralized gravity, the development progresses so that embryo-plantlets are obtained.

  12. The use of morphokinetic parameters to select all embryos with full capacity to implant

    OpenAIRE

    Chamayou, Sandrine; Patrizio, Pasquale; Storaci, Giorgia; Tomaselli, Venera; Alecci, Carmelita; Ragolia, Carmen; Crescenzo, Claudia; Guglielmino, Antonino

    2013-01-01

    Purpose Embryo kinetics analysis is an emerging tool for selecting embryo(s) for transfer. The aim of the present study was to determine morphokinetic parameters easily usable in the laboratory and predictive of embryo development and, most importantly, of embryo competence in producing a clinical pregnancy after day 5 transfer. Methods A retrospective time-lapse monitoring analysis of morphokinetic parameters for 72 fully implanted embryos (group A) were compared to 106 non-implanted embryos...

  13. DDT-induced feminization of gull embryos

    International Nuclear Information System (INIS)

    Fry, D.M.; Toone, C.K.

    1981-01-01

    Injection of DDT [1, 1, 1-trichloro-2,2-bis(p-chlorophenyl)ethane] into gull eggs at concentrations comparable to those found in contaminated seabird eggs in 1970 induces abnormal development of ovarian tissue and oviducts in male embryos. Developmental feminization of males is associated with inability to breed as adults and may explain the highly skewed sex ratio and reduced number of male gulls breeding on Santa Barbara Island in southern California

  14. Developmental toxicity of cartap on zebrafish embryos.

    Science.gov (United States)

    Zhou, Shengli; Dong, Qiaoxiang; Li, Shaonan; Guo, Jiangfeng; Wang, Xingxing; Zhu, Guonian

    2009-12-13

    Cartap is a widely used insecticide which belongs to a member of nereistoxin derivatives and acts on nicotinic acetylcholine receptor site. Its effects on aquatic species are of grave concern. To explore the potential developmental toxicity of cartap, zebrafish embryos were continually exposed, from 0.5 to 144h post-fertilization, to a range of concentrations of 25-1000microg/l. Results of the experiment indicated that cartap concentrations of 100microg/l and above negatively affected embryo survival and hatching success. Morphological analysis uncovered a large suite of abnormalities such as less melanin pigmentation, wavy notochord, crooked trunk, fuzzy somites, neurogenesis defects and vasculature defects. The most sensitive organ was proved to be the notochord which displayed defects at concentrations as low as 25microg/l. Both sensitivity towards exposure and localization of the defect were stage specific. To elucidate mechanisms concerning notochord, pigmentation, and hatching defects, enzyme assay, RT Q-PCR, and different exposure strategies were performed. For embryos with hatching failure, chorion was verified not to be digested, while removing cartap from exposure at early pre-hatching stage could significantly increase the hatching success. However, cartap was proved, via vitro assay, to have no effect on proteolytic activity of hatching enzyme. These findings implied that the secretion of hatching enzyme might be blocked. We also revealed that cartap inhibited the activity of melanogenic enzyme tyrosinase and matrix enzyme lysyl oxidase and induced expression of their genes. These suggested that cartap could impaired melanin pigmentation of zebrafish embryos through inhibiting tyrosinase activity, while inhibition of lysyl oxidase activity was responsible for notochord undulation, which subsequently caused somite defect, and at least partially responsible for defects in vasculature and neurogenesis.

  15. Role of melatonin in embryo fetal development.

    Science.gov (United States)

    Voiculescu, S E; Zygouropoulos, N; Zahiu, C D; Zagrean, A M

    2014-01-01

    Melatonin is an indoleamine produced by the pineal gland and secreted in a circadian manner. In the past few decades, research over this topic has been enhanced. Melatonin has many important roles in the human physiology: regulator of the circadian rhythms, sleep inducer, antioxidant, anticarcinogenic. This paper reviews the involvement of melatonin in embryo fetal development. The pineal gland develops completely postpartum, so both the embryo and the fetus are dependent on the maternal melatonin provided transplacentally. Melatonin appears to be involved in the normal outcome of pregnancy beginning with the oocyte quality and finishing with the parturition. Its pregnancy night-time concentrations increase after 24 weeks of gestation, with significantly high levels after 32 weeks. Melatonin receptors are widespread in the embryo and fetus since early stages. There is solid evidence that melatonin is neuroprotective and has a positive effect on the outcome of the compromised pregnancies. In addition, chronodisruption leads to a reproductive dysfunction. Thus, the influence of melatonin on the developing human fetus may not be limited to the entertaining of circadian rhythmicity, but further studies are needed.

  16. Human embryo cloning prohibited in Hong Kong.

    Science.gov (United States)

    Liu, Athena

    2005-12-01

    Since the birth of Dolly (the cloned sheep) in 1997, debates have arisen on the ethical and legal questions of cloning-for-biomedical-research (more commonly termed "therapeutic cloning") and of reproductive cloning using human gametes. Hong Kong enacted the Human Reproductive Technology Ordinance (Cap 561) in 2000. Section 15(1)(e) of this Ordinance prohibits the "replacing of the nucleus of a cell of an embryo with a nucleus taken from any other cell," i.e., nucleus substitution. Section 15(1)(f) prohibits the cloning of any embryo. The scope of the latter, therefore, is arguably the widest, prohibiting all cloning techniques such as cell nucleus replacement, embryo splitting, parthenogenesis, and cloning using stem cell lines. Although the Human Reproductive Technology Ordinance is not yet fully operative, this article examines how these prohibitions may adversely impact on basic research and the vision of the Hong Kong scientific community. It concludes that in light of recent scientific developments, it is time to review if the law offers a coherent set of policies in this area.

  17. Addition of ascorbate during cryopreservation stimulates subsequent embryo development.

    Science.gov (United States)

    Lane, Michelle; Maybach, Jeffery M; Gardner, David K

    2002-10-01

    Embryo development following cryopreservation is reduced compared with fresh embryos. One of the traumas that cryopreservation imparts on embryos is an increase in oxidative stress. Therefore, this study investigated the effects of the addition of the antioxidant ascorbate to the cryopreservation solutions on subsequent embryo development. Mouse embryos at the 2-cell and blastocyst stages were either slow-frozen or vitrified in solutions containing either no ascorbate or 0.1 or 0.5 mmol/l ascorbate. The effects on the levels of hydrogen peroxide and subsequent embryo development and physiology were assessed. Addition of ascorbate to the cryopreservation solutions reduced the levels of hydrogen peroxide in embryos. Furthermore, addition of 0.1 mmol/l ascorbate significantly enhanced inner cell mass development in blastocysts. Embryos cryopreserved with ascorbate had significantly lower levels of lactate dehydrogenase leakage, and increased rates of metabolism compared with those cryopreserved in the absence of ascorbate. The benefits of ascorbate were significantly greater in embryos that were slow-frozen compared with those that were vitrified. These data indicate that the addition of 0.1 mmol/l ascorbate to the cryopreservation solutions for the mammalian embryo would be of significant value.

  18. Parents' conceptualization of their frozen embryos complicates the disposition decision

    Science.gov (United States)

    Nachtigall, Robert D.; Becker, Gay; Friese, Carrie; Butler, Anneliese; MacDougall, Kirstin

    2009-01-01

    Objective To ascertain what couples think about their embryos and how they approach making a decision about disposition in light of the fact that the disposition of unused frozen embryos has significant implications for medical research and embryo donation. Design Ethnographic qualitative interview study. Setting Academic research environment. Patient(s) Fifty-eight couples who had conceived using a donor oocyte and had at least one frozen embryo in storage. Main Outcome Measure(s) Tape-recorded interviews with 58 wives and 37 husbands were transcribed and analyzed for emergent themes. Result(s) With an average of 7.1 embryos per couple, after an average of 4.2 years of storage, 72% of couples with frozen embryos had not reached a disposition decision. Most couples had not anticipated or appreciated the consequences of having surplus embryos. Parents variously conceptualized frozen embryos as biologic tissue, living entities, “virtual” children having interests that must be considered and protected, siblings of their living children, genetic or psychological “insurance policies,” and symbolic reminders of their past infertility. Conclusion(s) The disposition decision is not only a significant and frequently unresolved issue for couples with stored frozen embryos, but their deeply personal conceptualizations of their embryos contributes to their ambivalence, uncertainty, and difficulty in reaching a decision. PMID:16084886

  19. Cryopreservation of preimplantation embryos of cattle, sheep, and goats.

    Science.gov (United States)

    Youngs, Curtis R

    2011-08-05

    Preimplantation embryos from cattle, sheep, and goats may be cryopreserved for short- or long-term storage. Preimplantation embryos consist predominantly of water, and the avoidance of intracellular ice crystal formation during the cryopreservation process is of paramount importance to maintain embryo viability. Embryos are placed into a hypertonic solution (1.4 - 1.5 M) of a cryoprotective agent (CPA) such as ethylene glycol (EG) or glycerol (GLYC) to create an osmotic gradient that facilitates cellular dehydration. After embryos reach osmotic equilibrium in the CPA solution, they are individually loaded in the hypertonic CPA solution into 0.25 ml plastic straws for freezing. Embryos are placed into a controlled rate freezer at a temperature of -6°C. Ice crystal formation is induced in the CPA solution surrounding the embryo, and crystallization causes an increase in the concentration of CPA outside of the embryo, causing further cellular dehydration. Embryos are cooled at a rate of 0.5°C/min, enabling further dehydration, to a temperature of -34°C before being plunged into liquid nitrogen (-196°C). Cryopreserved embryos must be thawed prior to transfer to a recipient (surrogate) female. Straws containing the embryos are removed from the liquid nitrogen dewar, held in room temperature air for 3 to 5 sec, and placed into a 37°C water bath for 25 to 30 sec. Embryos cryopreserved in GLYC are placed into a 1 M solution of sucrose for 10 min for removal of the CPA before transfer to a recipient (surrogate) female. Embryos cryopreserved in EG, however, may be directly transferred to the uterus of a recipient.

  20. Preimplantation development of embryos in women of advanced maternal age

    Directory of Open Access Journals (Sweden)

    O. V. Chaplia

    2014-04-01

    Full Text Available In order to reveal the influence of genetic component on the early embryo development, the retrospective study of morphokinetic characteristics of 717 embryos subjected to preimplantation genetic testing was conducted. Blastomere biopsy for FISH-based preimplantation genetic screening of 7 chromosomes was performed on the third day of culture, while embryo developmental potential and morphological features at the cleavage and blastulation stage were studied regarding maternal age particularly in the group of younger women and patients older than 36. Results of genetic testing revealed that euploid embryos rate gradually decreased with maternal age comprising 39.9% in young women group and 25.3% of specimen belonging to elder patients. At the cleavage stage, morphological characteristics of aneuploid and euploid embryos didn’t differ significantly regardless of the age of patients that could be accounted for the transcriptional silence of embryo genome till the third day of its development. However, in case of prolonged culture chromosomally balanced embryos rarely faced developmental arrest (in 7.9% and formed blastocysts half more frequently compared to aberrant embryos (respectively 75.6 versus 49.8%. Nevertheless, no substantial difference was found between blastocyst formation rate among embryos with similar genetic component regardless of the maternal age. Taking into consideration high rate of chromosomally unbalanced embryos specific to patients of advanced maternal age, the relative proportion of aneuplouid blastocysts was significantly higher in this group of embryos. Thus, without genetic screening there is a possibility of inaccurate selection of embryos for women of advanced reproductive age for transfer procedure even in case of prolonged culture. Consequently, increase of aneuploid embryos frequency associated with permanent preimplantation natural selection effectiveness along with the postimplantation natural selection failure

  1. Cryopreservation of Preimplantation Embryos of Cattle, Sheep, and Goats

    Science.gov (United States)

    Youngs, Curtis R.

    2011-01-01

    Preimplantation embryos from cattle, sheep, and goats may be cryopreserved for short- or long-term storage. Preimplantation embryos consist predominantly of water, and the avoidance of intracellular ice crystal formation during the cryopreservation process is of paramount importance to maintain embryo viability. Embryos are placed into a hypertonic solution (1.4 – 1.5 M) of a cryoprotective agent (CPA) such as ethylene glycol (EG) or glycerol (GLYC) to create an osmotic gradient that facilitates cellular dehydration. After embryos reach osmotic equilibrium in the CPA solution, they are individually loaded in the hypertonic CPA solution into 0.25 ml plastic straws for freezing. Embryos are placed into a controlled rate freezer at a temperature of -6°C. Ice crystal formation is induced in the CPA solution surrounding the embryo, and crystallization causes an increase in the concentration of CPA outside of the embryo, causing further cellular dehydration. Embryos are cooled at a rate of 0.5°C/min, enabling further dehydration, to a temperature of -34°C before being plunged into liquid nitrogen (-196°C). Cryopreserved embryos must be thawed prior to transfer to a recipient (surrogate) female. Straws containing the embryos are removed from the liquid nitrogen dewar, held in room temperature air for 3 to 5 sec, and placed into a 37°C water bath for 25 to 30 sec. Embryos cryopreserved in GLYC are placed into a 1 M solution of sucrose for 10 min for removal of the CPA before transfer to a recipient (surrogate) female. Embryos cryopreserved in EG, however, may be directly transferred to the uterus of a recipient. PMID:21847080

  2. Embryo quality and impact of specific embryo characteristics on ongoing implantation in unselected embryos derived from modified natural cycle in vitro fertilization

    NARCIS (Netherlands)

    Pelinck, Marie-Jose; Hoek, Annemieke; Simons, Arnold H. M.; Heineman, Maas Jan; van Echten-Arends, Janny; Arts, Eus G. J. M.

    Objective: To study the implantation potential of unselected embryos derived from modified natural cycle IVF according to their morphological characteristics. Design: Cohort study. Setting: Academic department of reproductive medicine. Patient(S): A series of 449 single embryo transfers derived from

  3. Can Chlamydia abortus be transmitted by embryo transfer in goats?

    Science.gov (United States)

    Oseikria, M; Pellerin, J L; Rodolakis, A; Vorimore, F; Laroucau, K; Bruyas, J F; Roux, C; Michaud, S; Larrat, M; Fieni, F

    2016-10-01

    The objectives of this study were to determine (i) whether Chlamydia abortus would adhere to or penetrate the intact zona pellucida (ZP-intact) of early in vivo-derived caprine embryos, after in vitro infection; and (ii) the efficacy of the International Embryo Transfer Society (IETS) washing protocol for bovine embryos. Fifty-two ZP-intact embryos (8-16 cells), obtained from 14 donors were used in this experiment. The embryos were randomly divided into 12 batches. Nine batches (ZP-intact) of five embryos were incubated in a medium containing 4 × 10(7)Chlamydia/mL of AB7 strain. After incubation for 18 hours at 37 °C in an atmosphere of 5% CO2, the embryos were washed in batches in 10 successive baths of a phosphate buffer saline and 5% fetal calf serum solution in accordance with IETS guidelines. In parallel, three batches of ZP-intact embryos were used as controls by being subjected to similar procedures but without exposure to C. abortus. The 10 wash baths were collected separately and centrifuged for 1 hour at 13,000 × g. The washed embryos and the pellets of the 10 centrifuged wash baths were frozen at -20 °C before examination for evidence of C. abortus using polymerase chain reaction. C. abortus DNA was found in all of the infected batches of ZP-intact embryos (9/9) after 10 successive washes. It was also detected in the 10th wash fluid for seven batches of embryos, whereas for the two other batches, the last positive wash bath was the eighth and the ninth, respectively. In contrast, none of the embryos or their washing fluids in the control batches were DNA positive. These results report that C. abortus adheres to and/or penetrates the ZP of in vivo caprine embryos after in vitro infection, and that the standard washing protocol recommended by the IETS for bovine embryos, failed to remove it. The persistence of these bacteria after washing makes the embryo a potential means of transmission of the bacterium during embryo transfer from

  4. In vitro culture of individual mouse preimplantation embryos: the role of embryo density, microwells, oxygen, timing and conditioned media.

    Science.gov (United States)

    Kelley, Rebecca L; Gardner, David K

    2017-05-01

    Single embryo culture is suboptimal compared with group culture, but necessary for embryo monitoring, and culture systems should be improved for single embryos. Pronucleate mouse embryos were used to assess the effect of culture conditions on single embryo development. Single culture either before or after compaction reduced cell numbers (112.2 ± 3.1; 110.2 ± 3.5) compared with group culture throughout (127.0 ± 3.4; P media volume from 20 µl to 2 µl increased blastocyst cell numbers in single embryos cultured in 5% oxygen (84.4 ± 3.2 versus 97.8 ± 2.8; P Culture in microwell plates for the EmbryoScope and Primo Vision time-lapse systems changed cleavage timings and increased inner cell mass cell number (24.1 ± 1.0; 23.4 ± 1.2) compared with a 2 µl microdrop (18.4 ± 1.0; P media to single embryos increased hatching rate and blastocyst cell number (91.5 ± 4.7 versus 113.1 ± 4.4; P culture before or after compaction is therefore detrimental; oxygen, media volume and microwells influence single embryo development; and embryo-conditioned media may substitute for group culture. Copyright © 2017 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

  5. Comparison between Conventional Blind Embryo Transfer and Embryo Transfer Based on Previously Measured Uterine Length

    Directory of Open Access Journals (Sweden)

    Nasrin Saharkhiz

    2014-11-01

    Full Text Available Background: Embryo transfer (ET is one of the most important steps in assisted reproductive technology (ART cycles and affected by many factors namely the depth of embryo deposition in uterus. In this study, the outcomes of intracytoplasmic sperm injection (ICSI cycles after blind embryo transfer and embryo transfer based on previously measured uterine length using vaginal ultrasound were compared. Materials and Methods: This prospective randomised clinical trial included one hundred and forty non-donor fresh embryo transfers during January 2010 to June 2011. In group I, ET was performed using conventional (blind method at 5-6cm from the external os, and in group II, ET was done at a depth of 1-1.5 cm from the uterine fundus based on previously measured uterine length using vaginal sonography. Appropriate statistical analysis was performed using Student’s t test and Chi-square or Fisher’s exact test. The software that we used was PASW statistics version 18. A p value <0.05 was considered statistically significant. Results: Chemical pregnancy rate was 28.7% in group I and 42.1% in group II, while the difference was not statistically significant (p=0.105. Clinical pregnancy, ongoing pregnancy and implantation rates for group I were 21.2%, 17.7%, and 12.8%, while for group II were 33.9%, 33.9%, and 22.1, respectively. In group I and group II, abortion rates were 34.7% and 0%, respectively, indicating a statistically significant difference (p<0.005. No ectopic pregnancy occurred in two groups. Conclusion: The use of uterine length measurement during treatment cycle in order to place embryos at depth of 1-1.5cm from fundus significantly increases clinical and ongoing pregnancy and implantation rates, while leads to a decrease in abortion rate (Registration Number: IRCT2014032512494N1.

  6. Effects of embryo-derived exosomes on the development of bovine cloned embryos.

    Science.gov (United States)

    Qu, Pengxiang; Qing, Suzhu; Liu, Ruiqi; Qin, Hongyu; Wang, Weiwei; Qiao, Fang; Ge, Hui; Liu, Jun; Zhang, Yong; Cui, Wei; Wang, Yongsheng

    2017-01-01

    The developmental competence of in vitro cultured (IVC) embryos is markedly lower than that of their in vivo counterparts, suggesting the need for optimization of IVC protocols. Embryo culture medium is routinely replaced three days after initial culture in bovine, however, whether this protocol is superior to continuous nonrenewal culture method under current conditions remains unclear. Using bovine somatic cell nuclear transfer (SCNT) embryos as the model, our results showed that compared with routine renewal treatment, nonrenewal culture system significantly improved blastocyst formation, blastocyst quality (increased total cell number, decreased stress and apoptosis, enhanced Oct-4 expression and ratio of ICM/TE), as well as following development to term. Existence and function of SCNT embryo-derived exosomes were then investigated to reveal the cause of impaired development induced by culture medium replacement. Exosomes were successfully isolated through differential centrifugation and identified by both electron microscopy and immunostaining against exosomal membrane marker CD9. Supplementation of extracted exosomes into freshly renewed medium significantly rescued not only blastocyst formation and quality (in vitro development), but also following growth to term (in vivo development). Notably, ratio of ICM/TE and calving rate were enhanced to a similar level as that in nonrenewal group. In conclusion, our results for the first time indicate that 1: bovine SCNT embryos can secrete exosomes into chemically defined culture medium during IVC; 2: secreted exosomes are essential for SCNT blastocyst formation, blastocyst quality, and following development to term; 3: removal of exosomes induced by culture medium replacement impairs SCNT embryo development, which can be avoided by nonrenewal culture procedure or markedly recovered by exosome supplementation.

  7. Vibrational properties characterization of mouse embryo during microinjection

    Directory of Open Access Journals (Sweden)

    Hedrih Anđelka N.

    2013-01-01

    Full Text Available To determine the vibration characteristics (natural frequencies and mode shapes of a mouse embryo during microinjection the modal analysis is used. The spherical mouse embryo 60 μm in diameter is modeled as elastic finite elements biostructure consisting of 6μm thick micromembrane and 38 μm in diameter nucleus. Embryo modeling and modal analysis were based on the use of the finite elements method in the modal analysis system of ANSYS software. The modal analysis was carried out for first six modes of embryo natural frequencies. The numerical analysis of dependence of embryo own frequencies on the boundary conditions and external loads are presented. The relevant illustrations of the typical variations of the shape, deformation and particle velocities of vibrating embryo are given. [Projekat Ministarstva nauke Republike Srbije, br. OI174001: Dynamics of hybrid systems with complex structures: Mechanics of materials

  8. Digital microfluidic processing of mammalian embryos for vitrification.

    Science.gov (United States)

    Pyne, Derek G; Liu, Jun; Abdelgawad, Mohamed; Sun, Yu

    2014-01-01

    Cryopreservation is a key technology in biology and clinical practice. This paper presents a digital microfluidic device that automates sample preparation for mammalian embryo vitrification. Individual micro droplets manipulated on the microfluidic device were used as micro-vessels to transport a single mouse embryo through a complete vitrification procedure. Advantages of this approach, compared to manual operation and channel-based microfluidic vitrification, include automated operation, cryoprotectant concentration gradient generation, and feasibility of loading and retrieval of embryos.

  9. Digital microfluidic processing of mammalian embryos for vitrification.

    Directory of Open Access Journals (Sweden)

    Derek G Pyne

    Full Text Available Cryopreservation is a key technology in biology and clinical practice. This paper presents a digital microfluidic device that automates sample preparation for mammalian embryo vitrification. Individual micro droplets manipulated on the microfluidic device were used as micro-vessels to transport a single mouse embryo through a complete vitrification procedure. Advantages of this approach, compared to manual operation and channel-based microfluidic vitrification, include automated operation, cryoprotectant concentration gradient generation, and feasibility of loading and retrieval of embryos.

  10. Efficient Production and Cellular Characterization of Sheep Androgenetic Embryos

    OpenAIRE

    Zacchini, Federica; Czernik, Marta; Iuso, Domenico; Toschi, Paola; di Egidio, Fiorella; Scapolo, Pier Augusto; Loi, Pasqualino; Ptak, Grazyna

    2011-01-01

    The production of androgenetic embryos in large animals is a complex procedure. Androgenetic embryos have been produced so far only in cattle and sheep using pronuclear transfer (PT) between zygotes derived from in vitro fertilization (IVF) of previously enucleated oocytes. PT is required due to the poor developmental potential of androgenotes derived from IVF of enucleated oocytes. Here we compare the developemt to blastocyst of androgenetic embryos produced by the standard pronuclear transf...

  11. Cryopreservation of somatic embryos of paradise tree (Melia azedarach L.).

    Science.gov (United States)

    Scocchi, Adriana; Vila, Silvia; Mroginski, Luis; Engelmann, Florent

    2007-01-01

    In paradise tree (Melia azedarach L.), immature zygotic embryos sampled from immature fruits are the starting material for the production of somatic embryos. These somatic embryos are employed for freezing experiments. Immature fruits could be stored at 25 degrees C for up to 80 days without impairing the embryogenic potential of zygotic embryos, which represents a four-fold increase in immature fruit storage duration, compared with previous studies. Among the three cryopreservation techniques tested for freezing paradise tree somatic embryos, namely desiccation, encapsulation-dehydration and pregrowth-dehydration, only encapsulation-dehydration and pregrowth-dehydration led to successful results. The optimal protocol was the following: i) somatic embryos (encapsulated or not) pretreated in liquid Murashige & Skoog medium with daily increasing sucrose concentration (0.5 M/0.75 M/1.0 M); ii) dehydrated with silica gel to 21 - 26% moisture content (fresh weight basis), for encapsulation-dehydration, or to 19% moisture content, for pregrowth-dehydration; iii) frozen at 1 degree C/min from 20 degrees C to -30 degrees C with a programmable freezing apparatus; iv) rapid immersion in liquid nitrogen. The highest recovery achieved was 36% with encapsulation-dehydration and 30% with pregrowth-dehydration. Regrowth of frozen embryos was direct in most cases, as secondary embryogenesis originating from the root pole was observed on only around 10% of cryopreserved somatic embryos. Plants recovered from cryopreserved embryos presented the same phenotypic traits as non-frozen control plants.

  12. Cooling strategies for brazilian flounder Paralichthys orbignyanus embryos.

    Science.gov (United States)

    Varela, A S; Cardoso, T F; Fernandes E Silva, E; Goularte, K L; Okamoto, M H; Sampaio, L A; Jardim, R D; Corcini, C D

    Paralichthys orbignyanus is the species of the greatest potential for marine and estuarine fish farming in southern Brazil. Consequently, embryo cryopreservation becomes an important tool for increasing their production. To evaluate the effects of cooling protocols on the viability of embryos of P. orbignyanus at two stages of development (neurula and early differentiation of the tail). Control embryos were maintained at 23 degree C and treated embryos were cooled to 15 degree C, 10 degree C and 5 degree C at rapid, moderate and slow cooling rates. Then embryos were maintained at these different temperatures for 30, 60 and 90 min and the loss of viability assessed as hatching rates (HR) and morphologically normal larvae (MNL). The average HR for embryos following cooling was higher for those at the tail stage compared to the neurula stage (Pcooling. Also for tail stage embryos, there was no difference between MNL of control and rapidly cooled embryos. As first steps in the development of cryopreservation methods for P. orbignyanus embryos, the use of a rapid cooling and holding at 5 degree C for 30 min are recommended.

  13. Ultrastructural studies of Biomphalaria glabrata (Say, 1818) embryo

    International Nuclear Information System (INIS)

    Kikuchi, O.K.; Okazaki, K.; Kawano, T.; Ribeiro, A.A.G.F.C.

    1988-09-01

    Ultrastructural studies of Biomphalaria glabrata embryos (MOllusca: Gastropoda), and important snail vector of schistosomiasis has not been explored. In the present work it was evaluated a suitable electron microscopical technique for embryos processing. Promising results was obtained with double fixation in 1% glutaraldehyde plus 1% osmium tetroxide in 0.05 M cacodylate buffer (pH 7.4), preliminary staining overnight in 1% uranyl acetate and embedding in EPON or Polylite under vacuum. It was used embryos at young trochophore stage wich is characterized by active organogenesis. Some ultrastructural aspects of B. glabrata embryos cells are presented. (author) [pt

  14. Obtaining barley haploid embryos and seedlings using anther culture technique

    International Nuclear Information System (INIS)

    Arabi, M.I.E.; Al-Safadi, B.; Mir Ali, N.

    2000-01-01

    The effect of three barley genotypes (Igri, Arabi abiad, and Taqa 76), three irradiation doses (0, 5, and 10 Gy), and two media (FW, modified FW), on the number of formed embryos, and the ratio between regenerated embryos to green seedlings and albinos, were studied using anther culture. Also the study involved the compatibility between seedling morphology and chromosome number. results indicated significant differences among the genotypes, and media in callus and embryos formation and also in the ratio and albino seedlings. However, the effect of gamma rays dose was significant only on embryos regeneration. A high percentage of compatibility (90%) was obtained between the seedling morphology and chromosome number. (author)

  15. The impact of preimplantation genetic diagnosis on human embryos

    Directory of Open Access Journals (Sweden)

    García-Ferreyra J.

    2016-12-01

    Full Text Available Chromosome abnormalities are extremely common in human oocytes and embryos and are associated with a variety of negative outcomes for both natural cycles and those using assisted reproduction techniques. Aneuploidies embryos may fail to implant in the uterus, miscarry, or lead to children with serious medical problems (e.g., Down syndrome. Preimplantation genetic diagnosis (PGD is a technique that allows the detection of aneuploidy in embryos and seeks to improve the clinical outcomes od assisted reproduction treatments, by ensuring that the embryos chosen for the transfer are chromosomally normal.

  16. Embryo fossilization is a biological process mediated by microbial biofilms.

    Science.gov (United States)

    Raff, Elizabeth C; Schollaert, Kaila L; Nelson, David E; Donoghue, Philip C J; Thomas, Ceri-Wyn; Turner, F Rudolf; Stein, Barry D; Dong, Xiping; Bengtson, Stefan; Huldtgren, Therese; Stampanoni, Marco; Chongyu, Yin; Raff, Rudolf A

    2008-12-09

    Fossilized embryos with extraordinary cellular preservation appear in the Late Neoproterozoic and Cambrian, coincident with the appearance of animal body fossils. It has been hypothesized that microbial processes are responsible for preservation and mineralization of organic tissues. However, the actions of microbes in preservation of embryos have not been demonstrated experimentally. Here, we show that bacterial biofilms assemble rapidly in dead marine embryos and form remarkable pseudomorphs in which the bacterial biofilm replaces and exquisitely models details of cellular organization and structure. The experimental model was the decay of cleavage stage embryos similar in size and morphology to fossil embryos. The data show that embryo preservation takes place in 3 distinct steps: (i) blockage of autolysis by reducing or anaerobic conditions, (ii) rapid formation of microbial biofilms that consume the embryo but form a replica that retains cell organization and morphology, and (iii) bacterially catalyzed mineralization. Major bacterial taxa in embryo decay biofilms were identified by using 16S rDNA sequencing. Decay processes were similar in different taphonomic conditions, but the composition of bacterial populations depended on specific conditions. Experimental taphonomy generates preservation states similar to those in fossil embryos. The data show how fossilization of soft tissues in sediments can be mediated by bacterial replacement and mineralization, providing a foundation for experimentally creating biofilms from defined microbial species to model fossilization as a biological process.

  17. Cryopreservation of Byrsonima intermedia embryos followed by room temperature thawing

    Directory of Open Access Journals (Sweden)

    Luciano Coutinho Silva

    2014-07-01

    Full Text Available Byrsonima intermedia is a shrub from the Brazilian Cerrado with medicinal properties. The storage of biological material at ultra-low temperatures (-196°C is termed cryopreservation and represents a promising technique for preserving plant diversity. Thawing is a crucial step that follows cryopreservation. The aim of this work was to cryopreserve B. intermedia zygotic embryos and subsequently thaw them at room temperature in a solution rich in sucrose. The embryos were decontaminated and desiccated in a laminar airflow hood for 0-4 hours prior to plunging into liquid nitrogen. The embryo moisture content (% MC during dehydration was assessed. Cryopreserved embryos were thawed in a solution rich in sucrose at room temperature, inoculated in a germination medium and maintained in a growth chamber. After 30 days, the embryo germination was evaluated. No significant differences were observed between the different embryo dehydration times, where they were dehydrated for at least one hour. Embryos with a MC between 34.3 and 20.3% were germinated after cryopreservation. In the absence of dehydration, all embryos died following cryopreservation. We conclude that B. intermedia zygotic embryos can be successfully cryopreserved and thawed at room temperature after at least one hour of dehydration in a laminar airflow bench.

  18. In vitro production of embryos in South American camelids.

    Science.gov (United States)

    Trasorras, V; Giuliano, S; Miragaya, M

    2013-01-10

    Studies in reproductive biotechnology techniques have been minimal in South American camelids (SAC). Complex reproductive characteristics of these species contribute to slow progress. Nevertheless, some techniques, such as in vitro fertilization, intracytoplasmic sperm injection and nuclear transfer have been applied and have produced advances in knowledge on embryo environment and in vitro conditions necessary for development. Embryo production may have a high impact in both domestic and wild camelids population. Studies addressed to improve in vitro embryo production and oocyte collection could be a potential key to develop IVF and embryo production as a routine procedure in camelids. Copyright © 2012 Elsevier B.V. All rights reserved.

  19. Effect of ambient light exposure of media and embryos on development and quality of porcine parthenogenetically activated embryos

    DEFF Research Database (Denmark)

    Li, Rong; Liu, Ying; Callesen, Henrik

    2015-01-01

    was tested in two experiments: (1) ambient light on medium subsequently used for embryo in vitro development; and (2) ambient light exposure on activated oocytes before in vitro development. The results from Experiment 1 showed that exposure of culture medium to both types of ambient light decreased......Light exposure is a common stress factor during in vitro handling of oocytes and embryos that originates from both microscope and ambient light. In the current study, the effect of two types of ambient light (daylight and laboratory light) on porcine parthenogenetically activated (PA) embryos....... In conclusion, exposure to ambient light can be harmful to embryo development, both when medium is exposed for a long period of time and, to a greater extent, when the embryo itself is exposed for >1 h. In practice, it is therefore recommended to protect both culture medium and porcine embryos against ambient...

  20. Excised Embryo Rambutan (Nephelium Lappaceum L.) Dan Kepekaannya Terhadap Pengeringan*[desiccation Sensitivity of Rambutan (Nephelium Lappaceum L.) Excised Embryo

    OpenAIRE

    Soetisna, Usep; Zebua, Lisye Iriana

    2005-01-01

    A study on desiccation sensitivity of rambutan (Nephelium lappaceum L.)excised embryo has been conducted.Excised embryos were desiccated into target moisture content of 30%, 25%, 20%, 15% and 10% respectively.Results shown that the critical level for moisture content was 20% with 60% viability and 7.431% per etmal growth rate and 929ms electric conduction Further study is needed as to investigate on how long the embryo will withstand subsequent storage treatments.

  1. Diseases of amphibian eggs and embryos

    Science.gov (United States)

    Green, D.E.; Converse, K.A.; Majumdar, S.K.; Huffman, J.E.; Brenner, F.J.; Panah, A.I.

    2005-01-01

    Amphibians generally are prolific egg producers. In tropical and semi-tropical regions, deposition of eggs may occur year-round or may coincide with rainy seasons, while in temperate regions, deposition of eggs usually occurs immediately after emergence from hibernation. Numbers of eggs produced by each species may vary from a few dozen to thousands. Accordingly, some eggs may be infertile and wastage of embryos is to be expected. Fertility, viability and decomposition of eggs and embryos must be considered before it is assumed that diseases are present. An important consideration in the evaluation of egg masses is the fact that some will contain infertile and non-viable eggs. These infertile and nonviable eggs will undergo decomposition and they may appear similar to eggs that are infected by a pathogen. Evaluation of egg masses and embryos for the presence of disease may require repeated observations in a given breeding season as well as continued monitoring of egg masses during their growth and development and over successive breeding seasons. Amphibian eggs rarely are subjected to a comprehensive health (diagnostic) examination; hence, there is scant literature on the diseases of this life stage. Indeed, the eggs of some North American amphibians have yet to be described. Much basic physiology and normal biomedical baseline data on amphibian eggs is lacking. For example, it is known that the aquatic eggs of some species of shrimp quickly are coated by a protective and commensal bacterium that effectively impedes invasion of the eggs by other environmental organisms and potential pathogens. In the absence of this bacterium, shrimp eggs are rapidly killed by other bacteria and fungi (Green, 2001). The possibility that amphibian eggs also have important symbiotic or commensal bacteria needs to be investigated. Furthermore, the quantity and types of chemicals in the normal gelatinous capsules of amphibian eggs have scarcely been examined. Abnormalities of the

  2. Effect of culture medium volume and embryo density on early mouse embryonic development: tracking the development of the individual embryo.

    Science.gov (United States)

    Dai, Shan-Jun; Xu, Chang-Long; Wang, Jeffrey; Sun, Ying-Pu; Chian, Ri-Cheng

    2012-07-01

    To determine the optimal volume or density of embryos for the well-of-the-well (WOW) system in order to track the development of individual embryos and to determine whether the WOW system can reverse the negative impact of culturing embryos singly. (1) Mouse embryos (groups of nine at the 2-cell stage) were cultured in 6.25 μl, 12.50 μl, 25.00 μl and 50.00 μl of droplets of culture medium under paraffin oil; (2) Groups of three, six, nine and twelve embryos at the 2-cell stage were cultured in 50 μl of droplet of culture medium under paraffin oil; (3) Groups of nine embryos at the 2-cell stage were cultured in 50 μl of droplet under paraffin oil with or without nine micro-wells made on the bottom of the Petri dish into each of which were placed one of the nine embryos (WOW system). Also single 2-cell stage embryos was cultured individually in 5.5 μl of droplet of culture medium under paraffin oil with or without a single micro-well made on the bottom of the Petri dish (WOW system for single culture). At the end of culture, the percentages of blastocyst development, hatching and hatched blastocysts were compared in each group. The blastocysts were fixed for differential staining. The blastocyst development was significantly higher (P WOW system. The blastocyst development was not improved when single embryo cultured individually in a micro-well was compared to single embryo cultured individually without micro-well. The total cell numbers of blastocysts were significantly higher in group embryo culture than single embryo culture regardless of whether the WOW system was used. In addition, the total cell numbers of blastocysts were significantly higher (P WOW system than without. Group embryo culture is superior to single embryo culture for blastocyst development. The WOW system with 50 μl of droplet of culture medium can be used to track the individual development of embryo cultured in groups while preserving good embryonic development. The reduced

  3. Phytohemagglutinin facilitates the aggregation of blastomere pairs from Day 5 donor embryos with Day 4 host embryos for chimeric bovine embryo multiplication.

    Science.gov (United States)

    Simmet, Kilian; Reichenbach, Myriam; Reichenbach, Horst-Dieter; Wolf, Eckhard

    2015-12-01

    Multiplication of bovine embryos by the production of aggregation chimeras is based on the concept that few blastomeres of a donor embryo form the inner cell mass (ICM) and thus the embryo proper, whereas cells of a host embryo preferentially contribute to the trophectoderm (TE), the progenitor cells of the embryonic part of the placenta. We aggregated two fluorescent blastomeres from enhanced green fluorescent protein (eGFP) transgenic Day 5 morulae with two Day 4 embryos that did not complete their first cleavage until 27 hours after IVF and tested the effect of phytohemagglutinin-L (PHA) on chimeric embryo formation. The resulting blastocysts were characterized by differential staining of cell lineages using the TE-specific factor CDX2 and confocal laser scanning microscopy to facilitate the precise localization of eGFP-positive cells. The proportions of blastocyst development of sandwich aggregates with (n = 99) and without PHA (n = 46) were 85.9% and 54.3% (P multiplication of genetically valuable donor embryos. Copyright © 2015 Elsevier Inc. All rights reserved.

  4. A new direct transfer protocol for cryopreserved IVF embryos.

    Science.gov (United States)

    Sanches, Bruno Valente; Lunardelli, Paula Alvares; Tannura, Juliana Hayashi; Cardoso, Bruna Lopes; Pereira, Marcos Henrique Colombo; Gaitkoski, Douglas; Basso, Andrea Cristina; Arnold, Daniel Robert; Seneda, Marcelo Marcondes

    2016-04-01

    The global demand for in vitro-produced (IVP) embryos of determined sex has greatly increased over the last decade. Efficient protocols for the direct transfer of IVP embryos are lacking. This study aimed to compare the pregnancy rates for fresh, vitrified, or frozen/directly transferred IVP dairy cow embryos. Oocytes (n = 3171) recovered by ovum pickup (n = 112) from Girolando (Holstein-Gir) females (n = 36) were selected and submitted to IVM for 24 hours at 38.5 °C with 5% CO2 in air with saturated humidity. In vitro fertilization was performed with the thawed, sexed semen from 5 Holstein bulls. After IVF, presumptive zygotes were denuded and cultured for 7 days under the same IVM and IVF conditions of temperature and humidity, except with 5% CO2 and 5% O2. Grade I blastocysts were randomly assigned for either the transferred fresh, vitrified/thawing, or frozen/directly embryo transfer into previously synchronized recipient females. Conception rates were analyzed by binomial logistic regression, and a probability level of P < 0.05 was considered significant. The conception rates were 51.35 ± 1.87% (133/259) for the fresh embryos, 35.89 ± 3.87% (84/234) for the vitrified embryos, and 40.19 ± 4.65% (125/311) for the frozen directly transferred embryos. These data demonstrate that IVP embryos with sexed semen could be directly transferred into recipient cows with similar conception rates to vitrified embryos. The comparison found that the use of frozen embryos in direct transfer provides easier logistics and a more practical approach for the transfer of IVP embryos on dairy farms. Copyright © 2016 Elsevier Inc. All rights reserved.

  5. Radioactive marking of proteins in cultured mouse embryos

    International Nuclear Information System (INIS)

    Nowak, J.

    1984-01-01

    The purpose of this work was to build an in vitro test system, with which on the one hand postimplantation embryos of the mouse could be cultured without morphological of physiological damage and on the other hand their protein could be as highly marked as possible. With this radioactively marked proteins were to be won, which are optimally suited for a high separation by two-dimensional electrophoresis. In addition incubation and preparation methods were found for the ages of day 10, 11 and 12 of the embryonic development. With the use of 3 H-marked amino acids in the culture medium it was determined that embryos without embryonic membranes, so-called N-embryos, built in more radioactivity into their proteins than the embryos with embryonic membranes, the so-called DAO-embryos or the DO-embryos. On the contrary, the embryos with intact blood circulation (DO-embryos) showed an even distribution of radioactive marker in their bodies. Since an even distribution of the marker in the embryo is a necessary prerequisite for a representative presentation of the proteins by 2DE, the DO-preparation was considered the best suited method. In order to increase the amount of radioactivity incorporated into the proteins of the DO-embryos, the concentration of the used isotope or the incubation length could be increased. A combination of both proved to be the best method. A 14 C-marked amino acid mixture of 20 μCi/corresponds to 20 μl instead of the usual 150 μCi 3 H-marked amino acids in a culture medium proved to be equally suitable. Notable changes which would have indicated a damaging affect of the used radioactivity or the in vitro culturing were not observed. The achieved methodical conditions were used for the presentation of the embryo proteins by two-dimensional electrophoresis and fluorography. (orig./MG) [de

  6. Culture of bovine embryos on a polydimethylsiloxane (PDMS) microwell plate.

    Science.gov (United States)

    Akagi, Satoshi; Hosoe, Misa; Matsukawa, Kazutsugu; Ichikawa, Akihiko; Tanikawa, Tamio; Takahashi, Seiya

    2010-08-01

    We fabricated a polydimethylsiloxane (PDMS)-based microwell plate (PDMS-MP) containing 100 microwells with a rounded bottom and examined whether it can be used for culture of individual in vitro fertilized (IVF) embryos or parthenogenetically activated zona-free embryos in cattle. In Experiment 1, we examined the in vitro developmental ability of IVF embryos cultured individually on PDMS-MP. After IVF, 20 embryos were transferred into 100 microl drops on PDMS-MP and cultured individually in each well of PDMS-MP (PDMS group). After 7 days of culture, the embryos in the PDMS group developed to the blastocyst stage at the same rate of those in the control group cultured in a group of 20 embryos without PDMS-MP. There were no differences in total number of cells and the ratio of inner cell mass to total cells between the PDMS and control groups. In Experiment 2, we examined the in vitro developmental ability of parthenogenetically activated zona-free bovine embryos cultured individually on PDMS-MP. The zona-free embryos were cultured individually in each well of a PDMS-MP or in each well produced by pressing a darning needle onto the bottom of a culture dish (WOW group). After 7 days of culture, the blastocyst formation rate and cell number of blastocysts in the PDMS group did not differ from those of the zona-intact embryos in the control group. Also, there were no differences in the blastocyst formation rate and cell number of blastocysts between the WOW and PDMS groups. These results suggest that the culture system using PDMS-MP is useful for individual embryos or zona-free embryos in cattle.

  7. Precocious germination and its regulation in embryos of triticale caryopses

    Directory of Open Access Journals (Sweden)

    Stanisław Weidner

    2014-01-01

    Full Text Available Triticale var. Lasko embryos, isolated from grain gathered at milk ripeness, the beginning of wax ripeness and at full ripeness, were allowed to germinate for 48 h on agar with glucose. The highest incorporation of tritiated adenosine into polyribosomal RNA during germination was found in the ribosome fractions from embryos of grain gathered at full ripeness, lower incorporation was in preparations from embryos of milk ripe grain and the lowest in preparations from embryos of wax ripe grain. Different tendencies were observed in respect to the synthesis of ribosomal proteins. The highest incorporation of 14C-amino acids into ribosomal proteins was found in preparations of ribosome fractions from embryos of milk ripe grain, lower in preparations of embryos from fully ripe grain, the lowest in preparations of embryos from wax ripe grain. ABA (10-4 M completely inhibited the external symptoms of germination of immature embryos, while its inhibition of the synthesis of polyribosomal RNA and ribosomal proteins was greater the more mature the embryos that were germinated. The greatest stimulation of precocious germination by exogenous BA and GA3 was demonstrated in the least mature embryos isolated from milk ripe grain. Under the influence of both stimulators, an increase of the proportion of polyribosomes in the total ribosome fraction occurred in this sample, as did a rise in the intensity of ribosomal protein synthesis. The incorporation of 3H-adenosine into polyribosomal RNA, however, was lower than in the control sample. The results obtained suggest that the regulation of precocious germination of triticale embryos by phyto-hormones is not directly related to transcription.

  8. Cryopreservation of coffee zygotic embryos: dehydration and osmotic rehydration

    Directory of Open Access Journals (Sweden)

    Maísa de Siqueira Pinto

    Full Text Available ABSTRACT Conservation of plant genetic resources is important to prevent genetic erosion. Seed banks are the most common method of ex situ conservation; however, coffee seeds can not be stored by conventional methods. Cryopreservation is a viable alternative for long-term conservation of species that produce intermediate or recalcitrant seeds, as coffee. The aim of this work was to cryopreserve Coffea arabica L. cv Catuaí Vermelho IAC 144 zygotic embryos, and analyse the effects of dehydration prior cryopreservation and osmotic rehydration after thawing, in embryos germination and seedlings formation after cryopreservation. Prior to cryopreservation, different dehydration times (0, 15, 30, 60 and 120 min were tested. Dehydrated embryos were cryopreserved in liquid nitrogen for 1 hour, and after thawing were rehydrated by osmotic solutions. Dehydrated and non-cryopreserved embryos were also analysed. The test with 2,3,5 triphenyl tetrazolium chloride (TTC was used to evaluate the embryos viability. Non-dehydrated embryos did not survive after freezing. Embryos that were dehydrated until 20% of the moisture content did not germinate when osmotic rehydration was not performed. In contrast, cryopreserved embryos with the same moisture content presented 98% germination when they were rehydrated slowly in osmotic solution. According to tetrazolium tests, embryos presented maximum viability (75% after dehydration for 60 minutes (23% moisture content. Therefore, coffee zygotic embryos (Coffea arabica L. cv. Catuaí Vermelho can be successfully cryopreserved using physical dehydration in silica gel for 60 minutes (23% moisture content, followed by osmotic rehydration after thawing. This method allowed a germination of 98% of cryopreserved zygotic embryos.

  9. Studies of In Vitro Embryo Culture of Guppy (Poecilia reticulata).

    Science.gov (United States)

    Liu, LiLi; Lee, Ki-Young

    2014-09-01

    Different with other fishes, the guppies (Poecilia reticulata) is ovoviviparity, which retain their fertilized eggs within the follicle throughout gestation. The synchronously growing diplotene oocytes store nutrients in droplets and yolk, before their maturation and fertilization. The lecithotrophic strategy of development entails the provisioning of embryos with resources from the maternal yolk deposit rather than from a placenta, it allows the extracorporeal culture of guppy embryo. Studies on their early development of live bearers like the guppy including lineage tracing and genetic manipulations, have been limited. Therefore, to optimize conditions of embryo in vitro culture, explanted embryos from pregnant females were incubated in embryo medium (L-15 medium, supplemented with 5, 10, 15, 20% fetal bovine serum, respectively). We investigated whether the contents of FBS in vitro culture medium impact the development of embryos, and whether they would hatch in vitro. Our study found that in 5% of FBS of the medium, although embryos developed significantly slower in vitro than in the ovary, it was impossible to exactly quantify the developmental delay in culture, due to the obvious spread in developmental stage within each batch of eggs, and embryos can only be maintained until the early-eyed. And although in culture with 20% FBS the embryos can sustain rapid development of early stage, but cannot be cultured for the entire period of their embryonic development and ultimately died. In the medium with 10% and 15% FBS, the embryos seems well developed, even some can continue to grow after follicle ruptures until it can be fed. We also observed that embryonic in these two culture conditions were significantly different in development speed, in 15% it is faster than 10%. But 10% FBS appears to be more optimizing condition than 15% one on development process of embryos and survival rate to larvae stage.

  10. Nucleologenesis in the Caenorhabditis elegans embryo.

    Directory of Open Access Journals (Sweden)

    Darina Korčeková

    Full Text Available In the Caenorhabditis elegans nematode, the oocyte nucleolus disappears prior to fertilization. We have now investigated the re-formation of the nucleolus in the early embryo of this model organism by immunostaining for fibrillarin and DAO-5, a putative NOLC1/Nopp140 homolog involved in ribosome assembly. We find that labeled nucleoli first appear in somatic cells at around the 8-cell stage, at a time when transcription of the embryonic genome begins. Quantitative analysis of radial positioning showed the nucleolus to be localized at the nuclear periphery in a majority of early embryonic nuclei. At the ultrastructural level, the embryonic nucleolus appears to be composed of a relatively homogenous core surrounded by a crescent-shaped granular structure. Prior to embryonic genome activation, fibrillarin and DAO-5 staining is seen in numerous small nucleoplasmic foci. This staining pattern persists in the germline up to the ∼100-cell stage, until the P4 germ cell divides to give rise to the Z2/Z3 primordial germ cells and embryonic transcription is activated in this lineage. In the ncl-1 mutant, which is characterized by increased transcription of rDNA, DAO-5-labeled nucleoli are already present at the 2-cell stage. Our results suggest a link between the activation of transcription and the initial formation of nucleoli in the C. elegans embryo.

  11. Macroevolutionary developmental biology: Embryos, fossils, and phylogenies.

    Science.gov (United States)

    Organ, Chris L; Cooper, Lisa Noelle; Hieronymus, Tobin L

    2015-10-01

    The field of evolutionary developmental biology is broadly focused on identifying the genetic and developmental mechanisms underlying morphological diversity. Connecting the genotype with the phenotype means that evo-devo research often considers a wide range of evidence, from genetics and morphology to fossils. In this commentary, we provide an overview and framework for integrating fossil ontogenetic data with developmental data using phylogenetic comparative methods to test macroevolutionary hypotheses. We survey the vertebrate fossil record of preserved embryos and discuss how phylogenetic comparative methods can integrate data from developmental genetics and paleontology. Fossil embryos provide limited, yet critical, developmental data from deep time. They help constrain when developmental innovations first appeared during the history of life and also reveal the order in which related morphologies evolved. Phylogenetic comparative methods provide a powerful statistical approach that allows evo-devo researchers to infer the presence of nonpreserved developmental traits in fossil species and to detect discordant evolutionary patterns and processes across levels of biological organization. © 2015 Wiley Periodicals, Inc.

  12. Twinning of amphibian embryos by centrifugation

    Science.gov (United States)

    Black, S. D.

    1984-01-01

    In the frog Xenopus laevis, the dorsal structures of the embryonic body axis normally derive from the side of the egg opposite the side of sperm entry. However, if the uncleaved egg is inclined at lg or centrifuged in an inclined position, this topographic relationship is overridden: the egg makes its dorsal axial structures according to its orientation in the gravitational/centrifugal field, irrespective of the position of sperm entry. Certain conditions of centrifugation cause eggs to develop into conjoined twins with two sets of axial structures. A detailed analysis of twinning provided some insight into experimental axis orientation. First, as with single-axis embryos, both axes in twins are oriented according to the direction of centrifugation. One axis forms at the centripetal side of the egg and the other forms at the centrifugal side, even when the side of sperm entry is normal to the centrifugal force vector. Second, if eggs are centrifuged to give twins, but are inclined at lg to prevent post-centrifugation endoplasmic redistributions, only single-axis embryos develop. Thus, a second redistribution is required for high-frequency secondary axis formation. This can be accomplished by lg (as in the single centrifugations) or by a second centrifugation directed along the egg's animal-vegetal axis.

  13. Efficient somatic embryo production of Limau madu (Citrus ...

    African Journals Online (AJOL)

    fbn

    concentrations for producing cell suspension and somatic embryos of Limau madu (Citrus suhuiensis ... Citrus. The successful performance of all the stages of somatic embryogenesis has been reported, from the growth of embryogenic cells to embryo maturation in ..... line I and Cleopatra, line II and C. limonia L. Osbeck, cv.

  14. Microspore derived embryo formation and doubled haploid plant ...

    African Journals Online (AJOL)

    The optimal conditions for microspore embryo formation differed depending on genotype. Microspore-derived embryos (MDE) formation was influenced by the strength of the NLN medium, the microelement and sugar concentration, and the heat shock temperature and period. The 0.5XNLN liquid medium was the most ...

  15. Superovulation Response and In vivo Embryo Production Potential ...

    African Journals Online (AJOL)

    to a search dish containing holding medium and grouped in to embryos and UFO under a stereomicroscope (MotisSMZ 140/143®, Roanoke, USA). The embryos were graded for their developmental stage (from stage 1 = 1-cell to stage 9 = expanded hatched blastocyst) and quality (from quality 1 = excellent to quality 4.

  16. Embryo rescue of crosses between diploid and tetraploid grape ...

    African Journals Online (AJOL)

    ajl yemi

    2011-12-19

    Dec 19, 2011 ... embryo rescue from interspecific hybridization between diploid and tetraploid grape species. Wakana et al. (2003) and Motosugi et al. (2003) studied the formation and developments of hybrid seeds from cross between diploid and tetraploid, and then obtained triploid progenies through embryo rescue.

  17. Triazole-induced gene expression changes in the zebrafish embryo

    NARCIS (Netherlands)

    Hermsen, S.A.B.; Pronk, T.; van den Brandhof, E.J.; van der Ven, L.T.; Piersma, A.H.|info:eu-repo/dai/nl/071276947

    2012-01-01

    The zebrafish embryo is considered to provide a promising alternative test model for developmental toxicity testing. Most systems use morphological assessment of the embryos, however, microarray analyses may increase sensitivity and predictability of the test by detecting more subtle and detailed

  18. Early embryo development in Fucus distichus is auxin sensitive

    Science.gov (United States)

    Basu, Swati; Sun, Haiguo; Brian, Leigh; Quatrano, Ralph L.; Muday, Gloria K.

    2002-01-01

    Auxin and polar auxin transport have been implicated in controlling embryo development in land plants. The goal of these studies was to determine if auxin and auxin transport are also important during the earliest stages of development in embryos of the brown alga Fucus distichus. Indole-3-acetic acid (IAA) was identified in F. distichus embryos and mature tissues by gas chromatography-mass spectroscopy. F. distichus embryos accumulate [(3)H]IAA and an inhibitor of IAA efflux, naphthylphthalamic acid (NPA), elevates IAA accumulation, suggesting the presence of an auxin efflux protein complex similar to that found in land plants. F. distichus embryos normally develop with a single unbranched rhizoid, but growth on IAA leads to formation of multiple rhizoids and growth on NPA leads to formation of embryos with branched rhizoids, at concentrations that are active in auxin accumulation assays. The effects of IAA and NPA are complete before 6 h after fertilization (AF), which is before rhizoid germination and cell division. The maximal effects of IAA and NPA are between 3.5 and 5 h AF and 4 and 5.5 h AF, respectively. Although, the location of the planes of cell division was significantly altered in NPA- and IAA-treated embryos, these abnormal divisions occurred after abnormal rhizoid initiation and branching was observed. The results of this study suggest that auxin acts in the formation of apical basal patterns in F. distichus embryo development.

  19. The gastrocoel roof plate in embryos of different frogs.

    Science.gov (United States)

    Sáenz-Ponce, Natalia; Santillana-Ortiz, Juan-Diego; del Pino, Eugenia M

    2012-02-01

    The morphology of the gastrocoel roof plate and the presence of cilia in this structure were examined in embryos of four species of frogs. Embryos of Ceratophrys stolzmanni (Ceratophryidae) and Engystomops randi (Leiuperidae) develop rapidly, provide comparison for the analysis of gastrocoel roof plate development in the slow-developing embryos of Epipedobates machalilla (Dendrobatidae) and Gastrotheca riobambae (Hemiphractidae). Embryos of the analyzed frogs develop from eggs of different sizes, and display different reproductive and developmental strategies. In particular, dorsal convergence and extension and archenteron elongation begin during gastrulation in embryos of rapidly developing frogs, as in Xenopus laevis. In contrast, cells that involute during gastrulation are stored in the large circumblastoporal collar that develops around the closed blastopore in embryos of slow-developing frogs. Dorsal convergence and extension only start after blastopore closure in slow-developing frog embryos. However, in the neurulae, a gastrocoel roof plate develops, despite the accumulation of superficial mesodermal cells in the circumblastoporal collar. Embryos of all four species develop a ciliated gastrocoel roof plate at the beginning of neurulation. Accordingly, fluid-flow across the gastrocoel roof plate is likely the mechanism of left-right asymmetry patterning in these frogs, as in X. laevis and other vertebrates. A ciliated gastrocoel roof plate, with a likely origin as superficial mesoderm, is conserved in frogs belonging to four different families and with different modes of gastrulation. Copyright © 2011 International Society of Differentiation. Published by Elsevier B.V. All rights reserved.

  20. Fruit, seed and embryo development of different cassava (Manihot ...

    African Journals Online (AJOL)

    Fruit, seed and embryo developments of different cassava (Manihot esculenta Crantz) genotypes, as well as embryo rescue, were investigated. The fruits of three genotypes after uncontrolled open pollination presented the same progressive development with similar sizes at different stages. There are large differences in ...

  1. The role of growth regulators, embryo age and genotypes on ...

    African Journals Online (AJOL)

    Administrator

    2011-06-06

    Jun 6, 2011 ... One of the most important problem of tomato breeders is lengthy seed to seed cycle in a breeding program. In vitro techiques provide a lot of advantages for breeders. The objective of this work was to determine the effect of growth regulators and immature embryo age on embryo germination and rapid.

  2. Patients' Preference for Number of Embryos Transferred During IVF ...

    African Journals Online (AJOL)

    Background: The Human Fertilization and Embryology Authority is considering limiting the number of embryos that can be transferred to single embryo per cycle as has been done in several European countries, with the aim of reducing the rate of multiple pregnancies and its attendant complications following in vitro ...

  3. The role of growth regulators, embryo age and genotypes on ...

    African Journals Online (AJOL)

    One of the most important problem of tomato breeders is lengthy seed to seed cycle in a breeding program. In vitro techiques provide a lot of advantages for breeders. The objective of this work was to determine the effect of growth regulators and immature embryo age on embryo germination and rapid generation ...

  4. Permeability barriers to embryo cryopreservation of Pectinophora gossypiella (Lepidoptera: Gelechiidae)

    Science.gov (United States)

    The aim of this study was to develop a method to cryopreserve the embryos of the pink bollworm moth, Pectinophora gossypiella (Saunders). Previously developed dipteran cryopreservation protocols were not directly adaptable to use with the embryos of this lepidopteran species. Physiochemical and ele...

  5. The using of gibberellic acid hormone on cotton mature embryo ...

    African Journals Online (AJOL)

    Jane

    2011-08-22

    Aug 22, 2011 ... sperm and maternal tissues (ovule) there is no genetic or chromosomal proportionality. This is because endosperm tissue is only the fundamental feeding, hormone excitants and physiological source for embryo (Gill and Bajaj,. 1987). Successful embryo development in the first step depends on the natural ...

  6. Development of the ventral body wall in the human embryo

    NARCIS (Netherlands)

    Mekonen, Hayelom K.; Hikspoors, Jill P. J. M.; Mommen, Greet; Köhler, S. Eleonore; Lamers, Wouter H.

    2015-01-01

    Migratory failure of somitic cells is the commonest explanation for ventral body wall defects. However, the embryo increases ~ 25-fold in volume in the period that the ventral body wall forms, so that differential growth may, instead, account for the observed changes in topography. Human embryos

  7. Closure of the vertebral canal in human embryos and fetuses

    NARCIS (Netherlands)

    Mekonen, Hayelom K.; Hikspoors, Jill P. J. M.; Mommen, Greet; Kruepunga, Nutmethee; Köhler, S. Eleonore; Lamers, Wouter H.

    2017-01-01

    The vertebral column is the paradigm of the metameric architecture of the vertebrate body. Because the number of somites is a convenient parameter to stage early human embryos, we explored whether the closure of the vertebral canal could be used similarly for staging embryos between 7 and 10weeks of

  8. Storage oil breakdown during embryo development of Brassica napus (L.).

    Science.gov (United States)

    Chia, Tansy Y P; Pike, Marilyn J; Rawsthorne, Stephen

    2005-05-01

    In this study it is shown that at least 10% of the major storage product of developing embryos of Brassica napus (L.), triacylglycerol, is lost during the desiccation phase of seed development. The metabolism of this lipid was studied by measurements of the fate of label from [1-(14)C]decanoate supplied to isolated embryos, and by measurements of the activities of enzymes of fatty acid catabolism. Measurements on desiccating embryos have been compared with those made on embryos during lipid accumulation and on germinating seedlings. Enzymes of beta-oxidation and the glyoxylate cycle, and phosphoenolpyruvate carboxykinase were present in embryos during oil accumulation, and increased in activity and abundance as the seeds matured and became desiccated. Although the activities were less than those measured during germination, they were at least comparable to the in vivo rate of fatty acid synthesis in the embryo during development. The pattern of labelling, following metabolism of decanoate by isolated embryos, indicated a much greater involvement of the glyoxylate cycle during desiccation than earlier in oil accumulation, and showed that much of the (14)C-label from decanoate was released as CO(2) at both stages. Sucrose was not a product of decanoate metabolism during embryo development, and therefore lipid degradation was not associated with net gluconeogenic activity. These observations are discussed in the context of seed development, oil yield, and the synthesis of novel fatty acids in plants.

  9. Successful pregnancy following trans-myometrial embryo transfer ...

    African Journals Online (AJOL)

    ... factor infertility (poor motility). Routine mock embryo transfer indicated severe cervical stenosis which was confirmed at hysteroscopy. She subsequently had trans-myometrial embryo transfer. Blood pregnancy test at 2 weeks was positive and interval transvaginal ultrasound confirmed 2 viable intrauterine foetal poles.

  10. Chromosomal mosaicism in human preimplantation embryos : a systematic review

    NARCIS (Netherlands)

    van Echten-Arends, Jannie; Mastenbroek, Sebastiaan; Sikkema-Raddatz, Birgit; Korevaar, Johanna C.; Heineman, Maas Jan; van der Veen, Fulco; Repping, Sjoerd

    2011-01-01

    BACKGROUND: Although chromosomal mosaicism in human preimplantation embryos has been described for almost two decades, its exact prevalence is still unknown. The prevalence of mosaicism is important in the context of preimplantation genetic screening in which the chromosomal status of an embryo is

  11. Optimal developmental stage for vitrification of parthenogenetically activated porcine embryos

    DEFF Research Database (Denmark)

    Li, Rong; Li, Juan; Kragh, Peter

    2012-01-01

    -lapse monitored for 24 h or analyzed by diffential staining. After warming, the embryos had to be cultured for at least 8 h before their survival rates were stabilized. Both the survival rate and 8 h and the hatching rate at 24 h of Day 4 embryos were significantly higher than those vitrified on Day5 or Day 6 (P...

  12. In vitro bulblet regeneration from immature embryos of Muscari ...

    African Journals Online (AJOL)

    A high frequency bulblet regeneration was achieved for endemic and endangered ornamental plant Muscari azureum using immature embryos. Immature embryos of M. azureum were cultured on callus induction medium consisting of N6 mineral salts and vitamins, 400 mg/L casein + 40 g/L sucrose + 2 g/l L-proline, 2 mg/L ...

  13. Effects of embryo induction media and pretreatments in isolated ...

    African Journals Online (AJOL)

    chemical + heat and also the effects of 5 embryo induction media (NPB-99, C17, W14, CHB-2 and P2) on embryo production, percentage of total and green plant regeneration in isolated microspore culture of an Iranian spring bread wheat, cultivar 'Falat'. The results showed that combination of 21 days cold (4°C) with ...

  14. Factors affecting conception rates in cattle following embryo transfer ...

    African Journals Online (AJOL)

    Embryo Transfer Technology (ETT) plays an important role in improving productivity of dairy cattle (Bos indicus). Embryo Transfer Technology allows top quality female livestock to improve a herd or flock in much the same way that artificial insemination has allowed greater use of superior sires. The technology hastens ...

  15. Protein synthesis in the embryo of Pinus thunbergii seed, 2

    International Nuclear Information System (INIS)

    Yamamoto, Naoaki; Sasaki, Satohiko.

    1977-01-01

    14 C-Amino acid incorporating activity in the absence of exogenous mRNA was found in a cell-free system from embryos of light-germinated Pinus thunbergii seeds, but not in that from dark-imbibed seed embryos. Template activity in the cell-free system from the light-germinated seed embryos was observed in the ribosome fraction, especially the polyribosome fraction, but not in the 100,000 x g supernatant fraction (s100). These facts suggest that the nature of the block in protein synthesis during the imbibition of seeds in the dark is due to the lack or inactivity of mRNA. The s100 from light-germinated seed embryos was found to be less active in amino acid incorporation than that from dark-imbibed seed embryos. (auth.)

  16. Detection of programmed cell death in plant embryos.

    Science.gov (United States)

    Filonova, Lada H; Suárez, María F; Bozhkov, Peter V

    2008-01-01

    Programmed cell death (PCD) is an integral part of embryogenesis. In plant embryos, PCD functions during terminal differentiation and elimination of the temporary organ, suspensor, as well as during establishment of provascular system. Embryo abortion is another example of embryonic PCD activated at pathological situations and in polyembryonic seeds. Recent studies identified the sequence of cytological events leading to cellular self-destruction in plant embryos. As in most if not all the developmental cell deaths in plants, embryonic PCD is hallmarked by autophagic degradation of the cytoplasm and nuclear disassembly that includes breakdown of the nuclear envelope and DNA fragmentation. The optimized setup of terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) allows the routine in situ analysis of nuclear DNA fragmentation in plant embryos. This chapter provides step-by-step procedure of how to process embryos for TUNEL and how to combine TUNEL with immunolocalization of the protein of interest.

  17. Cryopreservation of coconut (Cocos nucifera L.) zygotic embryos by vitrification.

    Science.gov (United States)

    Sajini, K K; Karun, A; Amamath, C H; Engelmann, F

    2011-01-01

    The present study investigates the effect of preculture conditions, vitrification and unloading solutions on survival and regeneration of coconut zygotic embryos after cryopreservation. Among the seven plant vitrification solutions tested, PVS3 was found to be the most effective for regeneration of cryopreserved embryos. The optimal protocol involved preculture of embryos for 3 days on medium with 0.6 M sucrose, PVS3 treatment for 16 h, rapid cooling and rewarming and unloading in 1.2 M sucrose liquid medium for 1.5 h. Under these conditions, 70-80 survival (corresponding to size enlargement and weight gain) was observed with cryopreserved embryos and 20-25 percent of the plants regenerated (showing normal shoot and root growth) from cryopreserved embryos were established in pots.

  18. The effect of insecticide Deltamethrin on development of chick embryos

    International Nuclear Information System (INIS)

    Al-Naal, R.; Bassal, M. Osman, M.

    1997-04-01

    This study was conducted to evaluate the cyto and the embryo toxicity of Deltamethrin and its commercial formulation DECIS 50 EC in chick embryo during its critical embryonic development period before and in the organogenesis. The embryos were incubated in well closed plastic caps containing the complete egg composition at 38 o. the Deltamethrin and DECIS were found to cause histological and morphological malformations, specially in the brain, also they reduced the majority of the synthetic activities of the DNA, RNA, and proteins in the embryonic and the vascular areas. The flow cytometric analysis showed alterations in frequency of cells in both embryonic and vascular areas in the treated embryo during the cell cycle phases. Our study also showed that the DECIS had greater cyto and embryo toxicity than the Seltamethrin for analysis (author). 149 refs., 36 figs., 16 tabs

  19. Sex determination of duck embryos: observations on syrinx development

    Science.gov (United States)

    Wilson, Robert E.; Sonsthagen, Sarah A.; Franson, J. Christian

    2013-01-01

    Ducks exhibit sexual dimorphism in vocal anatomy. Asymmetrical ossification of the syrinx (bulla syringealis) is discernable at about 10 days of age in male Pekin duck (Anas platyrhynchos domestica) embryos, but information is lacking on the early development of the bulla in wild ducks. To evaluate the reliability of this characteristic for sexing developing embryos, we examined the syrinx of dead embryos and compared results with molecular sexing techniques in high arctic nesting Common Eiders (Somateria mollissima). Embryos 8 days or older were accurately (100%) sexed based on the presence/absence of a bulla, 2 days earlier than Pekin duck. The use of the tracheal bulla can be a valuable technique when sex identification of embryos or young ducklings is required.

  20. Cryopreservation of mouse embryos by ethylene glycol-based vitrification.

    Science.gov (United States)

    Mochida, Keiji; Hasegawa, Ayumi; Taguma, Kyuichi; Yoshiki, Atsushi; Ogura, Atsuo

    2011-11-18

    Cryopreservation of mouse embryos is a technological basis that supports biomedical sciences, because many strains of mice have been produced by genetic modifications and the number is consistently increasing year by year. Its technical development started with slow freezing methods in the 1970s(1), then followed by vitrification methods developed in the late 1980s(2). Generally, the latter technique is advantageous in its quickness, simplicity, and high survivability of recovered embryos. However, the cryoprotectants contained are highly toxic and may affect subsequent embryo development. Therefore, the technique was not applicable to certain strains of mice, even when the solutions are cooled to 4°C to mitigate the toxic effect during embryo handling. At the RIKEN BioResource Center, more than 5000 mouse strains with different genetic backgrounds and phenotypes are maintained(3), and therefore we have optimized a vitrification technique with which we can cryopreserve embryos from many different strains of mice, with the benefits of high embryo survival after vitrifying and thawing (or liquefying, more precisely) at the ambient temperature(4). Here, we present a vitrification method for mouse embryos that has been successfully used at our center. The cryopreservation solution contains ethylene glycol instead of DMSO to minimize the toxicity to embryos(5). It also contains Ficoll and sucrose for prevention of devitrification and osmotic adjustment, respectively. Embryos can be handled at room temperature and transferred into liquid nitrogen within 5 min. Because the original method was optimized for plastic straws as containers, we have slightly modified the protocol for cryotubes, which are more easily accessible in laboratories and more resistant to physical damages. We also describe the procedure of thawing vitrified embryos in detail because it is a critical step for efficient recovery of live mice. These methodologies would be helpful to researchers and

  1. Transferring embryos with indeterminate PGD results: the ethical implications.

    Science.gov (United States)

    Insogna, Iris G; Ginsburg, Elizabeth

    2016-01-01

    In vitro fertilization (IVF) and pre-implantation genetic diagnosis (PGD) are becoming increasingly common techniques to select embryos that are unaffected by a known genetic disorder. Though IVF-PGD has high success rates, 7.5 % of blastocysts have inconclusive results after testing. A recent case involving a known BRCA-1 carrier was brought before our Assisted Reproductive Technology Ethics Committee in order to gain a better appreciation for the ethical implications surrounding the transfer of embryos with indeterminate testing. Thirty-nine year old G0 BRCA-1 carrier requiring IVF for male factor infertility. The couple elected for PGD to select against BRCA-1 gene carrier embryos. However, several embryos were returned with inconclusive results. The couple wished to proceed with the transfer of embryos with an unknown carrier status. The case was presented before our Assisted Reproductive Technology Ethics Committee. Many considerations were explored, including the physician's duty to protect patient autonomy, the physician's duty to act in the best interest of the future child, and the physician's duty towards society. Transferring both embryos with unknown carrier status and known-carrier status was debated. Ultimately, the transfer of inconclusive embryos was felt to be ethically permissible in most cases if patients had been adequately counseled. However, the re-biopsy of embryos with inconclusive testing results was encouraged. The transfer of known-carrier embryos was felt to be unethical for certain disease-states, depending on the severity of illness and timing of disease onset. We strongly encourage physicians to create an action plan in advance with their patients, prior to testing, in the event that embryos are returned with inconclusive PGD results. The committee's decision, though helpful in guiding practice, should not overshadow the individual physician-patient relationship, and the need for thorough counseling.

  2. Preimplantation exposure to bisphenol A (BPA) affects embryo transport, preimplantation embryo development, and uterine receptivity in mice

    Science.gov (United States)

    Xiao, Shuo; Diao, Honglu; Smith, Mary Alice; Song, Xiao; Ye, Xiaoqin

    2011-01-01

    To investigate the effects of bisphenol A (BPA) on embryo and uterine factors in embryo implantation, timed pregnant C57BL6 females were treated subcutaneously with 0, 0.025, 0.5, 10, 40, and 100 mg/kg/day BPA from gestation days 0.5 to 3.5. In 100 mg/kg/day BPA-treated females, no implantation sites were detected on day 4.5 but retention of embryos in the oviduct and delayed embryo development were detected on day 3.5. When untreated healthy embryos were transferred to pseudopregnant females treated with 100 mg/kg/day BPA, no implantation sites were detected on day 4.5. In 40 mg/kg/day BPA-treated females, delayed implantation and increased perinatal lethality of their offspring were observed. Implantation seemed normal in the rest BPA-treated groups or the female offspring from 40 mg/kg/day BPA-treated group. These data demonstrate the adverse effects of high doses of BPA on processes critical for embryo implantation: embryo transport, preimplantation embryo development, and establishment of uterine receptivity. PMID:21907787

  3. The influence of the type of embryo culture medium on neonatal birthweight after single embryo transfer in IVF

    NARCIS (Netherlands)

    Vergouw, C.G.; Kostelijk, E.H.; Doejaaren, E.; Hompes, P.G.A.; Lambalk, C.B.; Schats, R.

    2012-01-01

    STUDY QUESTION Does the type of medium used to culture fresh and frozenthawed embryos influence neonatal birthweight after single embryo transfer (SET) in IVF? SUMMARY ANSWER A comparison of two commercially available culture media showed no significant influence on mean birthweight and mean

  4. [Study on embryo toxicity of Cinnabaris].

    Science.gov (United States)

    Liang, Aihua; Li, Chunying; Liu, Ting; Zhao, Yong; Cao, Chunyu; Xun, Baoyun; Wang, Jinhua

    2009-11-01

    To observe the effect of Cinnabaris on mouse embryos after pregnant mice were treated by Cinnabaris in different periods of pregnancy. Two separate experiments were performed: First, Cinnabaris was orally given into pregnant mice at the doses of 0.08, 0.4, 4.0 g x kg(-1) from D6 to D19 after pregnancy; Second, Cinnabaris was orally given into mice at the same doses mentioned above from D14 prior to pregnancy until D19 after pregnancy. All animals were sacrificed on D 20 of pregnancy by caesarean section. The numbers of survival, dead and absorbed fetuses were calculated and the survival fetus weight was measured. The survival fetuses were treated by two methods: One third survival fetuses were fixed and stained by Bouin solution for organ examination and the remaining two thirds fetuses were stained for skeleton examination. No obvious embryo toxicity was observed in the first experiment at Cinnabaris dose levels of 0.08, 0.4, or 4 g x kg(-1) x d(-1). There was no significant effect on embryonic development and the numbers of the survival, dead and absorbed fetus. No obvious malformations on appearance, organ, and skeleton examination of fetuses were found. The second experiment showed that the rates of abortion and absorbed fetus in 0.4, 4 g x kg(-1) x d(-1) Cinnabaris group were higher but without statistical significance compared with control group. Appearance and organ examination of Cinnabaris groups fetus showed no obvious malformation, but skeleton malformation was found in 0.4, 4 g x kg(-1) x d(-1) groups (the rates of skeleton malformation were 46.7% and 77.8%, respectively). No obvious embryonic development toxicity was observed when Cinnabaris was orally given in intermediate and late pregnant period, but the embryos in the early stage of pregnancy was more sensitive to Cinnabaris. When Cinnabaris was given prior to pregnancy until the whole period of pregnancy, it may be harmful for the fetuses at above the dose level 0.08 g x kg(-1) x d(-1) (equivalent

  5. Supplementation of bovine embryo culture medium with L-arginine improves embryo quality via nitric oxide production.

    Science.gov (United States)

    Santana, Priscila Di Paula Bessa; Silva, Thiago Velasco Guimarães; da Costa, Nathália Nogueira; da Silva, Bruno Barauna; Carter, Timothy Frederick; Cordeiro, Marcela da Silva; da Silva, Bruno José Martins; Santos, Simone do Socorro Damasceno; Herculano, Anderson Manoel; Adona, Paulo Roberto; Ohashi, Otávio Mitio; Miranda, Moysés dos Santos

    2014-10-01

    Nitric oxide (NO) is a cell-signaling molecule that regulates a variety of molecular pathways. We investigated the role of NO during preimplantation embryonic development by blocking its production with an inhibitor or supplementing in vitro bovine embryo cultures with its natural precursor, L-arginine, over different periods. Endpoints evaluated included blastocyst rates, development kinetics, and embryo quality. Supplementation with the NO synthase inhibitor N-Nitro-L-arginine-methyl ester (L-NAME) from Days 1 to 8 of culture decreased blastocyst (P L-arginine decreased blastocyst rates (P L-arginine improved embryo hatching rates (P supplementing the embryo culture medium with L-arginine favors preimplantation development of bovine embryos. © 2014 Wiley Periodicals, Inc.

  6. Natural selection of human embryos: decidualizing endometrial stromal cells serve as sensors of embryo quality upon implantation.

    Directory of Open Access Journals (Sweden)

    Gijs Teklenburg

    2010-04-01

    Full Text Available Pregnancy is widely viewed as dependent upon an intimate dialogue, mediated by locally secreted factors between a developmentally competent embryo and a receptive endometrium. Reproductive success in humans is however limited, largely because of the high prevalence of chromosomally abnormal preimplantation embryos. Moreover, the transient period of endometrial receptivity in humans uniquely coincides with differentiation of endometrial stromal cells (ESCs into highly specialized decidual cells, which in the absence of pregnancy invariably triggers menstruation. The role of cyclic decidualization of the endometrium in the implantation process and the nature of the decidual cytokines and growth factors that mediate the crosstalk with the embryo are unknown.We employed a human co-culture model, consisting of decidualizing ESCs and single hatched blastocysts, to identify the soluble factors involved in implantation. Over the 3-day co-culture period, approximately 75% of embryos arrested whereas the remainder showed normal development. The levels of 14 implantation factors secreted by the stromal cells were determined by multiplex immunoassay. Surprisingly, the presence of a developing embryo had no significant effect on decidual secretions, apart from a modest reduction in IL-5 levels. In contrast, arresting embryos triggered a strong response, characterized by selective inhibition of IL-1beta, -6, -10, -17, -18, eotaxin, and HB-EGF secretion. Co-cultures were repeated with undifferentiated ESCs but none of the secreted cytokines were affected by the presence of a developing or arresting embryo.Human ESCs become biosensors of embryo quality upon differentiation into decidual cells. In view of the high incidence of gross chromosomal errors in human preimplantation embryos, cyclic decidualization followed by menstrual shedding may represent a mechanism of natural embryo selection that limits maternal investment in developmentally impaired pregnancies.

  7. Effect of embryo age and recipient asynchrony on pregnancy rates in a commercial equine embryo transfer program.

    Science.gov (United States)

    Jacob, J C F; Haag, K T; Santos, G O; Oliveira, J P; Gastal, M O; Gastal, E L

    2012-04-01

    In the present study, 809 uterine flushes and 454 embryo transfers performed in mares over a 4-yr interval were examined to evaluate the effects of: (1) the day of embryo collection on recovery rates; (2) the degree of synchrony between donor and recipient mares on pregnancy rates; (3) the recipient day post ovulation on pregnancy rates; and (4) the age of the embryo at recovery on pregnancy rates at 60 days. Uterine flushes were performed on Days 6, 7, 8, 9, and 10 (Day 0 = ovulation) and embryos were transferred to recipients with degrees of synchrony varying between +1 to -6 (recipient ovulated 1 day before through 6 days after the donor). Recipient mares ranged from 2 to 8 days post ovulation. Embryo recovery rates were similar for flushes performed on Day 7 (61%), Day 8 (66%), Day 9 (59%), and Day 10 (56%), but the embryo recovery rate was lower (P recipient mares on Day 2 (33%) compared with mares on Day 3 (66%), Day 4 (66%), Day 5 (62%), Day 6 (55%), Day 7 (58%), and Day 8 (56%). Pregnancy rate was higher (P recipient mares does not need to be as restricted as previously reported in horses. Acceptable pregnancy rates (e.g., 70%, 99/142) were obtained even when recipient mares ovulated 4 to 5 days after the donors; (3) similar pregnancy rates were obtained when recipient mares received embryos within a large range of days post ovulation (Days 3 to 8); and (4) Day 7 embryos produced higher pregnancy rates when compared with Days 8 and 9 embryos. In clinical terms, the application of these new findings will be beneficial to large equine embryo transfer operations in producing more pregnancies per season. Copyright © 2012 Elsevier Inc. All rights reserved.

  8. Nano-nutrition of chicken embryos

    DEFF Research Database (Denmark)

    Grodzik, Marta; Sawosz, Filip; Sawosz, Ewa

    2013-01-01

    -toxic carbonic structures, and we hypothesized that bio-complexes of diamond nanoparticles with L-glutamine may affect molecular responses in breast muscle. The objective of the investigation was to evaluate the effect of diamond nanoparticle (ND) and L-glutamine (Gln) on expression of growth and differentiation...... factors of chicken embryo pectoral muscles. ND, Gln, and Gln/ND solutions (50 mg/L) were injected into fertilized broiler chicken eggs at the beginning of embryogenesis. Muscle tissue was dissected at day 20 of incubation and analysed for gene expression of FGF2, VEGF-A, and MyoD1. ND and especially Gln...... and differentiation dominated over proliferation. These preliminary results suggest that the bio-complex of glutamine and diamond nanoparticles may accelerate growth and maturation of muscle cells....

  9. [Cultural diversity in gamete and embryos donation].

    Science.gov (United States)

    Epelboin, S

    2014-09-01

    Through gamete and embryo donation have successively emerged new ways of designing individuals who, in turn, have generated mutations in the concept of parenthood. A debate is open to the society, which often raises ideological cleavages. Indeed, donation practices mobilize the conflicting interests of donor of gametes, the recipient couple, child, whose origins are complex, although his filiation is legally clear. Its place in the family genealogy can be examined in relation to other societies, which admit plural concepts called "classificatory" kinship. They set up role partition between parents and educators. Setting anthropological perspective provides a broadening of the reflection to answer questions from the donation practices, including genealogical questions of revelation to the child of his conception, his incorporation in family and social group and the importance of compensation of donation. Copyright © 2014. Published by Elsevier SAS.

  10. Selection for rapid embryo development correlates with embryo exposure to maternal androgens among passerine birds.

    Science.gov (United States)

    Schwabl, Hubert; Palacios, Maria G; Martin, Thomas E

    2007-08-01

    Greater offspring predation favors evolution of faster development among species. We hypothesized that greater offspring predation exerts selection on mothers to increase levels of anabolic androgens in egg yolks to achieve faster development. Here, we tested whether (1) concentrations of yolk androgens in passerine species were associated with offspring predation and (2) embryo and nestling development rates were associated with yolk androgen concentrations. We examined three androgens that increase in potency along the synthesis pathway: androstenedione (A(4)) to testosterone (T) to 5 alpha -dihydrotestosterone (5 alpha -DHT). Concentrations of none of these steroids were related to clutch size; only A(4) was allometrically related to egg volume. Species that experience greater predation showed higher yolk concentrations of T and 5 alpha -DHT. Higher concentrations of T and particularly 5 alpha -DHT were strongly correlated with faster development during the embryo period and less so during the nestling period. Development rates were most strongly correlated with 5 alpha -DHT, suggesting that potency increases along the androgen synthesis pathway and that effects are mediated by the androgen receptor pathway. These results are consistent with the hypothesis that selection for faster development by time-dependent offspring mortality may be achieved epigenetically by varying embryo exposure to maternal anabolic steroids.

  11. Selection for rapid embryo development correlates with embryo exposure to maternal androgens among passerine birds

    Science.gov (United States)

    Schwabl, H.; Palacios, M.G.; Martin, T.E.

    2007-01-01

    Greater offspring predation favors evolution of faster development among species. We hypothesized that greater offspring predation exerts selection on mothers to increase levels of anabolic androgens in egg yolks to achieve faster development. Here, we tested whether (1) concentrations of yolk androgens in passerine species were associated with offspring predation and (2) embryo and nestling development rates were associated with yolk androgen concentrations. We examined three androgens that increase in potency along the synthesis pathway: androstenedione (A4) to testosterone (T) to 5??- dihydrotestosterone (5??-DHT). Concentrations of none of these steroids were related to clutch size; only A4 was allometrically related to egg volume. Species that experience greater predation showed higher yolk concentrations of T and 5??-DHT. Higher concentrations of T and particularly 5??-DHT were strongly correlated with faster development during the embryo period and less so during the nestling period. Development rates were most strongly correlated with 5??-DHT, suggesting that potency increases along the androgen synthesis pathway and that effects are mediated by the androgen receptor pathway. These results are consistent with the hypothesis that selection for faster development by time-dependent offspring mortality may be achieved epigenetically by varying embryo exposure to maternal anabolic steroids. ?? 2007 by The University of Chicago. All rights reserved.

  12. Immunoanalysis of dehydrins in Araucaria angustifolia embryos.

    Science.gov (United States)

    Farias-Soares, Francine Lunardi; Burrieza, Hernán Pablo; Steiner, Neusa; Maldonado, Sara; Guerra, Miguel Pedro

    2013-08-01

    The aim of this study was to describe the dehydrin content of mature Araucaria angustifolia embryos, a species of endangered and economically important conifers, native to southern Brazil, northeastern Argentina, and eastern Paraguay. The A. angustifolia seeds have been categorized as recalcitrant. Dehydrins were studied by western blot analysis and in situ immunolocalization microscopy using antibodies raised against the K segment, a highly conserved lysine-rich 15-amino acid sequence extensively used to recognize proteins immunologically related to the dehydrin family. Western blot analysis of the heat-stable protein fraction, as estimated by 15 % SDS-PAGE, revealed three main bands of approximately 20-, 26-, and 29-kDa; when 17.5 % SDS-PAGE was used, each band resolved into two other bands. Two thermosensitive dehydrin bands of around 16 and 35 kDa were common to the axis and cotyledons, and another thermosensitive band, with molecular mass of approximately 10 kDa, was present in the cotyledons only. Following alkaline phosphatase (AP) treatment, a gel mobility shift was detected for each one of the four main bands that can be due to phosphorylation. Dehydrins were detected in all axis and cotyledon tissues using in situ immunolocalization microscopy. At the subcellular level, dehydrins were immunolocalized in the nuclei, protein bodies, and microbodies. In the nucleus, dehydrins were found to be associated with chromatin. We concluded that the gel mobility shift for the four main bands (probably due to phosphorylation), the presence of thermosensitive bands, and the specific localizations in nuclei and protein bodies provide key starting points to understand the function of dehydrins in the embryo cells of this species.

  13. Automatic Blastomere Recognition from a Single Embryo Image

    Directory of Open Access Journals (Sweden)

    Yun Tian

    2014-01-01

    Full Text Available The number of blastomeres of human day 3 embryos is one of the most important criteria for evaluating embryo viability. However, due to the transparency and overlap of blastomeres, it is a challenge to recognize blastomeres automatically using a single embryo image. This study proposes an approach based on least square curve fitting (LSCF for automatic blastomere recognition from a single image. First, combining edge detection, deletion of multiple connected points, and dilation and erosion, an effective preprocessing method was designed to obtain part of blastomere edges that were singly connected. Next, an automatic recognition method for blastomeres was proposed using least square circle fitting. This algorithm was tested on 381 embryo microscopic images obtained from the eight-cell period, and the results were compared with those provided by experts. Embryos were recognized with a 0 error rate occupancy of 21.59%, and the ratio of embryos in which the false recognition number was less than or equal to 2 was 83.16%. This experiment demonstrated that our method could efficiently and rapidly recognize the number of blastomeres from a single embryo image without the need to reconstruct the three-dimensional model of the blastomeres first; this method is simple and efficient.

  14. Hormetic effect induced by depleted uranium in zebrafish embryos

    Energy Technology Data Exchange (ETDEWEB)

    Ng, C.Y.P. [Department of Physics and Materials Science, City University of Hong Kong (Hong Kong); Cheng, S.H., E-mail: bhcheng@cityu.edu.hk [Department of Biomedical Sciences, City University of Hong Kong (Hong Kong); State Key Laboratory in Marine Pollution, City University of Hong Kong (Hong Kong); Yu, K.N., E-mail: peter.yu@cityu.edu.hk [Department of Physics and Materials Science, City University of Hong Kong (Hong Kong); State Key Laboratory in Marine Pollution, City University of Hong Kong (Hong Kong)

    2016-06-15

    Highlights: • Studied hormetic effect induced by uranium (U) in embryos of zebrafish (Danio rerio). • Hormesis observed at 24 hpf for exposures to 10 μg/l of depleted U (DU). • Hormesis not observed before 30 hpf for exposures to 100 μg/l of DU. • Hormetic effect induced in zebrafish embryos in a dose-and time-dependent manner. - Abstract: The present work studied the hormetic effect induced by uranium (U) in embryos of zebrafish (Danio rerio) using apoptosis as the biological endpoint. Hormetic effect is characterized by biphasic dose-response relationships showing a low-dose stimulation and a high-dose inhibition. Embryos were dechorionated at 4 h post fertilization (hpf), and were then exposed to 10 or 100 μg/l depleted uranium (DU) in uranyl acetate solutions from 5 to 6 hpf. For exposures to 10 μg/l DU, the amounts of apoptotic signals in the embryos were significantly increased at 20 hpf but were significantly decreased at 24 hpf, which demonstrated the presence of U-induced hormesis. For exposures to 100 μg/l DU, the amounts of apoptotic signals in the embryos were significantly increased at 20, 24 and 30 hpf. Hormetic effect was not shown but its occurrence between 30 and 48 hpf could not be ruled out. In conclusion, hormetic effect could be induced in zebrafish embryos in a concentration- and time-dependent manner.

  15. Shaping the norms that regulate international commerce of embryos.

    Science.gov (United States)

    Gard, Julie A; Stringfellow, David A

    2014-01-01

    As various embryo technologies in livestock were developed and evolved to a state of usefulness over the past 40 years, scientists with a specific interest in infectious diseases sought to determine the epidemiologic consequences of movement, especially international movement, of increasing numbers of embryos. Many of the foundational studies in this area were reported in Theriogenology, beginning in the 1970s and especially throughout the 1980s and 1990s. Unquestionably, Theriogenology has been a widely used venue for dissemination of basic information on this subject, which ultimately led to the development of the now universally accepted techniques for certification of embryo health. Today it is well-recognized that movement in commerce of embryos, especially in vivo-derived embryos, is a very low-risk method for exchange of animal germ plasm. This paper chronicles the evolution of strategies for health certification of embryos. An overview is provided of the calculated efforts of practitioners, scientists, and regulators to organize, forge necessary partnerships, stimulate needed research, provide purposeful analysis of the results, and, through these processes, guarantee the universal acceptance of efficient protocols for certifying the health of embryos intended for movement in international commerce. Copyright © 2014 Elsevier Inc. All rights reserved.

  16. Role of microRNAs in embryo implantation

    Directory of Open Access Journals (Sweden)

    Jingjie Liang

    2017-11-01

    Full Text Available Abstract Failure of embryo implantation is a major limiting factor in early pregnancy and assisted reproduction. Determinants of implantation include the embryo viability, the endometrial receptivity, and embryo-maternal interactions. Multiple molecules are involved in the regulation of implantation, but their specific regulatory mechanisms remain unclear. MicroRNA (miRNA, functioning as the transcriptional regulator of gene expression, has been widely reported to be involved in embryo implantation. Recent studies reveal that miRNAs not only act inside the cells, but also can be released by cells into the extracellular environment through multiple packaging forms, facilitating intercellular communication and providing indicative information associated with physiological and pathological conditions. The discovery of extracellular miRNAs shed new light on implantation studies. MiRNAs provide new mechanisms for embryo-maternal communication. Moreover, they may serve as non-invasive biomarkers for embryo selection and assessment of endometrial receptivity in assisted reproduction, which improves the accuracy of evaluation while reducing the mechanical damage to the tissue. In this review, we discuss the involvement of miRNAs in embryo implantation from several aspects, focusing on the role of extracellular miRNAs and their potential applications in assisted reproductive technologies (ART to promote fertility efficiency.

  17. [Research progress of whole embryo culture tool and its application].

    Science.gov (United States)

    Han, Jiayin; Liang, Aihua

    2010-03-01

    Whole embryo culture (WEC) is an experimental tool, which is made use of embryos in vitro to replace whole animals to investigate the growth and development of early organs, the embryo toxicity of chemical materials and the mechanism of the occurrence of embryo toxicity. Compared with experiment with whole animals, WEC could reduce the number of experimental animals, shorten experimental time, decrease experimental expenses, eliminate disturbing factors and control dosage more exactly. So it is generally received that WEC tool is a good experimental method to match the principles of replacement, reduction, refinement and responsibility. This article is a review of the WEC tool of rat and mouse, including the development of this tool, announcements, and the application in the development of organs, the embryo toxicity of environmental pollution and heavy metal, safety evaluation of medicine and the embryo toxicity of traditional Chinese medicine and its mechanism. There is also a discussion of the application of this tool in the investigation of the embryo toxicity of traditional Chinese medicine.

  18. Embryo technology in conservation efforts for endangered felids.

    Science.gov (United States)

    Pope, C E

    2000-01-01

    Most of the 36 species of wild cats are classified as threatened, vulnerable or endangered due to poaching and habitat loss. The important role of assisted reproduction techniques (ART) as part of a multifaceted captive breeding program for selected wild cat species is gradually gaining acceptance. This recognition is a result of the progress made during the last decade in which the feasibility of oocyte recovery from gonadotropin-treated females, in vitro fertilization, embryo cryopreservation and embryo transfer (ET) was demonstrated in the domestic cat (Felis catus). Additionally, embryos have been produced in vitro from oocytes matured in vitro after recovery from ex situ ovaries of both domestic and non-domestic cat species and domestic kittens have been born following transfer of these embryos. In vitro fertilization has been successful in at least one-third of wild cat species and kittens were born after transfer of Indian desert cat (Felis sylvestris ornata) embryos into a domestic cat and con-specific transfer of tiger (Panthera tigris) embryos. The domestic cat is not only a valuable model for development of in vitro techniques but may serve as a recipient of embryos from several species of small wild cats.

  19. Experimental taphonomy shows the feasibility of fossil embryos.

    Science.gov (United States)

    Raff, Elizabeth C; Villinski, Jeffrey T; Turner, F Rudolf; Donoghue, Philip C J; Raff, Rudolf A

    2006-04-11

    The recent discovery of apparent fossils of embryos contemporaneous with the earliest animal remains may provide vital insights into the metazoan radiation. However, although the putative fossil remains are similar to modern marine animal embryos or larvae, their simple geometric forms also resemble other organic and inorganic structures. The potential for fossilization of animals at such developmental stages and the taphonomic processes that might affect preservation before mineralization have not been examined. Here, we report experimental taphonomy of marine embryos and larvae similar in size and inferred cleavage mode to presumptive fossil embryos. Under conditions that prevent autolysis, embryos within the fertilization envelope can be preserved with good morphology for sufficiently long periods for mineralization to occur. The reported fossil record exhibits size bias, but we show that embryo size is unlikely to be a major factor in preservation. Under some conditions of death, fossilized remains will not accurately reflect the cell structure of the living organism. Although embryos within the fertilization envelope have high preservation potential, primary larvae have negligible preservation potential. Thus the paleo-embryological record may have strong biases on developmental stages preserved. Our data provide a predictive basis for interpreting the fossil record to unravel the evolution of ontogeny in the origin of metazoans.

  20. Seed architecture shapes embryo metabolism in oilseed rape.

    Science.gov (United States)

    Borisjuk, Ljudmilla; Neuberger, Thomas; Schwender, Jörg; Heinzel, Nicolas; Sunderhaus, Stephanie; Fuchs, Johannes; Hay, Jordan O; Tschiersch, Henning; Braun, Hans-Peter; Denolf, Peter; Lambert, Bart; Jakob, Peter M; Rolletschek, Hardy

    2013-05-01

    Constrained to develop within the seed, the plant embryo must adapt its shape and size to fit the space available. Here, we demonstrate how this adjustment shapes metabolism of photosynthetic embryo. Noninvasive NMR-based imaging of the developing oilseed rape (Brassica napus) seed illustrates that, following embryo bending, gradients in lipid concentration became established. These were correlated with the local photosynthetic electron transport rate and the accumulation of storage products. Experimentally induced changes in embryo morphology and/or light supply altered these gradients and were accompanied by alterations in both proteome and metabolome. Tissue-specific metabolic models predicted that the outer cotyledon and hypocotyl/radicle generate the bulk of plastidic reductant/ATP via photosynthesis, while the inner cotyledon, being enclosed by the outer cotyledon, is forced to grow essentially heterotrophically. Under field-relevant high-light conditions, major contribution of the ribulose-1,5-bisphosphate carboxylase/oxygenase-bypass to seed storage metabolism is predicted for the outer cotyledon and the hypocotyl/radicle only. Differences between in vitro- versus in planta-grown embryos suggest that metabolic heterogeneity of embryo is not observable by in vitro approaches. We conclude that in vivo metabolic fluxes are locally regulated and connected to seed architecture, driving the embryo toward an efficient use of available light and space.

  1. Radial extracorporeal shock wave treatment harms developing chicken embryos

    Science.gov (United States)

    Kiessling, Maren C.; Milz, Stefan; Frank, Hans-Georg; Korbel, Rüdiger; Schmitz, Christoph

    2015-01-01

    Radial extracorporeal shock wave treatment (rESWT) has became one of the best investigated treatment modalities for cellulite, including the abdomen as a treatment site. Notably, pregnancy is considered a contraindication for rESWT, and concerns have been raised about possible harm to the embryo when a woman treated with rESWT for cellulite is not aware of her pregnancy. Here we tested the hypothesis that rESWT may cause serious physical harm to embryos. To this end, chicken embryos were exposed in ovo to various doses of radial shock waves on either day 3 or day 4 of development, resembling the developmental stage of four- to six-week-old human embryos. We found a dose-dependent increase in the number of embryos that died after radial shock wave exposure on either day 3 or day 4 of development. Among the embryos that survived the shock wave exposure a few showed severe congenital defects such as missing eyes. Evidently, our data cannot directly be used to draw conclusions about potential harm to the embryo of a pregnant woman treated for cellulite with rESWT. However, to avoid any risks we strongly recommend applying radial shock waves in the treatment of cellulite only if a pregnancy is ruled out. PMID:25655309

  2. Radial extracorporeal shock wave treatment harms developing chicken embryos.

    Science.gov (United States)

    Kiessling, Maren C; Milz, Stefan; Frank, Hans-Georg; Korbel, Rüdiger; Schmitz, Christoph

    2015-02-06

    Radial extracorporeal shock wave treatment (rESWT) has became one of the best investigated treatment modalities for cellulite, including the abdomen as a treatment site. Notably, pregnancy is considered a contraindication for rESWT, and concerns have been raised about possible harm to the embryo when a woman treated with rESWT for cellulite is not aware of her pregnancy. Here we tested the hypothesis that rESWT may cause serious physical harm to embryos. To this end, chicken embryos were exposed in ovo to various doses of radial shock waves on either day 3 or day 4 of development, resembling the developmental stage of four- to six-week-old human embryos. We found a dose-dependent increase in the number of embryos that died after radial shock wave exposure on either day 3 or day 4 of development. Among the embryos that survived the shock wave exposure a few showed severe congenital defects such as missing eyes. Evidently, our data cannot directly be used to draw conclusions about potential harm to the embryo of a pregnant woman treated for cellulite with rESWT. However, to avoid any risks we strongly recommend applying radial shock waves in the treatment of cellulite only if a pregnancy is ruled out.

  3. Production, Preservation, and Transfer of South American Camelid Embryos

    Directory of Open Access Journals (Sweden)

    Virginia L. Trasorras

    2017-11-01

    Full Text Available The current review summarizes progress in the field of in vitro and in vivo production of South American Camelid embryos. Both methods require ovarian superstimulation (with FSH and eCG to obtain multiple ovulations (in vivo embryo production or to induce follicle growth for oocyte collection (in vitro embryo production. Moreover, superstimulation entails prior administration of hormones that inhibit follicular growth (progesterone, progestagens, and estrogens. Cumulus-oocyte complexes obtained must mature in vivo (buserelin administration or in vitro to then be subjected to in vitro fertilization or intracytoplasmic sperm injection. All these techniques also require morphologically normal, motile spermatozoa to achieve fertilization. Methods used to decrease semen viscosity and to select the best spermatozoa (Percoll®; Androcoll-ETM are described. Additionally, nuclear transfer or cloning has been applied in llamas. Up to now, embryo deep-freezing and vitrification have progressed slowly but are at the height of development. Embryos that are obtained by any of these techniques, either in vivo or in vitro, need to be transferred to synchronized recipient females. The best results are achieved after transfer to the left uterine horn with an ipsilateral ovulation. No live offspring have been obtained after the transfer of cryopreserved embryos. Applying reproductive biotechnologies, such as those described, will permit the expansion of genetically selected animals in the population and also that of wild camelid species, vicunas, and guanacos, whose embryos could then be transferred to the uterus of domestic species.

  4. In vitro regeneration and morphogenesis of somatic embryos of cassava

    Directory of Open Access Journals (Sweden)

    Ádila Melo Vidal

    Full Text Available In this study, the somatic embryogenesis and regeneration of Cigana Preta plants from shoot apices and immature leaves taken from plants cultivated in vitro were examined. To embryo induction the explants were cultivated in Murashige and Skoog (MS medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D or 4-amino-3,5,6-trichloropicolinic acid (Picloram at concentrations of 8.0 and 12 mg L-1. To development of embryos two media cultures with different concentration of BAP (D1 or D2 were evaluated. Embryos in the cotyledonary stage were incubated in germination medium containing MS salts and vitamins, 2.0 µM copper sulfate, 2.4 g L-1 of Phytagel® and 1.77 µM BAP. The highest frequency of calluses and the greatest number of embryos per explant were obtained using the auxin picloram at a concentration of 8.0 mg L-1. The plants regenerated in the picloram treatment exhibited normal development and were transferred to a multiplication medium after a minimum of four weeks. The histological sections of the malformed embryos from foliar explants cultured in the presence of 2,4-D demonstrated that the origin of the cotyledonary structures was independent of the formation of the shoot apical meristem, which was not formed in the embryos, and the majority of the embryos were classified as cornet-shaped. This study demonstrates that in cassava, the use of different auxins provides different conditions for the formation of somatic embryos, and the low rate of conversion into plants results from abnormalities in the embryos.

  5. Danio rerio embryos on Prozac - Effects on the detoxification mechanism and embryo development.

    Science.gov (United States)

    Cunha, V; Rodrigues, P; Santos, M M; Moradas-Ferreira, P; Ferreira, M

    2016-09-01

    In the past decade the presence of psychopharmaceuticals, including fluoxetine (FLU), in the aquatic environment has been associated with the increasing trend in human consumption of these substances. Aquatic organisms are usually exposed to chronic low doses and, therefore, risk assessments should evaluate the effects of these compounds in non-target organisms. Teleost fish possess an array of active defence mechanisms to cope with the deleterious effects of xenobiotics. These include ABC transporters, phase I and II of cellular detoxification and oxidative stress enzymes. Hence, the present study aimed at characterising the effect of FLU on embryo development of the model teleost zebrafish (Danio rerio) concomitantly with changes in the detoxification mechanisms during early developmental phases. Embryos were exposed to different concentrations of FLU (0.0015, 0.05, 0.1, 0.5 and 0.8μM) for 80hours post fertilization. Development was screened and the impact in the transcription of key genes, i.e., abcb4, abcc1, abcc2, abcg2, cyp1a, cyp3a65, gst, sod, cat, ahr, pxr, pparα, pparβ, pparγ, rxraa, rxrab, rxrbb, rxrga, rxrgb, raraa, rarab, rarga evaluated. In addition, accumulation assays were performed to measure the activity of ABC proteins and antioxidant enzymes (CAT and Cu/ZnSOD) after exposure to FLU. Embryo development was disrupted at the lowest FLU concentration tested (0.0015μM), which is in the range of concentrations found in WWTP effluents. Embryos exposed to higher concentrations of FLU decreased Cu/Zn SOD, and increased CAT (0.0015 and 0.5μM) enzymatic activity. Exposure to higher concentrations of FLU decreased the expression of most genes belonging to the detoxification system and upregulated cat at 0.0015μM of FLU. Most of the tested concentrations downregulated pparα, pparβ, pparγ, and raraa, rxraa, rxrab, rxrbb rxrgb and ahr gene expression while pxr was significantly up regulated at all tested concentrations. In conclusion, this study

  6. Synthetic seed production from somatic embryos of Pinus radiata.

    Science.gov (United States)

    Aquea, Felipe; Poupin, María Josefina; Matus, José Tomás; Gebauer, Marlene; Medina, Consuelo; Arce-Johnson, Patricio

    2008-10-01

    Pinus radiata is one of the most important forestry species in the southern hemisphere. This work describes the regeneration of this plant via somatic embryogenesis from immature zygotic embryos. To improve this process, somatic embryogenic cell suspensions were established in liquid media for the generation of material for embryo maturation. Each developmental stage of these suspensions was characterized by microscopy and their growth phases quantified. An alginate-containing medium was used as an encapsulation method for the somatic embryos that were then germinated as artificial seeds in vitro. The protocols described in this work are both useful and reliable for industrial purposes.

  7. [Cholinergic receptors in early (pre-nervous) sea urchin embryos].

    Science.gov (United States)

    Buznikov, G A; Rakic, L

    1998-10-01

    Agonists of nicotinic acetylcholine receptors (nAChR) nicotine and 1-acetyl-4-methylpiperazine do not act on the early sea urchin embryogenesis but evoke calcium shock in both oocytes and early embryos under certain conditions. Many nAChR ligands protect both oocytes and embryos against this shock. There seem to exist putative nAChR on the cell surface of the early sea urchin oocytes and early embryos. Pre-nervous acetylcholine seems to be functionally coupled via these receptors with the second messengers, endogenous activators of the protein kinase C.

  8. Dose estimation in embryo or fetus in external fields

    International Nuclear Information System (INIS)

    Gregori, Beatriz N.

    2001-01-01

    The embryo or the fetus can be irradiated as result of radiological procedures of diagnosis of therapy in where the beam effects directly on the same one or in tissues or peripherical organs. Some authors have suggested that in the first stages of the pregnancy the dose in ovaries can be the good estimated of the dose in embryo or fetus. In advanced conditions of the development, probably also in the early stage, is more appropriated to specify the dose in the embryo or fetus equal of the uterus. The dose in the uterus is a good estimated so much for external irradiation as for radionuclides incorporation

  9. Radial extracorporeal shock wave treatment harms developing chicken embryos

    OpenAIRE

    Kiessling, Maren C.; Milz, Stefan; Frank, Hans-Georg; Korbel, Rüdiger; Schmitz, Christoph

    2015-01-01

    Radial extracorporeal shock wave treatment (rESWT) has became one of the best investigated treatment modalities for cellulite, including the abdomen as a treatment site. Notably, pregnancy is considered a contraindication for rESWT, and concerns have been raised about possible harm to the embryo when a woman treated with rESWT for cellulite is not aware of her pregnancy. Here we tested the hypothesis that rESWT may cause serious physical harm to embryos. To this end, chicken embryos were expo...

  10. File list: Pol.Emb.10.AllAg.Early_embryo [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.Emb.10.AllAg.Early_embryo ce10 RNA polymerase Embryo Early embryo SRX495119,SRX...495120,SRX043866,SRX043864,SRX043865,SRX043863 http://dbarchive.biosciencedbc.jp/kyushu-u/ce10/assembled/Pol.Emb.10.AllAg.Early_embryo.bed ...

  11. File list: Pol.Emb.20.AllAg.Early_embryo [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.Emb.20.AllAg.Early_embryo ce10 RNA polymerase Embryo Early embryo SRX495120,SRX...495119,SRX043864,SRX043866,SRX043863,SRX043865 http://dbarchive.biosciencedbc.jp/kyushu-u/ce10/assembled/Pol.Emb.20.AllAg.Early_embryo.bed ...

  12. File list: NoD.Emb.50.AllAg.Whole_embryo [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available NoD.Emb.50.AllAg.Whole_embryo mm9 No description Embryo Whole embryo SRX658532,SRX6...X658531 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/NoD.Emb.50.AllAg.Whole_embryo.bed ...

  13. File list: InP.Emb.10.AllAg.Whole_embryo [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.Emb.10.AllAg.Whole_embryo mm9 Input control Embryo Whole embryo SRX1123797,SRX1...123798 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/InP.Emb.10.AllAg.Whole_embryo.bed ...

  14. File list: Pol.Emb.50.AllAg.Mixed_embryo [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.Emb.50.AllAg.Mixed_embryo ce10 RNA polymerase Embryo Mixed embryo SRX208772,SRX...208773,SRX208774,SRX208771 http://dbarchive.biosciencedbc.jp/kyushu-u/ce10/assembled/Pol.Emb.50.AllAg.Mixed_embryo.bed ...

  15. Cost-effectiveness of single versus double embryo transfer in IVF in relation to female age

    NARCIS (Netherlands)

    van Loendersloot, Laura L.; Moolenaar, Lobke M.; van Wely, Madelon; Repping, Sjoerd; Bossuyt, Patrick M.; Hompes, Peter G. A.; van der Veen, Fulco; Mol, Ben Willem J.

    2017-01-01

    Objective: To evaluate the cost-effectiveness of single embryo transfer followed by an additional frozen thawed single embryo transfer, if more embryos are available, as compared to double embryo transfer in relation to female age. Study design: We used a decision tree model to evaluate the costs

  16. Proteomics of desiccation tolerance during development and germination of maize embryos

    DEFF Research Database (Denmark)

    Huang, Hui; Møller, Ian Max; Song, Song-Quan

    2012-01-01

    Maize seeds were used to identify the key embryo proteins involved in desiccation tolerance during development and germination. Immature maize embryos (28N) during development and mature embryos imbibed for 72 h (72HN) are desiccation sensitive. Mature maize embryos (52N) during development...

  17. From callus to embryo: a proteomic view on the development and maturation of somatic embryos in Cyclamen persicum.

    Science.gov (United States)

    Rode, Christina; Lindhorst, Kathrin; Braun, Hans-Peter; Winkelmann, Traud

    2012-05-01

    In this study, the proteome structures following the pathway in somatic embryogenesis of Cyclamen persicum were analysed via high-resolution 2D-SDS-PAGE with two objectives: (1) to identify the significant physiological processes during somatic embryogenesis in Cyclamen and (2) to improve the maturation of somatic embryos. Therefore, the effects of maturation-promoting plant growth regulator abscisic acid (ABA) and high sucrose levels on torpedo-shaped embryos were investigated. In total, 108 proteins of differential abundance were identified using a combination of tandem mass spectrometry and a digital proteome reference map. In callus, enzymes related to energy supply were especially distinct, most likely due to energy demand caused by fast growth and cell division. The switch from callus to globular embryo as well as from globular to torpedo-shaped embryo was associated with controlled proteolysis via the ubiquitin-26S proteasome pathway. Storage compound accumulation was first detected 21 days after transfer to plant growth regulator (PGR)-free medium in early torpedo-shaped embryos. Increase in abundance of auxin-amidohydrolase during embryogenesis suggests a possible increase in auxin release in the late embryo stages of Cyclamen. A development-specific isoelectric point switch of catalases has been reported for the first time for somatic embryogenesis. Several proteins were identified to represent markers for the different developmental stages analysed. High sucrose levels and ABA treatment promoted the accumulation of storage compounds in torpedo-shaped embryos. Additionally, proteins of the primary metabolic pathways were decreased in the proteomes of ABA-treated embryos. Thus, ABA and high sucrose concentration in the culture medium improved maturation and consequently the quality of somatic embryos in C. persicum.

  18. Non-invasive metabolomic profiling of embryo culture media and morphology grading to predict implantation outcome in frozen-thawed embryo transfer cycles.

    Science.gov (United States)

    Li, Xiong; Xu, Yan; Fu, Jing; Zhang, Wen-Bi; Liu, Su-Ying; Sun, Xiao-Xi

    2015-11-01

    Assessment of embryo viability is a crucial component of in vitro fertilization and currently relies largely on embryo morphology and cleavage rate. Because morphological assessment remains highly subjective, it can be unreliable in predicting embryo viability. This study investigated the metabolomic profiling of embryo culture media using near-infrared (NIR) spectroscopy for predicting the implantation potential of human embryos in frozen-thawed embryo transfer (FET) cycles. Spent embryo culture media was collected on day 4 after thawed embryo transfer (n = 621) and analysed using NIR spectroscopy. Viability scores were calculated using a predictive multivariate algorithm of fresh embryos with known pregnancy outcomes. The mean viability indices of embryos resulting in clinical pregnancy following FET were significantly higher than those of non-implanted embryos and differed between the 0, 50, and 100 % implantation groups. Notably, the 0 % group index was significantly lower than the 100 % implantation group index (-0.787 ± 0.382 vs. 1.064 ± 0.331, P  0.05). NIR metabolomic profiling of thawed embryo culture media is independent of morphology and correlates with embryo implantation potential in FET cycles. The viability score alone or in conjunction with morphologic grading is a more objective marker for implantation outcome in FET cycles than morphology alone.

  19. In vitro testing of defense reactions in zygotic and somatic embryos of Abies numidica

    Directory of Open Access Journals (Sweden)

    Jiří Hřib

    2011-01-01

    Full Text Available Defense of desiccated cotyledonary somatic embryos and mature zygotic embryos of Abies numidica was tested in vitro by dual cultures with tester, fungus Phaeolus schweinitzii. Both types of embryos expressed defense reactions manifested by inhibited growth of fungal tester towards the embryos. Mycelial growth was described by logistic sigmoid growth model with a single asymptote. Mutual comparisons of mycelial growth in presence of zygotic and somatic embryos showed significant differences in parameters of mycelium growth curves towards the embryos. Larger defense reactions were observed in zygotic embryos relative to somatic embryos and unlimited control cultivations without embryo. The possible role of auxin in the defense response of plant embryos is discussed.

  20. Microinjection of A. aegypti Embryos to Obtain Transgenic Mosquitoes

    OpenAIRE

    Jasinskiene, Nijole; Juhn, Jennifer; James, Anthony A.

    2007-01-01

    In this video, Nijole Jasinskiene demonstrates the methodology employed to generate transgenic Aedes aegypti mosquitoes, which are vectors for dengue fever. The techniques for correctly preparing microinjection needles, dessicating embryos, and performing microinjection are demonstrated.

  1. Microinjection of A. aegypti embryos to obtain transgenic mosquitoes.

    Science.gov (United States)

    Jasinskiene, Nijole; Juhn, Jennifer; James, Anthony A

    2007-01-01

    In this video, Nijole Jasinskiene demonstrates the methodology employed to generate transgenic Aedes aegypti mosquitoes, which are vectors for dengue fever. The techniques for correctly preparing microinjection needles, desiccating embryos, and performing microinjection are demonstrated.

  2. In Silico Dynamics: computer simulation in a Virtual Embryo (SOT)

    Science.gov (United States)

    Abstract: Utilizing cell biological information to predict higher order biological processes is a significant challenge in predictive toxicology. This is especially true for highly dynamical systems such as the embryo where morphogenesis, growth and differentiation require preci...

  3. Effect of zona pellucida on porcine parthenogenetically activated embryos

    DEFF Research Database (Denmark)

    Li, Rong; Liu, Ying; Li, Juan

    2011-01-01

    and survival rates were analysed by chi-squared test. Exp. 1: after activation, 42 blastocysts formed on Day 6, during which the timing of development was monitored (Table 1). PAZF embryos developed faster than PAZI, especially during the first 3 cell cycles. Exp. 2: after activation, 212 and 197 blastocysts......The need for zona pellucida (ZP) during pre-implantation embryo development is still debated. In porcine parthenogenetically activated (PA) embryos, we have previously shown a different distribution in cell numbers on Day 6 blastocysts cultured with or without ZP (Li et al. 2010 Reprod. Fertil. Dev....... 22, 234). In the present study, we expanded this study to include also the timing of early development and the resulting quality and robustness (for vitrification) of porcine PA embryos. Parthenogenetic activation was made first by an electric pulse (1.26 kV cm–1, 80 μs) and then by incubation with 5...

  4. Homocysteine interference in neurulation: a chick embryo model.

    NARCIS (Netherlands)

    Afman, L.A.; Blom, H.J.; Put, N.M.J. van der; Straaten, H.W.M. van

    2003-01-01

    BACKGROUND: Periconceptional folic acid supplementation reduces the occurrence and recurrence risk of neural tube defects (NTD). Mothers of children with NTD have elevated plasma homocysteine levels. Administering homocysteine to chick embryos is reported to cause 27% NTD. Therefore, elevated plasma

  5. Oviduct: roles in fertilization and early embryo development.

    Science.gov (United States)

    Li, Shuai; Winuthayanon, Wipawee

    2017-01-01

    Animal oviducts and human Fallopian tubes are a part of the female reproductive tract that hosts fertilization and pre-implantation development of the embryo. With an increasing understanding of roles of the oviduct at the cellular and molecular levels, current research signifies the importance of the oviduct on naturally conceived fertilization and pre-implantation embryo development. This review highlights the physiological conditions within the oviduct during fertilization, environmental regulation, oviductal fluid composition and its role in protecting embryos and supplying nutrients. Finally, the review compares different aspects of naturally occurring fertilization and assisted reproductive technology (ART)-achieved fertilization and embryo development, giving insight into potential areas for improvement in this technology. © 2017 Society for Endocrinology.

  6. Morphometric analysis of human embryos to predict developmental competence

    DEFF Research Database (Denmark)

    Ziebe, Søren

    2013-01-01

    , but rather choosing and prioritising between the available embryos. Data suggest that only approximately 5% of aspirated human oocytes have the competence to implant and develop into a child and that, in most treatment cycles, there is no oocyte capable of implanting. The most likely outcome is a negative...... pregnancy test, no matter what we choose in the laboratory. Still, both with the increasing complexity of infertile patients treated today and the important focus on reducing multiple pregnancies, it becomes increasingly important to improve our ability to predict the developmental competence of each embryo......Morphometric and morphokinetic approaches toward embryo quality assessment have for many years been difficult due to technical limitations. Today, with improvements in laboratory techniques and subsequent quality, we have a better understanding of the morphometric and kinetics of embryo development...

  7. Cell adhesion in zebrafish embryos is modulated by March 8.

    Science.gov (United States)

    Kim, Mi Ha; Rebbert, Martha L; Ro, Hyunju; Won, Minho; Dawid, Igor B

    2014-01-01

    March 8 is a member of a family of transmembrane E3 ubiquitin ligases that have been studied mostly for their role in the immune system. We find that March 8 is expressed in the zebrafish egg and early embryo, suggesting a role in development. Both knock-down and overexpression of March 8 leads to abnormal development. The phenotype of zebrafish embryos and Xenopus animal explants overexpressing March 8 implicates impairment of cell adhesion as a cause of the effect. In zebrafish embryos and in cultured cells, overexpression of March 8 leads to a reduction in the surface levels of E-cadherin, a major cell-cell adhesion molecule. Experiments in cell culture further show that E-cadherin can be ubiquitinated by March 8. On the basis of these observations we suggest that March 8 functions in the embryo to modulate the strength of cell adhesion by regulating the localization of E-cadherin.

  8. Fruit maturation and in vitro germination of macaw palm embryos ...

    African Journals Online (AJOL)

    -industrial potential. Seed dormancy in palm species may be due to embryo immaturity, which could result from delayed embryogenesis. We evaluated the correspondence between the visual characteristics of maturing fruits and their ...

  9. Targeted mutagenesis in sea urchin embryos using TALENs.

    Science.gov (United States)

    Hosoi, Sayaka; Sakuma, Tetsushi; Sakamoto, Naoaki; Yamamoto, Takashi

    2014-01-01

    Genome editing with engineered nucleases such as zinc-finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs) has been reported in various animals. We previously described ZFN-mediated targeted mutagenesis and insertion of reporter genes in sea urchin embryos. In this study, we demonstrate that TALENs can induce mutagenesis at specific genomic loci of sea urchin embryos. Injection of TALEN mRNAs targeting the HpEts transcription factor into fertilized eggs resulted in the impairment of skeletogenesis. Sequence analyses of the mutations showed that deletions and/or insertions occurred at the HpEts target site in the TALEN mRNAs-injected embryos. The results suggest that targeted gene disruption using TALENs is feasible in sea urchin embryos. © 2013 The Authors Development, Growth & Differentiation © 2013 Japanese Society of Developmental Biologists.

  10. Chapter 1 Historical Background on Gamete and Embryo Cryopreservation.

    Science.gov (United States)

    Ali, Jaffar; AlHarbi, Naif H; Ali, Nafisa

    2017-01-01

    This chapter describes the development of the science of cryopreservation of gametes and embryos of various species including human. It attempts to record in brief the main contributions of workers in their attempts to cryopreserve gametes and embryos. The initial difficulties faced and subsequent developments and triumphs leading to present-day state of the art are given in a concise manner. The main players and their contributions are mentioned and the authors' aim is to do justice to them. This work also attempts to ensure that credit is correctly attributed for significant advances in gamete and embryo cryopreservation. In general this chapter has tried to describe the historical development of the science of cryopreservation of gametes and embryos as accurately as possible without bias or partiality.

  11. Haeckel's Embryos and Evolution: Setting the Record Straight.

    Science.gov (United States)

    Wells, Jonathan

    1999-01-01

    Argues that Ernst Haeckel's drawings of vertebrate embryos (1891), which have been widely used in biology textbooks to illustrate his "Biogenetic Law", are factually flawed. Discusses the problems with Haeckel's drawings and his theory. Contains 14 references. (WRM)

  12. Identification of abnormal gene expression in bovine transgenic somatic cell nuclear transfer embryos

    OpenAIRE

    Cho, Jongki; Kang, Sungkeun; Lee, Byeong Chun

    2014-01-01

    This study was conducted to investigate the expression of three genes related to early embryonic development in bovine transgenic cloned embryos. To accomplish this, development of bovine transgenic somatic cell nuclear transfer (SCNT) embryos was compared with non-transgenic embryos. Next, mRNA transcription of three specific genes (DNMT1, Hsp 70.1, and Mash2) related to early embryo development in transgenic SCNT embryos was compared between transgenic and non-transgenic SCNTs, parthenogene...

  13. Global transcriptome profiles of Italian Mediterranean buffalo embryos with normal and retarded growth.

    Directory of Open Access Journals (Sweden)

    Maria Strazzullo

    Full Text Available The transcriptome profiles were compared for buffalo embryos with normal growth and embryos with retarded growth on Day 25 after mating. Embryos with retarded growth on Day 25 after mating have a reduced likelihood of undergoing attachment to the uterine endometrium and establishing a pregnancy. Italian Mediterranean buffaloes were mated by AI and on Day 25 underwent trans-rectal ultrasonography to ascertain embryo development. Embryos with an embryonic width (EW>2.7 mm were classed as normal embryos and embryos with an EW<2.7 mm were classed as retarded embryos. Three buffaloes with embryos of the largest EW (3.7, 3.7 and 3.9 mm and three buffaloes with embryos of the smallest EW (1.5, 1.6 and 1.9 mm were slaughtered on Day 27 to recover embryos for transcriptome analysis using a bovine custom designed oligo array. A total of 1,047 transcripts were differentially expressed between embryos with normal growth and embryos with retarded growth. Retarded embryos showed 773/1,047 (74% transcripts that were down-regulated and 274/1,047 (26% transcripts that were up-regulated relative to normal embryos; in silico analyses focused on 680/1,047 (65% of the differentially expressed transcripts. The most altered transcripts observed in retarded embryos were associated with membrane structure and function and with metabolic and homeostasis maintenance functions. Other notable functions altered in retarded embryos were developmental processes and in particular nervous system differentiation and function. Specific biochemical pathways such as the complement cascade and coagulation were also altered in retarded embryos. It was concluded from the findings that buffalo embryos with retarded growth on Day 25 after mating show altered gene expression compared with normal embryos, and some de-regulated functions are associated with attachment to the uterine endometrium.

  14. ECTOPIC PREGNANCY AFTER SEQUENTIAL EMBRYO TRANSFER: REVIEW OF 22 CASES

    OpenAIRE

    Nadkarni Purnima K, Nadkarni Kishore, Singh Pooja P, Singh Prabhakar , Nadkarni Aditi A , Agarwal Neha R

    2015-01-01

    Objective: To assess the prevalence of ectopic pregnancy among women who conceived with assisted reproductive technology and to see if there is increased risk after sequential embryo transfer. Methods: The ectopic pregnancy rate for ART pregnancies was calculated among women who conceived and had ectopic pregnancy after ICSI followed by Sequential embryo transfer from an ART centre. Variation in ectopic risk by patient and ART treatment factors was assessed including Sequential transfer, risk...

  15. Emergence and development of gut motility in the chicken embryo

    OpenAIRE

    Chevalier, N. R.; Fleury, V.; Dufour, S.; Proux-Gillardeaux, V.; Asnacios, A.

    2017-01-01

    The gastrointestinal tract transports the food bolus by peristalsis. Gut motility starts at an early age in the developing embryo, well before it is required for nutrition of the organism. We present a comprehensive kinematic study of the emergence and physiological development of gut motility in all regions of the lower digestive tract of the chicken embryo from embryonic days E5 through E9. We characterized motility emergence time, propagation patterns, speed, frequency and amplitude of per...

  16. [The human embryo after Dolly: new practices for new times].

    Science.gov (United States)

    de Miguel Beriain, Iñigo

    2008-01-01

    The possiblity of cloning human beings introduced a lot of issues in our ethical and legal frameworks. In this paper, we will put the focus into the necessary changes in the concept of embryo that our legal systems will have to implement in order to face the new situation. The description of the embryo as a group of cells able to develop into a human being will be defended here as the best way of doing so.

  17. Expression profiling of the mouse early embryo: Reflections and Perspectives

    OpenAIRE

    Ko, Minoru S. H.

    2006-01-01

    Laboratory mouse plays important role in our understanding of early mammalian development and provides invaluable model for human early embryos, which are difficult to study for ethical and technical reasons. Comprehensive collection of cDNA clones, their sequences, and complete genome sequence information, which have been accumulated over last two decades, have provided even more advantages to mouse models. Here the progress in global gene expression profiling in early mouse embryos and, to ...

  18. Study Progress on Tissue Culture of Maize Mature Embryo

    Science.gov (United States)

    Wang, Hongzhen; Cheng, Jun; Cheng, Yanping; Zhou, Xioafu

    It has been paid more and more attention on maize tissue culture as it is a basic work in maize genetic transformation, especially huge breakthrough has been made in maize tissue culture utilizing mature embryos as explants in the recent years. This paper reviewed the study progress on maize tissue culture and plant regeneration utilizing mature embryos as explants from callus induction, subculture, plant regeneration and browning reduction and so on.

  19. Experimental taphonomy shows the feasibility of fossil embryos

    OpenAIRE

    Raff, Elizabeth C.; Villinski, Jeffrey T.; Turner, F. Rudolf; Donoghue, Philip C. J.; Raff, Rudolf A.

    2006-01-01

    The recent discovery of apparent fossils of embryos contemporaneous with the earliest animal remains may provide vital insights into the metazoan radiation. However, although the putative fossil remains are similar to modern marine animal embryos or larvae, their simple geometric forms also resemble other organic and inorganic structures. The potential for fossilization of animals at such developmental stages and the taphonomic processes that might affect preservation before mineralization ha...

  20. Embryo mechanics: balancing force production with elastic resistance during morphogenesis.

    Science.gov (United States)

    Davidson, Lance A

    2011-01-01

    Morphogenesis requires the spatial and temporal control of embryo mechanics, including force production and mechanical resistance to those forces, to coordinate tissue deformation and large-scale movements. Thus, biomechanical processes play a key role in directly shaping the embryo. Additional roles for embryo mechanics during development may include the patterning of positional information and to provide feedback to ensure the success of morphogenetic movements in shaping the larval body and organs. To understand the multiple roles of mechanics during development requires familiarity with engineering principles of the mechanics of structures, the viscoelastic properties of biomaterials, and the integration of force and stress within embryonic structures as morphogenesis progresses. In this chapter, we review the basic engineering principles of biomechanics as they relate to morphogenesis, introduce methods for quantifying embryo mechanics and the limitations of these methods, and outline a formalism for investigating the role of embryo mechanics in birth defects. We encourage the nascent field of embryo mechanics to adopt standard engineering terms and test methods so that studies of diverse organisms can be compared and universal biomechanical principles can be revealed. Copyright © 2011 Elsevier Inc. All rights reserved.

  1. A cutin fluorescence pattern in developing embryos of some angiosperms

    Directory of Open Access Journals (Sweden)

    Ewa Szczuka

    2014-01-01

    Full Text Available A cuticle visualized by auramine O fluorescence appears on the developing embryos of 9 species belonging to Cruciferae, Caryophyllaceae, Plantaginaceae, Linaceae and Papilionaceae. In the investigated species the formation and extent of fluorescing and non-fluorescing embryonic areas follow a similar pattern. At first the cutin fluorescing layer is formed on the apical part of the proembryo without delimited protoderm. This layer extends and at the late globular stage envelops the embryo proper, except for a cell adjoining the suspensor. Fluorescing cutin persists during the heart stage but disappears from the torpedo embryo. During these stages there is no cutine fluorescence on suspensorial cells. Continuous cutin fluorescence appears again on the surface of the whole embryo by the late torpedo stage. Then fluorescence disappears from the radicular part of U-shaped embryos, but persists on the shoot apex, cotyledons and at least on the upper part of hypocotyl. It is assumed that polarization and nutrition of the embryo may be influenced by cuticular changes.

  2. Acute drug treatment in the early C. elegans embryo.

    Directory of Open Access Journals (Sweden)

    Ana Carvalho

    Full Text Available Genetic and genome-wide RNAi approaches available in C. elegans, combined with tools for visualizing subcellular events with high-resolution, have led to increasing adoption of the early C. elegans embryo as a model for mechanistic and functional genomic analysis of cellular processes. However, a limitation of this system has been the impermeability of the embryo eggshell, which has prevented the routine use of small molecule inhibitors. Here, we present a method to permeabilize and immobilize embryos for acute inhibitor treatment in conjunction with live imaging. To identify a means to permeabilize the eggshell, we used a dye uptake assay to screen a set of 310 candidate genes defined by a combination of bioinformatic criteria. This screen identified 20 genes whose inhibition resulted in >75% eggshell permeability, and 3 that permeabilized embryos with minimal deleterious effects on embryo production and early embryonic development. To mount permeabilized embryos for acute drug addition in conjunction with live imaging, we combined optimized inhibition of one of these genes with the use of a microfabricated chamber that we designed. We demonstrate that these two developments enable the temporally controlled introduction of inhibitors for mechanistic studies. This method should also open new avenues of investigation by allowing profiling and specificity-testing of inhibitors through comparison with genome-wide phenotypic datasets.

  3. In vitro culture of coconut (Cocos nucifera L.) zygotic embryos.

    Science.gov (United States)

    Engelmann, Florent; Malaurie, Bernard; N'Nan, Oulo

    2011-01-01

    Coconut is a very important crop for millions of people in tropical countries. With coconut, in vitro culture protocols have been developed with two main objectives, viz. the large scale production of particular types of coconuts and the international exchange and conservation of coconut germplasm. The methods described in this chapter have been developed in the framework of collaborative activities between research institutes in Côte d'Ivoire and France. Two coconut embryo in vitro collecting protocols have been established, one consisting of storing the disinfected embryos in a KCl solution until they are brought back to the laboratory, where they are re-disinfected and inoculated in vitro under sterile conditions, and the other including in vitro inoculation of the embryos in the field. For international germplasm exchange, zygotic embryos inoculated in vitro in plastic test tubes or endosperm cylinders containing embryos in plastic bags are used. For in vitro culture, embryos are inoculated on semi-solid medium supplemented with sucrose and activated charcoal and placed in the dark, and then transferred to light conditions with the same (solid or liquid) medium once the first true leaf is visible and the root system has started developing.

  4. Brooding fathers, not siblings, take up nutrients from embryos

    Science.gov (United States)

    Sagebakken, Gry; Ahnesjö, Ingrid; Mobley, Kenyon B.; Gonçalves, Inês Braga; Kvarnemo, Charlotta

    2010-01-01

    It is well known that many animals with placenta-like structures provide their embryos with nutrients and oxygen. However, we demonstrate here that nutrients can pass the other way, from embryos to the parent. The study was done on a pipefish, Syngnathus typhle, in which males brood fertilized eggs in a brood pouch for several weeks. Earlier research has found a reduction of embryo numbers during the brooding period, but the fate of the nutrients from these ‘reduced’ embryos has been unknown. In this study, we considered whether (i) the brooding male absorbs the nutrients, (ii) siblings absorb them, or (iii) a combination of both. Males were mated to two sets of females, one of which had radioactively labelled eggs (using 14C-labelled amino acids), such that approximately half the eggs in the brood pouch were labelled. This allowed us to trace nutrient uptake from these embryos. We detected that 14C-labelled amino acids were transferred to the male brood pouch, liver and muscle tissue. However, we did not detect any significant 14C-labelled amino-acid absorption by the non-labelled half-siblings in the brood pouch. Thus, we show, to our knowledge, for the first time, that males absorb nutrients derived from embryos through their paternal brood pouch. PMID:19939847

  5. Selfed embryo death in Pinus taeda: a phenotypic profile.

    Science.gov (United States)

    Williams, Claire G

    2008-01-01

    Selective elimination of selfed embryos, or inbreeding depression, is shared among many members of the Pinaceae but it has not been fully characterized at the phenotypic level. Here, two death pattern model hypotheses are tested using 10 621 Pinus taeda embryos sampled in two cohorts. Cones from a single pedigree based on selfed, outbred, parent-offspring and offspring-parent matings were destructively sampled weekly before, during and after fertilization. Selfed embryo deaths adhered to two patterns over the course of development: death was linear with respect to days from fertilization; and a stage-specific death peak occurred during the early embryogeny stage. This death peak occurred from 23 to 36 d after fertilization in the 2004 cohort and from 27 to 34 d after fertilization in the 2006 cohort. Of those selfed embryos that died, 64-83% died at stages where a single dominant embryo was elongating inside the female gametophyte. Additional genetic models are needed to account for the stage-specific death component of selfed P. taeda embryos.

  6. [Characteristics of morphogenesis of the Japanese quail embryos during microgravity

    Science.gov (United States)

    Dadasheva, O. A.; Gur'eva, T. S.; Sychev, V. N.; Jehns, G.; Jahns, G. (Principal Investigator)

    1998-01-01

    Experiments performed in the period of 1995-1996 cooperatively with US investigators within the MIR/SHUTTLE and MIR/NASA space science projects continued exploration of avian embryogenesis in microgravity. Evaluation of Japanese quail embryos incubated in spaceflight microgravity showed that for the most part they were normally developed and compliant with duration of incubation. One of the major morphometric characteristics of embryo are its mass and size. Comparative analysis of body mass values in the space and laboratory and synchronous control groups pointed to a slight retardation. Body length of space embryos mimicked their mass curve. Data on the dynamics of mass and length of Japanese quail embryos support the well-known theory according to which growth and formation are distinguished by equifinality. No differences were revealed by the investigations of individual parts of embryonic bodies in the space and control groups. However, this finding was true only with regard to the embryos that had no developmental abnormalities. A part of embryos had defective eyes (microphtalmia), limbs (twisted fingers), and beaks.

  7. Fluorescence imaging techniques for studying Drosophila embryo development.

    Science.gov (United States)

    Mavrakis, Manos; Rikhy, Richa; Lilly, Mary; Lippincott-Schwartz, Jennifer

    2008-06-01

    This unit describes fluorescence-based techniques for noninvasive imaging of development in living Drosophila embryos, discussing considerations for fluorescent imaging within living embryos and providing protocols for generation of flies expressing fluorescently tagged proteins and for preparation of embryos for fluorescent imaging. The unit details time-lapse confocal imaging of live embryos and discusses optimizing image acquisition and performing three-dimensional imaging. Finally, the unit provides a variety of specific methods for optical highlighting of specific subsets of fluorescently tagged proteins and organelles in the embryo, including fluorescence recovery after photobleaching (FRAP), fluorescence loss in photobleaching (FLIP), and photoactivation techniques, permitting analysis of specific movements of fluorescently tagged proteins within cells. These protocols, together with the relative ease of generating transgenic animals and the ability to express tagged proteins in specific tissues or at specific developmental times, provide powerful means for examining in vivo behavior of any tagged protein in embryos in myriad mutant backgrounds. Copyright 2008 by John Wiley & Sons, Inc.

  8. High dose progesterone effects the growth of early chick embryo

    International Nuclear Information System (INIS)

    Iqbal, I.; Qamar, K.

    2014-01-01

    Objective: To find out the effect of high dose progesterone on the development of early chick embryo. Study Design: Lab based randomized controlled trial. Place and Duration of study: This study was carried out in Army Medical College and Post Graduate Institute of Poultry Sciences, Rawalpindi from June 2010 - December 2010. Material and Methods: Forty five specific pathogen free, fertile, eggs of Fyoumi species of chick were selected at zero hour of incubation. They were incubated at 37.5oC and 75% relative humidity for 26 hrs until the embryos reached stage 8 of the development. Then on stage 8 the eggs were divided into three groups consisting of 15 eggs per group. The first group (GI) was incubated without any operation. The second (G2) and third groups (G3) were injected with two and twenty times more than physiologic does of progesterone respectively. After 48 hours of incvbation, all embryos were examined for their development under light microscopy. Results: All the embryos of G1 and G2 showed normal development according to their stage of development, while 4 out of 11 embryos of G3 were under developed and their survival rate was also less. Conclusion: Exogenous progesterone at levels twenty times above its physiologic range effects the development of chick embryos. Further studies are needed to explain the mechanisms of this effect. (author)

  9. Action of uranium on pre implanted mouse embryos

    International Nuclear Information System (INIS)

    Kundt, Miriam S.

    2001-01-01

    The cultured preimplantation embryos are normally employed to evaluate the effects of environmental pollutants specially metals. Embryos were obtained from hybrid females CBA x C57 Bl following induction of super ovulation. They were incubated from 1 cell stage during 120 hs. in M16 cultured medium. Three different experiments were carried out: A, B and C using uranyl nitrate UO 2 (NO 3 ) 2 6H 2 O as source of uranium. In experiment 'A' the embryos were cultivated in the same culture dish containing final U concentrations of 13, 26, 52, 104 and 208 μgU/ml. In experiment 'B' embryos in a one cell stage were placed in culture medium with uranyl nitrate with final U concentrations of 26, 52, 104 μgU/ml. After 24 hours those embryos which had reached the two-cell stage were transferred to another culture dish to which fresh solutions of uranyl nitrate were added, maintaining the same concentrations of the previous one. In experiment 'C' the embryos were cultivated containing final U concentrations of 26, 52 and 104 μgU/ml and they were transferred to another culture dish every day to which fresh solutions of uranyl nitrate were added. Different embryos parameters were analyzed: 1) Development grade; 2) Number of cell per embryo and metaphases index; and 3) Embryo ploidy. 1) Embryos were observed each 24 hs. to evaluate development grade: 2, 4 and 8 cell stage, morula, early -expanded- hatched blastocysts and atresic embryos. No significant differences were observed in the proportion of embryos arrested either in the one-cell or in the two cell stages in control culture medium regarding different concentrations of U, in a total of 4388 embryos analyzed. From 2 cell stage, moment that the embryo begins to synthesize its own ARNm, the delay in embryonic development increased dose dependent. On the other hand, the toxicological effects in the same concentration are increase from 'A' treatment to 'C' treatment. Embriotoxicology effects are evidenced by an increment in

  10. Dosimetry of radioiodine for embryo and fetus

    Energy Technology Data Exchange (ETDEWEB)

    Berkovski, V.; Eckerman, K.F.; Phipps, A.W.; Nosske, D

    2003-07-01

    This paper discusses the biokinetic and dosimetric models adopted in ICRP Publication 88 for the evaluation of fetal doses resulting from maternal intakes of radioiodine. The biokinetic model is used to simulate the behaviour of iodine in both the mother and the fetus. Such simulations provide the basis for the estimation of the dose to the embryo and determine the distribution of maternal iodine at the beginning of the fetal period. The model considers iodine to accumulate in the fetal thyroid from the 11th week. The dose to the fetus delivered following birth is evaluated with the biokinetic and dosimetric models described in ICRP Publication 67. Although a substantial fraction of the emitted energy of electrons and photons is less than 10 keV, conventionally assumed to be non-penetrating radiation, these emissions can escape the small fetal thyroid. Absorbed fractions for both self-dose and crossfire were evaluated for the requirements of radioiodine dosimetry in ICRP Publication 88. (author)

  11. Neutron induced bystander effect among zebrafish embryos

    Science.gov (United States)

    Ng, C. Y. P.; Kong, E. Y.; Kobayashi, A.; Suya, N.; Uchihori, Y.; Cheng, S. H.; Konishi, T.; Yu, K. N.

    2015-12-01

    The present paper reported the first-ever observation of neutron induced bystander effect (NIBE) using zebrafish (Danio rerio) embryos as the in vivo model. The neutron exposure in the present work was provided by the Neutron exposure Accelerator System for Biological Effect Experiments (NASBEE) facility at the National Institute of Radiological Sciences (NIRS), Chiba, Japan. Two different strategies were employed to induce NIBE, namely, through directly partnering and through medium transfer. Both results agreed with a neutron-dose window (20-50 mGy) which could induce NIBE. The lower dose limit corresponded to the threshold amount of neutron-induced damages to trigger significant bystander signals, while the upper limit corresponded to the onset of gamma-ray hormesis which could mitigate the neutron-induced damages and thereby suppress the bystander signals. Failures to observe NIBE in previous studies were due to using neutron doses outside the dose-window. Strategies to enhance the chance of observing NIBE included (1) use of a mono-energetic high-energy (e.g., between 100 keV and 2 MeV) neutron source, and (2) use of a neutron source with a small gamma-ray contamination. It appeared that the NASBEE facility used in the present study fulfilled both conditions, and was thus ideal for triggering NIBE.

  12. Generation of transgenic Hydra by embryo microinjection.

    Science.gov (United States)

    Juliano, Celina E; Lin, Haifan; Steele, Robert E

    2014-09-11

    As a member of the phylum Cnidaria, the sister group to all bilaterians, Hydra can shed light on fundamental biological processes shared among multicellular animals. Hydra is used as a model for the study of regeneration, pattern formation, and stem cells. However, research efforts have been hampered by lack of a reliable method for gene perturbations to study molecular function. The development of transgenic methods has revitalized the study of Hydra biology(1). Transgenic Hydra allow for the tracking of live cells, sorting to yield pure cell populations for biochemical analysis, manipulation of gene function by knockdown and over-expression, and analysis of promoter function. Plasmid DNA injected into early stage embryos randomly integrates into the genome early in development. This results in hatchlings that express transgenes in patches of tissue in one or more of the three lineages (ectodermal epithelial, endodermal epithelial, or interstitial). The success rate of obtaining a hatchling with transgenic tissue is between 10% and 20%. Asexual propagation of the transgenic hatchling is used to establish a uniformly transgenic line in a particular lineage. Generating transgenic Hydra is surprisingly simple and robust, and here we describe a protocol that can be easily implemented at low cost.

  13. Fundamental cryobiology of mouse ova and embryos

    Energy Technology Data Exchange (ETDEWEB)

    Leibo, S. P.

    1977-01-01

    An increasing fraction of mouse ova and embryos are killed as the rate at which they are cooled to -196/sup 0/C is increased. The survival of these cells depends not only on cooling rate, but also on the minimum subzero temperature to which the cells are cooled. Low temperature microscopy demonstrates that lethal cooling rates are coincident with those that produce intracellular ice formation, and that the lethal temperature appears to be that at which intracellular ice forms. Furthermore, the microscopy shows that ova do not dehydrate when cooled at rates that produce intracellular ice and cell death, but undergo substantial shrinkage when cooled at rates that produce little intracellular ice and high survival. Measurements of the water permeability of mouse ova and the temperature coefficient of that permeability can be used to test a mathematical model formulated to describe the kinetics of water loss at subzero temperatures from a hypothetical cell. The observed dehydration of ova cooled to subzero temperatures at given rates is approximately predicted by the mathematical model, although there is some quantitative discrepancy between the observed and calculated responses.

  14. Fusion of blastomeres in mouse embryos under the action of femtosecond laser radiation. Efficiency of blastocyst formation and embryo development

    Energy Technology Data Exchange (ETDEWEB)

    Osychenko, A A; Zalesskii, A D; Krivokharchenko, A S; Zhakhbazyan, A K; Nadtochenko, V A [N N Semenov Institute of Chemical Physics, Russian Academy of Sciences, Moscow (Russian Federation); Ryabova, A V [A M Prokhorov General Physics Institute, Russian Academy of Sciences, Moscow (Russian Federation)

    2015-05-31

    Using the method of femtosecond laser surgery we study the fusion of two-cell mouse embryos under the action of tightly focused femtosecond laser radiation with the fusion efficiency reaching 60%. The detailed statistical analysis of the efficiency of blastomere fusion and development of the embryo up to the blastocyst stage after exposure of the embryos from different mice to a femtosecond pulse is presented. It is shown that the efficiency of blastocyst formation essentially depends on the biological characteristics of the embryo, namely, the strain and age of the donor mouse. The possibility of obtaining hexaploid embryonal cells using the methods of femtosecond laser surgery is demonstrated. (extreme light fields and their applications)

  15. Antioxidants improve mouse preimplantation embryo development and viability.

    Science.gov (United States)

    Truong, Thi T; Soh, Yu May; Gardner, David K

    2016-07-01

    What is the effect of three antioxidants (acetyl-L-carnitine, N-acetyl-L-cysteine and α-lipoic acid), when used individually and in combination, on mouse embryo development in culture, and subsequent fetal development post-transfer? A combination of antioxidants resulted in significant increases in blastocyst cell number, maintained intracellular glutathione (GSH) levels, supported earlier cleavage times from 5-cell stage to expanded blastocyst, and improved fetal developmental irrespective of incubator oxygen concentration. Acetyl-L-carnitine, N-acetyl-L-cysteine and α-lipoic acid have been shown to have beneficial effects individually in several tissues, and most recently on developing embryos, in the presence of oxidative stress. Morphokinetics of mouse embryos were quantitated using time-lapse imaging. GSH levels in pronucleate oocytes were measured. Blastocysts underwent differential nuclear staining for inner cell mass and trophectoderm cells or were transferred to recipient females to assess implantation and fetal development. Pronucleate oocytes from F1 mice were cultured in 5 or 20% oxygen either individually or in groups of 10, in media G1/G2, in the presence or absence of 10 µM acetyl-L-carnitine /10 µM N-acetyl-L-cysteine /5 µM α-lipoic acid, either individually or in combination. Controls were embryos cultured without antioxidants. Intracellular levels of reduced glutathione were quantitated in pronucleate oocytes. Embryo development and viability were analysed through time-lapse microscopy and embryo transfers. Antioxidants significantly increased mouse blastocyst cell numbers compared with control when used individually (Pantioxidants resulted in faster development rates to 5-cell cleavage stage, which continued until the expanded blastocyst stage when cultured in 20% oxygen. The beneficial effects of combining the antioxidants were greater for embryos cultured individually as opposed to in groups of 10 and for those embryos cultured in 20

  16. Effect of embryo density on in vitro development and gene expression in bovine in vitro-fertilized embryos cultured in a microwell system.

    Science.gov (United States)

    Sugimura, Satoshi; Akai, Tomonori; Hashiyada, Yutaka; Aikawa, Yoshio; Ohtake, Masaki; Matsuda, Hideo; Kobayashi, Shuji; Kobayashi, Eiji; Konishi, Kazuyuki; Imai, Kei

    2013-01-01

    To identify embryos individually during in vitro development, we previously developed the well-of-the-well (WOW) dish, which contains 25 microwells. Here we investigated the effect of embryo density (the number of embryos per volume of medium) on in vitro development and gene expression of bovine in vitro-fertilized embryos cultured in WOW dishes. Using both conventional droplet and WOW culture formats, 5, 15, and 25 bovine embryos were cultured in 125 μl medium for 168 h. The blastocysts at Day 7 were analyzed for number of cells and expression of ten genes (CDX2, IFN-tau, PLAC8, NANOG, OCT4, SOX2, AKR1B1, ATP5A1, GLUT1 and IGF2R). In droplet culture, the rates of formation of >4-cell cleavage embryos and blastocysts were significantly lower in embryos cultured at 5 embryos per droplet than in those cultured at 15 or 25 embryos per droplet, but not in WOW culture. In both droplet and WOW culture, developmental kinetics and blastocyst cell numbers did not differ among any groups. IFN-tau expression in embryos cultured at 25 embryos per droplet was significantly higher than in those cultured at 15 embryos per droplet and in artificial insemination (AI)-derived blastocysts. Moreover, IGF2R expression was significantly lower in the 25-embryo group than in the 5-embryo group and in AI-derived blastocysts. In WOW culture, these expressions were not affected by embryo density and were similar to those in AI-derived blastocysts. These results suggest that, as compared with conventional droplet culture, in vitro development and expression of IFN-tau and IGF2R in the microwell system may be insensitive to embryo density.

  17. Arrested human embryos are more likely to have abnormal chromosomes than developing embryos from women of advanced maternal age

    OpenAIRE

    Qi, Shu-Tao; Liang, Li-Feng; Xian, Ye-Xing; Liu, Jian-Qiao; Wang, Weihua

    2014-01-01

    Background Aneuploidy is one of the major factors that result in low efficiency in human infertility treatment by in vitro fertilization (IVF). The development of DNA microarray technology allows for aneuploidy screening by analyzing all 23 pairs of chromosomes in human embryos. All chromosome screening for aneuploidy is more accurate than partial chromosome screening, as errors can occur in any chromosome. Currently, chromosome screening for aneuploidy is performed in developing embryos, mai...

  18. Improving loblolly pine somatic embryo maturation: comparison of somatic and zygotic embryo morphology, germination, and gene expression.

    Science.gov (United States)

    Pullman, G S; Johnson, S; Peter, G; Cairney, J; Xu, N

    2003-04-01

    Clonal production of loblolly pine ( Pinus taeda L.) through somatic embryogenesis has the potential to meet the increasing industrial demands for high-quality uniform raw materials. A major barrier to the commercialization of this technology is the low quality of the resulting embryos. Twenty-five newly initiated loblolly pine genotypes were followed through the process of liquid culture establishment, embryo maturation, germination, and retrieval from cryogenic storage. A maturation medium, capable of promoting the development of loblolly pine somatic embryos that can germinate, is presented that combines 1/2 P6 modified salts, 2% maltose, 13% polyethylene glycol 8000 (PEG), 5 mg/l abscisic acid (ABA), and 2.5 g/l Gelrite. A procedure for converting and acclimating germinants to growth in soil and greenhouse conditions is also described. A set of somatic seedlings, produced from the maturation medium, showed 100% survival when planted in a field setting. Somatic seedlings showed normal yearly growth relative to standard seedlings from natural seed. The quality of the resulting embryos was examined and compared to that of zygotic embryos using such parameters as morphology, dry weight, germination performance, and gene expression. All of the observations that were made support the conclusion that even with the new maturation medium somatic embryos grow approximately only halfway through the normal sequence of development and then prematurely discontinue growth.

  19. Artificial intelligence techniques for embryo and oocyte classification.

    Science.gov (United States)

    Manna, Claudio; Nanni, Loris; Lumini, Alessandra; Pappalardo, Sebastiana

    2013-01-01

    One of the most relevant aspects in assisted reproduction technology is the possibility of characterizing and identifying the most viable oocytes or embryos. In most cases, embryologists select them by visual examination and their evaluation is totally subjective. Recently, due to the rapid growth in the capacity to extract texture descriptors from a given image, a growing interest has been shown in the use of artificial intelligence methods for embryo or oocyte scoring/selection in IVF programmes. This work concentrates the efforts on the possible prediction of the quality of embryos and oocytes in order to improve the performance of assisted reproduction technology, starting from their images. The artificial intelligence system proposed in this work is based on a set of Levenberg-Marquardt neural networks trained using textural descriptors (the local binary patterns). The proposed system was tested on two data sets of 269 oocytes and 269 corresponding embryos from 104 women and compared with other machine learning methods already proposed in the past for similar classification problems. Although the results are only preliminary, they show an interesting classification performance. This technique may be of particular interest in those countries where legislation restricts embryo selection. One of the most relevant aspects in assisted reproduction technology is the possibility of characterizing and identifying the most viable oocytes or embryos. In most cases, embryologists select them by visual examination and their evaluation is totally subjective. Recently, due to the rapid growth in our capacity to extract texture descriptors from a given image, a growing interest has been shown in the use of artificial intelligence methods for embryo or oocyte scoring/selection in IVF programmes. In this work, we concentrate our efforts on the possible prediction of the quality of embryos and oocytes in order to improve the performance of assisted reproduction technology

  20. Baseline report - tall upland shrubland at the Rocky Flats Environmental Technology Site

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1997-03-01

    Rocky Flats Environmental Technology Site (Site) is located on the Colorado Piedmont east of the Front Range between Boulder and Golden. At an elevation of approximately 6,000 feet, the Site contains a unique ecotonal mixture of mountain and prairie plant species, resulting from the topography and close proximity to the mountain front. The Buffer Zone surrounding the Industrial Area is one of the largest remaining undeveloped areas of its kind along the Colorado Piedmont. A number of plant communities at the Site have been identified as increasingly rare and unique by Site ecologists and the Colorado Natural Heritage Program (CNHP). These include the xeric tallgrass prairie, tall upland shrubland, wetlands, and Great Plains riparian woodland communities. Many of these communities support populations of increasingly rare animals as well, including the Preble`s meadow jumping mouse, grasshopper sparrow, loggerhead shrike, Merriam`s shrew, black crowned night heron, and Hops blue and Argos skipper butterflies. One of the more interesting and important plant communities at the Site is the tall upland shrubland community. It has been generally overlooked by previous Site ecological studies, probably due to its relatively small size; only 34 acres total. Although mentioned in a plant community ordination study conducted by Clark et al. and also in the Site baseline ecological study, few data were available on this plant community before the present study.

  1. DDE in birds' eggs: Comparisons of two methods for estimating critical levels

    Science.gov (United States)

    Blus, L.J.

    1984-01-01

    The sample egg technique and eggshell thickness-residue regression analysis were comparatively evaluated as tools in estimating critical levels of DDE in birds? eggs that seriously affect reproductive successa nd population starts....In comparing critical values of DDE that were derived from the two methods, the estimates were lower using the sample egg technique for both the Brown Pelican (3 ?g/g vs 8 ?g/g) and the Black-crowned Night-Heron (12?g/g/ vs 54 ?g/g) assuming a critical value of eggshell thinning at 20%....Extension of the regression line beyond the eggshell thickness-DDE residue data base is likely to result in spurious critical values of DDE. When sufficient thickness and residue data are available for estimating critical values of DDE from the regression equation, the estimates are meaningful but are likely to be inflated because adverse effects unrelated to eggshell thinning such as parental behavior and embryotoxicity unrelated to eggshell deficiencies are not taken into account.....Establishing critical levels of pollutants in eggs and tissues is a necessary procedure in assessing effects of these chemicals on individuals and populations of sensitive species. There are inherent difficulties in quantifying the effects of any pollutant on population trends and declines in productivity. The sample egg technique is apparently a more sensitive method for estimating critical levels of DDE, but some subjective interpretation is required for results obtained by both methods.

  2. Baseline report - tall upland shrubland at the Rocky Flats Environmental Technology Site

    International Nuclear Information System (INIS)

    1997-03-01

    Rocky Flats Environmental Technology Site (Site) is located on the Colorado Piedmont east of the Front Range between Boulder and Golden. At an elevation of approximately 6,000 feet, the Site contains a unique ecotonal mixture of mountain and prairie plant species, resulting from the topography and close proximity to the mountain front. The Buffer Zone surrounding the Industrial Area is one of the largest remaining undeveloped areas of its kind along the Colorado Piedmont. A number of plant communities at the Site have been identified as increasingly rare and unique by Site ecologists and the Colorado Natural Heritage Program (CNHP). These include the xeric tallgrass prairie, tall upland shrubland, wetlands, and Great Plains riparian woodland communities. Many of these communities support populations of increasingly rare animals as well, including the Preble's meadow jumping mouse, grasshopper sparrow, loggerhead shrike, Merriam's shrew, black crowned night heron, and Hops blue and Argos skipper butterflies. One of the more interesting and important plant communities at the Site is the tall upland shrubland community. It has been generally overlooked by previous Site ecological studies, probably due to its relatively small size; only 34 acres total. Although mentioned in a plant community ordination study conducted by Clark et al. and also in the Site baseline ecological study, few data were available on this plant community before the present study

  3. Live embryo imaging to follow cell cycle and chromosomes stability after nuclear transfer.

    Science.gov (United States)

    Balbach, Sebastian T; Boiani, Michele

    2015-01-01

    Nuclear transfer (NT) into mouse oocytes yields a transcriptionally and functionally heterogeneous population of cloned embryos. Most studies of NT embryos consider only embryos at predefined key stages (e.g., morula or blastocyst), that is, after the bulk of reprogramming has taken place. These retrospective approaches are of limited use to elucidate mechanisms of reprogramming and to predict developmental success. Observing cloned embryo development using live embryo cinematography has the potential to reveal otherwise undetectable embryo features. However, light exposure necessary for live cell cinematography is highly toxic to cloned embryos. Here we describe a protocol for combined bright-field and fluorescence live-cell imaging of histone H2b-GFP expressing mouse embryos, to record cell divisions up to the blastocyst stage. This protocol, which can be adapted to observe other reporters such as Oct4-GFP or Nanog-GFP, allowed us to quantitatively analyze cleavage kinetics of cloned embryos.

  4. Timing of oocyte retrieval and embryo transfer with pregnancy duration.

    Science.gov (United States)

    Beydoun, Hind A; Ugwu, Bethrand; Indika, Sathish; Stadtmauer, Laurel; Bocca, Silvina; Oehninger, Sergio

    2011-11-01

    Recent studies have suggested that assisted reproductive technology (ART) may be associated with a shorter pregnancy duration, possibly due to various aspects of the ART procedure. The purpose of this study was to examine whether pregnancy duration is affected by timing of oocyte retrieval and embryo transfer with respect to the first day of the last menstrual period (LMP) among pregnancies achieved through in vitro fertilization with or without intracytoplasmic sperm injection. A retrospective study was conducted at an academic center in Norfolk, Virginia, with analyses based on 294 ART cycles. Median and interquartile range for pregnancy duration was estimated at 38.2 ± 3.4 weeks. Similarly, median and interquartile ranges for days between LMP and day of oocyte retrieval (27.0 ± 2.0) and between LMP and embryo transfer (29.8 ± 2.2) differed significantly from the standard of 14 days. Timing of oocyte retrieval and embryo transfer with respect to LMP were accelerated among multiple compared with single gestations. For single gestations, pregnancy duration was positively associated with time duration between LMP and embryo transfer (β=0.14, p=0.036). The number of days between oocyte retrieval and embryo transfer was marginally associated with a shorter pregnancy duration in women with multiple gestations (β=3.70, p=0.083). Controlling for patient characteristics, timing of oocyte retrieval and embryo transfer were not significantly associated with pregnancy duration. With few exceptions, timing of oocyte retrieval or embryo transfer did not affect pregnancy duration among ART-conceived live births.

  5. Shared and unique patterns of embryo development in extremophile poeciliids.

    Directory of Open Access Journals (Sweden)

    Rüdiger Riesch

    Full Text Available Closely related lineages of livebearing fishes have independently adapted to two extreme environmental factors: toxic hydrogen sulphide (H(2S and perpetual darkness. Previous work has demonstrated in adult specimens that fish from these extreme habitats convergently evolved drastically increased head and offspring size, while cave fish are further characterized by reduced pigmentation and eye size. Here, we traced the development of these (and other divergent traits in embryos of Poecilia mexicana from benign surface habitats ("surface mollies" and a sulphidic cave ("cave mollies", as well as in embryos of the sister taxon, Poecilia sulphuraria from a sulphidic surface spring ("sulphur mollies". We asked at which points during development changes in the timing of the involved processes (i.e., heterochrony would be detectible.Data were extracted from digital photographs taken of representative embryos for each stage of development and each type of molly. Embryo mass decreased in convergent fashion, but we found patterns of embryonic fat content and ovum/embryo diameter to be divergent among all three types of mollies. The intensity of yellow colouration of the yolk (a proxy for carotenoid content was significantly lower in cave mollies throughout development. Moreover, while relative head size decreased through development in surface mollies, it increased in both types of extremophile mollies, and eye growth was arrested in mid-stage embryos of cave mollies but not in surface or sulphur mollies.Our results clearly demonstrate that even among sister taxa convergence in phenotypic traits is not always achieved by the same processes during embryo development. Furthermore, teleost development is crucially dependent on sufficient carotenoid stores in the yolk, and so we discuss how the apparent ability of cave mollies to overcome this carotenoid-dependency may represent another potential mechanism explaining the lack of gene flow between surface and

  6. Shared and unique patterns of embryo development in extremophile poeciliids.

    Science.gov (United States)

    Riesch, Rüdiger; Schlupp, Ingo; Langerhans, R Brian; Plath, Martin

    2011-01-01

    Closely related lineages of livebearing fishes have independently adapted to two extreme environmental factors: toxic hydrogen sulphide (H(2)S) and perpetual darkness. Previous work has demonstrated in adult specimens that fish from these extreme habitats convergently evolved drastically increased head and offspring size, while cave fish are further characterized by reduced pigmentation and eye size. Here, we traced the development of these (and other) divergent traits in embryos of Poecilia mexicana from benign surface habitats ("surface mollies") and a sulphidic cave ("cave mollies"), as well as in embryos of the sister taxon, Poecilia sulphuraria from a sulphidic surface spring ("sulphur mollies"). We asked at which points during development changes in the timing of the involved processes (i.e., heterochrony) would be detectible. Data were extracted from digital photographs taken of representative embryos for each stage of development and each type of molly. Embryo mass decreased in convergent fashion, but we found patterns of embryonic fat content and ovum/embryo diameter to be divergent among all three types of mollies. The intensity of yellow colouration of the yolk (a proxy for carotenoid content) was significantly lower in cave mollies throughout development. Moreover, while relative head size decreased through development in surface mollies, it increased in both types of extremophile mollies, and eye growth was arrested in mid-stage embryos of cave mollies but not in surface or sulphur mollies. Our results clearly demonstrate that even among sister taxa convergence in phenotypic traits is not always achieved by the same processes during embryo development. Furthermore, teleost development is crucially dependent on sufficient carotenoid stores in the yolk, and so we discuss how the apparent ability of cave mollies to overcome this carotenoid-dependency may represent another potential mechanism explaining the lack of gene flow between surface and cave

  7. Fertilization and Embryo Development of Fresh and Cryopreserved Sibling Oocytes

    Directory of Open Access Journals (Sweden)

    Robert F. Casper

    2010-01-01

    Full Text Available Background: Oocyte cryopreservation is potentially the best way to preserve female fertility forunmarried women or young girls at risk of losing ovarian function. The aim of this study was tocompare fertilization and embryo development in frozen-thawed oocytes to their fresh siblings inwomen undergoing in vitro fertilization (IVF and embryo transfer (ET.Materials and Methods: Eleven infertile women undergoing infertility treatment, between theages of 24 to 37 years (mean ± SD = 31.6 ± 3.5, were included in this study. Mature oocytesfrom each patient were randomized into cryopreserved and fresh groups prior to intracytoplasmicsperm injection (ICSI. One hundred and thirty nine oocytes were retrieved, of which 105 were atmetaphase II (MII. Forty- five fresh MII oocytes were kept in culture whereas their sibling 60 MIIoocytes were cryopreserved using a slow cooling protocol. The frozen oocytes remained in LN2for 2 hours before thawing. ICSI was performed 1-2 hours after thawing for frozen oocytes and 4-5hours after retrieval for fresh oocytes. Fertilization and embryo development were compared.Results: Following thawing, 31 oocytes (51.6 % survived and 22 fertilized (79% while 32 freshoocytes fertilized upon ICSI (71%. The mean ± SE scores for embryos developing from frozenthawedoocytes were significantly lower at 48 and 72 hours post-ICSI than for embryos resultingfrom fresh oocytes (p<0.05.Conclusion: Our data demonstrated that oocyte freezing resulted in acceptable survival ratesfollowing cryopreservation, and similar fertilization rates following ICSI as compared to the freshsibling oocytes. However the number of blastomeres and the embryo quality on day three wassuperior in embryos from fresh oocytes when compared to the frozen oocytes.

  8. Neutron irradiation of rat embryos in utero

    International Nuclear Information System (INIS)

    Vogel, H.H. Jr.

    1978-01-01

    In the rat radiation is most effective in producing congenital anomalies during the organ-forming period (days 9 to 13), which is approximately equivalent to the 14th to 50th days of human pregnancy. We have exposed female Sprague--Dawley rats on the 18th day of pregnancy to single whole-body doses of fission neutrons (20 to 150 rads). After 20 rads there was a small decrease in body weight which lasted from birth to weaning. During this period 9% of the irradiated rats died compared with 4% of the controls. After 50 rads, 65/275 (23.6%) of the rats died between birth and weaning, and the body-weight loss of the survivors was increased. After 100 rads, 62/133 (47%) died at birth or day 1 and 103/133 (77.4%) died before weaning. A large and significant decrease in body weight persisted in the survivors. After 150 rads of fission neutrons, all 95 rats died within 48 hr of birth. From cross-fostering experiments, we believe this is a direct effect of radiation on the embryos and not an indirect action through the mother or her milk. The LD 50 for the period from birth to weaning is approximately 75 rads of fission neutrons. Studies of organ weight were conducted daily for the first week after birth in an attempt to find the cause of radiation mortality. Body weight of the irradiated animals averaged only about one-half that of the controls. The liver, kidney, brain, and testes of the neutron-irradiated rats weighed significantly less than those of the controls. The weights of the spleen, lungs, duodenum, and stomach were decreased but not significantly. The bone marrow appeared depleted in the irradiated long bones, but the spleen maintained active hematopoiesis 1 to 2 months after neutron exposure

  9. The effect of flurbiprofen on the development of anencephaly in early stage chicken embryos.

    Science.gov (United States)

    Özeren, Ersin; Er, Uygur; Güvenç, Yahya; Demirci, Adnan; Arıkök, Ata Türker; Şenveli, Engin; Ergün, Rüçhan Behzat

    2015-04-01

    The study investigated the effect of flurbiprofen on the development of anencephaly in early stage chicken embryos. We looked at four groups with a total of 36 embryos. There was a control group, a normal saline group, a normal-dose group and a high-dose group with ten, ten, eight and eight eggs with embryo respectively. Two embryos in the control group, studied with light microscopy at 48 h, were consistent with 28-29 hours' incubation in the Hamburger-Hamilton System. They had open neural tubes. The other embryos in this group were considered normal. One embryo in the normal saline group was on the occlusion stage at 48 h. One embryo showed an open neural tube. They were compatible with 28-29 hours' incubation in the Hamburger-Hamilton system. The remaining eight embryos showed normal development. In the normal dose group, one embryo showed underdevelopment of the embryonic disc and the embryo was dead. In four embryos, the neural tubes were open. One cranial malformation was found that was complicated with anencephaly in one embryo. In two embryos the neural tubes were closed, as they showed normal development, and they reached their expected stages according to the Hamburger-Hamilton classification. There was no malformation or growth retardation. Four experimental embryos were anencephalic in the high dose group, and three embryos had open neural tubes. One embryo exhibited both anencephaly and a neural tube closure defect. None of the embryos in this group showed normal development. Even the usual therapeutic doses of flurbiprofen increased the risk of neural tube defect. Flurbiprofen was found to significantly increase the risk of anencephaly. The provision of improved technical materials and studies with larger sample sizes will reveal the stage of morphological disruption during the development of embryos.

  10. Creating and selling embryos for "donation": ethical challenges.

    Science.gov (United States)

    Klitzman, Robert; Sauer, Mark V

    2015-02-01

    The commercial creation and sale of embryos has begun, which poses a series of ethical questions that have received little scholarly attention. Some of the concerns that arise are similar to those posed by the sale of gametes, while other issues differ markedly. Questions emerge, first, regarding the rights of the unborn children and their ability to know their biological parents. Companies that create human embryos de novo may wish to keep gamete providers anonymous. Many of these offspring thus will never learn that their parents are not their biologic parents. Yet, such disclosures, regarding not only one but both of these biologic parents, may be important for these individuals; and a lack of this knowledge may impede their physical and psychological health. Second, questions surface regarding the fees that providers should charge for embryos and whether these amounts should vary based on the traits of 1 or both of the gamete donors. Some prospective parents may seek specific traits in a baby (eg, height or eye/hair coloring), which prompts the creation of embryos from 2 gamete donors who possess these characteristics. Third, ownership of embryos created without an advanced directive by patients poses dilemmas (eg, disposition of any remaining embryos). Fourth, guidelines do not yet exist to limit the number of embryos sold from each pair of gamete donors. Hence, unbeknownst to each other, full siblings could potentially meet, get married, and procreate. This discussion has several critical implications for future practice and professional education and policy. Patients with diseases associated with genetic tests may well ask obstetricians, gynecologists, and other physicians about these techniques and practices. Clinicians can refer such patients to assisted reproductive technology specialists; however, familiarity with the basic aspects of the issues and complexities involved could aid these providers and their patients Several of these issues can be

  11. Use of "excess" human embryos for stem cell research: protecting women's rights and health.

    Science.gov (United States)

    Cohen, C B

    2000-01-01

    Proposed National Institutes of Health guidelines for stem cell research are too narrowly drawn and do not adequately protect the freedom of choice and health of women who donate embryos. They need to be expanded to cover not only the point of embryo donation, but also that of embryo creation. Guidelines are provided to ensure that donors undergoing hyperstimulation and egg retrieval gave voluntary informed consent to the production of embryos that might later prove in excess. A standard for determining when embryos have been overproduced is presented to address the possibility that additional embryos will be created for stem cell research in violation of the guidelines and at risk to women's health.

  12. Causes and consequences of chromosome segregation error in preimplantation embryos.

    Science.gov (United States)

    Vázquez-Diez, Cayetana; FitzHarris, Greg

    2018-01-01

    Errors in chromosome segregation are common during the mitotic divisions of preimplantation development in mammalian embryos, giving rise to so-called 'mosaic' embryos possessing a mixture of euploid and aneuploid cells. Mosaicism is widely considered to be detrimental to embryo quality and is frequently used as criteria to select embryos for transfer in human fertility clinics. However, despite the clear clinical importance, the underlying defects in cell division that result in mosaic aneuploidy remain elusive. In this review, we summarise recent findings from clinical and animal model studies that provide new insights into the fundamental mechanisms of chromosome segregation in the highly unusual cellular environment of early preimplantation development and consider recent clues as to why errors should commonly occur in this setting. We furthermore discuss recent evidence suggesting that mosaicism is not an irrevocable barrier to a healthy pregnancy. Understanding the causes and biological impacts of mosaic aneuploidy will be pivotal in the development and fine-tuning of clinical embryo selection methods. © 2018 Society for Reproduction and Fertility.

  13. Establishing some Correlations between Certain Morphometric Parameters and Embryo Quality

    Directory of Open Access Journals (Sweden)

    Nicolae Păcală

    2011-05-01

    Full Text Available The aim of this paper was to establish some correlations between certain morphometric parameters and embryo quality. The morphometric parameters taken into consideration were: zona pellucida thickness, outer and inner diameter, and outer and inner perimeter. For experiments we used embryos recovered at 24 hours from mouse females superovulated with gonadotrope hormones (eCG and hCG. The embryos recovered were cultivated in KSOM media, supplemented with amino acids, and during the in vitro cultivation they were measured at different time intervals for establishing morphometric parameters. The data obtained were statistically analyzed using Minitab 15, using Fitted Line Plot regression that allows testing of the linear and polynomial regression of one variable. After statistical analyze of the data we found that the thickness of the zona pellucida can constitute a morphometric parameter that can be used as an indicator of subsequent development of the 2 cell embryos to morula and blastocyst stage respectively. The other morphometric parameters studied (outer and inner diameter, and outer and inner perimeter cannot be used as indicators of the embryo development.

  14. Current status and future direction of cryopreservation of camelid embryos.

    Science.gov (United States)

    Herrid, M; Vajta, G; Skidmore, J A

    2017-02-01

    Over the past 3 decades, and similar to the horse industry, fresh embryo transfer has been widely practiced on large commercial scales in different camelid species, especially the dromedary camel and alpaca. However, the inability to cryopreserve embryos significantly reduces its broader application, and as such limits the capacity to utilize elite genetic resources internationally. In addition, cryopreservation of the semen of camelids is also difficult, suggesting an extreme sensitivity of the germplasm to cooling and freezing. As a result, genetic resources of camelids must continue to be maintained as living collections of animals. Due to concerns over disease outbreaks such as that of the highly pathogenic Middle East Respiratory Syndrome in the Middle East and Asia, there is an urgent need to establish an effective gene banking system for camelid species, especially the camel. The current review compares and summarizes recent progress in the field of camelid embryo cryopreservation, identifying four possible reasons for the slow development of an effective protocol and describing eight future directions to improve the current protocols. At the same time, the results of a recent dromedary camel embryo transfer study which produced a high morphologic integrity and survival rate of Open Pulled Straw-vitrified embryos are also discussed. Copyright © 2016 Elsevier Inc. All rights reserved.

  15. Preparation of rat serum suitable for mammalian whole embryo culture.

    Science.gov (United States)

    Takahashi, Masanori; Makino, Sayaka; Kikkawa, Takako; Osumi, Noriko

    2014-08-03

    Mammalian whole embryo culture (WEC) is a widely used technique for examining pharmacological toxicity in developing mouse and rat embryos and for investigating the mechanisms of developmental processes. Immediately centrifuged (IC) rat serum is commonly used for WEC and is essential for the growth and development of cultured mouse and rat embryos ex vivo. For the culture of midgestation embryos (i.e., E8.0-12.5 for the mouse, and E10.0-14.5 for the rat), 100% rat serum is the best media for supporting the growth of the embryo ex vivo. To prepare rat serum suitable for WEC, the collected blood should be centrifuged immediately to separate the blood cells from the plasma fraction. After centrifugation, the fibrin clot forms in the upper layer; this clot should be squeezed gently using a pair of sterile forceps and subsequently centrifuged to completely separate the blood cells from the serum. In this video article, we demonstrate our standard protocol for the preparation of optimal IC rat serum, including blood collection from the abdominal aorta of male rats and extraction of the serum by centrifugation.

  16. A Review of the Teratogenic Factors Effect on Embryo

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    Manzarbanoo Shojaei fard

    2017-02-01

    Full Text Available Background & Objectives: Teratology is a branch of embryology science that studies causes, mechanisms and abnormal pattern development. Embryo growth traumatic factors during pregnancy are called teratogens that some teratogens pass the placental barrier and cause adverse effect during development stages and malformation, however a drug may improve general health of the mother, but it might be poisonous for embryo and cause diverse malformation. Since study of embryo health and risk factor in this stage is important, the aim of this review article was the investigation of some types of teratosgens (such as radiation, infectious agents, heat disorders, maternal conditions and particularly the effect of teratogenic drugs on embryo including some legal drugs (such as acetaminophen, thalidomide, acyclovir, sedatives and anticonvulsants and illegal drugs (such as nicotine, alcohol, cocaine and marijuana. Conclusion: In general, teratogens depending on the type and duration of exposure in pregnancyperiod, adversely affect embryo and cause various disorders. A better understanding of these teratogens can contribute to prevent these defects, since many other drugs with similar effects and lower teratogenicity can be used to improve mothers’ health.

  17. Double-effect reasoning and the conception of human embryos.

    Science.gov (United States)

    Murphy, Timothy F

    2013-08-01

    Some commentators argue that conception signals the onset of human personhood and that moral responsibilities toward zygotic or embryonic persons begin at this point, not the least of which is to protect them from exposure to death. Critics of the conception threshold of personhood ask how it can be morally consistent to object to the embryo loss that occurs in fertility medicine and research but not object to the significant embryo loss that occurs through conception in vivo. Using that apparent inconsistency as a starting point, they argue that if that embryo loss is tolerable as a way of conceiving children, it should be tolerable in fertility medicine and human embryonic research. Double-effect reasoning shows, by contrast, that conception in vivo is justified even if it involves the death of persons because the motives for wanting children are not inherently objectionable, because the embryo loss that occurs in unassisted conception is not the means by which successful conception occurs, and because the effect of having children is proportionate to the loss involved. A similar outcome holds true for in vitro fertilisation in fertility medicine but not for in vitro fertilisation for research involving human embryos.

  18. The Early Stages of Heart Development: Insights from Chicken Embryos

    Directory of Open Access Journals (Sweden)

    Johannes G. Wittig

    2016-04-01

    Full Text Available The heart is the first functioning organ in the developing embryo and a detailed understanding of the molecular and cellular mechanisms involved in its formation provides insights into congenital malformations affecting its function and therefore the survival of the organism. Because many developmental mechanisms are highly conserved, it is possible to extrapolate from observations made in invertebrate and vertebrate model organisms to humans. This review will highlight the contributions made through studying heart development in avian embryos, particularly the chicken. The major advantage of chick embryos is their accessibility for surgical manipulation and functional interference approaches, both gain- and loss-of-function. In addition to experiments performed in ovo, the dissection of tissues for ex vivo culture, genomic, or biochemical approaches is straightforward. Furthermore, embryos can be cultured for time-lapse imaging, which enables tracking of fluorescently labeled cells and detailed analysis of tissue morphogenesis. Owing to these features, investigations in chick embryos have led to important discoveries, often complementing genetic studies in mice and zebrafish. As well as including some historical aspects, we cover here some of the crucial advances made in understanding early heart development using the chicken model.

  19. Uterine secretomics: a window on the maternal-embryo interface.

    Science.gov (United States)

    Cheong, Ying; Boomsma, Carolien; Heijnen, Cobi; Macklon, Nick

    2013-03-15

    The analysis of endometrial secretions offers a window on human peri-implantation events that have hitherto been difficult to study noninvasively. Uterine secretomic analysis provides a sensitive means of interrogating the contents of secretions, which have been shown to play a key role in determining endometrial receptivity and embryo-endometrial signaling, and in providing a nourishing environment to the preimplantation embryo. Compared with other means of assessing the endometrium with genomics or proteomics, secretomics offers a nondisruptive approach, allowing analysis during conception cycles. It also provides information on the downstream molecular profile directly encountered by the embryo, offering an integrated review of the complete endometrial surface rather than information representative of only a discrete biopsy site. In this article, recent data derived from uterine secretomics are reviewed. In addition to in vivo studies, recent data from in vitro studies that are changing our understanding of the role of the endometrium in embryo selection are reviewed. Finally, the role of uterine secretions in nutrition and early development programming of the embryo is considered. Copyright © 2013 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  20. Proteomic analysis of embryo development in rice (Oryza sativa).

    Science.gov (United States)

    Xu, Hong; Zhang, Weiping; Gao, Yi; Zhao, Yong; Guo, Lin; Wang, Jianbo

    2012-04-01

    Although embryo development is a major subject in plant growth and development research, a number of aspects of the mechanism of this development process remain unknown. Rice (Oryza sativa) is an excellent monocot plant model for studying embryogenesis with a known genome sequence. Here, we conducted proteomic analysis of embryo development in rice (O. sativa L. ssp. indica cv. 9311). The aim was to investigate and characterize the changes in the protein expression profile during embryo development. For this purpose, the proteome of developing embryos was characterized by two-dimensional gel electrophoresis and nano liquid chromatography/mass spectrometry/mass spectrometry. Proteomic analyses identified 275 differentially expressed proteins throughout the 5 sequential developmental stages from 5 to 30 days after pollination. Most of these proteins were classified into eight functional categories: metabolism, protein synthesis/destination, disease and defense, transporter, transcription, signal transduction, cell growth/division, and storage proteins, which were involved in different cellular and metabolic processes. Hierarchical clustering analyses of protein expression profiles showed that highly expressed proteins in early stages were involved in metabolism, protein synthesis/destination, and most of the other cellular functions, whereas the proteins highly expressed in later stages functioned in the desiccation and dormancy of the embryo.

  1. Influence of the radiation (Co60) in pre-implants rabbit embryos: effect on atypic mitotic index and embryo pole development

    International Nuclear Information System (INIS)

    Approbato, Mario S.; Oliveira Moura, Katia K.V. de; Souza Florencio, Rodopiano de; Garcia, Ricardo; Faria, Renato S.; Benedetti, Leonardo N.; Goulart, Flamarion B.

    1995-01-01

    We studied the effect of ionizing irradiation on 12 New Zealand rabbits (65 embryos), at three different times: at match time (zero hour), two days after and four days after, with two different irradiation doses: five c Gy and ten c Gy. Six rabbits (36 blastocysts) were used as controls. the matching instant was the zero hour. Exactly six days after (± 60 minutes) the embryos of each rabbit was picked up by flushing the uterus with culture media. the embryos were fixed in methanol for 48 hours, and colored with acid Mayer hematoxylin. The following embryo parameters were studied: embryo pole development; percentage of abnormal mitotic figures. irradiation time was associated with lower scores of embryo pole development, but not with irradiation dose. There were no gross abnormalities of embryo pole. The abnormal mitotic cells was affected both by the time and dose of irradiation. (author)

  2. Beneficial effect of two culture systems with small groups of embryos on the development and quality of in vitro-produced bovine embryos.

    Science.gov (United States)

    Cebrian-Serrano, A; Salvador, I; Silvestre, M A

    2014-02-01

    Currently, in vitro-produced embryos derived by ovum pick up (OPU) and in vitro fertilization (IVF) technologies represent approximately one-third of the embryos worldwide in cattle. Nevertheless, the culture of small groups of embryos from an individual egg donor is an issue that OPU-IVF laboratories have to face. In this work, we tested whether the development and quality of the preimplantation embryos in vitro cultured in low numbers (five embryos) could be improved by the addition of epidermal growth factor, insulin, transferrin and selenium (EGF-ITS) or by the WOW system. With this aim, immature oocytes recovered from slaughtered heifers were in vitro matured and in vitro fertilized. Presumptive zygotes were then randomly cultured in four culture conditions: one large group (LG) (50 embryos/500 μl medium) and three smaller groups [five embryos/50 μl medium without (control) or with EGF-ITS (EGF-ITS) and five embryos per microwell in the WOW system (WOW)]. Embryos cultured in LG showed a greater ability to develop to blastocyst stage than embryos cultured in smaller groups, while the blastocyst rate of WOW group was significantly higher than in control. The number of cells/blastocyst in LG was higher than control or WOW, whereas the apoptosis rate per blastocyst was lower. On the other hand, the addition of EGF-ITS significantly improved both parameters compared to the control and resulted in similar embryo quality to LG. In conclusion, the WOW system improved embryo development, while the addition of EGF-ITS improved the embryo quality when smaller groups of embryos were cultured. © 2013 Blackwell Verlag GmbH.

  3. Endometrial signals improve embryo outcome: functional role of vascular endothelial growth factor isoforms on embryo development and implantation in mice.

    Science.gov (United States)

    Binder, N K; Evans, J; Gardner, D K; Salamonsen, L A; Hannan, N J

    2014-10-10

    Does vascular endothelial growth factor (VEGF) have important roles during early embryo development and implantation? VEGF plays key roles during mouse preimplantation embryo development, with beneficial effects on time to cavitation, blastocyst cell number and outgrowth, as well as implantation rate and fetal limb development. Embryo implantation requires synchronized dialog between maternal cells and those of the conceptus. Following ovulation, secretions from endometrial glands increase and accumulate in the uterine lumen. These secretions contain important mediators that support the conceptus during the peri-implantation phase. Previously, we demonstrated a significant reduction of VEGFA in the uterine cavity of women with unexplained infertility. Functional studies demonstrated that VEGF significantly enhanced endometrial epithelial cell adhesive properties and embryo outgrowth. Human endometrial lavages (n = 6) were obtained from women of proven fertility. Four-week old Swiss mice were superovulated and mated with Swiss males to obtain embryos for treatment with VEGF in vitro. Preimplantation embryo development was assessed prior to embryo transfer (n = 19-30/treatment group/output). Recipient F1 female mice (8-12 weeks of age) were mated with vasectomized males to induce pseudopregnancy and embryos were transferred. On Day 14.5 of pregnancy, uterine horns were collected for analysis of implantation rates as well as placental and fetal development (n = 14-19/treatment). Lavage fluid was assessed by western immunoblot analysis to determine the VEGF isoforms present. Mouse embryos were treated with either recombinant human (rh)VEGF, or VEGF isoforms 121 and 165. Preimplantation embryo development was quantified using time-lapse microscopy. Blastocysts were (i) stained for cell number, (ii) transferred to wells coated with fibronectin to examine trophoblast outgrowth or (iii) transferred to pseudo pregnant recipients to analyze implantation rates, placental and

  4. Transcriptomic analysis of neurulation and early organogenesis in rat embryos: an in vivo and ex vivo comparison

    NARCIS (Netherlands)

    Robinson, J.F.; Verhoef, A.; Piersma, A.H.

    2012-01-01

    Cultured embryos mimic the morphological developmental progression of embryos (in vivo) undergoing neurulation and early organogenesis. Using available genomics technologies, comparative molecular-based assessments between cultured embryos and in vivo models may further clarify commonalities and

  5. Equine cloning: in vitro and in vivo development of aggregated embryos.

    Science.gov (United States)

    Gambini, Andrés; Jarazo, Javier; Olivera, Ramiro; Salamone, Daniel F

    2012-07-01

    The production of cloned equine embryos remains highly inefficient. Embryo aggregation has not yet been tested in the equine, and it might represent an interesting strategy to improve embryo development. This study evaluated the effect of cloned embryo aggregation on in vitro and in vivo equine embryo development. Zona-free reconstructed embryos were individually cultured in microwells (nonaggregated group) or as 2- or 3-embryo aggregates (aggregated groups). For in vitro development, they were cultured until blastocyst stage and then either fixed for Oct-4 immunocytochemical staining or maintained in in vitro culture where blastocyst expansion was measured daily until Day 17 or the day on which they collapsed. For in vivo assays, Day 7-8 blastocysts were transferred to synchronized mares and resultant vesicles, and cloned embryos were measured by ultrasonography. Embryo aggregation improved blastocyst rates on a per well basis, and aggregation did not imply additional oocytes to obtain blastocysts. Embryo aggregation improved embryo quality, nevertheless it did not affect Day 8 and Day 16 blastocyst Oct-4 expression patterns. Equine cloned blastocysts expanded and increased their cell numbers when they were maintained in in vitro culture, describing a particular pattern of embryo growth that was unexpectedly independent of embryo aggregation, as all embryos reached similar size after Day 7. Early pregnancy rates were higher using blastocysts derived from aggregated embryos, and advanced pregnancies as live healthy foals also resulted from aggregated embryos. These results indicate that the strategy of aggregating embryos can improve their development, supporting the establishment of equine cloned pregnancies.

  6. Efficacy of postal communication with patients who have cryopreserved pre-embryos.

    Science.gov (United States)

    Brzyski, R G

    1998-11-01

    To compare the characteristics of patients who did and did not respond to a request for information regarding their cryopreserved pre-embryos. Mail survey. Academic-assisted reproductive technology program. One hundred thirty-six patients with cryopreserved pre-embryos. Patients were surveyed by first-class mail regarding their plans for their cryopreserved pre-embryos and their interest in embryo donation. Age, number of stored pre-embryos, and duration of storage of responders and nonresponders at 6 weeks after mailing. Eighty-three patients (62%) did not respond to the survey. Compared with responders, nonresponders were significantly older at the time of embryo cryopreservation, had fewer pre-embryos cryopreserved, and had the pre-embryos cryopreserved for a longer duration. Five responders (9%) expressed an interest in embryo donation. Three patients requested disposal of pre-embryos. Sixteen surveys (12%) were returned as undeliverable. As a group, these patients had the fewest pre-embryos cryopreserved and had the longest duration of storage. A disturbing number of patients with cryopreserved pre-embryos ignored efforts by our program to maintain contact. Older patients with few cryopreserved pre-embryos may require special attention to avoid abandonment.

  7. Embryo production in dogs: from in vitro fertilization to cloning.

    Science.gov (United States)

    Luvoni, G C; Chigioni, S; Beccaglia, M

    2006-08-01

    Increased availability of canine embryos would be desirable to develop research and to apply assisted reproductive technologies in the treatment of infertility and in the improvement of reproductive performances in valuable Canids, both domestic and non-domestic. Embryo production through in vitro fertilization and nuclear transfer has been technically achieved in the dog, and the transfer of cloned embryos has recently resulted in the birth of puppies. However, the efficiency of these technologies is still very limited. This is mainly because of the peculiar characteristics of the canine oocyte and the lack of its full acquisition of developmental competence in vitro. This paper discusses the latest results and aspects on which further research should be focused to provide advances in the field.

  8. Phenotype classification of zebrafish embryos by supervised learning.

    Directory of Open Access Journals (Sweden)

    Nathalie Jeanray

    Full Text Available Zebrafish is increasingly used to assess biological properties of chemical substances and thus is becoming a specific tool for toxicological and pharmacological studies. The effects of chemical substances on embryo survival and development are generally evaluated manually through microscopic observation by an expert and documented by several typical photographs. Here, we present a methodology to automatically classify brightfield images of wildtype zebrafish embryos according to their defects by using an image analysis approach based on supervised machine learning. We show that, compared to manual classification, automatic classification results in 90 to 100% agreement with consensus voting of biological experts in nine out of eleven considered defects in 3 days old zebrafish larvae. Automation of the analysis and classification of zebrafish embryo pictures reduces the workload and time required for the biological expert and increases the reproducibility and objectivity of this classification.

  9. Phenotype classification of zebrafish embryos by supervised learning.

    Science.gov (United States)

    Jeanray, Nathalie; Marée, Raphaël; Pruvot, Benoist; Stern, Olivier; Geurts, Pierre; Wehenkel, Louis; Muller, Marc

    2015-01-01

    Zebrafish is increasingly used to assess biological properties of chemical substances and thus is becoming a specific tool for toxicological and pharmacological studies. The effects of chemical substances on embryo survival and development are generally evaluated manually through microscopic observation by an expert and documented by several typical photographs. Here, we present a methodology to automatically classify brightfield images of wildtype zebrafish embryos according to their defects by using an image analysis approach based on supervised machine learning. We show that, compared to manual classification, automatic classification results in 90 to 100% agreement with consensus voting of biological experts in nine out of eleven considered defects in 3 days old zebrafish larvae. Automation of the analysis and classification of zebrafish embryo pictures reduces the workload and time required for the biological expert and increases the reproducibility and objectivity of this classification.

  10. Partridge embryo pathology in relation to gentamicin-induced lesions

    Directory of Open Access Journals (Sweden)

    Hadi Tavakkoli

    2016-10-01

    Full Text Available Objective: To determine the macroscopic and microscopic lesions of various dosages of gentamicin in the partridge embryo. Methods: Fertile chukar partridge eggs were allocated into four groups. Group 1: salineinjected group whose individuals were administered by sterile physiological saline solution of 0.2 mL/egg inserted into yolk sac. Groups 2, 3 and 4 whose individuals were similarly administered by gentamicin sulfate at a dosage of 80 mg/kg egg-weight once, twice and three times, respectively. Results: Results showed that the embryos were congested and stunted in the gentamicininjected groups. Defects in feet, wings and feather development were accompanied by microscopic lesions in brain, meninges, heart, lungs, liver and kidneys. Histopathological lesions were noticed as edema, undeveloped tissues, necrosis and degeneration in the affected organs. Conclusions: Based on acquired results, it is concluded that gentamicin at above-described dosages causes toxicopathological effects to the partridge embryo in a dose dependent manner.

  11. Use of the Chick Embryo Model in Uveal Melanoma

    Science.gov (United States)

    Kalirai, Helen; Shahidipour, Haleh; Coupland, Sarah E.; Luyten, Gregorius

    2015-01-01

    Animal models play a crucial role in basic and translational oncology research. Conventional rodent experiments, however, face ethical, practical and technical issues that limit their use. The chick embryo represents an accessible and economical in vivo model, which has long been used in developmental biology and for the study of angiogenesis. It is also a recognised xenograft model, and because of its lack of immune system in early development, the chick embryo has established itself as a key model system for cancer research, with which to study various steps in the metastatic process. In this chapter, we review the chick embryo model and the technical approaches adopted by cancer biologists, including advances in real-time imaging, and discuss how this has been or can be applied to improve our understanding of the biological events during uveal melanoma development and metastasis. PMID:27171889

  12. Pine somatic embryogenesis using zygotic embryos as explants.

    Science.gov (United States)

    Pullman, Gerald S; Bucalo, Kylie

    2011-01-01

    Somatic embryogenesis (SE) has the potential to be the lowest-cost method to rapidly produce large numbers of high-value somatic seedlings with desired characteristics for plantation forestry. At least 24 of the 115-120 known Pinus species can undergo SE. Initiation for most species works best with immature megagametophytes as starting material, although a few pines can initiate SE cultures from isolated mature seed embryos. Successful initiation depends heavily on explant type, embryo developmental stage, and medium salt base. Most first reports of initiation used 2,4-D and BAP or a combination of cytokinins. More recent reports have optimized initiation for many Pinus spp., but still use mostly the combinations of auxin and cytokinins. Initiation can be stimulated with medium supplements including abscisic acid (ABA), brassinosteroids, ethylene inhibitors, gibberellin inhibitors, organic acids, putrescine, specific sugar types (maltose, galactose, D-chiro-inositol, and D-xylose), triacontanol, vitamins (B12, biotin, vitamin E, and folic acid), or manipulation of environmental factors including pH, water potential, cone cold storage, gelling agent concentration, and liquid medium. Embryo development and maturation usually occur best on medium containing ABA along with water potential reduction (with sugars and polyethylene glycol) or water availability reduction (with raised gelling agent increasing gel-strength). Activated carbon and maltose may also improve embryo maturation. The main issues holding SE technology back are related to the high cost of producing a somatic seedling, incurred from low initiation percentages for recalcitrant species, culture loss, and decline after initiation and poor embryo maturation resulting in no or poor germination. Although vast progress has been made in pine SE technology over the past 24 years, fundamental studies on seed and embryo physiology, biochemistry, and gene expression are still needed to help improve the technology

  13. Embryo quality predictive models based on cumulus cells gene expression

    Directory of Open Access Journals (Sweden)

    Devjak R

    2016-06-01

    Full Text Available Since the introduction of in vitro fertilization (IVF in clinical practice of infertility treatment, the indicators for high quality embryos were investigated. Cumulus cells (CC have a specific gene expression profile according to the developmental potential of the oocyte they are surrounding, and therefore, specific gene expression could be used as a biomarker. The aim of our study was to combine more than one biomarker to observe improvement in prediction value of embryo development. In this study, 58 CC samples from 17 IVF patients were analyzed. This study was approved by the Republic of Slovenia National Medical Ethics Committee. Gene expression analysis [quantitative real time polymerase chain reaction (qPCR] for five genes, analyzed according to embryo quality level, was performed. Two prediction models were tested for embryo quality prediction: a binary logistic and a decision tree model. As the main outcome, gene expression levels for five genes were taken and the area under the curve (AUC for two prediction models were calculated. Among tested genes, AMHR2 and LIF showed significant expression difference between high quality and low quality embryos. These two genes were used for the construction of two prediction models: the binary logistic model yielded an AUC of 0.72 ± 0.08 and the decision tree model yielded an AUC of 0.73 ± 0.03. Two different prediction models yielded similar predictive power to differentiate high and low quality embryos. In terms of eventual clinical decision making, the decision tree model resulted in easy-to-interpret rules that are highly applicable in clinical practice.

  14. Ionic channels underlying the ventricular action potential in zebrafish embryo.

    Science.gov (United States)

    Alday, Aintzane; Alonso, Hiart; Gallego, Monica; Urrutia, Janire; Letamendia, Ainhoa; Callol, Carles; Casis, Oscar

    2014-06-01

    Over the last years zebrafish has become a popular model in the study of cardiac physiology, pathology and pharmacology. Recently, the application of the 3Rs regulation and the characteristics of the embryo have reduced the use of adult zebrafish use in many studies. However, the zebrafish embryo cardiac physiology is poorly characterized since most works have used indirect techniques and direct recordings of cardiac action potential and ionic currents are scarce. In order to optimize the zebrafish embryo model, we used electrophysiological, pharmacological and immunofluorescence tools to identify the characteristics and the ionic channels involved in the ventricular action potentials of zebrafish embryos. The application of Na(+) or T-type Ca(+2) channel blockers eliminated the cardiac electrical activity, indicating that the action potential upstroke depends on Na(+) and T-type Ca(+2) currents. The plateau phase depends on L-type Ca(+2) channels since it is abolished by specific blockade. The direct channel blockade indicates that the action potential repolarization and diastolic potential depends on ERG K(+) channels. The presence in the embryonic heart of the Nav1.5, Cav1.2, Cav3.2 and ERG channels was also confirmed by immunofluorescence, while the absence of effect of specific blockers and immunostaining indicate that two K(+) repolarizing currents present in human heart, Ito and IKs, are absent in the embryonic zebrafish heart. Our results describe the ionic channels present and its role in the zebrafish embryo heart and support the use of zebrafish embryos to study human diseases and their use for drug testing. Copyright © 2014 Elsevier Ltd. All rights reserved.

  15. Bessel beam fluorescence lifetime tomography of live embryos (Conference Presentation)

    Science.gov (United States)

    Xu, Dongli; Peng, Leilei

    2016-03-01

    Optical tomography allows isotropic 3D imaging of embryos. Scanning-laser optical tomography (SLOT) has superior light collecting efficiency than wide-field optical tomography, making it ideal for fluorescence imaging of live embryos. We previously reported an imaging system that combines SLOT with a novel Fourier-multiplexed fluorescence lifetime imaging (FmFLIM) technique named FmFLIM-SLOT. FmFLIM-SLOT performs multiplexed FLIM-FRET readout of multiple FRET sensors in live embryos. Here we report a recent effort on improving the spatial resolution of the FmFLIM-SLOT system in order to image complex biochemical processes in live embryos at the cellular level. Optical tomography has to compromise between resolution and the depth of view. In SLOT, the commonly-used focused Gaussian beam diverges quickly from the focal plane, making it impossible to achieve high resolution imaging in a large volume specimen. We thus introduce Bessel beam laser-scanning tomography, which illuminates the sample with a spatial-light-modulator-generated Bessel beam that has an extended focal depth. The Bessel beam is scanned across the whole specimen. Fluorescence projection images are acquired at equal angular intervals as the sample rotates. Reconstruction artifacts due to annular-rings of the Bessel beam are removed by a modified 3D filtered back projection algorithm. Furthermore, in combination of Fourier-multiplexing fluorescence lifetime imaging (FmFLIM) method, the Bessel FmFLIM-SLOT system is capable of perform 3D lifetime imaging of live embryos at cellular resolution. The system is applied to in-vivo imaging of transgenic Zebrafish embryos. Results prove that Bessel FmFLIM-SLOT is a promising imaging method in development biology research.

  16. The Roles of Glutathione Peroxidases during Embryo Development

    Science.gov (United States)

    Ufer, Christoph; Wang, Chi Chiu

    2011-01-01

    Embryo development relies on the complex interplay of the basic cellular processes including proliferation, differentiation, and apoptotic cell death. Precise regulation of these events is the basis for the establishment of embryonic structures and the organ development. Beginning with fertilization of the oocyte until delivery the developing embryo encounters changing environmental conditions such as varying levels of oxygen, which can give rise to reactive oxygen species (ROS). These challenges are met by the embryo with metabolic adaptations and by an array of anti-oxidative mechanisms. ROS can be deleterious by modifying biological molecules including lipids, proteins, and nucleic acids and may induce abnormal development or even embryonic lethality. On the other hand ROS are vital players of various signaling cascades that affect the balance between cell growth, differentiation, and death. An imbalance or dysregulation of these biological processes may generate cells with abnormal growth and is therefore potentially teratogenic and tumorigenic. Thus, a precise balance between processes generating ROS and those decomposing ROS is critical for normal embryo development. One tier of the cellular protective system against ROS constitutes the family of selenium-dependent glutathione peroxidases (GPx). These enzymes reduce hydroperoxides to the corresponding alcohols at the expense of reduced glutathione. Of special interest within this protein family is the moonlighting enzyme glutathione peroxidase 4 (Gpx4). This enzyme is a scavenger of lipophilic hydroperoxides on one hand, but on the other hand can be transformed into an enzymatically inactive cellular structural component. GPx4 deficiency – in contrast to all other GPx family members – leads to abnormal embryo development and finally produces a lethal phenotype in mice. This review is aimed at summarizing the current knowledge on GPx isoforms during embryo development and tumor development with an emphasis

  17. A computational model for BMP movement in sea urchin embryos.

    Science.gov (United States)

    van Heijster, Peter; Hardway, Heather; Kaper, Tasso J; Bradham, Cynthia A

    2014-12-21

    Bone morphogen proteins (BMPs) are distributed along a dorsal-ventral (DV) gradient in many developing embryos. The spatial distribution of this signaling ligand is critical for correct DV axis specification. In various species, BMP expression is spatially localized, and BMP gradient formation relies on BMP transport, which in turn requires interactions with the extracellular proteins Short gastrulation/Chordin (Chd) and Twisted gastrulation (Tsg). These binding interactions promote BMP movement and concomitantly inhibit BMP signaling. The protease Tolloid (Tld) cleaves Chd, which releases BMP from the complex and permits it to bind the BMP receptor and signal. In sea urchin embryos, BMP is produced in the ventral ectoderm, but signals in the dorsal ectoderm. The transport of BMP from the ventral ectoderm to the dorsal ectoderm in sea urchin embryos is not understood. Therefore, using information from a series of experiments, we adapt the mathematical model of Mizutani et al. (2005) and embed it as the reaction part of a one-dimensional reaction-diffusion model. We use it to study aspects of this transport process in sea urchin embryos. We demonstrate that the receptor-bound BMP concentration exhibits dorsally centered peaks of the same type as those observed experimentally when the ternary transport complex (Chd-Tsg-BMP) forms relatively quickly and BMP receptor binding is relatively slow. Similarly, dorsally centered peaks are created when the diffusivities of BMP, Chd, and Chd-Tsg are relatively low and that of Chd-Tsg-BMP is relatively high, and the model dynamics also suggest that Tld is a principal regulator of the system. At the end of this paper, we briefly compare the observed dynamics in the sea urchin model to a version that applies to the fly embryo, and we find that the same conditions can account for BMP transport in the two types of embryos only if Tld levels are reduced in sea urchin compared to fly. Copyright © 2014 Elsevier Ltd. All rights

  18. Wnt signaling in the early sea urchin embryo.

    Science.gov (United States)

    Kumburegama, Shalika; Wikramanayake, Athula H

    2008-01-01

    Wnt signaling regulates a remarkably diverse array of cellular and developmental events during animal embryogenesis and homeostasis. The crucial role that Wnt signaling plays in regulating axial patterning in early embryos has been particularly striking. Recent work has highlighted the conserved role that canonical Wnt signaling plays in patterning the animal-vegetal (A-V) axis in sea urchin and sea anemone embryos. In sea urchin embryos, the canonical Wnt signaling pathway is selectively turned on in vegetal cells as early as the 16-cell stage embryo, and signaling through this pathway is required for activation of the endomesodermal gene regulatory network. Loss of nuclear beta-catenin signaling animalizes the sea urchin embryo and blocks pattern formation along the entire A-V axis. Nuclear entry of beta-catenin into vegetal cells is regulated cell autonomously by maternal information that is present at the vegetal pole of the unfertilized egg. Analysis of Dishevelled (Dsh) regulation along the A-V axis has revealed the presence of a cytoarchitectural domain at the vegetal pole of the unfertilized sea urchin egg. This vegetal cortical domain appears to be crucial for the localized activation of Dsh at the vegetal pole, but the precise mechanisms are unknown. The elucidation of how Dsh is selectively activated at the vegetal cortical domain is likely to provide important insight into how this enigmatic protein is regulated during canonical Wnt signaling. Additionally, this information will shed light on the origins of embryonic polarity during animal evolution. This chapter examines the roles played by the canonical Wnt signaling pathway in the specification and patterning of the A-V axis in the sea urchin. These studies have led to the identification of a novel role for canonical Wnt signaling in regulating protein stability, and continued studies of Wnt signaling in this model system are likely to reveal additional roles for this pathway in regulating early

  19. Dealing of embryo/fetal exposures in a medical setting

    International Nuclear Information System (INIS)

    Miller, K.L.

    1991-01-01

    The medical environment presents two uniquely different situations involving potential radiation exposure to the embryo or fetus. On the one hand we have the potential for radiation exposure to the embryo of fetus of the occupationally exposed employee where the recommendations are well defined. On the other hand we have the pregnant or potentially pregnant patient requiring diagnostic radiological examinations or therapeutic procedures where the recommendations are not as well defined. This paper provides an overview of both situations, a discussion of potential exposures and a review of methods for estimating fetal doses when the exposed individual is the patient

  20. Status of the human embryo: Philosophical Foundations from Phenomenology

    Directory of Open Access Journals (Sweden)

    Maria Emilia de Oliveira Schpallir Silva

    2017-10-01

    Full Text Available Given the difficulty in demonstrating the moment of ontogenesis in which personalization takes place, we sought to define, from a philosophic point of view, the nature of the human embryo regarding its individuality, using Phenomenology, specifically reflections of philosophers Bourghet and Merleau-Ponty on the embryo. Although the statement of their individuality does not entail ethical content in itself, from the point of view of ethical responsibility, it is an extremely important fact to be considered in the bioethical reflection about the moment of ontogeny from which human life must (ethical duty be protected.

  1. ‘Vanishing embryo syndrome’ in IVF/ICSI

    DEFF Research Database (Denmark)

    Hvidtjørn, Dorte; Grove, Jakob; Schendel, Diana

    2005-01-01

    BACKGROUND: In a Danish population-based cohort study assessing the risk of cerebral palsy in children bornafter IVF, we made some interesting observations regarding ‘vanishing co-embryos’. METHODS andRESULTS: All live-born children born in Denmark from 1 January 1995 to 31 December 2000 were...... found indications of an increasedrisk of cerebral palsy in those children resulting from pregnancies, where the number of embryos transferred washigher than the number of children born. CONCLUSIONS: The association between vanishing embryo syndromeand incidence of cerebral palsy following IVF requires...

  2. Improving production of Zebra Fish Embryos in the lab

    DEFF Research Database (Denmark)

    Thomsen, Jens Peter; Adu, Robert Ohene

    2011-01-01

    in the laboratory. Culture conditions were maintained in the aquaria as stipulated in the OECD draft proposal for a new guideline on fish embryo tests. Furthermore, a sequence of steps were adopted and followed to improve upon previous work done in the lab in 2006. About 200 eggs were produced in one spawn trap...... within an hour of onset of light, an improvement over the 50-60 eggs produced in the previous work. This result demonstrates that with the right culture conditions and proper optimisation of procedure the required number of embryos needed for toxicity testing can be obtained....

  3. [Effect of hyperglycemia on rat embryos in vitro].

    Science.gov (United States)

    He, W G

    1993-10-01

    Using whole embryo culture, the effect of hyperglycemia on the embryogenesis of rat was studied. Hyperglycemia could cause death, malformation and intrauterine growth retardation of embryos. The effect of hyperglycemia was more obvious in the early embryogenesis of rat in vitro, and showed a dose related effect. The results suggested that the blood glucose level of diabetic women should be controlled strictly during pregnancy, particularly in the first trimester of pregnancy. It may play an important role in preventing death, congenital malformation and intrauterine growth retardation of fetus in diabetic women.

  4. 'New embryos' - new challenges for the ethics of stem cell research.

    Science.gov (United States)

    Holm, Søren

    2008-01-01

    Among the many ethical issues raised by human embryonic stem cell research (in the following all references to 'stem cells' should be read as references to human embryonic stem cells), two have gained specific prominence: (1) whether stem cell research is ethically problematic because it entails the destruction of human embryos and (2) what kind of control embryo donors should have over the stem cell lines derived from their embryos. In the present paper, I will analyse how these two issues are engaged by various attempts to derive stem cells from anomalous embryos (e.g. embryos in cleavage arrest, embryos not implanted following pre-implantation genetic diagnosis or embryos created by altered nuclear transfer) or in ways that are claimed to be non-destructive for the embryo (e.g. blastocyst or blastomere biopsy). Copyright 2008 S. Karger AG, Basel.

  5. [Chapter 8. The embryo and the fœtus in criminal law].

    Science.gov (United States)

    Roujou de Boubée, Gabriel

    2018-03-07

    In criminal law the embryo is not assimilated to a person. It is therefore not protected in relation to offences against persons. However, the embryo does enjoy a certain, very relative and unapplied, protection through specific offences.

  6. Characteristics of pregnancies and offspring following transfer of bovine in vivo embryos assessed by nanorespirometry

    DEFF Research Database (Denmark)

    Lopes, Ana Sofia; Madsen, S E; Greve, Torben

    2008-01-01

    .05), with quality I embryos associated with heavier calves when compared to quality III embryos. Furthermore, light traction was more frequently used to retrieve heavier animals (P male calves were 3.2 kg heavier than females (P

  7. Effect of chilling on the development of in vitro produced bovine embryos at various cleavage stages

    OpenAIRE

    Balasubramanian, S.; Rho, Gyu-Jin

    2006-01-01

    Purpose: Bovine embryos and zygotes are known to be sensitive to “temperature shock” when cooled to temperatures near 0°C. The effect of chilling on in vitro derived embryos at various cleavage stages was investigated.

  8. The influence of the type of embryo culture medium on neonatal birthweight after single embryo transfer in IVF.

    Science.gov (United States)

    Vergouw, Carlijn G; Kostelijk, E Hanna; Doejaaren, Els; Hompes, Peter G A; Lambalk, Cornelis B; Schats, Roel

    2012-09-01

    Does the type of medium used to culture fresh and frozen-thawed embryos influence neonatal birthweight after single embryo transfer (SET) in IVF? A comparison of two commercially available culture media showed no significant influence on mean birthweight and mean birthweight adjusted for gestational age, gender and parity (z-scores) of singletons born after a fresh or frozen-thawed SET. Furthermore, we show that embryo freezing and thawing cycles may lead to a significantly higher mean birthweight. Animal studies have shown that culture media constituents are responsible for changes in birthweight of offspring. In human IVF, there is still little knowledge of the effect of medium type on birthweight. Until now, only a small number of commercially available culture media have been investigated (Vitrolife, Cook(®) Medical and IVF online medium). Our study adds new information: it has a larger population of singleton births compared with the previously published studies, it includes outcomes of other media types (HTF and Sage(®)), not previously analysed, and it includes data on frozen-thawed SETs. This study was a retrospective analysis of birthweights of singleton newborns after fresh (Day 3) or frozen-thawed (Day 5) SET cycles, using embryos cultured in either of two different types of commercially available culture media, between 2008 and 2011. Before January 2009, a single-step culture medium was used: human tubal fluid (HTF) with 4 mg/ml human serum albumin. From January 2009 onwards, a commercially available sequential medium was introduced: Sage(®), Quinn's advantage protein plus medium. Singletons born after a fresh SET (99 embryos cultured in HTF and 259 in Sage(®)) and singletons born after a frozen-thawed SET (32 embryos cultured in HTF only, 41 in HTF and Sage(®) and 86 in Sage(®) only) were analysed. Only patients using autologous gametes without the use of a gestational carrier were considered. Also excluded were (vanishing) twins, triplets

  9. To delay or not to delay a frozen embryo transfer after a failed fresh embryo transfer attempt?

    Science.gov (United States)

    Santos-Ribeiro, Samuel; Siffain, Johannie; Polyzos, Nikolaos P; van de Vijver, Arne; van Landuyt, Lisbet; Stoop, Dominic; Tournaye, Herman; Blockeel, Christophe

    2016-05-01

    To evaluate if increasing the interval between a failed fresh embryo transfer and a subsequent frozen embryo transfer (FET) cycle has any effect on clinical pregnancy rates (CPRs). Retrospective cohort study. University-based tertiary referral center. Women who underwent at least one FET after ovarian stimulation for in vitro fertilization (IVF) and a failed fresh embryo transfer attempt from January 2010 to November 2014. We divided our sample according to the "timing" of the first FET (TF-FET), defined by the interval between oocyte retrieval and the FET cycle start date. The start of the FET was classified as either immediate (≤22 days after oocyte retrieval) or delayed (>22 days after oocyte retrieval). None. CPR after the first FET. A total of 1,183 FET cycles (performed in 1,087 women) were included in our study. No significant differences were found between the immediate and delayed FET groups regarding age, number of oocytes retrieved, number of good-quality embryos produced, embryo developmental stage at FET, and number of frozen embryos transferred. Most importantly, the CPRs of the first FET did not differ significantly according to the TF-FET (32.5% after immediate FET vs. 31.7% after delayed FET), even after adjusting for potential confounding with the use of multivariable logistic regression. FETs performed immediately after fresh IVF cycles had CPRs similar to those postponed to a later time. Therefore, deferring FETs may unnecessarily prolong time to pregnancy. Copyright © 2016 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  10. Application of Green Fluorescent Protein as a Marker for Selection of Transgenic Mouse Embryos before Implantation

    OpenAIRE

    BAĞIŞ, Haydar

    2001-01-01

    In this study, we evaluated the application of green fluorescent protein (GFP) from the jellyfish Aequorea victoria, for efficient selection of possible transgenic and non-mosaic mouse embryos after microinjection. We injected 353 one-cell mouse zygotes with a DNA fragment carrying Gfp gene under the control of b-actin gene promoter. Eighty-seven per cent (307 embryos) of the injected embryos survived after microinjection. The surviving embryos were cultured for an additional day and then an...

  11. EMBRYOSPLITTING - A WAY TO INCREASE THE EFFICIENCY OF EMBRYO - TRANSFER IN SHEEP

    OpenAIRE

    Vintila, I.; Benscik, I.; Pacala, N.; Corin, N.; Babusik, I.; Kulickova, L.

    1996-01-01

    In has been compared the gestation results and the number of lambs obtained from 40 unsplitted integral embryos collected from 31 superovulated sheep, to the same parameters in the situation when 40 demi -embryos were used, in order to estimate the embryosplitting advantage to increase the efficiency or embryo- transfer in sheep. The gestation percentage obtained by the recipient sheep in the case of integral embryos, was 57,5% and in case of demiembryos was 47,5%, only. Also, the proportion ...

  12. Climatic factors affecting quantity and quality grade of in vivo derived embryos of cattle.

    Science.gov (United States)

    Chinchilla-Vargas, Josué; Jahnke, Marianna M; Dohlman, Tyler M; Rothschild, Max F; Gunn, Patrick J

    2018-02-17

    The present study investigated the effects of climatic variables on the quality grade and quantity of in vivo derived cattle embryos in the Midwestern United States. Climatic information included greatest and least daily temperature, average daily wind speed and average temperature-humidity index for each of the 765 records. The response variables included the number of ovarian structures, viable embryos, quality grade 1 embryos, quality grade 2 embryos, quality grade 3 embryos, freezable embryos (sum of quality grade 1 and quality grade 2 embryos), transferable embryos (sum of quality grade 1-3 embryos), degenerate embryos and unfertilized ova. Measures for variables among the breeds of donors and sires grouped by geographical origin were compared. A negative effect of greater temperatures during the early embryonic development stage tended (P climatic variables have significant effects on the in vivo production of cattle embryos and that wind speed should be considered in future analyses of factors affecting embryo quality. Copyright © 2018 Elsevier B.V. All rights reserved.

  13. Influence of recipient cytoplasm cell stage on transcription in bovine nucleus transfer embryos

    DEFF Research Database (Denmark)

    Smith, S. D.; Soloy, E.; Kanka, J.

    1996-01-01

    Nucleus transfer for the production of multiple embryos derived from a donor embryo relies upon the reprogramming of the donor nucleus so that it behaves similar to a zygotic nucleus. One indication of nucleus reprogramming is the RNA synthetic activity. In normal bovine embryogenesis, the embryo...

  14. Polyamine levels during the development of zygotic and somatic embryos of Pinus radiata

    Science.gov (United States)

    Rakesh Minocha; Dale R. Smith; Cathie Reeves; Kevin D. Steele; Subhash C. Minocha

    1999-01-01

    Changes in the cellular content of three polyamines (putrescine, spermidine and spermine) were compared at different stages of development in zygotic and somatic embryos of Pinus radiata D. Don. During embryo development, both the zygotic and the somatic embryos showed a steady increase in spermidine content, with either a small decrease or no...

  15. In vivo assay for the developmental competence of embryo-derived zebrafish cell lines

    NARCIS (Netherlands)

    Speksnijder, JE; Hage, WJ; Lanser, PH; Collodi, P; Zivkovic, D

    We have produced chimeric zebrafish embryos by transplanting permanent embryo-derived cell lines into blastula-stage embryos. Furthermore, we have established a fluorescent in vivo assay to monitor the developmental effects and fate of such transplanted cells using confocal laser scanning

  16. A microfluidic system supports single mouse embryo culture leading to full-term development

    NARCIS (Netherlands)

    Esteves, Telma Cristina; van Rossem, F.; Nordhoff, Verena; Schlatt, Stefan; Boiani, Michele; le Gac, Severine

    2013-01-01

    The present study demonstrates the feasibility of application of a microfluidic system for in vitro culture of pre-implantation mouse embryos, with subsequent development to full-term upon embryo transfer. Specifically, embryos cultured in groups in nL volume chambers achieve pre-implantation

  17. Differences in gene expression profiles between human preimplantation embryos cultured in two different IVF culture media

    NARCIS (Netherlands)

    Kleijkers, Sander H. M.; Eijssen, Lars M. T.; Coonen, Edith; Derhaag, Josien G.; Mantikou, Eleni; Jonker, Martijs J.; Mastenbroek, Sebastiaan; Repping, Sjoerd; Evers, Johannes L. H.; Dumoulin, John C. M.; van Montfoort, Aafke P. A.

    2015-01-01

    Is gene expression in human preimplantation embryos affected by the medium used for embryo culture in vitro during an IVF treatment? Six days of in vitro culture of human preimplantation embryos resulted in medium-dependent differences in expression level of genes involved in apoptosis, protein

  18. Differences in gene expression profiles between human preimplantation embryos cultured in two different IVF culture media

    NARCIS (Netherlands)

    Kleijkers, S.H.M.; Eijssen, L.M.T.; Coonen, E.; Derhaag, J.G.; Mantikou, E.; Jonker, M.J.; Mastenbroek, S.; Repping, S.; Evers, J.L.H.; Dumoulin, J.C.M.; van Montfoort, A.P.A.

    2015-01-01

    STUDY QUESTION: Is gene expression in human preimplantation embryos affected by the medium used for embryo culture in vitro during an IVF treatment? SUMMARY ANSWER: Six days of in vitro culture of human preimplantation embryos resulted in medium-dependent differences in expression level of genes

  19. Polyamines and their biosynthetic enzymes during somatic embryo development in red spruce (Picea rubens Sarg.)

    Science.gov (United States)

    Rakesh Minocha; Subhash C. Minocha; Stephanie Long

    2004-01-01

    The major objective of this study was to determine if the observed changes in polyamines and their biosynthetic enzymes during somatic embryo development were specifically related to either the stage of the embryo development or to the duration of time spent on the maturation medium. Somatic embryos of red spruce (Picea rubens) at different...

  20. White communicating rami in human embryos at the end of the fifth week.

    Science.gov (United States)

    Bruska, M; Łupicka, J; Pytel, A; Woźniak, W

    2010-05-01

    White communicating rami were traced in 8 human embryos of developmental stages 14 and 15 (aged 33 and 36 postovulatory days, respectively). In embryos at stage 14 the white communicating rami were found in the spinal nerves T1 to T9. In embryos at stage 15 the white communicating rami were present at the spinal cord levels T1 to L3.

  1. Cold-induced imbibition damage of lettuce embryos: A study using cryo-scanning electron microscopy

    NARCIS (Netherlands)

    Nijsse, J.; Walther, P.; Hoekstra, F.

    2004-01-01

    The impact of rehydration on a multicellular organism was studied in lettuce (Lactuca sativa L.) embryos, using cryo-scanning electron microscopy (cryo-SEM). Naked embryos were sensitive to imbibitional stress, whereas embryos with an intact, thick-walled endosperm were not. Imbibitional injury to

  2. ECTOPIC PREGNANCY AFTER SEQUENTIAL EMBRYO TRANSFER: REVIEW OF 22 CASES

    Directory of Open Access Journals (Sweden)

    Nadkarni Purnima K, Nadkarni Kishore, Singh Pooja P, Singh Prabhakar , Nadkarni Aditi A , Agarwal Neha R

    2015-07-01

    Full Text Available Objective: To assess the prevalence of ectopic pregnancy among women who conceived with assisted reproductive technology and to see if there is increased risk after sequential embryo transfer. Methods: The ectopic pregnancy rate for ART pregnancies was calculated among women who conceived and had ectopic pregnancy after ICSI followed by Sequential embryo transfer from an ART centre. Variation in ectopic risk by patient and ART treatment factors was assessed including Sequential transfer, risk factor for ectopic pregnancy, tubal infertility and previous ectopic pregnancy. Results: Of 1960 women, who underwent ICSI, 1047(53.41% had positive pregnancy, 22(2.1% women had an ectopic pregnancy out of which 2(9% had heterotrophic pregnancy. The mean age was 30.4±4.33. Tubal factor contributed to 31.81% cases and 27.27% had previous history of ectopic pregnancy. Sequential transfer was done in all the patients with more than one embryo transferred. There was no significant increase in the ectopic pregnancy when Sequential embryo transfer was done. Conclusion: Tubal factor infertility and history of previous ectopic pregnancy contributed to risk factor for ectopic pregnancy in our study. Sequential transfer did not increase the risk of ectopic pregnancy.

  3. Single molecule transcription factor dynamics in the syncytial Drosophila embryo

    Science.gov (United States)

    Darzacq, Xavier

    During early development in the Drosophila embryo, cell fates are determined over the course of just 2 hours with exquisite spatio-temoral precision. One of the key regulators of this process is the transcription factor Bicoid which forms a concentration gradient across the long axis of the embryo. Although Bicoids' primary role is activation at the anterior, where concentrations are highest, it is also known to play a role in the posterior where there are only 100s of molecules per nucleus. Understanding how Bicoid can find its target at such low concentrations has remained intractable, largely due to the inability to perform single molecule imaging in the context of the developing embryo. Here we use lattice light sheet microscopy to overcome the technical barriers of sample thickness and auto-fluorescence to characterize the single molecule dynamics of Bicoid. We find that off-rates do not vary across the embryo and that instead the on-rates are modulated through the formation of clusters that enrich local concentration. This data is contrary to the current concentration dependent model of Bicoid function since local concentration within the nucleus is now a regulated parameter and suggests a previously unknown mechanism for regulation at extremely low concentrations.

  4. Development of HY1 hybrid embryos between a cultivar of ...

    African Journals Online (AJOL)

    Reciprocal crosses between Phaseolus vulgaris and P. coccineus frequently lead to embryos abortion in all the developmental stages. However, the P. vulgaris NI637 cultivar and the wild form NI1108 of P. coccineus present abilities for combination. The abortion rate of pods is 93.93% when NI1108 is the maternal parent ...

  5. Embryo rescue of crosses between diploid and tetraploid grape ...

    African Journals Online (AJOL)

    ajl yemi

    2011-12-19

    Dec 19, 2011 ... Five cross combinations Jumeigui×Xinghua No.1, 87-1×Kyoho, Kyoho×Muscat Hamburg, Jumeigui×. Hongqitezao and Red globle×Kyoho were used as the testing materials. Factors that affect embryo rescue from crossed seeds between diploid and tetraploid grape were studied applying L25(55).

  6. Embryo rescue of crosses between diploid and tetraploid grape ...

    African Journals Online (AJOL)

    Five cross combinations Jumeigui×Xinghua No.1, 87-1×Kyoho, Kyoho×Muscat Hamburg, Jumeigui× Hongqitezao and Red globle×Kyoho were used as the testing materials. Factors that affect embryo rescue from crossed seeds between diploid and tetraploid grape were studied applying L25(55) orthogonal experiment ...

  7. Embryo mortality and early post-oestrous cycle embryonic death ...

    African Journals Online (AJOL)

    An estimate of embryo mortality (cycles longer than 28 days) was obtained from milk progesterone analysis and delayed return rate in two dairy herds. Oestrus cycle lengths were measured, and cycles grouped according to whether artificial insemination (AI) had, or had not, been performed. Early post-oestrous cycle ...

  8. High frequency induction of somatic embryos and plantlet ...

    African Journals Online (AJOL)

    High frequency induction of somatic embryos and plantlet regeneration from nodal explants of Hygrophila spinosa T. Anders. A Varshney, A Shahzad, M Anis. Abstract. An efficient protocol is described for the rapid in vitro plant regeneration of a medicinally important plant, Hygrophila spinosa through direct somatic ...

  9. Factors affecting conception rates in cattle following embryo transfer

    African Journals Online (AJOL)

    ACSS

    production, sale of breeding stock, among others. New technologies can help achieve increased productivity, but need to be transferred to producers to cause impact. (Ehui and Shapiro, 2009; Yang and. Honaramooz, 2010). One such technology is whereby one high quality cow could be made to produce up to 32 embryos ...

  10. Miliary tuberculosis after in vitro fertilization and embryo ...

    African Journals Online (AJOL)

    Miliary tuberculosis after in vitro fertilization and embryo transplantation. Hongbo Liu, Li Zhao. Department of Respiratory Medicine, Shengjing Hospital of China Medical University, Shenyang,. Liaoning China. 110004. Abstract. Background: With the development of assisted reproductive technology, more patients with ...

  11. In vitro culture of mouse embryos amniotic fluid ID human

    African Journals Online (AJOL)

    1989-07-15

    Jul 15, 1989 ... Because human amniotic fluid is a physiological, balanced ultrafiltrate, it has been considered as an inexpensive alternative culture medium in. IVF. A study of the development of mouse embryos in human amniotic fluid was undertaken to assess the suitability of this as an optional culture medium in human ...

  12. Debating elective single embryo transfer after in vitro fertilization: a ...

    African Journals Online (AJOL)

    However, despite clinical recommendations and policy statements, patients in clinical practice frequently do request for the transfer of multiple embryos in order to have twins. Such requests conflict with policy guidelines and create an ethical dilemma for physicians: Should the physician do as the couple requests, and there ...

  13. A Marble Embryo: Meanings of a Portrait from 1900

    Science.gov (United States)

    Hopwood, Nick

    2012-01-01

    Portraits of scientists use attributes of discovery to construct identities; portraits that include esoteric accessories may fashion identities for these too. A striking example is a marble bust of the anatomist Wilhelm His by the Leipzig sculptor Carl Seffner. Made in 1900, it depicts the founder of modern human embryology looking down at a model embryo in his right hand. This essay reconstructs the design and viewing of this remarkable portrait in order to shed light on private and public relations between scientists, research objects and audiences. The bust came out of a collaboration to model the face of the composer Johann Sebastian Bach and embodies a shared commitment to anatomical exactitude in three dimensions. His’s research agendas and public character explain the contemplative pose and unprecedented embryo model, which he had laboriously constructed from material a midwife supplied. The early contexts of display in the His home and art exhibitions suggest interpretive resources for viewers and hence likely meanings. Seffner’s work remains exceptional, but has affinities to portraits of human embryologists and embryos produced since 1960. Embryo images have acquired such controversial prominence that the model may engage us more strongly now than it did exhibition visitors around 1900. PMID:22606754

  14. Intersensory Redundancy Educates Selective Attention in Bobwhite Quail Embryos

    Science.gov (United States)

    Lickliter, Robert; Bahrick, Lorraine E.; Markham, Rebecca G.

    2006-01-01

    We assessed whether exposure to amodal properties in bimodal stimulation (e.g. rhythm, rate, duration) could educate attention to amodal properties in subsequent unimodal stimulation during prenatal development. Bobwhite quail embryos were exposed to an individual bobwhite maternal call under several experimental and control conditions during the…

  15. High resolution multiplexed functional imaging in live embryos (Conference Presentation)

    Science.gov (United States)

    Xu, Dongli; Zhou, Weibin; Peng, Leilei

    2017-02-01

    Fourier multiplexed fluorescence lifetime imaging (FmFLIM) scanning laser optical tomography (FmFLIM-SLOT) combines FmFLIM and Scanning laser optical tomography (SLOT) to perform multiplexed 3D FLIM imaging of live embryos. The system had demonstrate multiplexed functional imaging of zebrafish embryos genetically express Foster Resonant Energy Transfer (FRET) sensors. However, previous system has a 20 micron resolution because the focused Gaussian beam diverges quickly from the focused plane, makes it difficult to achieve high resolution imaging over a long projection depth. Here, we present a high-resolution FmFLIM-SLOT system with achromatic Bessel beam, which achieves 3 micron resolution in 3D deep tissue imaging. In Bessel-FmFLIM-SLOT, multiple laser excitation lines are firstly intensity modulated by a Michelson interferometer with a spinning polygon mirror optical delay line, which enables Fourier multiplexed multi-channel lifetime measurements. Then, a spatial light modulator and a prism are used to transform the modulated Gaussian laser beam to an achromatic Bessel beam. The achromatic Bessel beam scans across the whole specimen with equal angular intervals as sample rotated. After tomography reconstruction and the frequency domain lifetime analysis method, both the 3D intensity and lifetime image of multiple excitation-emission can be obtained. Using Bessel-FmFLIM-SLOT system, we performed cellular-resolution FLIM tomography imaging of live zebrafish embryo. Genetically expressed FRET sensors in these embryo will allow non-invasive observation of multiple biochemical processes in vivo.

  16. Perflurooctanoic Acid Induces Developmental Cardiotoxicity in Chicken Embryos and Hatchlings

    Science.gov (United States)

    Perfluorooctanoic acid (PFOA) is a widespread environmental contaminant that is detectable in serum of the general U.S. population. PFOA is a known developmental toxicant that induces mortality in mammalian embryos and is thought to induce toxicity via interaction with the peroxi...

  17. Can repeated superovulation and embryo recovery in Boer goats ...

    African Journals Online (AJOL)

    LehloenyaKC

    The alternative method of transcervical embryo collection in goats has been limited in the past because of the difficulty of passing the catheter through the cervix (Ishwar & Memon, 1996; Suyadi et al., 2000). Even after the discovery that by injecting animals with hormones such as oxytocin, oestrogen and prostaglandin, ...

  18. Asymbiotic germination of immature embryos of a medicinally ...

    African Journals Online (AJOL)

    H.piri

    The immature embryos (28 weeks after pollination) were inoculated on M (Mitra et al., 1976), and PDA. (Potato Dextrose Agar) media, with and without different growth additives. The seeds showed positive germination response in both the nutrient media but the frequency and onset of germination response and associated ...

  19. Desiccation tolerance of embryos of Syagrus oleracea, a cerrado ...

    African Journals Online (AJOL)

    aghomotsegin

    2015-03-18

    Mar 18, 2015 ... There is great interest in seedling production of plant species with ornamental and food potential; however, there is great difficulty in propagating this species. A method of fruit drying was proposed with the goal of evaluating the desiccation tolerance of embryos of guariroba palm Syagrus oleracea.

  20. Successful whole embryo culture with commercially available reagents.

    Science.gov (United States)

    Glanville-Jones, Hannah C; Woo, Ngai; Arkell, Ruth M

    2013-01-01

    Since its development in the 1970's, whole embryo culture (WEC) has provided an important method of growing and observing murine embryos ex utero. During WEC, embryos are immersed in a combination of rat serum and cell culture media, and supplied with heat and appropriate mixtures of CO₂ and oxygen that mimic growth conditions in utero. One significant factor limiting the widespread use of WEC is the perception that commercially produced rat serum is inadequate to support normal rates of embryonic growth and development. Conversely, production of serum 'in-house' is technically demanding, time-consuming and expensive. The current study aimed to identify a WEC medium comprising commercially manufactured rat serum that would produce cultured embryos of comparable standard to those grown in utero. A mixed culture medium, composed of 50% commercial rat serum and 50% F12 Ham's cell culture medium with an N-2 neuronal cell growth supplement, was shown to support both a rate of growth, and the development of a range of features comparable to that which normally occur in vivo. Furthermore, the F12 (N-2) supplemented rat serum displayed a very low propensity to induce morphological abnormalities during the culture period. The study establishes a novel method of successful WEC using readily available commercial reagents and should enable the broader use of WEC.

  1. [Developmental toxicity of retrorsine on mouse embryos in vitro].

    Science.gov (United States)

    Han, Jiayin; Liang, Aihu; Yi, Yan

    2011-07-01

    To investigate the fetotoxicity of retrorsine. Mouse whole embryo culture (WEC) was applied. Post-implantation (8.5 d) mouse embryos were isolated from their mothers and put into the medium of immediately centrifuged serum (ICS) prepared from rats. Different concentrations of retrorsine (12.5, 25, 50, 100 mg x L(-1)) were added into the WEC culture. Development (yolk sac diameter, crown-rump length, head length, somite number) and organic morphodifferentiation (yolk sac circulation, allantois, embryonic flexion, heart, brain, optic-otic-olfactory organ, branchial arch, maxillary, mandible, bud) of embryos were observed at 48 h after treatment. Obvious fetotoxicity could be observed in various retrorsine treatment groups in a dose-dependent manner. Development of embryos was delayed significantly at dose 12.5-100 mg x L(-1). Malformations were shown in all organic morphodifferentiation indexes, especially in otic-olfactory organ, branchial arch, maxillary, mandible, bud. Retrorsine had obvious fetotoxicity in vitro WEC culture, indicating that exposure of pregnant mice to retrorsine may have potential risk on fetals.

  2. [Toxicity of monocrotaline on in vitro cultured mouse embryos].

    Science.gov (United States)

    Han, Jiayin; Liang, Aihua; Yi, Yan; Gao, Shuangrong; Nilsen, Odd Georg

    2011-02-01

    To investigate the fetotoxicity of monocrotaline. Mouse whole embryo culture (WEC) was applied. Post-implantation (8.5 d) mouse embryos were isolated from their mothers and put into the medium of immediately centrifuged serum (ICS) prepared from rats. Different concentrations of monocrotaline (100, 50, 25, 12.5 mg x L(-1)) were added into the WEC. Development (yolk sac diameter, crown-rump length, head length, somite number) and organic morphodifferentiation (yolk sac circulation, allantois, embryonic flexion, heart, brain, optic-otic-olfactory organ, branchial arch, maxillary, mandible, bud) of embryos were observed at 48 h after treatment. Obvious fetotoxicity could be observed in various monocrotaline treatment groups in a dose-dependent manner. Development of embryos was delayed significantly at dose 12.5-100 mg x L(-1). Malformations were shown in all organic morphodifferentiation indice, especially in opti-otic organ, mandible and bud. Monocrotaline had obvious fetotoxicity in vitro WEC, indicating that exposure of pregnant mice to monocrotaline may have potential risk on fetus.

  3. Gas exchange of the ostrich embryo during peak metabolism in ...

    African Journals Online (AJOL)

    Oxygen (O2) consumption and carbon dioxide (CO2) excretion of ostrich embryos were studied on 45 ostrich eggs in various stages of development. A closed respirometry system was used for eggs subjected to ????10 days of incubation, while an open flow system was used for older eggs. A total of 102 measurements ...

  4. Embryo implantation: Shedding light on the roles of ovarian ...

    African Journals Online (AJOL)

    Implantation is a crucial step in mammalian reproduction, as it is a gateway to further embryonic development and successful pregnancy. Successful implantation requires coordinated interactions between the blastocyst and uterus. Uterine receptivity for embryo implantation is regulated by the ovarian hormones estrogen ...

  5. Miliary tuberculosis after in vitro fertilization and embryo ...

    African Journals Online (AJOL)

    Background: With the development of assisted reproductive technology, more patients with infertility prefer to get pregnant by in vitro fertilization and embryo transplantation (IVF-ET). But the indications of IVF-ET must be strictly controlled by the clinicians. Case report: We described a case of a 29-year-old pregnant Chinese ...

  6. Germination response of coconut ( Cocos nucifera L.) zygotic embryo

    African Journals Online (AJOL)

    The study investigated the effects of liquid and solid media in the propagation of coconut (Cocos nucifera) zygotic embryos at initiation stage. Eeuwen's medium supplemented with growth hormones naphthalene acetic acid ( NAA) and indole butyric acid (IBA) at different concentrations (0.5, 1.0, 1.5, 2.0 and 2.5mg/l) were ...

  7. Superovulation response and in vivo embryo production potential of ...

    African Journals Online (AJOL)

    Boran (n=25) and Boran*Holstein (n=11) cows were superovullatedFSH with three doses level (300, 250 and 200IU) divided in to morning and afternoon decreasing doses over 4 daysto study the superovulatory response and embryo production potential. Time to estrus, duration of estrus, and CL count were used to ...

  8. Increasing twinning rate in beef cattle through embryo transfer ...

    African Journals Online (AJOL)

    Thirty-nine Limousin X Friesian heifers and forty-five Limousin X Friesian cows were randomly allocated to three treatments in a twin-induction breeding program. Animals were inseminated twice with Limousin semen (treatment A) or inseminated twice also with Hereford semen and each implanted with one embryo 7 days ...

  9. Efficient somatic embryo production of Limau madu ( Citrus ...

    African Journals Online (AJOL)

    Effects of N6-benzylaminopurine (BAP) concentration, initial cell density and carbon sources and concentrations for producing cell suspension and somatic embryos of Limau madu (Citrus suhuiensis Hort. ex Tanaka) were investigated using cell suspension culture. Cells were first inoculated into Murashige and Skoog (MS) ...

  10. Germination response of coconut (Cocos nucifera L.) zygotic embryo ...

    African Journals Online (AJOL)

    ADOWIE PERE

    ABSTRACT: The study investigated the effects of liquid and solid media in the propagation of coconut (Cocos nucifera) zygotic embryos at initiation stage. Eeuwen's medium supplemented with growth hormones naphthalene acetic acid ( NAA) and indole butyric acid (IBA) at different concentrations (0.5, 1.0, 1.5, 2.0 and ...

  11. PLANETARY EMBRYO BOW SHOCKS AS A MECHANISM FOR CHONDRULE FORMATION

    Energy Technology Data Exchange (ETDEWEB)

    Mann, Christopher R.; Boley, Aaron C. [Department of Physics and Astronomy University of British Columbia Vancouver, BC V6T 1Z1 (Canada); Morris, Melissa A. [Physics Department State University of New York at Cortland Cortland, NY 13045 (United States)

    2016-02-20

    We use radiation hydrodynamics with direct particle integration to explore the feasibility of chondrule formation in planetary embryo bow shocks. The calculations presented here are used to explore the consequences of a Mars-size planetary embryo traveling on a moderately excited orbit through the dusty, early environment of the solar system. The embryo’s eccentric orbit produces a range of supersonic relative velocities between the embryo and the circularly orbiting gas and dust, prompting the formation of bow shocks. Temporary atmospheres around these embryos, which can be created via volatile outgassing and gas capture from the surrounding nebula, can non-trivially affect thermal profiles of solids entering the shock. We explore the thermal environment of solids that traverse the bow shock at different impact radii, the effects that planetoid atmospheres have on shock morphologies, and the stripping efficiency of planetoidal atmospheres in the presence of high relative winds. Simulations are run using adiabatic and radiative conditions, with multiple treatments for the local opacities. Shock speeds of 5, 6, and 7 km s{sup −1} are explored. We find that a high-mass atmosphere and inefficient radiative conditions can produce peak temperatures and cooling rates that are consistent with the constraints set by chondrule furnace studies. For most conditions, the derived cooling rates are potentially too high to be consistent with chondrule formation.

  12. Transcervical embryo recovery in Saanen goats | Lima-Verde ...

    African Journals Online (AJOL)

    In order to study the embryo recovery rate using the transcervical technique in Saanen goats raised in the tropics, 20 donors were submitted to an oestrus synchronisation treatment using intravaginal progestagen sponges for 11 days. On the ninth day of treatment, goats received intramuscular injections of 50 µg ...

  13. Effect of nitrates on embryo induction efficiency in cotton ...

    African Journals Online (AJOL)

    Cotton (Gossypium hirsutum L.) cv Coker-312 callus culture was assessed in terms of its usefulness as a system for investigating the effect of nitrates from different chemical compounds of nitrogen on embryo induction percentage in calli as the plant growth and cell differentiation mainly based on nitrogen. Both sources and ...

  14. The contractility of elongated microvilli in early sea urchin embryos.

    Science.gov (United States)

    Spiegel, Evelyn; Howard, Louisa; Spiegel, Melvin

    1990-04-01

    Elongated microvilli attach the early sea urchin embryo to the fertilization envelope and support it in a concentric position within the perivitelline space. The contractility of the elongated microvilli was demonstrated in several ways. (1) During normal cleavage, these microvilli change their length to adapt to the change in shape and numbers of blastomeres. (2) When treated with calcium-free sea water, embryos become eccentrically located and the microvilli extend further than normal on one side; when returned to normal sea water, the embryos become centered again. (3) Several agents cause the fertilization envelope to become higher and thinner than normal and the elongated microvilli to extend correspondingly if treated within ten min after fertilization. In some cases, both elongated microvilli and fertilization envelope return to normal size when returned to normal sea water. (4) Fertilization in a papain solution causes the elongated microvilli and the fertilization envelope to contract to the surface of the embryo. (5) Refertilization after the papain-induced contraction can bring about the elongation of these microvilli and the elevation of the fertilization envelope a second time. It was also shown that elongated microvilli are extended immediately upon fertilization, at the same time as the short microvilli. The firm adherence of the tips of elongated microvilli to the fertilization envelope by means of extracellular matrix fibers is shown in a high voltage electron microscope stereoimage. This allows us to understand why it is that when the elongated microvilli extend or contract, the fertilization envelope also extends and contracts accordingly.

  15. Microspore embryogenesis: reprogramming cell fate from pollen to embryo development

    NARCIS (Netherlands)

    Hui Li,

    2014-01-01

    Microspore embryogenesis is an expression of plant cell totipotency that leads to the production of haploid embryos. Besides being a widely exploited plant breeding tool, microspore embryogenesis is also a fascinating system that can be used to obtain a deeper mechanistic understanding of plant

  16. Clipboard Functional embryo sac formation in Arabidopsis without ...

    Indian Academy of Sciences (India)

    Srinivas

    (C) Ovule after the completion of meiosis, showing the meiotic tetrad (mt) of reduced megaspores, outer integument (oi) and inner integument (ii). (D) Ovule at anthesis, containing an egg cell (ec) and fused central cell nucleus (ccn), indicated with arrows. The embryo sac is outlined with a dotted line. (E) Developing seed.

  17. Cardiac hypertrophy in chick embryos induced by hypothermia

    International Nuclear Information System (INIS)

    Boehm, C.; Johnson, T.R.; Caston, J.D.; Przybylski, R.J.

    1987-01-01

    A decrease in incubation temperature from 38 to 32 0 C elicits a decrease in chicken embryo size and weight with concomitant heart enlargement if done after day 10 of incubation. When assayed at day 18 of incubation with the hypothermia started on day 11 or 14, evidence is presented that the heart enlargement is an hypertrophy with no detectable hyperplasia. Supporting data are presented for various physical parameters showing increases in heart wet and dry weight, volume, area, wall thickness, and cell size. There was little difference in DNA content and nuclear [ 3 H]thymidine labeling index between hearts of control and hypothermic embryos. Hearts of hypothermic embryos showed a slight increase in water content and considerable increases in RNA, protein, and glycogen content per unit DNA. The average size of polysomes isolated from hypothermic hearts was larger than that of polysomes isolated from controls. Microscopic studies showed no obvious increase in amount of capillary beds, connective tissue, and myocardial cells. Annulate lamellae were found only in myocardial cells of hypothermic embryos in sparse amounts and low frequency but always associated with large deposits of glycogen

  18. Widespread Contamination of Arabidopsis Embryo and Endosperm Transcriptome Data Sets.

    Science.gov (United States)

    Schon, Michael A; Nodine, Michael D

    2017-04-01

    A major goal of global gene expression profiling in plant seeds has been to investigate the parental contributions to the transcriptomes of early embryos and endosperm. However, consistency between independent studies has been poor, leading to considerable debate. We have developed a statistical tool that reveals the presence of substantial RNA contamination from maternal tissues in nearly all published Arabidopsis thaliana endosperm and early embryo transcriptomes generated in these studies. We demonstrate that maternal RNA contamination explains the poor reproducibility of these transcriptomic data sets. Furthermore, we found that RNA contamination from maternal tissues has been repeatedly misinterpreted as epigenetic phenomena, which has resulted in inaccurate conclusions regarding the parental contributions to both the endosperm and early embryo transcriptomes. After accounting for maternal RNA contamination, no published genome-wide data set supports the concept of delayed paternal genome activation in plant embryos. Moreover, our analysis suggests that maternal and paternal genomic imprinting are equally rare events in Arabidopsis endosperm. Our publicly available software (https://github.com/Gregor-Mendel-Institute/tissue-enrichment-test) can help the community assess the level of contamination in transcriptome data sets generated from both seed and non-seed tissues. © 2017 American Society of Plant Biologists. All rights reserved.

  19. Development of the epaxial muscles in the human embryo

    NARCIS (Netherlands)

    Mekonen, Hayelom K.; Hikspoors, Jill P. J. M.; Mommen, Greet; Eleonore KÖhler, S.; Lamers, Wouter H.

    2016-01-01

    Although the intrinsic muscles of the back are defined by their embryological origin and innervation pattern, no detailed study on their development is available. Human embryos (5-10 weeks development) were studied, using Amira3D® reconstruction and Cinema4D® remodeling software for visualization.

  20. Analysis of embryo, cytoplasmic and maternal correlations for quality ...

    Indian Academy of Sciences (India)

    Home; Journals; Journal of Genetics; Volume 85; Issue 2. Analysis of embryo, cytoplasmic and maternal correlations for quality traits of rapeseed (Brassica napus L.) across environments. C. H. Shi H. Z. Zhang J. G. Wu. Research Note Volume 85 Issue 2 August 2006 pp 147-151 ...

  1. Synchrotron X-ray tomographic microscopy of fossil embryos.

    Science.gov (United States)

    Donoghue, Philip C J; Bengtson, Stefan; Dong, Xi-ping; Gostling, Neil J; Huldtgren, Therese; Cunningham, John A; Yin, Chongyu; Yue, Zhao; Peng, Fan; Stampanoni, Marco

    2006-08-10

    Fossilized embryos from the late Neoproterozoic and earliest Phanerozoic have caused much excitement because they preserve the earliest stages of embryology of animals that represent the initial diversification of metazoans. However, the potential of this material has not been fully realized because of reliance on traditional, non-destructive methods that allow analysis of exposed surfaces only, and destructive methods that preserve only a single two-dimensional view of the interior of the specimen. Here, we have applied synchrotron-radiation X-ray tomographic microscopy (SRXTM), obtaining complete three-dimensional recordings at submicrometre resolution. The embryos are preserved by early diagenetic impregnation and encrustation with calcium phosphate, and differences in X-ray attenuation provide information about the distribution of these two diagenetic phases. Three-dimensional visualization of blastomere arrangement and diagenetic cement in cleavage embryos resolves outstanding questions about their nature, including the identity of the columnar blastomeres. The anterior and posterior anatomy of embryos of the bilaterian worm-like Markuelia confirms its position as a scalidophoran, providing new insights into body-plan assembly among constituent phyla. The structure of the developing germ band in another bilaterian, Pseudooides, indicates a unique mode of germ-band development. SRXTM provides a method of non-invasive analysis that rivals the resolution achieved even by destructive methods, probing the very limits of fossilization and providing insight into embryology during the emergence of metazoan phyla.

  2. Somatic embryogenesis from zygotic embryos and thin cell layers ...

    African Journals Online (AJOL)

    Oil palm hybrid BRS Manicoré is important for plantations in the north of Brazil, as it is resistant to fatal yellowing and is compact. Seed germination is slow and reduced, so somatic embryogenesis is a promising alternative for its propagation. Two kinds of starting explants were used: Zygotic embryos (ZE) and thin cell layers ...

  3. Ex Ovo Model for Directly Visualizing Chick Embryo Development

    Science.gov (United States)

    Dorrell, Michael I.; Marcacci, Michael; Bravo, Stephen; Kurz, Troy; Tremblay, Jacob; Rusing, Jack C.

    2012-01-01

    We describe a technique for removing and growing chick embryos in culture that utilizes relatively inexpensive materials and requires little space. It can be readily performed in class by university, high school, or junior high students, and teachers of any grade level should be able to set it up for their students. Students will be able to…

  4. Human Embryo Editing: Opportunities and Importance of Transnational Cooperation.

    Science.gov (United States)

    Pei, Duanqing; Beier, David W; Levy-Lahad, Ephrat; Marchant, Gary; Rossant, Janet; Izpisua Belmonte, Juan Carlos; Lovell-Badge, Robin; Jaenisch, Rudolf; Charo, Alta; Baltimore, David

    2017-10-05

    A recent National Academies report articulates a path forward for research, ethics, and governance of clinical applications involving genome editing. In light of recent human embryo editing developments, scientists and stakeholders from all nations should cooperate to take advantage of this historic opportunity for medicine and also basic human biology. Copyright © 2017 Elsevier Inc. All rights reserved.

  5. In Vivo Nanotoxicity Testing using the Zebrafish Embryo Assay.

    Science.gov (United States)

    Rizzo, Larissa Y; Golombek, Susanne K; Mertens, Marianne E; Pan, Yu; Laaf, Dominic; Broda, Janine; Jayapaul, Jabadurai; Möckel, Diana; Subr, Vladimir; Hennink, Wim E; Storm, Gert; Simon, Ulrich; Jahnen-Dechent, Willi; Kiessling, Fabian; Lammers, Twan

    2013-06-10

    Nanoparticles are increasingly used for biomedical purposes. Many different diagnostic and therapeutic applications are envisioned for nanoparticles, but there are often also serious concerns regarding their safety. Given the fact that numerous new nanomaterials are being developed every day, and that not much is known about the long-term toxicological impact of exposure to nanoparticles, there is an urgent need to establish efficient methods for nanotoxicity testing. The zebrafish (Danio rerio) embryo assay has recently emerged as an interesting 'intermediate' method for in vivo nanotoxicity screening, enabling (semi-) high-throughput analyses in a system significantly more complex than cultured cells, but at the same time also less 'invasive' and less expensive than large-scale biocompatibility studies in mice or rats. The zebrafish embryo assay is relatively well-established in the environmental sciences, but it has not yet gained wide notice in the nanomedicine field. Using prototypic polymeric drug carriers, gold-based nanodiagnostics and nanotherapeutics, and iron oxide-based nanodiagnostics, we here show that toxicity testing using zebrafish embryos is easy, efficient and informative, and faithfully reflects, yet significantly extends, cell-based toxicity testing. We therefore expect that the zebrafish embryo assay will become a popular future tool for in vivo nanotoxicity screening.

  6. Influence of carbon nanotube length on toxicity to zebrafish embryos

    Directory of Open Access Journals (Sweden)

    Cheng J

    2012-07-01

    Full Text Available Jinping Cheng,1,2 Shuk Han Cheng11Department of Biology and Chemistry, City University of Hong Kong, Hong Kong; 2State Key Laboratory of Estuarine and Coastal Research, East China Normal University, Shanghai, ChinaAbstract: There is currently a large difference of opinion in nanotoxicology studies of nanomaterials. There is concern about why some studies have indicated that there is strong toxicity, while others have not. In this study, the length of carbon nanotubes greatly affected their toxicity in zebrafish embryos. Multiwalled carbon nanotubes (MWCNTs were sonicated in a nitric acid solution for 24 hours and 48 hours. The modified MWCNTs were tested in early developing zebrafish embryo. MWCNTs prepared with the longer sonication time resulted in severe developmental toxicity; however, the shorter sonication time did not induce any obvious toxicity in the tested developing zebrafish embryos. The cellular and molecular changes of the affected zebrafish embryos were studied and the observed phenotypes scored. This study suggests that length plays an important role in the in vivo toxicity of functionalized CNTs. This study will help in furthering the understanding on current differences in toxicity studies of nanomaterials.Keywords: length, carbon nanotubes, sonication, developmental toxicity, zebrafish

  7. Increase of uric acid synthesis in irradiated chicken's embryos

    International Nuclear Information System (INIS)

    Loyda, H.J.

    1984-01-01

    Several important intermediate and end products of uric acid metabolism as well as their corresponding enzymatic reactions were studied in 16 day-old chicken embryos which had been one or more times irradiated or respectively treated with ammonium chloride. After sublethal X-irradiation and at the time of the second irradiation with 800 R, the activity of the glutamine synthetase and the xanthin dehydrogenase in the kidneys of the embryos was increased. In contrast to this the glutamate dehydrogenase activity was moderately decreased. Two hours after the main irradiation the uric acid values as well as the amount of fixed nitrogen in the blood serum of previously-irradiated embryos are noticeably higher than the comparative data in non-previously irradiated animals. The glutamic acid values increase after the second irradiation, but still remain lower than with the non-previously irradiated animals. I achieved concuring ressults when I treated the embryos with ammonium chloride instead of radiation. (orig./MG) [de

  8. Maternal AP determinants in the Drosophila oocyte and embryo.

    Science.gov (United States)

    Ma, Jun; He, Feng; Xie, Gengqiang; Deng, Wu-Min

    2016-09-01

    An animal embryo cannot initiate its journey of forming a new life on its own. It must rely on maternally provided resources and inputs to kick-start its developmental process. In Drosophila, the initial polarities of the embryo along both the anterior-posterior (AP) and dorsal-ventral (DV) axes are also specified by maternal determinants. Over the past several decades, genetic and molecular studies have identified and characterized such determinants, as well as the zygotic genetic regulatory networks that control patterning in the early embryo. Extensive studies of oogenesis have also led to a detailed knowledge of the cellular and molecular interactions that control the formation of a mature egg. Despite these efforts, oogenesis and embryogenesis have been studied largely as separate problems, except for qualitative aspects with regard to maternal regulation of the asymmetric localization of maternal determinants. Can oogenesis and embryogenesis be viewed from a unified perspective at a quantitative level, and can that improve our understanding of how robust embryonic patterning is achieved? Here, we discuss the basic knowledge of the regulatory mechanisms controlling oogenesis and embryonic patterning along the AP axis. We explore properties of the maternal Bicoid gradient in relation to embryo size in search for a unified framework for robust AP patterning. WIREs Dev Biol 2016, 5:562-581. doi: 10.1002/wdev.235 For further resources related to this article, please visit the WIREs website. © 2016 Wiley Periodicals, Inc.

  9. Production of Buffalo Embryonic Stem Cell from HMC Embryos

    Directory of Open Access Journals (Sweden)

    M. Zandi

    2016-06-01

    Full Text Available Embryonic stem cells (ESCs are derived from the inner cell mass (ICM of blastocyst and differentiate into all three embryonic germ layers: ectoderm, endoderm, and mesoderm. In this study, ESCs are derived from Hand Made Cloning (HMG blastocysts and their efficiencies compared to ESCs derived from In Vitro Fertilization (IVF embryos. Feeder layer was used for ESCs culture, and culture medium consisting of Knockout- Dulbecco’s Modified Eagle’s Medium (Ko-DMEM supplemented with Knockout Serum Replacement (KSR, Leukemia Inhibitory Factor (LIF, Basic Fibroblast Growth Factor-2 (FGF-2, L-glutamine, nonessential amino acids and gentamicin. The cell surface antigens used for characterization were the SSEA-1, SSEA-4, TRA-1-60 and TRA-1-81 and the pluripotency markers were NANOG, OCT3/4 and SOX2. Results showed that, the growth rate of ESCs colonies in ESCs from IVF embryos was significantly higher than ESCs from HMG embryos (120% compared with 65%, respectively. Not only real-time PCR results revealed the same expression level of SOX2, OCT3/4 and cMYC between them, but also ESCs from HMG embryos resulted to higher expression of NANOG. Both of ESCs groups maintain in pluripotency state for more than two years and differentiated to the different types of cells like neuron, epithelial, lipid and muscle cells.

  10. Effect of nitrates on embryo induction efficiency in cotton (Gossypium ...

    African Journals Online (AJOL)

    Fred

    Cotton (Gossypium hirsutum L.) cv Coker-312 callus culture was assessed in terms of its usefulness as a system for investigating the effect of nitrates from different chemical compounds of nitrogen on embryo induction percentage in calli as the plant growth and cell differentiation mainly based on nitrogen. Both sources and ...

  11. Lipid composition and metabolism in embryos of Brassica napus

    International Nuclear Information System (INIS)

    Sparace, S.A.; Pomroy, M.K.

    1990-01-01

    Seven and 14-day old microspore-derived developing embryos of the low-erucate Brassica napus L. (cv. Topas) were analyzed for their acyl lipid composition and capacity to incorporate [ 14 C]acetate into lipid. The most significant changes in the lipid compositions of these ages of embryos are (1) increased total lipid from 2 to 5% of fresh weight; (2) increased proportion of TAG from 31 to 74%, and shifts in the fatty acid composition of TAG from 25 to 50% 18:1; 28 to 23% 18:2; and 24 to 13% 18:3. Lipids of 7-day embryos also consist of primarily 8% DAG, 2% MG, 12% FFA, 10% DGDG, 15% PA and approximately 5% each of PC, PE and PG. The levels of these lipids generally decrease as the embryos mature and accumulate TAG. [ 14 C]Acetate is incorporated into all lipids and fatty acids except 18:2 or 18:3. As much as 39, 59 and 34% of the fatty acid radioactivity of Mg was recovered in 20:0, 22:0 and 24:0, respectively

  12. In vitro culture of mouse embryos amniotic fluid ID human

    African Journals Online (AJOL)

    1989-07-15

    Jul 15, 1989 ... Human amniotic fluid was compared with Ham's F-10 culture medium as a possible alternative for use in in vitro fertilisation. The cleavage success of mouse embryos in human amniotic fluid (experimental group) was 92% compared with 86% in. Ham's F-10 medium. It is concluded that human amniotic.

  13. Characteristics of sugar uptake by immature maize embryos

    International Nuclear Information System (INIS)

    Griffith, S.M.; Jones, R.J.; Brenner, M.L.

    1986-01-01

    Characteristics of sugar uptake by immature maize embryos were determined in vitro utilizing a 14 C-sugar solution incubation method. Hexose uptake rates were greater than those for sucrose, however, all showed biphasic kinetics. Glucose and fructose saturable components were evidence at <50 mM and sucrose at <5 mM. Chemical inhibitors (CCCP, DNP, NaCN, and PCMBS) and low temperature reduced sugar uptake. Sucrose influx was pH dependent while glucose was not. Embryos maintained a high sucrose to hexose ratio throughout development. At 25 days after pollination sucrose levels exceeded 200 mM while hexose levels remained below 5 mM. Glucose was rapidly converted to sucrose upon transport into the embryo. These circumstantial data indicate that sugar uptake by immature maize embryos is metabolically dependent and carrier mediated. Furthermore, sucrose transport appears to occur against its concentration gradient involving a H+/sucrose cotransport mechanism, while glucose influx is driven by its concentration gradient and subsequent metabolism

  14. DNA methylation pattern in pig in vivo produced embryos

    Czech Academy of Sciences Publication Activity Database

    Fulka, Josef; Fulková, H.; Slavík, Tomáš; Okada, K.; Fulka Jr., J.

    2006-01-01

    Roč. 126, č. 2 (2006), s. 213-217 ISSN 0948-6143 R&D Projects: GA ČR GESTE/05/E004 Institutional research plan: CEZ:AV0Z50450515 Keywords : methylation * DNA * embryos Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.220, year: 2006

  15. Asymbiotic germination of immature embryos of a medicinally ...

    African Journals Online (AJOL)

    The immature embryos (28 weeks after pollination) were inoculated on M (Mitra et al., 1976), and PDA (Potato Dextrose Agar) media, with and without different growth additives. The seeds showed positive germination response in both the nutrient media but the frequency and onset of germination response and associated ...

  16. Local variation in embryo development rate in annual fish

    Czech Academy of Sciences Publication Activity Database

    Polačik, Matej; Reichard, Martin; Vrtílek, Milan

    (2018) ISSN 0022-1112 R&D Projects: GA ČR(CZ) GBP505/12/G112 Institutional support: RVO:68081766 Keywords : diapause * erratic development * escape embryo * killifish * Mozambique * secondary pool Subject RIV: EG - Zoology Impact factor: 1.519, year: 2016

  17. In vitro production of horse embryos: fundamental aspects

    NARCIS (Netherlands)

    López Tremoleda, Jordi

    2003-01-01

    Developments in assisted reproduction have provided valuable tools for sub-fertility treatment and for selective breeding in animals. In horses, techniques such as artificial insemination and embryo transfer are used successfully to aid genetic progress but the commercial application of other

  18. Embryo politics: ethics and policy in Atlantic democracies

    National Research Council Canada - National Science Library

    Banchoff, Thomas F

    2011-01-01

    ... States, the United Kingdom, Germany, and France have grappled with these questions so far. In setting out an argument about the intersection of politics, ethics, and policy, I focus on national bioethics committees, elected leaders, and their efforts to reconcile the moral status of the embryo and the imperative of biomedical progress in practice. In order to st...

  19. Multiple shoot induction from embryo derived callus cultures of ...

    African Journals Online (AJOL)

    Administrator

    The production of whole plant of cowpea (Vigna unguiculata) L. Walp from calli is an efficient, reliable ... practices on beans. Kobuyana et al. (1992) reported the use of embryo culture to generate mature plants in vitro from hybridization of three lines of the garden bean ... for the regeneration of shoots to fertile plants.

  20. Proteomic analysis of zebrafish embryos exposed to simulated-microgravity

    Science.gov (United States)

    Hang, Xiaoming; Ma, Wenwen; Wang, Wei; Liu, Cong; Sun, Yeqing

    Microgravity can induce a serial of physiological and pathological changes in human body, such as cardiovascular functional disorder, bone loss, muscular atrophy and impaired immune system function, etc. In this research, we focus on the influence of microgravity to vertebrate embryo development. As a powerful model for studying vertebrate development, zebrafish embryos at 8 hpf (hour past fertilization) and 24 hpf were placed into a NASA developed bioreac-tor (RCCS) to simulate microgravity for 64 and 48 hours, respectively. The same number of control embryos from the same parents were placed in a tissue culture dish at the same temper-ature of 28° C. Each experiment was repeated 3 times and analyzed by two-dimensional (2-D) gel electrophoresis. Image analysis of silver stained 2-D gels revealed that 64 from total 292 protein spots showed quantitative and qualitative variations that were significantly (Pmuscle B. Other protein spots showed significant expression alteration will be identified successively and the corresponding genes expression will also be measured by Q-PCR method at different development stages. The data presented in this study illustrate that zebrafish embryo can be significantly induced by microgravity on the expression of proteins involved in bone and muscle formation. Key Words: Danio rerio; Simulated-microgravity; Proteomics

  1. Ovulatory response and embryo quality in Katahdin ewes ...

    African Journals Online (AJOL)

    The aims of this study were to evaluate the superovulatory response (SR), embryo quality and embryonic development in Katahdin ewes that received a dietary supplement of palm oil (PO). Ten ewes were assigned to receive 35 g palm oil (n = 10, PO3) and 10 to receive 70 g palm oil (n = 10, PO6) for 25 days. On day 0, the ...

  2. Neurotoxic effect of the dithiocarbamate tecoram on the chick embryo

    NARCIS (Netherlands)

    Steenis, G. van; Logten, M.J. van

    Tecoram, when administered at doses of 0.01, 0.1, 1 or 10 mg per egg in propylene glycol or in saline to chick embryos caused paralysis, shortening of the extremities, muscular atrophy, dwarfing and death. Microscopically there were signs of peripheral neuropathy, mainly confined to the distal parts

  3. In vitro production of bovine embryos

    DEFF Research Database (Denmark)

    Stroebech, L.; Mazzoni, Gianluca; Pedersen, Hanne Skovsgaard

    2015-01-01

    In vitro production (IVP) of bovine embryos has become a widespread technology implemented in cattle breeding and production. The implementation of genomic selection and systems biology adds great dimensions to the impact of bovine IVP. The physical procedures included in the IVP process can stil...

  4. Survival of Macrobrachium amazonicum embryos submitted to cooling.

    Science.gov (United States)

    Ferreira, Arthur Vinícius Lourenço; Martins, Moisés Fernandes; Martins de Sousa, Míriam Luzia Nogueira; Soares Filho, Aldeney Andrade; Sampaio, Célia Maria de Souza

    2017-06-01

    Cooling techniques have several applications for reproduction in aquaculture. However, few studies have sought to create protocols for cooling and cryopreservation of Macrobrachium amazonicum embryos. Thus, the objective of this work was to verify the survival of M. amazonicum embryos and the correlation between embryonic volume and mortality of M. amazonicum embryos after cooling. Embryo pools were collected from three females and divided into two treatment groups: dimethyl sulfoxide (DMSO) 3% and ethylene glycol (EG) 0.5%, both of them associated with 2 M sucrose. Positive and negative control groups consisted of seawater 10%. Aliquots of 10 µg of embryos were placed in Falcon® tubes containing a cryoprotectant solution and submitted directly to the test temperature of 2°C for 2 and 6 h of cooling. Further analysis of survival and embryonic volume were performed under a stereoscopic microscope. Data were subjected to analysis of variance (ANOVA), and means were compared using the Tukey test at 5%. The highest embryonic survival rate was observed after the shortest storage time for both the DMSO 3% and the 0.5% EG groups, with survival rates of 84.8 ± 3.9 and 79.7 ± 2.8%, respectively. There was a reduction in survival after 24 h, with the DMSO 3% group presenting a survival rate of 71.7 ± 6.6%, and the EG 0.5% group, 66 ± 6.9%. Survival showed a statistically significant difference when compared with the positive controls after 2 h and 24 h of cooling, with 99 ± 0.5% and 95.8 ± 1.5% survival rates, respectively. There was no significant statistical difference in the embryonic volume, but it was possible to observe a change in the appearance of the embryos, from a translucent coloration to an opaque white or brownish coloration, after 24 h in incubators. Thus, it can be concluded that survival is inversely proportional to storage time and that, although there was no change in the embryonic volume after cooling, a change in the appearance of embryos could

  5. A Chemically Defined Medium for Rabbit Embryo Cryopreservation

    Science.gov (United States)

    Bruyère, Pierre; Baudot, Anne; Joly, Thierry; Commin, Loris; Pillet, Elodie; Guérin, Pierre; Louis, Gérard; Josson-Schramme, Anne; Buff, Samuel

    2013-01-01

    This study evaluates a new synthetic substitute (CRYO3, Ref. 5617, Stem Alpha, France) for animal-based products in rabbit embryo cryopreservation solutions. This evaluation was performed using two approaches: a thermodynamic approach using differential scanning calorimetry and a biological approach using rabbit embryo slow-freezing. During the experiment, foetal calf serum (FCS) was used as a reference. Because FCS varies widely by supplier, three different FCS were selected for the thermodynamic approach. The rabbit embryo slow-freezing solutions were made from Dulbecco's phosphate buffer saline containing 1.5 M Dimethyl Sulfoxide and 18% (v.v−1) of CRYO3 or 18% (v.v−1) of FCS. These solutions were evaluated using four characteristics: the end of melting temperature, the enthalpy of crystallisation (thermodynamic approach) and the embryo survival rates after culture and embryo transfer (biological approach). In the thermodynamic approach, the solutions containing one of the three different FCS had similar mean thermodynamic characteristics but had different variabilities in the overall data with aberrant values. The solution containing CRYO3 had similar thermodynamic properties when compared to those containing FCS. Moreover, no aberrant value was measured in the solution containing CRYO3. This solution appears to be more stable than the solutions containing a FCS. In the biological approach, the in vitro embryo survival rates obtained with the solution containing CRYO3 (73.7% and 81.3%) and with the solution containing a FCS (77.6% and 71.9%) were similar (p = 0.7). Nevertheless, during the in vivo evaluation, the implantation rate (21.8%) and the live-foetuses rate (18.8%) of the CRYO3 group were significantly higher than the implantation rate (7.1%, p = 0.0002) and the live-foetuses rate (5.3%, p = 0.0002) of the FCS group. The pregnancy rate was also higher in the CRYO3 group compared to the FCS group (81.3% and 43.8%, respectively, p = 0

  6. Influence of irradiation (Co60) in pre-implant rabbits embryos: effect on blastocyst diameters and embryos smaller than 2 mm

    International Nuclear Information System (INIS)

    Approbato, Mario S.; Oliveira Moura, Katia K.V. de; Souza Florencio, Rodopiano de; Cunha Junior, Carlos; Garcia, Ricardo; Faria, Renato S.; Benedetti, Leonardo N.; Goulart, Flamarion B.

    1995-01-01

    We studied the effect of ionizing irradiation on 12 New Zealand rabbits (65 embryos), in three different times: at match time (zero hour), two days after and four days after, with two different irradiation doses, 5 c Gy and 10 c Gy. Six rabbits (36 blastocysts) were used as controls. The matching instant was the zero hour. Exactly six days after (± 60 minutes) the embryos of each rabbit was picked up by flushing the uterus with culture media. The embryos were fixed in methanol for 48 hours, and colored with acid Mayer hematoxylin. The following embryos parameters were studied: diameter growth; percentage of embryos smaller than 2 mm. We observed that only the irradiation time influenced the blastocysts diameter (no irradiation dose). There was no relation between percentage of embryos smaller than 2 mm and the irradiation. (author)

  7. The effect of the number of transferred embryos, the interval between nuclear transfer and embryo transfer, and the transfer pattern on pig cloning efficiency.

    Science.gov (United States)

    Rim, Chol Ho; Fu, Zhixin; Bao, Lei; Chen, Haide; Zhang, Dan; Luo, Qiong; Ri, Hak Chol; Huang, Hefeng; Luan, Zhidong; Zhang, Yan; Cui, Chun; Xiao, Lei; Jong, Ui Myong

    2013-12-01

    To improve the efficiency of producing cloned pigs, we investigated the influence of the number of transferred embryos, the culturing interval between nuclear transfer (NT) and embryo transfer, and the transfer pattern (single oviduct or double oviduct) on cloning efficiency. The results demonstrated that transfer of either 150-200 or more than 200NT embryos compared to transfer of 100-150 embryos resulted in a significantly higher pregnancy rate (48 ± 16, 50 ± 16 vs. 29 ± 5%, pcloning efficiency is achieved by adjusting the number and in vitro culture time of reconstructed embryos as well as the embryo transfer pattern. Copyright © 2013 Elsevier B.V. All rights reserved.

  8. SOMATIC EMBRYOGENESIS AND MORPHOANATOMY OF Ocotea porosa SOMATIC EMBRYOS

    Directory of Open Access Journals (Sweden)

    Luciana Luiza Pelegrini

    2013-12-01

    Full Text Available http://dx.doi.org/10.5902/1980509812343Ocotea porosa seeds have strong tegument dormancy, recalcitrant behavior, low and irregular germinationand that makes its natural propagation difficult. The aim of this study was to establish a protocol ofregeneration of Ocotea porosa from somatic embryogenesis. Immature embryonic axes were inoculatedon WPM culture medium supplemented with 2.4-D (200 μM combined or not with hydrolyzed casein orglutamine (0.5 or 1 g l-1, during 90 days. The repetitive embryogenesis was induced on medium with 2.4-D(22.62 μM combined with 2-iP (2.46 μM followed by transfer to culture medium with hydrolyzed caseinor glutamine (1 g l-1 during 90 days. The maturation of somatic embryos was tested in culture mediumcontaining NAA (0.5 μM and 2-iP (5; 10 and 20 μM. The highest percentage of somatic embryos induction(8.3% was observed in WPM culture medium containing 200 μM 2.4-D and 1 g L-1 hydrolyzed casein andthe development of somatic embryos occurred indirectly. Repetitive somatic embryogenesis was promotedin WPM medium containing hydrolyzed casein or glutamine. However, the culture medium containinghydrolyzed casein promoted the maintenance of embryogenic capacity for more than two years. Duringthe maturity phase, there was a low progression of globular embryos to cordiform and torpedo stages.The different ontogenetic stages of somatic embryos of Ocotea porosa were characterized by histologicalstudies.

  9. Transcriptome analysis of zebrafish embryos exposed to deltamethrin.

    Science.gov (United States)

    Chueh, Tsung-Cheng; Hsu, Li-Sung; Kao, Chin-Ming; Hsu, Tung-Wei; Liao, Hung-Yu; Wang, Kuan-Yi; Chen, Ssu Ching

    2017-05-01

    Deltamethrin (DTM), a type II pyrethroid, is one of the most commonly used insecticides. The increased use of pyrethroid leads to potential adverse effects, particularly in sensitive populations such as children and pregnant women. None of the related studies was focused on the transcriptome responses in zebrafish embryos after treatment with DTM; therefore, RNA-seq, a high-throughput method, was performed to analyze the global expression of differential expressed genes (DEGs) in zebrafish embryos treated with DTM (40 and 80 μg/L) from fertilization to 48 h postfertilization (hpf) as compared with that in the control group (without DTM treatment). Two cDNA libraries were generated from treated embryos and one cDNA library from nontreated embryos, respectively. Over 92% of reads mapped to the reference in these three libraries. It was observed that many differential genes were expressed in comparison with embryos before and after DTM. The 20 most differentially expressed upregulated or downregulated genes were majorly involved in the signaling transduction. Validation of selected nine genes expression using qRT-PCR confirmed RNA-seq results. The transcriptome sequences were further subjected to gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, showing G-protein-coupled receptor signaling pathway and neuroactive ligand-receptor interaction, respectively, were most enriched. The data from this study contributed to a better understanding of the potential consequences of fish exposed to DTM, to an evaluation of the potential threat of DTM to fish populations in aquatic environments. © 2016 Wiley Periodicals, Inc. Environ Toxicol 32: 1548-1557, 2017. © 2016 Wiley Periodicals, Inc.

  10. Pollination and embryo development in Brassica rapa L. in microgravity

    Science.gov (United States)

    Kuang, A.; Popova, A.; Xiao, Y.; Musgrave, M. E.

    2000-01-01

    Plant reproduction under spaceflight conditions has been problematic in the past. In order to determine what aspect of reproductive development is affected by microgravity, we studied pollination and embryo development in Brassica rapa L. during 16 d in microgravity on the space shuttle (STS-87). Brassica is self-incompatible and requires mechanical transfer of pollen. Short-duration access to microgravity during parabolic flights on the KC-135A aircraft was used initially to confirm that equal numbers of pollen grains could be collected and transferred in the absence of gravity. Brassica was grown in the Plant Growth Facility flight hardware as follows. Three chambers each contained six plants that were 13 d old at launch. As these plants flowered, thin colored tape was used to indicate the date of hand pollination, resulting in silique populations aged 8-15 d postpollination at the end of the 16-d mission. The remaining three chambers contained dry seeds that germinated on orbit to produce 14-d-old plants just beginning to flower at the time of landing. Pollen produced by these plants had comparable viability (93%) with that produced in the 2-d-delayed ground control. Matched-age siliques yielded embryos of equivalent developmental stage in the spaceflight and ground control treatments. Carbohydrate and protein storage reserves in the embryos, assessed by cytochemical localization, were also comparable. In the spaceflight material, growth and development by embryos rescued from siliques 15 d after pollination lagged behind the ground controls by 12 d; however, in the subsequent generation, no differences between the two treatments were found. The results demonstrate that while no stage of reproductive development in Brassica is absolutely dependent upon gravity, lower embryo quality may result following development in microgravity.

  11. [Cryopreservation study on seeds and embryos in Dalbergia odorifera].

    Science.gov (United States)

    Zeng, Lin; He, Ming-Jun; Chen, Kui; Wei, Jian-He

    2014-06-01

    The mature seeds and excised embryos of Dalbergia odorifera were used as materials to study the effect of moisture content on their survival, as well as the effect of rapid freezing and vitrification freezing method on seeds and in vitro embryos cryopreservation. The results showed that the germination rate and vigor decreased from 82.67%, 85% to 18.35%, 25% respectively, when the seed moisture content decreased from 15.04% to 8.14%; and the germination rate decreased from 82.67% to 37.50%, 25.37% respectively by vitrification freezing method and rapid freezing method, when the seed moisture content decreased from 15.04% to 9.37%. Among all the moisture content gradient, 12.35% moisture reached the maximal germination rate, which were 63.58% and 50.45% respectively by vitrification freezing and rapid freezing; and when the embryo moisture content was 26.32%, the germination rate decreased from 95.67% to 58.31% and 33.82% respectively by vitrification freezing and rapid freezing. And when the moisture content was in the range of 14.17% -21.34%, the germination rate was a bit of decrease. The experiment results showed that the optimum conditions of seed cryopreservation were: moisture content 12.35%, vitrification freezing; and the optimum conditions of in vitro embryo cryopreservation were: moisture 15.04%, vitrification freezing. In conclusion, the effects of moisture content on germination rate after cryopreservation in D. odorifera seeds and embryo were significant, and vitrification freezing method is much better than rapid freezing method.

  12. Movement of the external ear in human embryo.

    Science.gov (United States)

    Kagurasho, Miho; Yamada, Shigehito; Uwabe, Chigako; Kose, Katsumi; Takakuwa, Tetsuya

    2012-02-01

    External ears, one of the major face components, show an interesting movement during craniofacial morphogenesis in human embryo. The present study was performed to see if movement of the external ears in a human embryo could be explained by differential growth. In all, 171 samples between Carnegie stage (CS) 17 and CS 23 were selected from MR image datasets of human embryos obtained from the Kyoto Collection of Human Embryos. The three-dimensional absolute position of 13 representative anatomical landmarks, including external and internal ears, from MRI data was traced to evaluate the movement between the different stages with identical magnification. Two different sets of reference axes were selected for evaluation and comparison of the movements. When the pituitary gland and the first cervical vertebra were selected as a reference axis, the 13 anatomical landmarks of the face spread out within the same region as the embryo enlarged and changed shape. The external ear did move mainly laterally, but not cranially. The distance between the external and internal ear stayed approximately constant. Three-dimensionally, the external ear located in the caudal ventral parts of the internal ear in CS 17, moved mainly laterally until CS 23. When surface landmarks eyes and mouth were selected as a reference axis, external ears moved from the caudal lateral ventral region to the position between eyes and mouth during development. The results indicate that movement of all anatomical landmarks, including external and internal ears, can be explained by differential growth. Also, when the external ear is recognized as one of the facial landmarks and having a relative position to other landmarks such as the eyes and mouth, the external ears seem to move cranially. © 2012 Kagurasho et al; licensee BioMed Central Ltd.

  13. Identification of emergent motion compartments in the amniote embryo.

    Science.gov (United States)

    Loganathan, Rajprasad; Little, Charles D; Joshi, Pranav; Filla, Michael B; Cheuvront, Tracey J; Lansford, Rusty; Rongish, Brenda J

    2014-01-01

    The tissue scale deformations (≥ 1 mm) required to form an amniote embryo are poorly understood. Here, we studied ∼400 μm-sized explant units from gastrulating quail embryos. The explants deformed in a reproducible manner when grown using a novel vitelline membrane-based culture method. Time-lapse recordings of latent embryonic motion patterns were analyzed after disk-shaped tissue explants were excised from three specific regions near the primitive streak: 1) anterolateral epiblast, 2) posterolateral epiblast, and 3) the avian organizer (Hensen's node). The explants were cultured for 8 hours-an interval equivalent to gastrulation. Both the anterolateral and the posterolateral epiblastic explants engaged in concentric radial/centrifugal tissue expansion. In sharp contrast, Hensen's node explants displayed Cartesian-like, elongated, bipolar deformations-a pattern reminiscent of axis elongation. Time-lapse analysis of explant tissue motion patterns indicated that both cellular motility and extracellular matrix fiber (tissue) remodeling take place during the observed morphogenetic deformations. As expected, treatment of tissue explants with a selective Rho-Kinase (p160ROCK) signaling inhibitor, Y27632, completely arrested all morphogenetic movements. Microsurgical experiments revealed that lateral epiblastic tissue was dispensable for the generation of an elongated midline axis- provided that an intact organizer (node) is present. Our computational analyses suggest the possibility of delineating tissue-scale morphogenetic movements at anatomically discrete locations in the embryo. Further, tissue deformation patterns, as well as the mechanical state of the tissue, require normal actomyosin function. We conclude that amniote embryos contain tissue-scale, regionalized morphogenetic motion generators, which can be assessed using our novel computational time-lapse imaging approach. These data and future studies-using explants excised from overlapping anatomical

  14. SOMATIC EMBRYOGENESIS AND MORPHOANATOMY OF Ocotea porosa SOMATIC EMBRYOS

    Directory of Open Access Journals (Sweden)

    Luciana Luiza Pelegrini

    2013-01-01

    Full Text Available Ocotea porosa seeds have strong tegument dormancy, recalcitrant behavior, low and irregular germination and that makes its natural propagation difficult. The aim of this study was to establish a protocol of regeneration of Ocotea porosa from somatic embryogenesis. Immature embryonic axes were inoculated on WPM culture medium supplemented with 2.4-D (200 μM combined or not with hydrolyzed casein or glutamine (0.5 or 1 g l-1, during 90 days. The repetitive embryogenesis was induced on medium with 2.4-D (22.62 μM combined with 2-iP (2.46 μM followed by transfer to culture medium with hydrolyzed casein or glutamine (1 g l-1 during 90 days. The maturation of somatic embryos was tested in culture medium containing NAA (0.5 μM and 2-iP (5; 10 and 20 μM. The highest percentage of somatic embryos induction (8.3% was observed in WPM culture medium containing 200 μM 2.4-D and 1 g L-1 hydrolyzed casein and the development of somatic embryos occurred indirectly. Repetitive somatic embryogenesis was promoted in WPM medium containing hydrolyzed casein or glutamine. However, the culture medium containing hydrolyzed casein promoted the maintenance of embryogenic capacity for more than two years. During the maturity phase, there was a low progression of globular embryos to cordiform and torpedo stages. The different ontogenetic stages of somatic embryos of Ocotea porosa were characterized by histological studies.

  15. Effect of the microenvironment and embryo density on developmental characteristics and gene expression profile of bovine preimplantative embryos cultured in vitro.

    Science.gov (United States)

    Hoelker, Michael; Rings, Franka; Lund, Qamaruddin; Ghanem, Nasser; Phatsara, Chirawath; Griese, Josef; Schellander, Karl; Tesfaye, Dawit

    2009-03-01

    The Well of the Well (WOW) system has been developed to culture embryos in small groups or to track the development of single embryos. In the present study, we aimed to examine the effects of the microenvironment provided by the WOW system and embryo density on developmental rates, embryo quality and preimplantative gene expression profile of the resulting embryos. Embryos cultured in a group of 16 reached the blastocyst stage at a significantly lower level than zygotes cultured in a group of 50 (22.2 vs 30.3%), whereas zygotes cultured in WOW were able to compensate against low embryo densities, reaching a blastocyst rate as high as embryos cultured in a group of 50 (31.3 vs 30.3%). Moreover, embryos derived from WOW culture did not differ in terms of differential cell counts and apoptotic cell index compared with controls. The gene expression analysis revealed 62 transcripts to be upregulated and 33 transcripts to be downregulated by WOW culture. Comparing the in vivo derived blastocysts with the blastocysts derived from WOW culture, and group culture, expression of ATP5A1, PLAC8 and KRT8 was more similar to the embryos derived from WOW culture, whereas expression of S100A10 and ZP3 genes was more similar to blastocysts cultured in a group. In conclusion, microenvironment as well as embryo density significantly affected developmental rates. While subsequent blastocysts did not differ in terms of differential cell counts and apoptotic cell index, significant differences were observed in terms of the relative abundance of transcripts in the resulting embryos.

  16. Pregnancy outcomes in a university hospital after legal requirement for single-embryo transfer.

    Science.gov (United States)

    Ercan, Cihangir Mutlu; Kerimoglu, Ozlem Secilmis; Sakinci, Mehmet; Korkmaz, Cem; Duru, Namik Kemal; Ergun, Ali

    2014-04-01

    To present our in vitro fertilization outcomes after implementation of the Turkish government policy limiting the number of transferred embryos to one, in women under the age of 35, and two, in women over age 35; and to demonstrate the feasibility of this policy. We retrospectively reviewed the records of 423 patients who underwent ovarian stimulation, oocyte pick-up, intracytoplasmic sperm injection and embryo transfer due to primary or secondary infertility over a 22-month time interval. Clinical pregnancy was defined as a fetal heartbeat in the seventh gestational week. Embryo transfers were carried out on day 3 or 5. Descriptive statistics are given as the number of subjects and percentages. A total of 353 embryo transfers were performed: 261 (73.9%) were single-embryo transfer (244 (69.1%) elective and 17 (4.8%) non-elective) and 92 (26.1%) were double-embryo transfer. Of the 244 elective single-embryo transfers, 6.6% (n=16) were performed using frozen-thawed embryos. The average patient age was 29.6±4.5 years in the single-embryo group and 36.5±2.4 years in the double-embryo group. The cumulative pregnancy rates per oocyte pick-up were similar in both groups: 41% (n=107) in the single-embryo group and 43.4% (n=40) in the double-embryo group (p=0.678). The cumulative live birth rate of the single-embryo group (32.1%) was not statistically different from the double-embryo group (35.8%) (p=0.518). The twin pregnancy rate after single-embro transfer was significantly lower than with double-embryo transfer (2.8% (n=3) vs. 32.5% (n=13); ppregnancies occurred in women 35 years or older who underwent double-embryo transfer. No significant difference in the spontaneous abortion rates was recorded between the single- and double-embryo transfer groups (16 (6.1%) vs. 6 (6.5%); p=0.894). Single-embryo transfer results in a pregnancy rate comparable to double-embryo transfer, with a significantly reduced multiple pregnancy rate. Double-embryo transfer in patients over

  17. Developmental competence and epigenetic profile of porcine embryos produced by two different cloning methods

    DEFF Research Database (Denmark)

    Liu, Ying; Lucas-Hahn, Andrea; Petersen, Bjoern

    2017-01-01

    on conventionally produced embryos. The goal of this study was to unravel putative differences between two cloning methods, with regard to developmental competence, expression profile of a panel of developmentally important genes and epigenetic profile of porcine cloned embryos produced by either CNT or HMC, either...... affect porcine embryo development and epigenetic profile. The two cloning methods essentially produce embryos of similar quality on D0 and after 5 days in vitro culture, but thereafter both histone acetylation and gene expression differ between the two types of cloned embryos....

  18. Using Surplus Embryos and Research Embryos in Stem Cell Research: Ethical Viewpoints of Buddhist, Hindu and Catholic Leaders in Malaysia on the Permissibility of Research.

    Science.gov (United States)

    Sivaraman, Mathana Amaris Fiona

    2018-02-01

    The sources of embryos for Embryonic Stem Cell Research (ESCR) include surplus embryos from infertility treatments, and research embryos which are created solely for an ESCR purpose. The latter raises more ethical concerns. In a multi-religious country like Malaysia, ethical discussions on the permissibility of ESCR with regard to the use surplus and research embryos are diversified. Malaysia has formulated guidelines influenced by the national fatwa ruling which allows the use of surplus embryos in ESCR. Input from other main religions is yet to be documented. In light of this, this study addresses (i) the ethical viewpoints of Buddhist, Hindu and Catholic leaders on the permissibility of using surplus and research embryos; and (ii) the moral standpoints of religious leaders towards attaining a consensus on the practice of ESCR in Malaysia. Responses from the religious leaders were obtained via semi-structured, face-to-face interviews. The findings show that generally the Buddhist and Hindu leaders approve the use of surplus embryos. Their responses on the creation of research embryos for ESCR are varied. Meanwhile, the Catholic leaders distinctively objected to ESCR regardless of the embryo sources, referring to it as the destruction of life. Taking into account the diverse views, this study explores the response of the religious leaders for a general consensus wherever possible. The ethical discourse surrounding ESCR in a multi-religious setting offers new perspective, which needs to be explored in a broader global community.

  19. Efficiency of porcine somatic cell nuclear transfer – a retrospective study of factors related to embryo recipient and embryos transferred

    Directory of Open Access Journals (Sweden)

    Yongye Huang

    2013-10-01

    The successful generation of pigs via somatic cell nuclear transfer depends on reducing risk factors in several aspects. To provide an overview of some influencing factors related to embryo transfer, the follow-up data related to cloned pig production collected in our laboratory was examined. (i Spring showed a higher full-term pregnancy rate compared with winter (33.6% vs 18.6%, P = 0.006. Furthermore, a regression equation can be drawn between full-term pregnancy numbers and pregnancy numbers in different months (y = 0.692x−3.326. (ii There were no significant differences detected in the number of transferred embryos between surrogate sows exhibiting full-term development compared to those that did not. (iii Non-ovulating surrogate sows presented a higher percentage of full-term pregnancies compared with ovulating sows (32.0% vs 17.5%, P = 0.004; respectively. (iv Abortion was most likely to take place between Day 27 to Day 34. (v Based on Life Table Survival Analysis, delivery in normally fertilized and surrogate sows is expected to be completed before Day 117 or Day 125, respectively. Additionally, the length of pregnancy in surrogate sows was negatively correlated with the average litter size, which was not found for normally fertilized sows. In conclusion, performing embryo transfer in appropriate seasons, improving the quality of embryos transferred, optimizing the timing of embryo transfer, limiting the occurrence of abortion, combined with ameliorating the management of delivery, is expected to result in the harvest of a great number of surviving cloned piglets.

  20. Characterization of the onset of embryonic control and early development in the bovine embryo

    International Nuclear Information System (INIS)

    Barnes, F.L.

    1988-01-01

    Bovine embryos were used to determine if morphological and molecular features of early development are similar to in vivo recovered bovine embryos and to determine at what level early bovine development is regulated. Radiolabeling of IVP embryos and in vivo recovered embryos with 35 S-methionine for SDS-polyacrylamide gel electrophoresis reveals that these embryos are equivalent. Few differences in protein profiles are observed between 1- and early 4-cell embryos. A change in protein profiles begins at the mid 4-cell stage and continues into the 8-cell stage. Few differences in protein profiles are observed between 1- and early 4-cell embryos. A change in protein profiles begins at the mid 4-cell stage and continues into the 8-cell stage. Few differences in protein profiles are observed between late 8-cells and morulae. This transition is α-amanitin sensitive therefore due to de novo embryonic transcription. Embryonic transcription is partially responsible for terminating the post-transcriptionally regulated period of early bovine development. Argentophillic nucleolar organizing regions (Ag-NORs) indicate onset of nucleolar activation. Ag-NORs were absent in 2- and 4-cell IVP embryos and rarely occurred in 8-cell IVP embryos cultured in vitro. IVP 1- and 2-cell embryos cultured to blastocysts in sheep oviducts demonstrated Ag-NORs. Thus the lack of nucleolar activation of IVP embryos cultured in vitro is culture induced between the 2- and 8-cell stage

  1. Somatic Embryos in Catharanthus roseus: A Scanning Electron Microscopic Study

    Directory of Open Access Journals (Sweden)

    Junaid ASLAM

    2014-06-01

    Full Text Available Catharanthus roseus (L. G. Don is an important medicinal plant as it contains several anti-cancerous compounds, like vinblastine and vincristine. Plant tissue culture technology (organogenesis and embryogenesis has currently been used in fast mass propagating raw materials for secondary metabolite synthesis. In this present communication, scanning electron microscopic (SEM study of somatic embryos was conducted and discussed. The embryogenic callus was first induced from hypocotyls of in vitro germinated seeds on which somatic embryos, differentiated in numbers, particularly on 2,4-D (1.0 mg/L Murashige and Skoog (MS was medium. To understand more about the regeneration method and in vitro formed embryos SEM was performed. The SEM study revealed normal somatic embryo origin and development from globular to heart-, torpedo- and then into cotyledonary-stage of embryos. At early stage, the embryos were clustered together in a callus mass and could not easily be detached from the parental tissue. The embryos were often long cylindrical structure with or without typical notch at the tip. Secondary embryos were also formed on primary embryo structure. The advanced cotyledonary embryos showed prominent roots and shoot axis, which germinated into plantlets. The morphology, structure and other details of somatic embryos at various stages were presented.

  2. Impact of the IVF laboratory environment on human preimplantation embryo phenotype.

    Science.gov (United States)

    Gardner, D K; Kelley, R L

    2017-08-01

    The phenotype of the human embryo conceived through in vitro fertilization (IVF), that is its morphology, developmental kinetics, physiology and metabolism, can be affected by numerous components of the laboratory and embryo culture system (which comprise the laboratory environment). The culture media formulation is important in determining embryo phenotype, but this exists within a culture system that includes oxygen, temperature, pH and whether an embryo is cultured individually or in a group, all of which can influence embryo development. Significantly, exposure of an embryo to one suboptimal component of the culture system of laboratory typically predisposes the embryo to become more vulnerable to a second stressor, as has been well documented for atmospheric oxygen and individual culture, as well as for oxygen and ammonium. Furthermore, the inherent viability of the human embryo is derived from the quality of the gametes from which it is created. Patient age, aetiology, genetics, lifestyle (as well as ovarian stimulation in women) are all known to affect the developmental potential of gametes and hence the embryo. Thus, as well as considering the impact of the IVF laboratory environment, one needs to be aware of the status of the infertile couple, as this impacts how their gametes and embryos will respond to an in vitro environment. Although far from straight forward, analysing the interactions that exist between the human embryo and its environment will facilitate the creation of more effective and safer treatments for the infertile couple.

  3. Influence of embryo handling and transfer method on pig cloning efficiency.

    Science.gov (United States)

    Shi, Junsong; Zhou, Rong; Luo, Lvhua; Mai, Ranbiao; Zeng, Haiyu; He, Xiaoyan; Liu, Dewu; Zeng, Fang; Cai, Gengyuan; Ji, Hongmei; Tang, Fei; Wang, Qinglai; Wu, Zhenfang; Li, Zicong

    2015-03-01

    The somatic cell nuclear transfer (SCNT) technique could be used to produce genetically superior or genetically engineered cloned pigs that have wide application in agriculture and bioscience research. However, the efficiency of porcine SCNT currently is very low. Embryo transfer (ET) is a key step for the success of SCNT. In this study, the effects of several ET-related factors, including cloned embryo culture time, recipient's ovulation status, co-transferred helper embryos and ET position, on the success rate of pig cloning were investigated. The results indicated that transfer of cloned embryos cultured for a longer time (22-24h vs. 4-6h) into pre-ovulatory sows decreased recipient's pregnancy rate and farrowing rate, and use of pre-ovulatory and post-ovulatory sows as recipients for SCNT embryos cultured for 22-24h resulted in a similar porcine SCNT efficiency. Use of insemination-produced in vivo fertilized, parthenogenetically activated and in vitro fertilized embryos as helper embryos to establish and/or maintain pregnancy of SCNT embryos recipients could not improve the success rate of porcine SCNT. Transfer of cloned embryos into double oviducts of surrogates significantly increased pregnancy rate as well as farrowing rate of recipients, and the developmental rate of transferred cloned embryos, as compared to unilateral oviduct transfer. This study provided useful information for optimization of the embryo handling and transfer protocol, which will help to improve the ability to generate cloned pigs. Copyright © 2015 Elsevier B.V. All rights reserved.

  4. Factors that affect infertility patients' decisions about disposition of frozen embryos.

    Science.gov (United States)

    Lyerly, Anne Drapkin; Steinhauser, Karen; Namey, Emily; Tulsky, James A; Cook-Deegan, Robert; Sugarman, Jeremy; Walmer, David; Faden, Ruth; Wallach, Edward

    2006-06-01

    To describe factors that affect infertility patients' decision making regarding their cryopreserved embryos. Forty-six semistructured in-depth interviews of individuals and couples participating in IVF programs. Two major southeastern academic medical centers. Fifty-three individuals, including 31 women, 8 men, and 7 couples. Qualitative analysis of interview transcripts. INTERVENTION (S): None. Seven broad themes informed participants' decisions about embryo disposition: family and personal issues, trust, definition of the embryo, prospective responsibility to the embryo, responsibility to society, adequacy of information, and lack of acceptable disposition options. Many wished for alternative options, such as a ceremony at the time of disposal or placement of embryos in the woman's body when pregnancy was unlikely. Recent debates regarding embryo disposition do not reflect the range of values that infertility patients consider when deciding about frozen embryos. In addition to questions about the embryo's moral status, decision making about embryos is informed by a range of factors in the lives of individuals who created them. These perspectives may have important implications for the content and timing of informed consent, facilitating embryo disposition, and advancing policy debates about the ethics of frozen embryo use.

  5. Factors influencing germination and growth of isolated embryos of Pinus heldreichii

    Directory of Open Access Journals (Sweden)

    Stojičić Dragana

    2008-01-01

    Full Text Available The effects of nutritional, hormonal, and physical factors on the germination and growth of isolated mature zygotic embryos of Pinus heldreichii were studied under in vitro conditions. The optimum medium for embryo germination and adequate conversion into seedlings was basal Gresshoff and Doy (1GD medium. All tested carbohydrates at the applied concentrations stimulated embryo growth, but only 3% sucrose increased embryo germination compared to the control, while maltose had an inhibitory effect. Among the applied plant growth regulators, only gibberelic acid (GA3 had a few benefits on the development of isolated embryos into plantlets, while the others had a negative effect. Embryos cultured on an agar-solidified medium grew better and the germination percentage was higher than in a liquid medium. Light treatment did not affect embryo germination, but generally stimulated seedling growth.

  6. Effect of different ovule isolation times on the embryo development of Campanula hybrids

    DEFF Research Database (Denmark)

    Röper, Anna Catharina; Lütken, Henrik Vlk; Hegelund, Josefine Nymark

    2012-01-01

    , hybridization between plant species is associated with many challenges to enable survival of the developing embryo. Here we present an optimised technique for embryo rescue via ovule isolation in selected intra- and interspecific Campanula hybrids. A frequent problem in embryo rescue is the malformation...... of the endosperm. To circumvent this, embryos were isolated and the optimal ovule isolation time and growth conditions were determined to increase embryo survival. Ovules were isolated one to four weeks after pollination and cultivated on a modified MS medium. When ovules were allowed to stay inside the ovary...... for 2-3 weeks the number of germinating embryos increased as compared to ovules isolated one week after pollination. Additionally, ovules isolated 2-3 weeks after pollination showed an increased embryo germination rate. Among the Campanula hybrids, produced here from both the intraspecific crosses...

  7. The efficacy of the well of the well (WOW) culture system on development of bovine embryos in a small group and the effect of number of adjacent embryos on their development.

    OpenAIRE

    Kang, Sung-Sik; Ofuji, Sosuke; Imai, Kei; Huang, Weiping; Koyama, Keisuke; Yanagawa, Yojiro; Takahashi, Yoshiyuki; Nagano, Masashi

    2014-01-01

    The aim of the present study was to clarify the efficacy of the well of the well (WOW) culture system for a small number of embryos and the effect of number of adjacent embryos in a WOW dish on blastocyst development. In conventional droplet culture, embryos in the small-number group (5-6 embryos/droplet) showed low blastocyst development compared with a control group (25-26 embryos/droplet). However, small and large numbers of embryos (5-6 and 25 embryos, respectively) in a WOW dish showed n...

  8. File list: ALL.Emb.10.AllAg.4-8h_embryos [Chip-atlas[Archive

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  9. HSPC117 deficiency in cloned embryos causes placental abnormality and fetal death

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Yingying [Department of Reproduction and Development, Kunming Institute of Zoology and Kunming Primate Research Center, Chinese Academy of Sciences, Kunming 650223 (China); State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080 (China); Graduate University of Chinese Academy of Sciences, Beijing 100049 (China); Hai, Tang; Liu, Zichuan; Zhou, Shuya; Lv, Zhuo [State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080 (China); Graduate University of Chinese Academy of Sciences, Beijing 100049 (China); Ding, Chenhui; Liu, Lei [State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080 (China); Niu, Yuyu [Department of Reproduction and Development, Kunming Institute of Zoology and Kunming Primate Research Center, Chinese Academy of Sciences, Kunming 650223 (China); Zhao, Xiaoyang; Tong, Man [State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080 (China); Graduate University of Chinese Academy of Sciences, Beijing 100049 (China); Wang, Liu [State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080 (China); Jouneau, Alice [INRA, UMR 1198, ENVA, CNRS, FRE 2857, Biologie du Developpement et Reproduction, Jouy en Josas F-78350 (France); Zhang, Xun [Neuroendocrine Unit, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114 (United States); Ji, Weizhi, E-mail: wji@mail.kiz.ac.cn [Department of Reproduction and Development, Kunming Institute of Zoology and Kunming Primate Research Center, Chinese Academy of Sciences, Kunming 650223 (China); Zhou, Qi, E-mail: qzhou@ioz.ac.cn [State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080 (China)

    2010-07-02

    Somatic cell nuclear transfer (SCNT) has been successfully used in many species to produce live cloned offspring, albeit with low efficiency. The low frequency of successful development has usually been ascribed to incomplete or inappropriate reprogramming of the transferred nuclear genome. Elucidating the genetic differences between normal fertilized and cloned embryos is key to understand the low efficiency of SCNT. Here, we show that expression of HSPC117, which encodes a hypothetical protein of unknown function, was absent or very low in cloned mouse blastocysts. To investigate the role of HSPC117 in embryo development, we knocked-down this gene in normal fertilized embryos using RNA interference. We assessed the post-implantation survival of HSPC117 knock-down embryos at 3 stages: E9 (prior to placenta formation); E12 (after the placenta was fully functional) and E19 (post-natal). Our results show that, although siRNA-treated in vivo fertilized/produced (IVP) embryos could develop to the blastocyst stage and implanted without any difference from control embryos, the knock-down embryos showed substantial fetal death, accompanied by placental blood clotting, at E12. Furthermore, comparison of HSPC117 expression in placentas of nuclear transfer (NT), intracytoplasmic sperm injection (ICSI) and IVP embryos confirmed that HSPC117 deficiency correlates well with failures in embryo development: all NT embryos with a fetus, as well as IVP and ICSI embryos, had normal placental HSPC117 expression while those NT embryos showing reduced or no expression of HSPC117 failed to form a fetus. In conclusion, we show that HSPC117 is an important gene for post-implantation development of embryos, and that HSPC117 deficiency leads to fetal abnormalities after implantation, especially following placental formation. We suggest that defects in HSPC117 expression may be an important contributing factor to loss of cloned NT embryos in vivo.

  10. Reprogrammed transcriptome in rhesus-bovine interspecies somatic cell nuclear transfer embryos.

    Directory of Open Access Journals (Sweden)

    Kai Wang

    Full Text Available Global activation of the embryonic genome (EGA, one of the most critical steps in early mammalian embryo development, is recognized as the time when interspecies somatic cell nuclear transfer (iSCNT embryos fail to thrive.In this study, we analyzed the EGA-related transcriptome of rhesus-bovine iSCNT 8- to 16-cell embryos and dissected the reprogramming process in terms of embryonic gene activation, somatic gene silencing, and maternal RNA degradation. Compared with fibroblast donor cells, two thousand and seven genes were activated in iSCNT embryos, one quarter of them reaching expression levels comparable to those found in in vitro fertilized (IVF rhesus embryos. This suggested that EGA in iSCNT embryos had partially recapitulated rhesus embryonic development. Eight hundred and sixty somatic genes were not silenced properly and continued to be expressed in iSCNT embryos, which indicated incomplete nuclear reprogramming. We compared maternal RNA degradation in bovine oocytes between bovine-bovine SCNT and iSCNT embryos. While maternal RNA degradation occurred in both SCNT and iSCNT embryos, we saw more limited overall degradation of maternal RNA in iSCNT embryos than in SCNT embryos. Several important maternal RNAs, like GPF9, were not properly processed in SCNT embryos.Our data suggested that iSCNT embryos are capable of triggering EGA, while a portion of somatic cell-associated genes maintain their expression. Maternal RNA degradation seems to be impaired in iSCNT embryos. Further understanding of the biological roles of these genes, networks, and pathways revealed by iSCNT may expand our knowledge about cell reprogramming, pluripotency, and differentiation.

  11. HSPC117 deficiency in cloned embryos causes placental abnormality and fetal death

    International Nuclear Information System (INIS)

    Wang, Yingying; Hai, Tang; Liu, Zichuan; Zhou, Shuya; Lv, Zhuo; Ding, Chenhui; Liu, Lei; Niu, Yuyu; Zhao, Xiaoyang; Tong, Man; Wang, Liu; Jouneau, Alice; Zhang, Xun; Ji, Weizhi; Zhou, Qi

    2010-01-01

    Somatic cell nuclear transfer (SCNT) has been successfully used in many species to produce live cloned offspring, albeit with low efficiency. The low frequency of successful development has usually been ascribed to incomplete or inappropriate reprogramming of the transferred nuclear genome. Elucidating the genetic differences between normal fertilized and cloned embryos is key to understand the low efficiency of SCNT. Here, we show that expression of HSPC117, which encodes a hypothetical protein of unknown function, was absent or very low in cloned mouse blastocysts. To investigate the role of HSPC117 in embryo development, we knocked-down this gene in normal fertilized embryos using RNA interference. We assessed the post-implantation survival of HSPC117 knock-down embryos at 3 stages: E9 (prior to placenta formation); E12 (after the placenta was fully functional) and E19 (post-natal). Our results show that, although siRNA-treated in vivo fertilized/produced (IVP) embryos could develop to the blastocyst stage and implanted without any difference from control embryos, the knock-down embryos showed substantial fetal death, accompanied by placental blood clotting, at E12. Furthermore, comparison of HSPC117 expression in placentas of nuclear transfer (NT), intracytoplasmic sperm injection (ICSI) and IVP embryos confirmed that HSPC117 deficiency correlates well with failures in embryo development: all NT embryos with a fetus, as well as IVP and ICSI embryos, had normal placental HSPC117 expression while those NT embryos showing reduced or no expression of HSPC117 failed to form a fetus. In conclusion, we show that HSPC117 is an important gene for post-implantation development of embryos, and that HSPC117 deficiency leads to fetal abnormalities after implantation, especially following placental formation. We suggest that defects in HSPC117 expression may be an important contributing factor to loss of cloned NT embryos in vivo.

  12. Spontaneous activity in the statoacoustic ganglion of the chicken embryo.

    Science.gov (United States)

    Jones, T A; Jones, S M

    2000-03-01

    Statoacoustic ganglion cells in the mature bird include neurons that are responsive to sound (auditory) and those that are not (nonauditory). Those that are nonauditory have been shown to innervate an otolith organ, the macula lagena, whereas auditory neurons innervate the basilar papilla. In the present study, single-unit recordings of statoacoustic ganglion cells were made in embryonic (E19, mean = 19.2 days of incubation) and hatchling (P6-P14, mean = 8.6 days posthatch) chickens. Spontaneous activity from the two age groups was compared with developmental changes. Activity was evaluated for 47 auditory, 11 nonauditory, and 6 undefined eighth nerve neurons in embryos and 29 auditory, 26 nonauditory, and 1 undefined neurons in hatchlings. For auditory neurons, spontaneous activity displayed an irregular pattern [discharge interval coefficient of variation (CV) was >0.5, mean CV for embryos was 1.46 +/- 0.58 and for hatchlings was 1.02 +/- 0.25; means +/- SD]. Embryonic discharge rates ranged from 0.05 to 97.6 spikes per second (sp/s) for all neurons (mean 18.6 +/- 16.9 sp/s). Hatchling spontaneous rates ranged from 1.2 to 185.2 sp/s (mean 66.5 +/- 39.6 sp/s). Discharge rates were significantly higher for hatchlings (P embryonic auditory neurons displayed long silent periods between irregular bursts of neural activity, a feature not seen posthatch. All regular bursting discharge patterns were correlated with heart rate in both embryos and hatchlings. Preferred intervals were visible in the time interval histograms (TIHs) of only one embryonic neuron in contrast to 55% of the neurons in posthatch animals. Generally, the embryonic auditory TIH displayed a modified quasi-Poisson distribution. Nonauditory units generally displayed regular (CV 0.5) activity and Gaussian and modified-Gaussian TIHs. Long silent periods or bursting patterns were not a characteristic of embryonic nonauditory neurons. CV varied systematically as a function of discharge rate in nonauditory

  13. Simple Genome Editing of Rodent Intact Embryos by Electroporation.

    Directory of Open Access Journals (Sweden)

    Takehito Kaneko

    Full Text Available The clustered regularly interspaced short palindromic repeat (CRISPR/CRISPR-associated (Cas system is a powerful tool for genome editing in animals. Recently, new technology has been developed to genetically modify animals without using highly skilled techniques, such as pronuclear microinjection of endonucleases. Technique for animal knockout system by electroporation (TAKE method is a simple and effective technology that produces knockout rats by introducing endonuclease mRNAs into intact embryos using electroporation. Using TAKE method and CRISPR/Cas system, the present study successfully produced knockout and knock-in mice and rats. The mice and rats derived from embryos electroporated with Cas9 mRNA, gRNA and single-stranded oligodeoxynucleotide (ssODN comprised the edited targeted gene as a knockout (67% of mice and 88% of rats or knock-in (both 33%. The TAKE method could be widely used as a powerful tool to produce genetically modified animals by genome editing.

  14. Ambient UV-B radiation causes deformities in amphibian embryos

    Science.gov (United States)

    Blaustein, A.R.; Kiesecker, J.M.; Chivers, D.P.; Anthony, R.G.

    1997-01-01

    There has been a great deal of recent attention on the suspected increase in amphibian deformities. However, most reports of amphibian deformities have been anecdotal, and no experiments in the field under natural conditions have been performed to investigate this phenomenon. Under laboratory conditions, a variety of agents can induce deformities in amphibians. We investigated one of these agents, UV-B radiation, in field experiments, as a cause for amphibian deformities. We monitored hatching success and development in long-toed salamanders under UV-B shields and in regimes that allowed UV-B radiation. Embryos under UV-B shields had a significantly higher hatching rate and fewer deformities, and developed more quickly than those exposed to UV-B. Deformities may contribute directly to embryo mortality, and they may affect an individual's subsequent survival after hatching.

  15. Recent microfluidic devices for studying gamete and embryo biomechanics.

    Science.gov (United States)

    Lai, David; Takayama, Shuichi; Smith, Gary D

    2015-06-25

    The technical challenges of biomechanic research such as single cell analysis at a high monetary cost, labor, and time for just a small number of measurements is a good match to the strengths of microfluidic devices. New scientific discoveries in the fertilization and embryo development process, of which biomechanics is a major subset of interest, is crucial to fuel the continual improvement of clinical practice in assisted reproduction. The following review will highlight some recent microfluidic devices tailored for gamete and embryo biomechanics where biomimicry arises as a major theme of microfluidic device design and function, and the application of fundamental biomechanic principles are used to improve outcomes of cryopreservation. Copyright © 2015 Elsevier Ltd. All rights reserved.

  16. Toxic effects of brominated indoles and phenols on zebrafish embryos.

    Science.gov (United States)

    Kammann, U; Vobach, M; Wosniok, W

    2006-07-01

    Organobromine compounds in the marine environment have been the focus of growing attention in past years. In contrast to anthropogenic brominated flame retardants, other brominated compounds are produced naturally, e.g., by common polychaete worms and algae. Brominated phenols and indoles assumed to be of biogenic origin have been detected in water and sediment extracts from the German Bight. These substances as well as some of their isomers have been tested with the zebrafish embryo test and were found to cause lethal as well as nonlethal malformations. The zebrafish test was able to detect a log K(OW)-related toxicity for bromophenols, suggesting nonpolar narcosis as a major mode of action. Different effect patterns could be observed for brominated indoles and bromophenols. The comparison of effective concentrations in the zebrafish embryo test with the concentrations determined in water samples suggests the possibility that brominated indoles may affect early life stages of marine fish species in the North Sea.

  17. Preventing Mitochondrial Diseases: Embryo-Sparing Donor-Independent Options.

    Science.gov (United States)

    Adashi, Eli Y; Cohen, I Glenn

    2018-05-01

    Mutant mitochondrial DNA gives rise to a broad range of incurable inborn maladies. Prevention may now be possible by replacing the mutation-carrying mitochondria of zygotes or oocytes at risk with donated unaffected counterparts. However, mitochondrial replacement therapy is being held back by theological, ethical, and safety concerns over the loss of human zygotes and the involvement of a donor. These concerns make it plain that the identification, validation, and regulatory adjudication of novel embryo-sparing donor-independent technologies remains a pressing imperative. This Opinion highlights three emerging embryo-sparing donor-independent options that stand to markedly allay theological, ethical, and safety concerns raised by mitochondrial replacement therapy. Copyright © 2018 Elsevier Ltd. All rights reserved.

  18. Composition of commercial media used for human embryo culture.

    Science.gov (United States)

    Morbeck, Dean E; Krisher, Rebecca L; Herrick, Jason R; Baumann, Nikola A; Matern, Dietrich; Moyer, Thomas

    2014-09-01

    To determine the composition of commercially available culture media and test whether differences in composition are biologically relevant in a murine model. Experimental laboratory study. University-based laboratory. Cryopreserved hybrid mouse one-cell embryos were used in experiments. Amino acid, organic acid, ions, and metal content were determined for two different lots of media from Cook, In Vitro Care, Origio, Sage, Vitrolife, Irvine CSC, and Global. To determine whether differences in the composition of these media are biologically relevant, mouse one-cell embryos were thawed and cultured for 120 hours in each culture media at 5% and 20% oxygen in the presence or absence of protein in an EmbryoScope time-lapse incubator. The compositions of seven culture media were analyzed for concentrations of 39 individual amino acids, organic acids, ions, and elements. Blastocyst rates and cell cycle timings were calculated at 96 hours of culture, and the experiments were repeated in triplicate. Of the 39 analytes, concentrations of glucose, lactate, pyruvate, amino acids, phosphate, calcium, and magnesium were present in variable concentrations, likely reflecting differences in the interpretation of animal studies. Essential trace elements, such as copper and zinc, were not detected. Mouse embryos failed to develop in one culture medium and were differentially affected by oxygen in two other media. Culture media composition varies widely, with differences in pyruvate, lactate, and amino acids especially notable. Blastocyst development was culture media dependent and showed an interaction with oxygen concentration and presence of protein. Copyright © 2014 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  19. Transferring embryos with indeterminate PGD results: the ethical implications

    OpenAIRE

    Insogna, Iris G.; Ginsburg, Elizabeth

    2016-01-01

    Background In vitro fertilization (IVF) and pre-implantation genetic diagnosis (PGD) are becoming increasingly common techniques to select embryos that are unaffected by a known genetic disorder. Though IVF-PGD has high success rates, 7.5?% of blastocysts have inconclusive results after testing. A recent case involving a known BRCA-1 carrier was brought before our Assisted Reproductive Technology Ethics Committee in order to gain a better appreciation for the ethical implications surrounding ...

  20. Oral Morphine Consumption Reduces Lens Development in Rat Embryos

    Directory of Open Access Journals (Sweden)

    Hossein Bahadoran

    2012-07-01

    Full Text Available Objective: Consumption of morphine, during pregnancy, in addition to inducing defects in the mother’s nervous system function, caused defects or delays in the formation and evolution of embryonic visual system. In the present study, changes in lens development was assessed in embryos exposed in utero to morphine. Material and Methods: Female Wistar rats (250-300 g were mated with male rats and pregnancy was determined by sperm observation in vaginal smear. This day was considered as embryonic day zero (E0. The females were then divided randomly into the experimental and the control groups. The control group received tap water and the experimental group received morphine (0.05 mg/ml in their water. On embryonic day 13 ( E13, blood samples were collected from the retro-orbital sinus of all animals for plasma corticosterone detection. On embryonic day 17(E17, the animals were killed by an overdose of chloroform and the embryos were taken out surgically. The embryos were fixed in 10% formalin for 30 days. At this time, the head of the embryos were removed for tissue processing and Hematoxylin- Eosin (H&E staining. The samples were evaluated using light microscope and MOTIC software. Results: Our data indicated that plasma corticosterone level was dramatically increased and the lens was thinner in the experimental group. (Although the proliferation of lens cells increased in the experiment group but that lens had delay in removing the proliferated and elongation cells with abnormal density in the lateral part of the lens in compare with control group. I have no idea what the authors are stating here. Moreover, the opening of the eyelids was delayed in the off springs of the mothers who received morphine. Conclusions: This study showed that morphine consumption during pregnancy leads to defects in fetal visual system development, particularly in the lens, and eyelids.

  1. Shared and unique patterns of embryo development in extremophile poeciliids

    OpenAIRE

    Riesch, Rüdiger; Schlupp, Ingo; Langerhans, R. Brian; Plath, Martin

    2011-01-01

    Background: Closely related lineages of livebearing fishes have independently adapted to two extreme environmental factors: toxic hydrogen sulphide (H2S) and perpetual darkness. Previous work has demonstrated in adult specimens that fish from these extreme habitats convergently evolved drastically increased head and offspring size, while cave fish are further characterized by reduced pigmentation and eye size. Here, we traced the development of these (and other) divergent traits in embryos of...

  2. Embryo reduction before day 11 in mares with twin conceptuses.

    Science.gov (United States)

    Ginther, O J; Bergfelt, D R

    1988-07-01

    Incidence of embryo reduction (natural elimination of one member of a twin set) before d 11 was studied by comparing the number of ultrasonically detected conceptuses per ovulation between single and double ovulators. Effect of unilateral (n = 24) vs bilateral (n = 26) double ovulations on the incidence of embryo reduction also was considered. Each of 50 double-ovulating mares was matched with two single ovulators yielding 100 ovulations, or potential embryos, per group. Frequency with which an ovulation resulted in a conceptus was greater for single ovulators (85%, P less than .01) and for bilateral double ovulators (77%, P less than .05) than for unilateral double ovulators (60%). The difference between single and bilateral ovulators was not significant. Expected frequencies for none, one or two conceptuses per mare for double ovulators were calculated using the d-11 pregnancy rate for the single ovulators (85%). For unilateral ovulators, but not for bilateral ovulators, observed frequencies of none, one or two conceptuses were different (P less than .05) from expected (observed and expected, respectively, for no conceptuses, 29% and 2%; one conceptus, 21% and 26%; two conceptuses, 50% and 72%). Results did not indicate the existence of an embryo reduction phenomenon before d 11 in bilateral ovulators; each ovum had the same chance (no significant difference) of developing into a d-11 conceptus as an ovum in single ovulators. In unilateral double ovulators, the lower d-11 pregnancy rate per ovum, compared with bilateral ovulators and single ovulators, was attributable to a greater frequency of mares with no embryonic vesicles rather than to a greater frequency of mares with one vesicle.(ABSTRACT TRUNCATED AT 250 WORDS)

  3. Secondary Cartilage Revealed in a Non-Avian Dinosaur Embryo

    OpenAIRE

    Bailleul, Alida M.; Hall, Brian K.; Horner, John R.

    2013-01-01

    The skull and jaws of extant birds possess secondary cartilage, a tissue that arises after bone formation during embryonic development at articulations, ligamentous and muscular insertions. Using histological analysis, we discovered secondary cartilage in a non-avian dinosaur embryo, Hypacrosaurus stebingeri (Ornithischia, Lambeosaurinae). This finding extends our previous report of secondary cartilage in post-hatching specimens of the same dinosaur species. It provides the first information ...

  4. Single-site neural tube closure in human embryos revisited.

    Science.gov (United States)

    de Bakker, Bernadette S; Driessen, Stan; Boukens, Bastiaan J D; van den Hoff, Maurice J B; Oostra, Roelof-Jan

    2017-10-01

    Since the multi-site closure theory was first proposed in 1991 as explanation for the preferential localizations of neural tube defects, the closure of the neural tube has been debated. Although the multi-site closure theory is much cited in clinical literature, single-site closure is most apparent in literature concerning embryology. Inspired by Victor Hamburgers (1900-2001) statement that "our real teacher has been and still is the embryo, who is, incidentally, the only teacher who is always right", we decided to critically review both theories of neural tube closure. To verify the theories of closure, we studied serial histological sections of 10 mouse embryos between 8.5 and 9.5 days of gestation and 18 human embryos of the Carnegie collection between Carnegie stage 9 (19-21 days) and 13 (28-32 days). Neural tube closure was histologically defined by the neuroepithelial remodeling of the two adjoining neural fold tips in the midline. We did not observe multiple fusion sites in neither mouse nor human embryos. A meta-analysis of case reports on neural tube defects showed that defects can occur at any level of the neural axis. Our data indicate that the human neural tube fuses at a single site and, therefore, we propose to reinstate the single-site closure theory for neural tube closure. We showed that neural tube defects are not restricted to a specific location, thereby refuting the reasoning underlying the multi-site closure theory. Clin. Anat. 30:988-999, 2017. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

  5. CanUreaplasma diversumbe transmitted from donor to recipient through the embryo? Two case reports outliningU. diversumlosses in bovine embryo pregnancies.

    Science.gov (United States)

    Crane, M Bronwyn; Hughes, Colleen A

    2018-01-01

    Two bovine embryo recovery results are outlined from different herds. Both cases involve significant late gestational loss from embryos relating back to a single donor. Ureaplasma diversum was confirmed in 3 of 4 cases submitted for postmortem examination. Natural infection originating from the donor and transmitted to the recipient has not previously been documented.

  6. A hyperactive sleeping beauty transposase enhances transgenesis in zebrafish embryos

    Directory of Open Access Journals (Sweden)

    Lardelli Michael

    2010-11-01

    Full Text Available Abstract Background Transposons are useful molecular tools for transgenesis. The 'sleeping beauty' transposon is a synthetic member of the Tc1/mariner transposon family. Davidson et al. (2003 previously described a vector for zebrafish transgenesis consisting of the inverted repeats of 'sleeping beauty' flanking the gene to be transposed. Subsequently, there have been attempts to enhance the transpositional activity of 'sleeping beauty' by increasing the activity of its transposase. Recently, Mates et al. (2009 generated a hyperactive transposase giving a 100-fold increased transposition rate in mouse embryos. Findings The aim of this experiment was to determine whether this novel hyperactive transposase enhances transgenesis in zebrafish embryos. Using our previously characterised mitfa-amyloidβ-GFP transgene, we observed an eight-fold enhancement in transient transgenesis following detection of transgene expression in melanophores by whole mount in-situ hybridisation. However, high rates of defective embryogenesis were also observed. Conclusion The novel hyperactive 'sleeping beauty' transposase enhances the rate of transgenesis in zebrafish embryos.

  7. Cleavage events and sperm dynamics in chick intrauterine embryos.

    Directory of Open Access Journals (Sweden)

    Hyung Chul Lee

    Full Text Available This study was undertaken to elucidate detailed event of early embryogenesis in chicken embryos using a noninvasive egg retrieval technique before oviposition. White Leghorn intrauterine eggs were retrieved from 95 cyclic hens aged up to 54-56 weeks and morphogenetic observation was made under both bright field and fluorescent image in a time course manner. Differing from mammals, asymmetric cleavage to yield preblastodermal cells was observed throughout early embryogenesis. The first two divisions occurred synchronously and four polarized preblastodermal cells resulted after cruciform cleavage. Then, asynchronous cleavage continued in a radial manner and overall cell size in the initial cleavage region was smaller than that in the distal area. Numerous sperms were visible, regardless of zygotic nuclei formation. Condensed sperm heads were present mainly in the perivitelline space and cytoplasm, and rarely in the yolk region, while decondensed sperm heads were only visible in the yolk. In conclusion, apparent differences in sperm dynamics and early cleavage events compared with mammalian embryos were detected in chick embryo development, which demonstrated polarized cleavage with penetrating supernumerary sperm into multiple regions.

  8. Toxic effects of silica nanoparticles on zebrafish embryos and larvae.

    Directory of Open Access Journals (Sweden)

    Junchao Duan

    Full Text Available Silica nanoparticles (SiNPs have been widely used in biomedical and biotechnological applications. Environmental exposure to nanomaterials is inevitable as they become part of our daily life. Therefore, it is necessary to investigate the possible toxic effects of SiNPs exposure. In this study, zebrafish embryos were treated with SiNPs (25, 50, 100, 200 µg/mL during 4-96 hours post fertilization (hpf. Mortality, hatching rate, malformation and whole-embryo cellular death were detected. We also measured the larval behavior to analyze whether SiNPs had adverse effects on larvae locomotor activity. The results showed that as the exposure dosages increasing, the hatching rate of zebrafish embryos was decreased while the mortality and cell death were increased. Exposure to SiNPs caused embryonic malformations, including pericardial edema, yolk sac edema, tail and head malformation. The larval behavior testing showed that the total swimming distance was decreased in a dose-dependent manner. The lower dose (25 and 50 µg/mL SiNPs produced substantial hyperactivity while the higher doses (100 and 200 µg/mL SiNPs elicited remarkably hypoactivity in dark periods. In summary, our data indicated that SiNPs caused embryonic developmental toxicity, resulted in persistent effects on larval behavior.

  9. [The oxygen consumption of ostrich embryos during incubation].

    Science.gov (United States)

    Reiner, G; Dzapo, V

    1995-02-01

    This work deals with the oxygen consumption of ostrich chicks during incubation. Brood eggs were incubated in a hermetic isolated acrylic-glass cylinder. Reduction of oxygen content in the air surrounding the egg was measured using an oxygen-sensitive electrode. A sigmoid curve could be drawn during incubation, with the steepest phase being around day 26. Maximum oxygen consumption was reached on day 36. It was slightly decreased until day 39, when the embryo switches to lung circulation, followed again by an increase until hatching. Average oxygen consumptions for the whole brood interval were calculated to 63.6 liters. Oxygen volumes consumed on day 36 result in a demand about to 240 liters of fresh air per egg and day. Oxygen consumption of the embryos on day 36 was significantly positive correlated with their vitality. Numb or less vital embryos could be clearly differentiated from others. The higher a chick's oxygen consumption, the earlier and shorter its hatching. Possible applications of the method in regard to the evaluation of incubation parameters or chicken constitution are discussed.

  10. Utilization of half-embryo test to identify irradiated beans

    Energy Technology Data Exchange (ETDEWEB)

    Villavicencio, Anna Lucia C.H. [Instituto de Pesquisas Energeticas e Nucleares (IPEN), Sao Paulo, SP (Brazil); Mancini-Filho, Jorge [Sao Paulo Univ., SP (Brazil). Faculdade de Ciencias Farmaceuticas; Delincee, Henry [Federal Research Centre for Nutrition - BFE, Karlsruhe (Germany)

    1996-07-01

    Germination tests were carried out in irradiated and non-irradiated bean seeds which allow to observe characteristically variations on the shoots and roots. The methodology used in this work, is based upon biological changes which occur in two Brazilian beans, Phaseolus vulgaris L., var. carioca and Vigna unguiculata (L.) Walp, var. macacar, irradiated in a {sup 60} Co source, with doses of 0,0.5, 1.0, 2.5, 5.0 and 10.0 kGy. The shoots and roots were observed during 3 days of culturing period under specified conditions. The differences observed in these two varieties were analysed immediately after irradiation and after 6 months of storage period at room temperature. Irradiated half-embryos showed markedly reduced root grow and almost totally retarded shoot elongation. Differences between irradiated and nonirradiated half-embryo could be observed after irradiation when different beans and storage time were varied. The shoots of half-embryos irradiated with more than 2.5 kGy did not undergo any elongation, whereas, the shoots of non-irradiated or those beans irradiated under 1.0 kGy elongated significantly within the 3 day test period. (author)

  11. Changes in RNA Splicing in Developing Soybean (Glycine max Embryos

    Directory of Open Access Journals (Sweden)

    Delasa Aghamirzaie

    2013-11-01

    Full Text Available Developing soybean seeds accumulate oils, proteins, and carbohydrates that are used as oxidizable substrates providing metabolic precursors and energy during seed germination. The accumulation of these storage compounds in developing seeds is highly regulated at multiple levels, including at transcriptional and post-transcriptional regulation. RNA sequencing was used to provide comprehensive information about transcriptional and post-transcriptional events that take place in developing soybean embryos. Bioinformatics analyses lead to the identification of different classes of alternatively spliced isoforms and corresponding changes in their levels on a global scale during soybean embryo development. Alternative splicing was associated with transcripts involved in various metabolic and developmental processes, including central carbon and nitrogen metabolism, induction of maturation and dormancy, and splicing itself. Detailed examination of selected RNA isoforms revealed alterations in individual domains that could result in changes in subcellular localization of the resulting proteins, protein-protein and enzyme-substrate interactions, and regulation of protein activities. Different isoforms may play an important role in regulating developmental and metabolic processes occurring at different stages in developing oilseed embryos.

  12. Utilization of half-embryo test to identify irradiated beans

    International Nuclear Information System (INIS)

    Villavicencio, Anna Lucia C.H.; Mancini-Filho, Jorge

    1996-01-01

    Germination tests were carried out in irradiated and non-irradiated bean seeds which allow to observe characteristically variations on the shoots and roots. The methodology used in this work, is based upon biological changes which occur in two Brazilian beans, Phaseolus vulgaris L., var. carioca and Vigna unguiculata (L.) Walp, var. macacar, irradiated in a 60 Co source, with doses of 0,0.5, 1.0, 2.5, 5.0 and 10.0 kGy. The shoots and roots were observed during 3 days of culturing period under specified conditions. The differences observed in these two varieties were analysed immediately after irradiation and after 6 months of storage period at room temperature. Irradiated half-embryos showed markedly reduced root grow and almost totally retarded shoot elongation. Differences between irradiated and nonirradiated half-embryo could be observed after irradiation when different beans and storage time were varied. The shoots of half-embryos irradiated with more than 2.5 kGy did not undergo any elongation, whereas, the shoots of non-irradiated or those beans irradiated under 1.0 kGy elongated significantly within the 3 day test period. (author)

  13. [Progress in proteomics of mammalian oocyte and early embryo].

    Science.gov (United States)

    Chen, Lingsheng; Xu, Ping; Shi, Deshun; Li, Xiangping

    2014-07-01

    The development of female germ cell is the cornerstone for animal reproduction. Mammalian oocyte and early embryo have many distinct phenomena and mechanisms during their growth and development, involving series dynamic changes of protein synthesis/degradation and phosphorylation. Research on the regulatory mechanism of oocyte division, maturation, and developmental principle of pre-implantation embryo is an important topic in the field of animal developmental biology. Proteomics using all of proteins expressed by a cell or tissue as research object, systematically identify, quantify and study the function of all these proteins. With the rapid development of protein separation and identification technology, proteomics provide some new methods and the research contents on fields of oogenesis, differentiation, maturation and quality control, such as protein quantification, modification, location and interaction important information which other omics technology can not provide. These information will contribute to uncover the molecular mechanisms of mammalian oocyte maturation and embryonic development. And it is great significant for improving the culture system of oocyte in vitro maturation, the efficiency of embryo production in vitro, somatic cell clone and transgenic animal production.

  14. Cryopreservation of cocoa (Theobroma cacao L.) somatic embryos by vitrification.

    Science.gov (United States)

    Adu-Gyamfi, Raphael; Wetten, Andy

    2012-01-01

    Losses of cultivated cocoa (Theobroma cacao L.) due to diseases and continued depletion of forests that harbour the wild progenitors of the crop make ex situ conservation of cocoa germplasm of paramount importance. In order to enhance security of in situ germplasm collections, 2-3 mm floral-derived secondary somatic embryos were cryopreserved by vitrification. This work demonstrates the most uncomplicated clonal cocoa cryopreservation. Optimal post-cryostorage survival (74.5 percent) was achieved by 5 d preculture of SSEs on 0.5 M sucrose medium followed by 60 min dehydration in cold PVS2. To minimise free radical related cryo-injury, cation sources were removed from the embryo development solution and/or the recovery medium, the former treatment resulting in a significant benefit. After optimisation with cocoa genotype AMAZ 15, the same protocol was effective across all five additional cocoa genotypes tested. For the multiplication of clones, embryos regenerated following cryopreservation were used as explant sources, and vitrification was found to maintain their embryogenic potential.

  15. Secondary cartilage revealed in a non-avian dinosaur embryo.

    Science.gov (United States)

    Bailleul, Alida M; Hall, Brian K; Horner, John R

    2013-01-01

    The skull and jaws of extant birds possess secondary cartilage, a tissue that arises after bone formation during embryonic development at articulations, ligamentous and muscular insertions. Using histological analysis, we discovered secondary cartilage in a non-avian dinosaur embryo, Hypacrosaurus stebingeri (Ornithischia, Lambeosaurinae). This finding extends our previous report of secondary cartilage in post-hatching specimens of the same dinosaur species. It provides the first information on the ontogeny of avian and dinosaurian secondary cartilages, and further stresses their developmental similarities. Secondary cartilage was found in an embryonic dentary within a tooth socket where it is hypothesized to have arisen due to mechanical stresses generated during tooth formation. Two patterns were discerned: secondary cartilage is more restricted in location in this Hypacrosaurus embryo, than it is in Hypacrosaurus post-hatchlings; secondary cartilage occurs at far more sites in bird embryos and nestlings than in Hypacrosaurus. This suggests an increase in the number of sites of secondary cartilage during the evolution of birds. We hypothesize that secondary cartilage provided advantages in the fine manipulation of food and was selected over other types of tissues/articulations during the evolution of the highly specialized avian beak from the jaws of their dinosaurian ancestors.

  16. Secondary cartilage revealed in a non-avian dinosaur embryo.

    Directory of Open Access Journals (Sweden)

    Alida M Bailleul

    Full Text Available The skull and jaws of extant birds possess secondary cartilage, a tissue that arises after bone formation during embryonic development at articulations, ligamentous and muscular insertions. Using histological analysis, we discovered secondary cartilage in a non-avian dinosaur embryo, Hypacrosaurus stebingeri (Ornithischia, Lambeosaurinae. This finding extends our previous report of secondary cartilage in post-hatching specimens of the same dinosaur species. It provides the first information on the ontogeny of avian and dinosaurian secondary cartilages, and further stresses their developmental similarities. Secondary cartilage was found in an embryonic dentary within a tooth socket where it is hypothesized to have arisen due to mechanical stresses generated during tooth formation. Two patterns were discerned: secondary cartilage is more restricted in location in this Hypacrosaurus embryo, than it is in Hypacrosaurus post-hatchlings; secondary cartilage occurs at far more sites in bird embryos and nestlings than in Hypacrosaurus. This suggests an increase in the number of sites of secondary cartilage during the evolution of birds. We hypothesize that secondary cartilage provided advantages in the fine manipulation of food and was selected over other types of tissues/articulations during the evolution of the highly specialized avian beak from the jaws of their dinosaurian ancestors.

  17. Gas exchange and energy expenditure in chicken embryos

    DEFF Research Database (Denmark)

    Chwalibog, André; Tauson, Anne-Helene; Ali, Abdalla

    . The pattern of curves for gas exchange was identical for RO and LA, but on a lower level for LA. The energy expenditure followed the pattern of curves for gas exchange, with a mean value around 50 J/h on day 10, increasing to 528 (RO) and 402 (LA) J/h on day 19 (Figure 1). The main source of EE was oxidized...... is independent of genetic origin of embryos. However, the embryos from the slow growing broiler line had a lower metabolic rate and oxidised less fat than the modern, fast growing line. The reduced utilization of yolk fat might be a tool for saving fat reserves for the immediate post-hatching period......) in this phase may be a crucial parameter predicting metabolic rate and consquently, growth performance of post-hatched chickens. The aim of this investigation was to determine EE in embryos of slow and fast growing lines of chickens. Taking advantage of the indirect calorimetry technique it was also possible...

  18. Early somatic embryo induction events in alfalfa callus cultures

    International Nuclear Information System (INIS)

    El-Bakry, A.A.; Hildebrand, D.F.

    1987-01-01

    High and low regenerating alfalfa Medicago sativa L. cv Regen S full sibs were isolated from a callus culture screen on modified Blaydes medium. The average number of embryos per ovary were thirty and zero for the high and low genotypes respectively after six weeks in culture. Proembryonic cell masses (4-8 celled) were observed after 4-5 days in culture and maximum meristematic activity was at 6-7 days in culture, for the high regenerating genotypes. Well formed globular embryos, both epidermal and subepidermal in origin, were observed after 2 weeks is culture. Samples in culture for 3, 6 and 14 days from the high and low regenerating genotypes were radiolabeled in vivo with 35 S-methionine and run both on one and two dimension gels. The results will be discussed in relation to differences in proteins between the high and low regenerating genotypes at the stage of maximum meristematic activity (day 6) and differences occurring relative to the appearance of globular stage embryos (day 14) will be presented

  19. Teratogenic efects of injected methylmercury on avian embryos

    Science.gov (United States)

    Heinz, Gary H.; Hoffman, David J.; Klimstra, Jon D.; Stebbins, Katherine R.; Kondrad, Shannon L.; Erwin, Carol A.

    2011-01-01

    Controlled laboratory studies with game farm mallards (Anas platyrhynchos) and chickens (Gallus gallus) have demonstrated that methylmercury can cause teratogenic effects in birds, but studies with wild species of birds are lacking. To address this need, doses of methylmercury chloride were injected into the eggs of 25 species of birds, and the dead embryos and hatched chicks were examined for external deformities. When data for controls were summed across all 25 species tested and across all types of deformities, 24 individuals out of a total of 1,533 (a rate of 1.57%) exhibited at least one deformity. In contrast, when data for all of the mercury treatments and all 25 species were summed, 188 deformed individuals out of a total of 2,292 (8.20%) were found. Some deformities, such as lordosis and scoliosis (twisting of the spine), misshapen heads, shortening or twisting of the neck, and deformities of the wings, were seldom observed in controls but occurred in much greater frequency in Hg-treated individuals. Only 0.59% of individual control dead embryos and hatchlings exhibited multiple deformities versus 3.18% for Hg-dosed dead embryos and hatchlings. Methylmercury seems to have a widespread teratogenic potential across many species of birds.

  20. Human embryo research and the language of moral uncertainty.

    Science.gov (United States)

    Cheshire, William P

    2004-01-01

    In bioethics as in the sciences, enormous discussions often concern the very small. Central to public debate over emerging reproductive and regenerative biotechnologies is the question of the moral status of the human embryo. Because news media have played a prominent role in framing the vocabulary of the debate, this study surveyed the use of language reporting on human embryo research in news articles spanning a two-year period. Terminology that devalued moral status-for example, the descriptors things, property, tissue, or experimental material -was found to outnumber fivefold those that affirmed any degree of moral status above that of inanimate cellular matter; for example, living, human life, or human being. A quarter of the articles failed to note that the embryos under discussion were human. These findings confirm that even among scientific and philosophical experts a diversity of opinion exists on society's moral obligations to nascent human life. The skewed linguistic distribution also indicates a distinct bias. Concerned readers should take notice when any category of humanity becomes subject to prejudicial and disparaging language and the value of vulnerable human life is trivialized alongside sensational assertions of anticipated medical cures. The responsibility for holding the media to a higher standard of truth and fairness falls to us all.