WorldWideScience
1

Fully functional HLA B27-restricted CD4+ as well as CD8+ T cell responses in TCR transgenic mice.  

OpenAIRE

The strong association of HLA B27 with spondyloarthropathies contrasts strikingly with most autoimmune diseases, which are HLA class II associated and thought to be mediated by CD4+ T lymphocytes. By introducing a human-derived HLA B27-restricted TCR into HLA B27 transgenic mice, we have obtained a functional TCR transgenic model, GRb, dependent on HLA B27 for response. Surprisingly, HLA B27 supported CD4+ as well as CD8+ T cell responses in vivo and in vitro. Further, HLA B27-restricted CD4+...

Roddis, M.; Carter, Rw; Sun, My; Weissensteiner, T.; Mcmichael, Aj; Bowness, P.; Bodmer, Hc

2004-01-01

2

Evidence for the prevention of enthesitis in HLA-B27/h?(2)m transgenic rats treated with a monoclonal antibody against TNF-?.  

Science.gov (United States)

Transgenic rats with high expression of HLA-B27 and human ?(2) -microglobulin (B27TR) develop a multisystem inflammatory disease resembling human inflammatory bowel disease (IBD) and spondyloarthropaties (SpA). Tumour necrosis factor ? (TNF-?) has a crucial role in sustaining chronic inflammation in the gut and joints. The aim of this work was to evaluate whether TNF-? blockade could prevent or reduce the inflammation of peripheral joints in B27TR. A first group of 9-week-old B27TR received an anti-TNF-? monoclonal antibody (mAb) or an isotypic IgG2a,k up to the age of 18 weeks. An untreated group was monitored up to the age of 18 weeks and then randomly assigned to a 9-week treatment with anti-TNF-? mAb or IgG2a,k. Each rat was monitored for clinical IBD and peripheral joint manifestations. After sacrifice the colon and hind paws were examined for macroscopical and microscopical pathological changes. Early TNF-? blockade prevented, and late treatment improved IBD signs in B27TR. Erythema, oedema, inflammatory infiltrate close to the tendons and enthesis, proliferating chondrocyte-like cells, signs of new endochondral bone ossification and bone erosion were observed in peripheral joints of four out of six IgG2a,k-treated B27TR, both at 18 and 27 weeks. Immunopositivity for phosphorylated Smad1/5/8 indicated that the process of joint remodelling was activated in B27TR. Some entheses showed chondroid nodules. Anti-TNF-? treatment reduced inflammation and preserved the enthesis organization in most animals. Occasional and transient erythema and oedema were still present in three of six of the late anti-TNF-?-treated animals. Smad1/5/8 signalling was not inhibited by late anti-TNF-? treatment. In B27TR, articular involvement follows IBD onset and develops at entheses. Early TNF-? blockade prevents the onset of IBD and consequently the development of enthesitis in peripheral joints in the B27TR model of human SpA. PMID:20015205

Milia, Anna Franca; Ibba-Manneschi, Lidia; Manetti, Mirko; Benelli, Gemma; Generini, Sergio; Messerini, Luca; Matucci-Cerinic, Marco

2011-02-01

3

Hla-b27.  

Science.gov (United States)

Possession of the human leukocyte antigen (HLA) class I molecule B27 is strongly associated with ankylosing spondylitis (AS), but the pathogenic role of HLA-B27 is unknown. Two broad theories most likely explain the role of HLA-B27 in AS pathogenesis. The first is based on the natural immunological function of HLA-B27 of presenting antigenic peptides to cytotoxic T cells. Thus, HLA-B27-restricted immune responses to self-antigens, or arthritogenic peptides, might drive immunopathology. B27 can also "behave badly," misfolding during assembly and leading to endoplasmic reticulum stress and autophagy responses. ?2m-free B27 heavy chain structures including homodimers (B272) can also be expressed at the cell surface following endosomal recycling of cell surface heterotrimers. Cell surface free heavy chains and B272 bind to innate immune receptors on T, NK, and myeloid cells with proinflammatory effects. This review describes the natural function of HLA-B27, its disease associations, and the current theories as to its pathogenic role. PMID:25861975

Bowness, Paul

2015-03-21

4

HLA-B27 Test  

Science.gov (United States)

... name: Human Leukocyte Antigen B27 Related tests: ESR ; C-Reactive Protein ; Rheumatoid Factor ; HLA Testing At a Glance Test ... an ESR (erythrocyte sedimentation rate) or a CRP (C-reactive protein) . HLA-B27 is sometimes ordered to help evaluate ...

5

Heart conduction disturbance: an HLA-B27 associated disease.  

OpenAIRE

In recent studies from Sweden an increased prevalence of HLA-B27 associated diseases and of HLA-B27 was found in an unselected group of men with permanently implanted pacemakers and with a heart block. Furthermore, a significantly increased prevalence of HLA-B27 was found in men with a pacemaker who had no clinical or radiological signs of HLA-B27 associated disease. To obtain more insight into the association between HLA-B27 and heart block, and the possible role of HLA-B27 in causing this b...

Peeters, A. J.; Ten Wolde, S.; Sedney, M. I.; Vries, R. R.; Dijkmans, B. A.

1991-01-01

6

HLA-B27 subtypes among the Chukotka native groups  

Energy Technology Data Exchange (ETDEWEB)

The purpose of this study was to assess the relative frequency of the known HLA-B27 subtypes in HLA-B27 positive Chukotka natives, which have higher frequencies of HLA-B27 (to 40%) and spondylarthropathies (to 2%) than the Russian Caucasian population. Using oligotyping of the polymerase-chain reaction amplified second and third exons of the HLA-B27 gene in 86 DNA samples from HLA-B27 positive individuals were successfully typed. All had HLA-B*2705, including 4 patients with Reiter`s syndrome and 5 with ankylosing spondyloarthritis, except one Eskimo who had HLA-B*2702. None had HLA-B*2704, a frequent subtype in Orientals. With respect to HLA-B27 subtypes the indigenous populations from the eastern part of the Chukotka Peninsula are genetically more closely related to Caucasians than to Orientals. (author). 18 refs, 1 fig., 2 tabs.

Krylov, M.Y.; Alexeeva, L.I.; Erdesz, S.; Benevolenskaya, L.I. [Akademiya Meditsinskikh Nauk SSSR, Moscow (Russian Federation). Inst. Revmatizma; Reveille, J.D.; Arnett, F.C. [Texas Univ., Houston, TX (United States). Health Science Center

1995-12-31

7

HLA-B27 subtypes among the Chukotka native groups  

International Nuclear Information System (INIS)

The purpose of this study was to assess the relative frequency of the known HLA-B27 subtypes in HLA-B27 positive Chukotka natives, which have higher frequencies of HLA-B27 (to 40%) and spondylarthropathies (to 2%) than the Russian Caucasian population. Using oligotyping of the polymerase-chain reaction amplified second and third exons of the HLA-B27 gene in 86 DNA samples from HLA-B27 positive individuals were successfully typed. All had HLA-B*2705, including 4 patients with Reiter's syndrome and 5 with ankylosing spondyloarthritis, except one Eskimo who had HLA-B*2702. None had HLA-B*2704, a frequent subtype in Orientals. With respect to HLA-B27 subtypes the indigenous populations from the eastern part of the Chukotka Peninsula are genetically more closely related to Caucasians than to Orientals. (author). 18 refs, 1 fig., 2 tabs

8

HLA-B27 and an electrocardiographic peculiarity  

OpenAIRE

INTRODUCTION: An increased cardiovascular mortality has been described in patients with spondyloarthropathies due to HLA-B27. Numerous cardiovascular afflictions are currently known to be associated with HLA-B27. These include aortic root dilation, aortic regurgitation, mitral regurgitation, myocarditis, heart failure, pericarditis, pericardial effusion, atrioventricular conduction block and more recently, the presence of J-waves. MATERIALS AND METHODS: 48 HLA-B27 positive patient...

Ker, James

2011-01-01

9

HLA-B27 typing in the categorisation of uveitis in a HLA-B27 rich population  

OpenAIRE

AIMS—To determine whether HLA-B27 typing helps the clinician in the diagnostic examination of uveitis in a HLA-B27 rich population and also whether the clinical picture of HLA-B27 positive unilateral acute or recurrent anterior uveitis (AAU) is distinguishable from the idiopathic negative form.?METHODS—During a 3 year period 220 consecutive patients with undetermined uveitis at onset were examined in the Helsinki University Eye Clinic. HLA-B27 antigen was tested for 85% of the patient...

Huhtinen, M.; Karma, A.

2000-01-01

10

Isolated HLA-B27 associated Achilles tendinitis.  

OpenAIRE

The case of a 37 year old man with a longstanding HLA-B27 associated bilateral Achilles tendinitis without seronegative spondyloarthropathy is reported. This case suggests that heel enthesopathy may for a long time be the only clinical manifestation of the HLA-B27 associated disease process.

Olivieri, I.; Gemignani, G.; Gherardi, S.; Grassi, L.; Ciompi, M. L.

1987-01-01

11

Bartonella henselae associated uveitis and HLA-B27  

OpenAIRE

AIM—To investigate the frequency of HLA-B27 in patients with presumed Bartonella henselae associated uveitis and to describe the clinical characteristics of HLA-B27 positive patients with uveitis and presumed ocular bartonellosis (POB).?METHODS—The diagnosis of POB was considered in 19 patients with unexplained uveitis (except for the HLA-B27 association) and high positive IgG (titre ?1:900) and/or IgM (titre ?1:250) antibodies against B henselae. In addition to B henselae serology...

Kerkhoff, F. T.; Rothova, A.

2000-01-01

12

Coexisting HLA-B27 positive spondyloarthritis and polyarteritis nodosa.  

OpenAIRE

A 38 year old woman presented with widespread polyarteritis nodosa a few years after the onset of HLA-B27 positive spondyloarthritis. The concomitant coexistence of these two disorders suggests a possible association in this genetically susceptible subject.

Sattar, M. A.

1992-01-01

13

Multiple joint tuberculosis presenting as HLA-B27 disease  

OpenAIRE

A case of multifocal osteoarticular tuberculosis in a young Caucasian male is presented. The diagnostic difficulty, compounded by slow progression of the disorder and the presence of the tissue antigen HLA-B27, is discussed.

Hopkins, G. O.

1983-01-01

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Panuveíte em artrite indiferenciada HLA-B27 positiva / Panuveitis in HLA-B27 positive undifferentiated arthritis  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: Portuguese Abstract in portuguese Entre os vários tipos de inflamação ocular associados às doenças reumatológicas, a uveíte anterior é particularmente comum nas espondiloartropatias, em especial quando associada à presença do genótipo HLA-B27. Relatou-se o caso de um paciente com artrite indiferenciada HLA-B27 positivo, complicado c [...] om panuveíte e vasculite da retina, refratária ao tratamento imunossupressor tradicional, que obteve boa resposta clínica ao uso de anti-TNF-alfa. Abstract in english Among the several types of ocular inflammation associated to the rheumatic diseases, anterior uveitis is particularly common in the spondyloarthropathies, especially when associated to the presence of the HLA-B27 genotype. We report the case of HLA-B27 positive patient with undifferentiated arthriti [...] s, complicated with panuveitis and retinal vasculitis, that was refractory to the traditional imunossupressive treatment, and had a good clinical response with anti-TNF-alpha therapy.

Mário Sérgio Ferreira, Santos; Vitor, Cortizo; Cícero Ricardo Torres da, Costa; Ronnielly Melo, Tavares; Ricardo Eric Barros, Lopes.

2008-10-01

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Panuveíte em artrite indiferenciada HLA-B27 positiva Panuveitis in HLA-B27 positive undifferentiated arthritis  

Directory of Open Access Journals (Sweden)

Full Text Available Entre os vários tipos de inflamação ocular associados às doenças reumatológicas, a uveíte anterior é particularmente comum nas espondiloartropatias, em especial quando associada à presença do genótipo HLA-B27. Relatou-se o caso de um paciente com artrite indiferenciada HLA-B27 positivo, complicado com panuveíte e vasculite da retina, refratária ao tratamento imunossupressor tradicional, que obteve boa resposta clínica ao uso de anti-TNF-alfa.Among the several types of ocular inflammation associated to the rheumatic diseases, anterior uveitis is particularly common in the spondyloarthropathies, especially when associated to the presence of the HLA-B27 genotype. We report the case of HLA-B27 positive patient with undifferentiated arthritis, complicated with panuveitis and retinal vasculitis, that was refractory to the traditional imunossupressive treatment, and had a good clinical response with anti-TNF-alpha therapy.

Mário Sérgio Ferreira Santos

2008-10-01

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Invasion and persistence of Salmonella in human fibroblasts positive or negative for endogenous HLA B27  

OpenAIRE

OBJECTIVE—Analysis of the interaction of enteropathogenic bacteria with HLA B27 transfected murine fibroblasts showed a specific influence of HLA B27 on microbial invasiveness. This possible novel mechanism for the action of HLA B27 should be verified by using endogenous HLA B27 positive and negative human fibroblasts as a model for the direct interaction of arthritogenic bacteria and host cells.?METHODS—Fibroblasts were obtained from healthy donors positive or negative for HLA B27; cul...

Huppertz, H.; Heesemann, J.

1997-01-01

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HLA-B27 frequency in a group of patients with psoriatic arthritis Freqüência de HLA-B27 em uma amostra de pacientes com artrite psoriática  

OpenAIRE

BACKGROUND: HLA-B27 is associated with spondyloarthritis, a group of diseases that includes psoriatic arthritis. OBJECTIVES: To describe the HLA-B27 frequency in a group of Brazilian patients with psoriatic arthritis and correlate its presence or absence with their clinical manifestations. METHODS: Cross-sectional study with 44 psoriatic arthritis patients of a Rheumatology clinic. Demographic and social data were recorded, as were skin and joints clinical examination. HLA-B27 was tested. All...

Danilo Garcia Ruiz; Mário Newton Leitão de Azevedo; Omar Lupi

2012-01-01

18

Acute anterior uveitis, ankylosing spondylitis, back pain, and HLA-B27.  

OpenAIRE

One hundred and sixty-nine patients with acute anterior uveitis were studied for the presence of HLA-B27 tissue type, radiological evidence of ankylosing spondylitis, and a history of back pain. 60% were male; 45% were HLA-B27+. The male:female ratio in the HLA-B27+ group was the same as in the whole group. 24% had radiological evidence of ankylosing spondylitis, and, of these, 83% were HLA-B27+ while 17% were HLA-B27-. There was a definite correlation between the severity of the ankylosing s...

Beckingsale, A. B.; Davies, J.; Gibson, J. M.; Rosenthal, A. R.

1984-01-01

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HLA-B*27 typing by sequence specific amplification without DNA extraction.  

OpenAIRE

HLA-B27 appears to play a direct role in the pathogenesis of ankylosing spondylitis and almost all patients with this disease have HLA-B27. Therefore, a diagnosis of ankylosing spondylitis can virtually be excluded in the absence of HLA-B27. Many techniques have been used for HLA-B*27 typing. Of these, molecular methods are the most sensitive and specific but require extracted DNA as the testing material. A technique where HLA-B*27 is amplified directly from whole blood using sequence specifi...

Sayer, D. C.; Cassell, H. S.; Christiansen, F. T.

1999-01-01

20

HLA B27 allele types in homogeneous groups of juvenile idiopathic arthritis patients in Latvia  

OpenAIRE

Abstract Juvenile idiopathic arthritis (JIA) is a heterogeneous condition and therapeutic strategies vary in different JIA types. The routinely accepted practice to start with Sulphasalazine (SS) as the first line treatment in patients with HLA B27 positive JIA proves to be ineffective in a large proportion of children. Objective to investigate HLA B27 positive JIA patients clinical characteristics, determined HLA B27 allele types and their connection with antirheumatic treatment in homogenou...

Guseinova Dinara; Lazareva Arina; Sochnevs Arturs; Zavadska Dace; Eglite Jelena; Stanevicha Valda; Shantere Ruta; Gardovska Dace

2010-01-01

21

Cell-surface expression and immune receptor recognition of HLA-B27 homodimers.  

OpenAIRE

OBJECTIVE: HLA-B27 is capable of forming in vitro a heavy-chain homodimer structure lacking beta(2)-microglobulin. We undertook this study to ascertain if patients with spondylarthritis express beta(2)-microglobulin-free HLA-B27 heavy chains in the form of homodimers and receptors for HLA-B27 homodimers. METHODS: Expression of HLA-B27 heavy chains by mononuclear cells was analyzed by fluorescence-activated cell sorter staining, Western blotting with the monoclonal antibody HC-10, and 2-dimens...

Kollnberger, S.; Bird, L.; Sun, My; Retiere, C.; Braud, Vm; Mcmichael, A.; Bowness, P.

2002-01-01

22

Klebsiella 'modifying factor': binding studies with HLA-B27+ and B27- lymphocytes.  

OpenAIRE

On the basis that extracts of some klebsiella organisms bind selectively to the lymphocytes of HLA-B27+ individuals and induce the appearance of new antigens, attempts were made to detect the binding of klebsiella products to HLA-B27+ and B27- lymphocytes by a number of different techniques. Firstly, blocking of the binding of two different HLA-B27 specific monoclonal antibodies to HLA-B27+ lymphocytes has been examined following exposure of the lymphocytes to a cell-free culture filtrate fro...

Trapani, J. A.; Mckenzie, I. F.

1985-01-01

23

Ankylosing spondylitis and HLA-B27: restriction fragment length polymorphism and sequencing of an HLA-B27 allele from a patient with ankylosing spondylitis  

OpenAIRE

Two groups of patients with ankylosing spondylitis (AS) from England and Poland were examined for restriction fragment length polymorphisms (RFLPs) associated with the disease. No preferential association was found between the 9.2 kb PvuII fragment in HLA-B27 positive patients with AS compared with HLA-B27 healthy subjects as had been previously reported. In the English group, however, a 14 kb PvuII fragment was more common in HLA-B27 positive subjects with AS than in normal controls. Also 4....

1993-01-01

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The arthritis-associated HLA-B*27:05 allele forms more cell surface B27 dimer and free heavy chain ligands for KIR3DL2 than HLA-B*27:09  

OpenAIRE

OBJECTIVES: HLA-B*27:05 is associated with AS whereas HLA-B*27:09 is not associated. We hypothesized that different interactions with KIR immune receptors could contribute to the difference in disease association between HLA-B*27:05 and HLAB*27:09. Thus, the objective of this study was to compare the formation of ?2m-free heavy chain (FHC) including B27 dimers (B272) by HLA-B*27:05 and HLA-B*27:09 and their binding to KIR immunoreceptors. METHODS: We studied the formation of HLA-B*27:05 and ...

Cauli, A.; Shaw, J.; Giles, J.; Hatano, H.; Rysnik, O.; Payeli, S.; Mchugh, K.; Dessole, G.; Porru, G.; Desogus, E.; Fiedler, S.; Ho?lper, S.; Carette, A.; Blanco-gelaz, Ma; Vacca, A.

2013-01-01

25

HLA-B27 frequency in a group of patients with psoriatic arthritis / Freqüência de HLA-B27 em uma amostra de pacientes com artrite psoriática  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in portuguese FUNDAMENTOS: O HLA-B27 está associado às espondiloartrites, grupo de doenças que engloba, entre outras, a artrite psoriásica. OBJETIVOS: Descrever a freqüência de HLA-B27 em uma amostra de pacientes brasileiros com artrite psoriásica e correlacionar sua presença ou ausência com as manifestações clín [...] icas dos mesmos. MÉTODOS: Estudo transversal avaliando 44 pacientes com artrite psoriásica de um ambulatório de Reumatologia. A avaliação consistia em registro de informações demográficas e sociais, exame clínico da pele e das articulações e pesquisa de HLA-B27. Os dados gerados foram tratados por meio de estatística descritiva e comparativa em Software apropriado. Foram utilizados testes paramétricos e não-paramétricos com significância estatística de 5%. RESULTADOS: O HLA-B27 resultou negativo em 32 dos 44 pacientes estudados (72,7%). A maioria dos pacientes era do sexo masculino, da raça branca, procedente do Rio de Janeiro, portador de psoríase em placas e com idade média de 52,9 anos. Houve associação estatisticamente significativa entre o HLA-B27 positivo e o sexo masculino (p=0,004). O HLA-B27 negativo teve tendência à correlação com artrite de mãos e punhos (p=0,07). Houve correlação inversa significativa entre os valores do HLA e do teste de Schöber (p=0,02). CONCLUSÃO: Apesar do HLA-B27 ser negativo na maioria dos pacientes estudados, esteve significativamente associado ao sexo masculino e inversamente correlacionado ao teste de Schöber. Abstract in english BACKGROUND: HLA-B27 is associated with spondyloarthritis, a group of diseases that includes psoriatic arthritis. OBJECTIVES: To describe the HLA-B27 frequency in a group of Brazilian patients with psoriatic arthritis and correlate its presence or absence with their clinical manifestations. METHODS: [...] Cross-sectional study with 44 psoriatic arthritis patients of a Rheumatology clinic. Demographic and social data were recorded, as were skin and joints clinical examination. HLA-B27 was tested. All data were processed descriptively and comparatively by appropriate software. Parametric and non parametric tests were used with 5% statistical significance. RESULTS: HLA-B27 was negative in 32 of the 44 patients (72,7%). Most of them were male, Caucasian, living in Rio de Janeiro, with plaque type psoriasis and average age of 52,9 years. There was statistical significant correlation between positive HLA-B27 and male gender (p=0,004). Negative HLA-B27 had a tendency to correlate with hands and wrists arthritis (p=0,07). There was an inverse significant correlation between HLA values and Schöber's test (p=0,02). CONCLUSION: Although HLA-B27 is negative in most of patients, it is significantly associated to male gender and inversely correlated with Schöber's test.

Danilo Garcia, Ruiz; Mário Newton Leitão de, Azevedo; Omar, Lupi.

2012-12-01

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What is the role of HLA-B27 in spondyloarthropathies?  

Science.gov (United States)

HLA-B27 (Human Leukocyte Antigen-B27) accounts approximately for the one third of the overall genetic susceptibility to spondylorthropathies (SpAs). Up to 70 HLA-B27 subtypes have been reported all over the world with a decreasing north-south gradient of its frequency, which is reverse to that of endemic malaria. In an attempt to explain the possible role of HLA-B27 in SpAs pathogenesis, several theories have been suggested [1. Arthritogenic peptide, 2. Misfolding, 3. Cell surface HLA-B27 homodimers, 4. ?2m (?2-microglobulin) deposition, 5. ?2m-free/peptide free heavy chains of HLA-I, 6. Enhanced survival of some microbes in HLA-B27 cells, 7. ERAP1/ERAP2 (endoplasmic reticulum aminopeptidases 1 and 2) and Tapasin function, 8. ?2m over-expression] each of which contributes up to a point to our understanding of HLA-B27 subtypes' role in SpAs manifestation. However, reviewing all the suggested hypotheses it seems logical to pass from a puzzle of distinct hypotheses to a global theory. This review summarizes the current knowledge of HLA-B27 aiming to understand its potential use in clinical practice of SpAs diagnosis and to direct its future studies. PMID:21296192

Chatzikyriakidou, Anthoula; Voulgari, Paraskevi V; Drosos, Alexandros A

2011-06-01

27

HLA B27 and ankylosing spondylitis in the Israeli population.  

Science.gov (United States)

The distribution of 24 HLA antigens of the A and B loci was investigated in 38 Israeli ankylosing spondylitis (AS) patients of various ethnic origins. This was compared with the distribution in rheumatoid arthritis (RA) and osteoarthritis (OA), as well as in 456 controls representing the Jewish population and 260 controls representing the Arab population. Included in the study were Ashkenazi Jews and non-Ashkenazi Jews, as well as Moslem and Christian Arabs. The frequency of HLA B27 among AS patients (79 per cent) was significantly greater (P less than 10(-10)) than among the controls (three per cent). Ashkenazi Jews showed a higher relative risk than non-Ashkenazi Jews and Arabs. Six of the AS patients were offspring of consanguineous marriages, but this was not higher than expected and therefore no indication for rare recessive genes contributing to the disease could be demonstrated. This study confirms the association between AS and B27, and extends our knowledge to the heterogeneous population of Israel not previously investigated. A significant but weak association of B27 with RA was noted. No correlation of other HLA antigens with RA or OA was observed. PMID:266593

Brautbar, C; Porat, S; Nelken, D; Gabriel, K R; Cohen, T

1977-01-01

28

Functional Interaction of the Ankylosing Spondylitis-associated Endoplasmic Reticulum Aminopeptidase 1 Polymorphism and HLA-B27 in Vivo*  

OpenAIRE

The association of ERAP1 with ankylosing spondylitis (AS)1 among HLA-B27-positive individuals suggests that ERAP1 polymorphism may affect pathogenesis by altering peptide-dependent features of the HLA-B27 molecule. Comparisons of HLA-B*27:04-bound peptidomes from cells expressing different natural variants of ERAP1 revealed significant differences in the size, length, and amount of many ligands, as well as in HLA-B27 stability. Peptide analyses suggested that the mechanism of ERAP1/HLA-B27 in...

Garci?a-medel, Noel; Sanz-bravo, Alejandro; Nguyen, Dung; Galocha, Begon?a; Go?mez-molina, Patricia; Marti?n-esteban, Adria?n; Alvarez-navarro, Carlos; Castro, Jose? A. Lo?pez

2012-01-01

29

Clinical Features and Prognosis of HLA-B27 Positive and Negative Anterior Uveitis in a Korean Population  

OpenAIRE

Clinical features and prognosis of HLA-B27 positive anterior uveitis (AU) were assessed compared with HLA-B27 negative AU in a Korean population, based on the medical records of AU patients seen at a university hospital. Twenty-seven HLA-B27 negative, idiopathic AU patients (group I) and 55 HLA-B27 positive AU patients (group II) were studied. HLA-B27 positive group was further divided into 29 with associated systemic disease (seronegative spondyloarthropathy) (group IIA) and 26 without assoc...

Park, Sung Chul; Ham, Don-il

2009-01-01

30

The solvent-inaccessible Cys67 residue of HLA-B27 contributes to T cell recognition of HLA-B27/peptide complexes.  

OpenAIRE

Crystallographic studies have suggested that the cysteine at position 67 (Cys(67)) in the B pocket of the MHC molecule HLA-B*2705 is of importance for peptide binding, and biophysical studies have documented altered thermodynamic stability of the molecule when Cys(67) was mutated to serine (Ser(67)). In this study, we used HLA-B27.Cys(67) and HLA-B27.Ser(67) tetramers with defined T cell epitopes to determine the contribution of this polymorphic, solvent-inaccessible MHC residue to T cell rec...

Appel, H.; Kuon, W.; Kuhne, M.; Hu?lsmeyer, M.; Kollnberger, S.; Kuhlmann, S.; Weiss, E.; Zeitz, M.; Wucherpfennig, K.; Bowness, P.; Sieper, J.

2004-01-01

31

An update on the genetics of HLA-B27 associated acute anterior uveitis  

OpenAIRE

The discovery of the association of HLA B27 with spondyloarthropathy led to more questions than answers about the role of this gene in disease susceptibility. The realization that HLA B27 was not responsible for all of the genetic effect helped to lay a foundation for further investigation into the genetics of uveitis. Over several decades, genetic findings have provided clues to advance the understanding of mechanisms of uveitis and to catalyze new research on diagnostics, animal models and ...

Martin, Tammy M.; Rosenbaum, James T.

2011-01-01

32

Finnish HLA studies confirm the increased risk conferred by HLA?B27 homozygosity in ankylosing spondylitis  

OpenAIRE

OBJECTIVE: To determine the influence of HLA-B27 homozygosity and HLA-DRB1 alleles in the susceptibility to, and severity of, ankylosing spondylitis in a Finnish population. METHODS: 673 individuals from 261 families with ankylosing spondylitis were genotyped for HLA-DRB1 alleles and HLA-B27 heterozygosity/homozygosity. The frequencies of HLA-B27 homozygotes in probands from these families were compared with the expected number of HLA-B27 homozygotes in controls under Hardy-Weinberg equilibri...

Jaakkola, E.; Herzberg, I.; Laiho, K.; Barnardo, Mc; Pointon, Jj; Kauppi, M.; Kaarela, K.; Tuomilehto-wolf, E.; Tuomilehto, J.; Wordsworth, Bp; Brown, Ma

2005-01-01

33

HLA-B27 allele diversity in Indians: impact of ethnic origin and the caste system.  

Science.gov (United States)

HLA-B27 is a serological specificity which encompasses an increasing number of subtypes that show varied racial/ethnic prevalence in the world. Here, data from 5129 Indians (4500 population and caste; 629 tribal) is compiled from the literature. In addition, HLA-B27 subtyping of 58 positive individuals from Maharastra is presented. Analysis revealed an increased B27 antigen frequency among the north Indian groups (>5%) compared to the south Indian groups (population groups from Maharastra, but these differed in their distribution among the caste and tribal groups studied. The study showed that more extensive subtyping in other Indian caste groups will be necessary to resolve the evolutionary implications of HLA-B27 subtypes and their relationship to disease association in the Indian context. PMID:14725340

Shankarkumar, U

2003-01-01

34

Genética, HLA-B27 y espondilitis anquilosante: 40 años / Genetics of ankylosing spondylitis  

Scientific Electronic Library Online (English)

Full Text Available [...] Abstract in english Ankylosing spondylitis (AS) is a prototypical inflammatory disease of the locomotor system affecting axial skeleton. It is part of the general group of spondyloarthopathies (SpA). Its strong association with histocompatibility antigen HLA-B27 is known since 1973. However, HLA-B27 contribution to AS [...] genetic risk is approximately 16%. Therefore, other genes are necessarily involved in the pathogenesis of the disease. Genomic development and the possibility of making genome wide screening have contributed enormously to the study of the disease. In this paper, we describe the actual knowledge about AS genetic risk, which has contributed to understand the influence of HLA-B27 on the etiology and pathogenesis of the disease. We also intend to foresee how these findings will result in an improvement of patients’ quality of life.

Patricia, Castro-Santos; Miguel A, Gutiérrez; Roberto, Díaz-Peña.

1165-11-01

35

Interaction between ERAP1 and HLA-B27 in ankylosing spondylitis implicates peptide handling in the mechanism for HLA-B27 in disease susceptibility.  

Science.gov (United States)

Ankylosing spondylitis is a common form of inflammatory arthritis predominantly affecting the spine and pelvis that occurs in approximately 5 out of 1,000 adults of European descent. Here we report the identification of three variants in the RUNX3, LTBR-TNFRSF1A and IL12B regions convincingly associated with ankylosing spondylitis (P CARD9 that show strong association across all our datasets (P < 5 × 10(-6) overall, with support in each of the three datasets studied). We also show that polymorphisms of ERAP1, which encodes an endoplasmic reticulum aminopeptidase involved in peptide trimming before HLA class I presentation, only affect ankylosing spondylitis risk in HLA-B27-positive individuals. These findings provide strong evidence that HLA-B27 operates in ankylosing spondylitis through a mechanism involving aberrant processing of antigenic peptides. PMID:21743469

Evans, David M; Spencer, Chris C A; Pointon, Jennifer J; Su, Zhan; Harvey, David; Kochan, Grazyna; Oppermann, Udo; Opperman, Udo; Dilthey, Alexander; Pirinen, Matti; Stone, Millicent A; Appleton, Louise; Moutsianas, Loukas; Moutsianis, Loukas; Leslie, Stephen; Wordsworth, Tom; Kenna, Tony J; Karaderi, Tugce; Thomas, Gethin P; Ward, Michael M; Weisman, Michael H; Farrar, Claire; Bradbury, Linda A; Danoy, Patrick; Inman, Robert D; Maksymowych, Walter; Gladman, Dafna; Rahman, Proton; Morgan, Ann; Marzo-Ortega, Helena; Bowness, Paul; Gaffney, Karl; Gaston, J S Hill; Smith, Malcolm; Bruges-Armas, Jacome; Couto, Ana-Rita; Sorrentino, Rosa; Paladini, Fabiana; Ferreira, Manuel A; Xu, Huji; Liu, Yu; Jiang, Lei; Lopez-Larrea, Carlos; Díaz-Peña, Roberto; López-Vázquez, Antonio; Zayats, Tetyana; Band, Gavin; Bellenguez, Céline; Blackburn, Hannah; Blackwell, Jenefer M; Bramon, Elvira; Bumpstead, Suzannah J; Casas, Juan P; Corvin, Aiden; Craddock, Nicholas; Deloukas, Panos; Dronov, Serge; Duncanson, Audrey; Edkins, Sarah; Freeman, Colin; Gillman, Matthew; Gray, Emma; Gwilliam, Rhian; Hammond, Naomi; Hunt, Sarah E; Jankowski, Janusz; Jayakumar, Alagurevathi; Langford, Cordelia; Liddle, Jennifer; Markus, Hugh S; Mathew, Christopher G; McCann, Owen T; McCarthy, Mark I; Palmer, Colin N A; Peltonen, Leena; Plomin, Robert; Potter, Simon C; Rautanen, Anna; Ravindrarajah, Radhi; Ricketts, Michelle; Samani, Nilesh; Sawcer, Stephen J; Strange, Amy; Trembath, Richard C; Viswanathan, Ananth C; Waller, Matthew; Weston, Paul; Whittaker, Pamela; Widaa, Sara; Wood, Nicholas W; McVean, Gilean; Reveille, John D; Wordsworth, B Paul; Brown, Matthew A; Donnelly, Peter

2011-08-01

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Use of HLA-B27 tetramers to identify low-frequency antigen-specific T cells in Chlamydia-triggered reactive arthritis.  

OpenAIRE

Reports of the use of HLA-B27/peptide tetrameric complexes to study peptide-specific CD8+ T cells in HLA-B27+-related diseases are rare. To establish HLA-B27 tetramers we first compared the function of HLA-B27 tetramers with HLA-A2 tetramers by using viral epitopes. HLA-B27 and HLA-A2 tetramers loaded with immunodominant peptides from Epstein–Barr virus were generated with comparable yields and both molecules detected antigen-specific CD8+ T cells. The application of HLA-B27 tetramers in HL...

Appel, H.; Kuon, W.; Kuhne, M.; Wu, P.; Kuhlmann, S.; Kollnberger, S.; Thiel, A.; Bowness, P.; Sieper, J.

2004-01-01

37

Association of HLA B27 with benign clinical course of nephropathia epidemica caused by Puumala hantavirus.  

Science.gov (United States)

Both the severe course of nephropathia epidemica (NE) caused by Puumala hantavirus, and the fast progression of human immunodeficiency virus (HIV) disease, are associated with the HLA B8 DRB1*0301 haplotype. As HLA B27, on the contrary, is associated with the slow progression of HIV disease, we wanted to test whether the same is true for NE. Only six (8%) NE patients, half the figure expected, had the HLA B27 allele in 74 randomly selected hospital-treated patients. All six had a benign overall clinical course of NE; none had any severe complications, the severity of renal failure was also mild, and the treatment time at the hospital was half that needed for HLA B27- patients (P = 0.004). Patients who were HLA B27 had maximal blood leucocyte count > 10.000 x 10(9)/L (P = 0.020) more often, probably reflecting differences in immune response. Thus, similar HLA associations can be found in both HIV infection and NE caused by Puumala virus. PMID:9519867

Mustonen, J; Partanen, J; Kanerva, M; Pietilä, K; Vapalahti, O; Pasternack, A; Vaheri, A

1998-03-01

38

Evaluation of 278 hla-b27 positive patients suspected of seronegative spondyloarthropathies  

International Nuclear Information System (INIS)

To determine HLA-B27 prevalence in patients suspected of Seronegative spondyloarthropathy referred to the Transplantation Department of Blood Transfusion Organization, and to evaluate clinical findings among HLA-B27 positive patients. One thousand six hundred ten patients having clinical manifestation of seronegative SpAs were screened for HLA typing by serological methods from January 1997 to June 2002 at Transplantation Department of Blood Transfusion Organization, Ahwaz, Iran. Serologic-based HLA typing using Antigen-specific sera to determine a person's HLA type was performed. Among these patients, individuals found HLA-B27 positive were investigated regarding clinical findings, age, and sex distribution. In this study the frequency of HLA-B27 antigen was 17.26% (278 cases). The minimum age in males was 10 years and the maximum age in female was 70 years. Median age with seronegative SpAs findings (34.2% including 28.42% females, 71.57% males) was 20-30 years. Based on our results, the most frequent clinical manifestation, was peripheral joints arthritis (58.7%; 34.35% females, 65.65 % males). There were no association between any of the major clinical manifestations and age or sex distribution. These findings confirm the strong association of the HLA B27 allele with various types of spondyloarthritis and suggests that HLA typing would help in the diagnosis of seronagative SpAs, specially ankylosing spondylitis with indeterminate clinical presentation and also in rminate clinical presentation and also in identifying at risk family members. (author)

39

Autoantibodies to HLA B27 in the sera of HLA B27 patients with ankylosing spondylitis and Reiter's syndrome. Molecular mimicry with Klebsiella pneumoniae as potential mechanism of autoimmune disease  

OpenAIRE

Ankylosing spondylitis (AS) and Reiter's syndrome (RS) both show a strong correlation with the HLA B27 haplotype. We studied whether sharing of homologous amino acid sequences in the HLA B27 antigen with an invading microbe might occur, and if so, what is the biological significance of such homology. In a computer search of the Dayhoff data bank, we found a homology of six consecutive amino acids between HLA B27.1 antigen residues 72-77 and Klebsiella pneumoniae nitrogenase residues 188-193. ...

1987-01-01

40

Association of HLA-B27 and ERAP1 with ankylosing spondylitis susceptibility in Beijing Han Chinese.  

Science.gov (United States)

This study investigated the genetic polymorphisms of HLA-B27, together with polymorphisms on endoplasmic reticulum aminopeptidase 1 (ERAP1), and susceptibility for ankylosing spondylitis (AS) in the Beijing Han population. A case-control study was carried out for 602 AS patient samples and 619 matched controls of Han Chinese. HLA-B27 genotyping was performed by polymerase chain reaction-sequence specific primers (PCR-SSP), and four ERAP1 SNPs (rs27037, rs27980, rs27582, and rs27434) were selected and genotyped on the Sequenom iPlex platform (Sequenom, San Diego, CA). Association analysis was performed using the likelihood ratio ?(2) test. This study identified four HLA-B27 alleles in Beijing Han AS patients, B*27:02, B*27:04, B*27:05, and B*27:07, of which B*27:05 was the most significant geographical different subtype among AS patients in Chinese. Our results confirmed that HLA-B27 was strongly associated with AS (P=1.9 × 10(-150) ), and the most strongly associated alleles were B*27:04, B*27:05, and B*27:02. Our study also confirmed a weak association between ERAP1 (rs27434) and AS. We also observed that for HLA-B*27:02 and HLA-B*27:04 positive AS patients, rs27434 and rs27582 were associated with AS. In contrast, for HLA-B27-negative and HLA-B*27:05-positive AS patients, this association was not observed. This is the first study to show that both B27 and ERAP1 are AS genetic susceptibility genes in Beijing Han. Interactions between ERAP1 and HLA-B*27:02 and B*27:04 may play an important role in the AS pathogenesis. PMID:24666027

Zhang, Z; Dai, D; Yu, K; Yuan, F; Jin, J; Ding, L; Hao, Y; Liang, F; Liu, N; Zhao, X; Long, J; Xi, Y; Sun, Y-Y

2014-05-01

41

Cutting edge: HLA-B27 can form a novel beta 2-microglobulin-free heavy chain homodimer structure.  

OpenAIRE

HLA-B27 has a striking association with inflammatory arthritis. We show that free HLA-B27 heavy chains can form a disulfide-bonded homodimer, dependent on residue Cys67 in their extracellular alpha 1 domain. Despite the absence of beta 2-microglobulin, HLA-B27 heavy chain homodimers (termed HC-B27) were stabilized by a known peptide epitope. HC-B27 complexes were recognized by the conformation-specific Ab W6/32, but not the ME1 Ab. Surface labeling and immunoprecipitation demonstrated the pre...

Allen, Rl; O Callaghan, Ca; Mcmichael, Aj; Bowness, P.

1999-01-01

42

Structural analysis of an HLA-B27 functional variant: identification of residues that contribute to the specificity of recognition by cytolytic T lymphocytes.  

OpenAIRE

The structure of a variant HLA-B27 antigen, B27.2, that is distinguished from the HLA-B27.1 and HLA-B27.3 subgroups by specific cytolytic T lymphocytes has been established by comparative peptide mapping and sequence analysis. There are only three amino acid substitutions between B27.1 and B27.2: aspartate-77, threonine-80, and leucine-81 in HLA-B27.1 are changed to asparagine-77, isoleucine-80, and alanine-81 in HLA-B27.2. These changes account for their single charge difference detectable b...

Vega, M. A.; Ezquerra, A.; Rojo, S.; Aparicio, P.; Bragado, R.; Lo?pez Castro, J. A.

1985-01-01

43

A prospective study of patients with spondyloarthropathy with special reference to HLA-B27 and to gut histology.  

Science.gov (United States)

Ileocolonoscopy was performed on 357 patients meeting the European Spondylarthropathy Study Group criteria for spondyloarthropathy. HLA loci A, B and C were determined in all patients; HLA-B27 was detected in 196 and was absent in the 161 remaining patients. A number of clinical, laboratory and radiological variables were determined before ileocolonoscopy and compared between the HLA-B27+ and HLA-B27- patients. The HLA-B27+ patients were mainly men, with significantly more family members with spondyloarthropathies. Clinical evidence of tendinitis and uveitis was more frequently found in these patients. Like several authors, we found that these patients were more severely affected since they presented more severe radiological involvement of the sacroiliac joints. Syndesmophytes, bamboo spine and erosive joint lesions were more frequent in this group, the hip involvement being of the concentric type. In HLA-B27+ patients ankylosing spondylitis was more prevalent, while in the B27- patients enterogenic, urogenital or undifferentiated spondyloarthropathy was diagnosed. The HLA-B27- patients experienced more episodes of diarrhea, and Crohn-like inflammatory gut lesions were more frequently seen on ileocolonoscopy. In this group a number of patients probably had a form of subclinical Crohn's disease of which the locomotor symptoms were the only clinical expression. PMID:8230018

Mielants, H; Veys, E M; Goemaere, S; Cuvelier, C; De Vos, M

1993-08-01

44

Clinical characteristics of patients with spondyloarthritides and HLA-B27 positive antigen.  

Science.gov (United States)

The aim of this study was to present our experiences in diagnosing spondyloarthritides (SpA), and to list the most common clinical features of HLA-B27 positive patients. The study included 65 HLA-B27 positive patients with confirmed diagnosis of ankylosing spondylitis (AS) and psoriatic arthritis (PsA) who were analyzed between 2009 and 2010 in Clinic of Internal Medicine in Osijek. The diagnosis of seronegative spondyloarthritides was based on the ASAS (Assessment in AS Working Group) classification criteria for axial and then supplemented with ASAS criteria for peripheral SpA and was confirmed by radiological techniques. For diagnosing the ankylosing spondylitis (AS), there have been applied the modified New York criteria. Radiological criteria for definite sacroiliitis according to the modified New York criteria is bilateral sacroiliitis, grade 2-4 (> or = 2) or unilateral sacroiliitis, grade 3-4. For diagnosing the psoriatic arthritis (PsA), there were used CASPAR diagnostic criteria. Other features of SpA are defined within the existing classification criteria. HLA-B27 antigen was determined by direct immune-fluorescence technique using flow cytometer. The average age of patients was 50.34 years, of whom 27 female (41.53%), 38 male (58.46%). Duration of illness was 15.79 years on average. With 75.38% of patients, there had been determined the diagnosis of AS; 24.62% of patients had the diagnosis of PsA. The most common clinical characteristics that patients had were: inflammatory back pain (pain Inflammation along the lumbosacral spine), peripheral arthritis, intermittent pain in the gluteus, sacroiliitis, enthesitis, uveitis, dactilitis. PMID:21755709

Glasnovi?, Marija; Bosnjak, Ivica; Sram, Miroslav; Vranjes, Zeljko; Vcev, Aleksandar; Dobrosevi?, Blazenka; Petricevi?, Jasminka Sinci?; Horvati?, Elizabeta; Orki?, Zelimir; Tadzi?, Refmir; Soldo, Anamarija; Sisljagi?, Dina; Dinjar, Kristijan

2011-06-01

45

Bilateral macular thickening in mild unilateral anterior uveitis: is HLA-B27 involved?  

OpenAIRE

Abstract Background Macular thickening (MT) without clinically recognized macular edema has been described in anterior uveitis (AU). Although fellow-eyes of patients have been used as controls in several studies, little is known about macular thickness in these eyes. We studied the rate and extent of MT in both AU-affected and quiescent fellow-eyes of phakic AU patients with good visual acuity (VA). We also assessed macular thickness related to HLA-B27 presence and to recurrence, since these ...

Wexler Alexandra; Sand Trond; Elsås Tor B

2012-01-01

46

Endogenous Processing and Presentation of T-cell Epitopes from Chlamydia trachomatis with Relevance in HLA-B27-associated Reactive Arthritis*  

OpenAIRE

Chlamydia trachomatis triggers reactive arthritis, a spondyloarthropathy linked to the human major histocompatibility complex molecule HLA-B27, through an unknown mechanism that might involve molecular mimicry between chlamydial and self-derived HLA-B27 ligands. Chlamydia-specific CD8+ T-cells are found in reactive arthritis patients, but the immunogenic epitopes are unknown. A previous screening of the chlamydial genome for putative HLA-B27 ligands predicted multiple peptides that were recog...

Cragnolini, Juan J.; Garci?a-medel, Noel; Lo?pez Castro, Jose? A.

2009-01-01

47

Multiple sclerosis and HLA-B27 negative sacroiliitis in a Crohn?s disease patient  

Directory of Open Access Journals (Sweden)

Full Text Available SUMMARY A relationship between inflammatory bowel disease and MS is supported by a higher than expected coexistence of these diseases among families and individuals. A 32 year-old male with Crohn?s disease of the terminal ileum diagnosed 4 years earlier and HLA-B27 bilateral sacroiliitis diagnosed two years earlier, was admitted to our hospital because of an acute episode of blurred vision. In addition the patient complained of urine incontinence. Before this admission the patient had been elsewhere administered three doses of Remicade and 16mg of methylprednisolone p.os. During admission the diagnosis of multiple sclerosis was made (MRI and IgG Index and Remicade was discontinued. The patient was started on therapy with interferon-beta for MS, oxybutynin hydrochloride (10mg/day for urine incontinence, prednizolone (10mg/day, methotrexate (10mg/week and azathioprine (100mg/day for Crohn?s disease and is now in excellent clinical status. To the best of our knowledge this is one of the very rare cases of Crohn?s disease with HLA-B27 negative sacroiliitis preceding multiple sclerosis diagnosis. Key words: Crohn?s disease, inflammatory bowel disease, ulcerative colitis, multiple sclerosis, Remicade

K.H. Katsanos, N. Tzambouras, E.V. Tsianos

2007-03-01

48

Usage of Conventional PCR Technology for the Detection of HLA-B27 Allele: A Significant Molecular Marker of Ankylosing Spondylitis  

OpenAIRE

Ankylosing spondylitis is a chronic inflammatory disease that has been linked to the human leukocyte antigen class I allele HLA-B27. More than 90 % of patients with ankylosing spondylitis possess the HLA-B27 allele, but only 1 % of people with HLA-B27 develop the disease. Ankylosing spondylitis predominately affects young males. The present study was planned to find out the involvement of HLA-B27 specific allele in relation to age and sex in symptomatic suspected patients of ankylosing spon...

Sharma, Narotam; Sharma, Veena; Masood, Tariq; Nautiyal, Satish Chandra; Sailwal, Shivani; Singh, Rajesh K.; Kushwaha, Rajeev K.; Singh, R. K.

2012-01-01

49

Solid-phase Edman degradation analysis of HLA-B27 molecules in healthy individuals and ankylosing spondylitis patiens.  

Czech Academy of Sciences Publication Activity Database

Rio de Janeiro : ImmunoRIO, 2007. ---. [Immuno RIO 2007, International Congrass of Immunology /13./. 21.08.2007-25.08.2007, Rio de Janeiro] Institutional research plan: CEZ:AV0Z50520514; CEZ:AV0Z50520701 Keywords : HLA-B27 * ankylosing spondylitis * Edman degradation analysis Subject RIV: EB - Genetics ; Molecular Biology

Ivašková, E.; Flieger, M.; Stod?lková, E.; Sedlá?ková, M.; Man, P.; Votruba, J.; Pohl, L.; ?apková, Jana; Ivanyi, P.

50

Comparación de la eficacia de la prednisolona y la rimexolona en el tratamiento de iridociclitis aguda en pacientes HLA-B27 positivos / Efficacy of prednisolone and rimexolone in HLA-B27 positive patients with acute anterior uveitis  

Scientific Electronic Library Online (English)

Full Text Available Objetivo: Comparar la eficacia y seguridad de las suspensiones oftálmicas de acetato de prednisolona al 1% y de rimexolona al 1%, en el tratamiento de la uveítis anterior aguda (UAA) en pacientes HLA B27+. Material y métodos: Se seleccionaron al azar 68 pacientes con UAA HLA -B27+ para tratamiento c [...] on acetato de prednisolona al 1% o rimexolona al 1%. En todos los pacientes la inflamación en cámara anterior era leve a moderada. La presión intraocular (PIO) y el grado de inflamación fueron evaluados semanalmentre durante seis semanas. Fue un estudio clínico prospectivo, aleatorio y doble ciego. Resultados: Al cuantificar las células en cámara anterior no se encontraron diferencias estadísticamente significativas entre uno y otro grupos. En el grupo de rimexolona el flare disminuyó desde la primera semana. En los grupos la presión intraocular se elevó con respecto a la basal desde la primera semana, la variación fue más significativa en el grupo de rimexolona. La PIO final fue menor en el grupo de rimexolona, siendo esta diferencia estadísticamente significativa . Conclusión: Para el tratamiento de la UAA HLA -B27+, leve a moderada, la rimexolona al 1% y la prednisolona al 1% tiene una eficiencia similar. En este estudio las variaciones de presión intraocular en los dos grupos no fueron clínicamente significativas. Abstract in english Purpose: To compare the efficacy and safety of prednisolone acetate 1 % vs. rimexolone 1 % ophthalmic suspension in the treatment of acute anterior uveitis (AAU) in HLA-B27+ patients. Methods: Sixty-eight AAU HLA-B27+ patients were randomly selected for treatment with prednisolone acetate 1% or Rime [...] xolone 1%. All patients showed mild to moderate anterior chamber inflammation. This was a prospective, randomized, double blind, clinical trial. Results: There was no statistically significant difference between both groups when anterior chamber cells were measured. In the rimexolone group, flare diminished since the first week. In both groups the intraocular pressure (IOP) raised since the first week; the increase washighly significant in the rimexolone group. Final intraocular pressure was higher in the prednisolone group. Conclusion: Rimexolone 1 % is as effective as prednisolone acetate 1% in the treatment of mild to moderate AAU HLA B27+. IOP increased in both groups, but this variation was not clinically significant.

Lourdes, Arellanes-García; Gonzalo, Padilla-Aguilar; Patricia, Navarro-López; Cynthia, Espinoza-Martínez.

2005-10-01

51

Antibody activity in ankylosing spondylitis sera to two sites on HLA B27.1 at the MHC groove region (within sequence 65-85), and to a Klebsiella pneumoniae nitrogenase reductase peptide (within sequence 181-199)  

OpenAIRE

74 overlapping peptides of varying lengths from Klebsiella pneumoniae nitrogenase reductase (residues 181-199) and from the HLA B27.1 molecule (residues 65-85) were synthesized and tested by ELISA against sera from HLA B27+ ankylosing spondylitis (AS) patients, and sera from HLA B27+ and HLA B27- healthy first-degree relatives. Antibody activity in AS sera to Klebsiella peptides of four to eight amino acids was maximal with the peptide NSRQTDR. Activity to HLA B27 peptides was maximal with th...

1990-01-01

52

Effect of HLA-B*27 and its subtypes on clinical manifestations and severity of ankylosing spondylitis in Iranian patients.  

Science.gov (United States)

The aim of this study was to assess the role of HLA-B*27 and it's subtypes in determining severity and clinical manifestations of ankylosing spondylitis (AS).A total of 163 AS patients were assessed for clinical manifestations and severity using structured questionnaires. HLA-B*27 screening and B*27 sub-typing were performed by PCR.One hundred twenty two patients (74.8%) were B*27 positive. The male to female ratio, peripheral arthritis, steroid use, intense dorsal kyphosis and decrease of cervical slope had a significantly higher frequency in B*27 positive patients compared to B*27 negative ones (p=0.01, 0.001, 0.01, 0.04 and 0.04, respectively). However, the age of diagnosis was significantly lower in B*27 positive patients (p=0.005). Trend in uveitis and some severity markers including: BASMI and ASQoL were toward higher values in B*27 positive group with no significant difference. After controlling confounding variables, significant relationship was found only between B*27 and BASMI (p=0.01). B*27 subtypes in patients were included B*2705: 48.4%, B*2702: 42.6%, B*2704: 5.7% and B*2707: 3.3%. No significant differences were seen for severity markers and clinical manifestations between subtypes; although trend toward lower values of severity markers, less intense dorsal kyphosis and less decrease of cervical slope were observed in B*2704 and B*2707 versus other polymorphisms.Clinical features and severity of AS is influenced by HLA-B*27. Trend toward higher severity markers in B*2705 and B*2702 versus other polymorphisms might be subject of interest for evaluation in other ethnicities with concentration to other novel susceptibility genes co-inherited in each B*27 subtype. PMID:23996708

Fallahi, Sasan; Mahmoudi, Mahdi; Nicknam, Mohammad Hossein; Gharibdoost, Farhad; Farhadi, Elham; Saei, Azad; Nourijelyani, Keramat; Ahmadzadeh, Nooshin; Jamshidi, Ahmad Reza

2013-12-01

53

Structural Basis for T Cell Alloreactivity among Three HLA-B14 and HLA-B27 Antigens*  

OpenAIRE

The existence of cytotoxic T cells (CTL) cross-reacting with the human major histocompatibility antigens HLA-B14 and HLA-B27 suggests that their alloreactivity could be due to presentation of shared peptides in similar binding modes by these molecules. We therefore determined the crystal structures of the subtypes HLA-B*1402, HLA-B*2705, and HLA-B*2709 in complex with a proven self-ligand, pCatA (peptide with the sequence IRAAPPPLF derived from cathepsin A (residues 2–10)), and of HLA-B*140...

Kumar, Pravin; Vahedi-faridi, Ardeschir; Saenger, Wolfram; Merino, Elena; Lo?pez Castro, Jose? A.; Uchanska-ziegler, Barbara; Ziegler, Andreas

2009-01-01

54

Persistence of HLA-B27 cross-reactive bacteria in bowel flora of patients with ankylosing spondylitis.  

OpenAIRE

Previous studies have shown that antisera raised in rabbits to certain enteric bacteria (cross-reactive bacteria) are capable of specifically lysing in a 51chromium-release lymphocytotoxicity test the lymphocytes of HLA-B27-positive (B27+) patients with ankylosing spondylitis (AS). This study investigated the clinical relevance of this finding by ascertaining whether Escherichia coli isolated from the rectal swabs of 20 B27+ AS patients (B27+ AS+) and 46 controls (35 B27- AS- and 11 B27+ AS-)...

Prendergast, J. K.; Mcguigan, L. E.; Geczy, A. F.; Kwong, T. S.; Edmonds, J. P.

1984-01-01

55

Molecular analysis of HLA-B27 haplotypes in Caucasoids. Frequencies of B27-Cw in Jewish and Spanish populations.  

Science.gov (United States)

PCR in combination with SSO probes was used to analyze the polymorphism in exons 2 and 3 of HLA-B27 subtypes and HLA-C-related alleles in two genetically distant Caucasian groups: Spanish and Jewish populations. AS patients and healthy B27 donors from both populations were analyzed in order to ascertain B27-Cw haplotypes. Three different ancestral haplotypes were found to be represented in both populations: B*2705/Cw*0102, B*2705/Cw*02022, and B*2702/Cw*02022. The B*2705 (92.5%) was the most frequent allele found in the Spanish population, carried by B*2705/Cw*0102 (60.9%) and B2705/Cw*02022 (30.4%) haplotypes. In contrast, B*2702 (59.4%) was the most prevalent allele found in the Jewish population and was carried by the B*2702/Cw*02022 (63.3%) haplotype. No different allelic and haplotypic distributions were among healthy and AS patients in either Spanish or Jewish populations. The differences found in the distribution of B27 haplotypes among Spanish and Jewish Caucasian populations are consistent with the genetic distance of these ethnic groups. When the Jewish population was subdivided into Ashkenazi (A) and non-Ashkenazi (NA) groups, no significant differences were observed in the distribution of B*2702/Cw*02022 haplotype. Minor differences were observed in the underrepresented B*2705 haplotypes. The present results reflect the ancestral affinities of A and NA Jewish populations. A possible HLA-B27 evolutive pathway in Caucasians is proposed according to the data available for the B27/Cw ancestral haplotypes in Spanish and Jewish groups. PMID:7860357

González-Roces, S; Brautbar, C; Peña, M; Dominguez, O; Coto, E; Alvarez, V; Segal, R; López-Larrea, C

1994-10-01

56

LC MALDI-TOF MS/MS and LC ESI FTMS analyses of HLA-B27 associated peptides isolated from peripheral blood cells.  

Czech Academy of Sciences Publication Activity Database

Ro?. 116, - (2008), s. 79-85. ISSN 0165-2478 R&D Projects: GA MZd 1A8631 Institutional research plan: CEZ:AV0Z50200510; CEZ:AV0Z50520514 Keywords : hla -b27 * healthy blood donors Subject RIV: EE - Microbiology, Virology Impact factor: 2.858, year: 2008

Stod?lková, Eva; Novák, Petr; Deininger, S.-O.; Man, Petr; ?apková, Jana; Kavan, Daniel; Ivašková, E.; Flieger, Miroslav

2008-01-01

57

Complete sequence of HLA-B27 cDNA identified through the characterization of structural markers unique to the HLA-A, -B, and -C allelic series  

Energy Technology Data Exchange (ETDEWEB)

Antigen HLA-B27 is a high-risk genetic factor with respect to a group of rheumatoid disorders, especially ankylosing spondylitis. A cDNA library was constructed from an autozygous B-cell line expressing HLA-B27, HLA-Cw1, and the previously cloned HLA-A2 antigen. Clones detected with an HLA probe were isolated and sorted into homology groups by differential hybridization and restriction maps. Nucleotide sequencing allowed the unambiguous assignment of cDNAs to HLA-A, -B, and -C loci. The HLA-B27 mRNA has the structure features and the codon variability typical of an HLA class I transcript but it specifies two uncommon amino acid replacements: a cysteine in position 67 and a serine in position 131. The latter substitution may have functional consequences, because it occurs in a conserved region and at a position invariably occupied by a species-specific arginine in humans and lysine in mice. The availability of the complete sequence of HLA-B27 and of the partial sequence of HLA-Cw1 allows the recognition of locus-specific sequence markers, particularly, but not exclusively, in the transmembrane and cytoplasmic domains.

Szoets, H.; Reithmueller, G.; Weiss, E.; Meo, T.

1986-03-01

58

Polymorphisms of HLA-A, -B, -Cw and DRB1 antigens in Moroccan patients with ankylosing spondylitis and a comparison of clinical features with frequencies of HLA-B*27.  

Science.gov (United States)

Ankylosing spondylitis (AS) is very often associated with human leukocyte antigen (HLA), particularly HLA-B*27. However, the strength of this association and clinical features may vary in different ethnic groups. Our study aims to assess the distribution of HLA-A, -B, -Cw and DRB1 alleles in Moroccan patients with AS and to compare the clinical features of AS and the frequencies of HLA-B27 in patients from Morocco with other series. Seventy-five patients diagnosed with AS and assessed for clinical manifestations were selected and compared to 100 healthy controls. HLA class I and II antigens were typed by polymerase chain reaction sequence-specific oligonucleotide. HLA-B27 subtypes were studied by polymerase chain reaction amplification with sequence-specific primers. HLA-B27 was found in 64% of patients. It was positively associated with younger age at disease onset, family history, and uveitis while it had a negative association with late onset. Six B*27 subtypes were identified in the AS group. HLA-B*2705 and B*2702 were the most common observed subtypes. Among other HLA genes, a significant increase in the prevalence of HLA-Cw*02 and HLA-DRB*15 was found in AS patients. HLA-B27 is involved in the predisposition of AS in the Moroccan population. HLA-B*2705 and B*2702 were the predominant subtypes supporting previous reports in Caucasian spondyloarthropathies. Other HLA genes, HLA-Cw*02 and HLA-DRB1*15, seem to confer predisposing effect to the disease. However, the lower frequency of HLA-B27 compared to the literature in our study suggests the existence of different genetic and/or environmental factors in Morocco. PMID:25626601

El Mouraghi, I; Ouarour, A; Ghozlani, I; Collantes, E; Solana, R; El Maghraoui, A

2015-02-01

59

Transgenic Rat Models of Huntington's Disease.  

Science.gov (United States)

Several animal models for Huntington's disease (HD) have been created in order to investigate mechanisms of disease, and to evaluate the potency of novel therapies. Here, we describe the characteristics of the two transgenic rat models: transgenic rat model of HD (fragment model) and the Bacterial Artificial Chromosome HD model (full-length model). We discuss their genetic, behavioural, neuropathological and neurophysiological features. PMID:24013873

Carreira, João Casaca; Jahanshahi, Ali; Zeef, Dagmar; Kocabicak, Ersoy; Vlamings, Rinske; von Hörsten, Stephan; Temel, Yasin

2013-09-01

60

Peptide-binding motifs associated with MHC molecules common in Chinese rhesus macaques are analogous to those of human HLA supertypes and include HLA-B27-like alleles  

DEFF Research Database (Denmark)

Chinese rhesus macaques are of particular interest in simian immunodeficiency virus/human immunodeficiency virus (SIV/HIV) research as these animals have prolonged kinetics of disease progression to acquired immunodeficiency syndrome (AIDS), compared to their Indian counterparts, suggesting that they may be a better model for HIV. Nevertheless, the specific mechanism(s) accounting for these kinetics remains unclear. The study of major histocompatibility complex (MHC) molecules, including their MHC/peptide-binding motifs, provides valuable information for measuring cellular immune responses and deciphering outcomes of infection and vaccine efficacy. In this study, we have provided detailed characterization of six prevalent Chinese rhesus macaque MHC class I alleles, yielding a combined phenotypic frequency of 29 %. The peptide-binding specificity of two of these alleles, Mamu-A2*01:02 and Mamu-B*010:01, as well as the previously characterized allele Mamu-B*003:01 (and Indian rhesus Mamu-B*003:01), was found tobe analogous to that of alleles in the HLA-B27 supertype family. Specific alleles in the HLA-B27 supertype family, including HLA-B*27:05, have been associated with long-term nonprogression to AIDS in humans. All six alleles characterized in the present study were found to have specificities analogous to HLA supertype alleles. These data contribute to the concept that Chinese rhesus macaque MHC immunogenetics is more similar to HLA than their Indian rhesus macaque counterparts and thereby warrants further studies to decipher the role of these alleles in the context of SIV infection.

Mothé, Bianca R.; Southwood, Scott

2013-01-01

61

HLA Class I-Mediated Control of HIV-1 in the Japanese Population, in Which the Protective HLA-B*57 and HLA-B*27 Alleles Are Absent  

OpenAIRE

We investigated the effect of HLA class I alleles on clinical parameters for HIV-1 disease progression in the Japanese population, where two strongly protective alleles, HLA-B*57 and HLA-B*27, are virtually nonexistent. HLA-B alleles showed a dominant role, primarily through HLA-B*67:01 and the HLA-B*52:01-C*12:02 haplotype. Neither a rare-allele nor a heterozygote advantage was found, suggesting that the effect of HLA alleles in the Japanese population is either different from those observed...

Naruto, Takuya; Gatanaga, Hiroyuki; Nelson, George; Sakai, Keiko; Carrington, Mary; Oka, Shinichi; Takiguchi, Masafumi

2012-01-01

62

Lessons Learned from the Transgenic Huntington's Disease Rats  

Science.gov (United States)

Huntington's disease (HD) is a fatal inherited disorder leading to selective neurodegeneration and neuropsychiatric symptoms. Currently, there is no treatment to slow down or to stop the disease. There is also no therapy to effectively reduce the symptoms. In the investigation of novel therapies, different animal models of Huntington's disease, varying from insects to nonhuman primates, have been created and used. Few years ago, the first transgenic rat model of HD, carrying a truncated huntingtin cDNA fragment with 51 CAG repeats under control of the native rat huntingtin promoter, was introduced. We have been using this animal model in our research and review here our experience with the behavioural, neurophysiological, and histopathological phenotype of the transgenic Huntington's disease rats with relevant literature. PMID:22852099

Vlamings, Rinske; Zeef, Dagmar H.; Janssen, Marcus L. F.; Oosterloo, Mayke; Schaper, Frederic; Jahanshahi, Ali; Temel, Yasin

2012-01-01

63

Immunodeficient Parameters in the HIV-1 Transgenic Rat Model  

Directory of Open Access Journals (Sweden)

Full Text Available Recently an HIV-1 transgenic (HIV-1Tg rat model was created that carries a gag-pol-deleted HIV-1 genome under the control of the HIV-1 viral promoter. However, other viral proteins are expressed in most organs and tissues, and are found in the circulating blood. Since HIV-1 targets the immune system in humans, we examined two immunological parameters, leukocyte-endothelial adhesion (LEA and inflammatory cytokine production, in 5 mo old HIV-1Tg rats to identify immune functions that may be impaired even before the onset of symptoms of HIV-1 infection. We administered a single injection (i.p. of the bacterial endotoxin, lipopolysaccharide (LPS, 250 ug/kg, to 5 mo old HIV-1Tg rats, age-matched transgenic control (Tg rats, and F344/NHsd (F344 control background strain rats. LPS induced an LEA response in both the Tg control and F344 control animals. However, in the HIV-1Tg rats, there was no LEA response to LPS. Following LPS administration, there was significantly greater serum levels of TNF-? and IL-1?, two pro-inflammatory cytokines, in the HIV-1Tg rats compared to the control animals. In contrast, the serum level of IL-10, an anti-inflammatory cytokine, was comparable in the HIV-1Tg, Tg control, and F344 control rats. Our data show that, in the HIV-1Tg rat, there is a negative correlation between the LEA response and the induction of pro-inflammatory cytokines in response to bacterial endotoxin. These findings suggest that the persistent presence of viral proteins may be, at least, partially responsible for the immunodeficiency that occurs with HIV-1 infection, and that the HIV-1Tg rat could be a valid rodent model in which to study various aspects of HIV-1 infection.

Sulie L. Chang

2007-01-01

64

A transgenic rat with ubiquitous expression of firefly luciferase gene  

Science.gov (United States)

In vivo imaging strategies provide cellular and molecular events in real time that helps us to understand biological processes in living animals. The development of molecular tags such as green fluorescent proteins and luciferase from the firefly Photinus pyralis has lead to a revolution in the visualization of complex biochemical processes. We developed a novel inbred transgenic rat strain containing firefly luciferase based on the transgenic (Tg) technique in rats. This Tg rat expressed the luciferase gene ubiquitously under control of the ROSA26 promoter. Cellular immune responsiveness against the luciferase protein was evaluated using conventional skin grafting and resulted in the long-term acceptance of Tg rat skin on wild-type rats. Strikingly, organ transplant with heart and small bowel demonstrated organ viability and graft survival, suggesting that cells from luciferase-Tg are transplantable to track their fate. Taking advantage of the less immunogenic luciferase, we also tested the role of hepatocyte-infusion in a liver injury model, and bone marrow-derived cells in a skin defect model. Employed in conjunction with modern advances in optical imaging, this luciferase-Tg rat system provides an innovative animal tool and a new means of facilitating biomedical research such as in the case of regeneration medicine.

Hakamata, Yoji; Murakami, Takashi; Kobayashi, Eiji

2006-02-01

65

Lessons Learned from the Transgenic Huntington's Disease Rats  

OpenAIRE

Huntington's disease (HD) is a fatal inherited disorder leading to selective neurodegeneration and neuropsychiatric symptoms. Currently, there is no treatment to slow down or to stop the disease. There is also no therapy to effectively reduce the symptoms. In the investigation of novel therapies, different animal models of Huntington's disease, varying from insects to nonhuman primates, have been created and used. Few years ago, the first transgenic rat model of HD, carrying a truncated hunti...

Rinske Vlamings; Zeef, Dagmar H.; Janssen, Marcus L. F.; Mayke Oosterloo; Frederic Schaper; Ali Jahanshahi; Yasin Temel

2012-01-01

66

Memory deficits in the transgenic rat model of Huntington's disease.  

Science.gov (United States)

Memory deficits are common in patients with Huntington's disease (HD) and have a substantial impact on the quality of life of patients and their relatives. A good model resembling the human memory deficits is needed for research purposes. In this study we investigated the memory function of the transgenic rat model of Huntington's disease (tgHD) in the object location (OLT) and the object recognition task (ORT). Several studies have shown that the recent developed tgHD rat model resembles the human phenotype of HD. Impairments of spatial and object recognition memory in the OLT and ORT, however, have to our knowledge not yet been reported in this transgenic model. Our findings show that in both early and late stages of the disease the tgHD rats have clear deficits for both visuospatial and visual object memory. Since HD patients are known to be impaired in both types of memory, these results confirm the validity of this tgHD rat as a model for the human HD phenotype. PMID:22101303

Zeef, Dagmar H; van Goethem, Nick P; Vlamings, Rinske; Schaper, Frédéric; Jahanshahi, Ali; Hescham, Sarah; von Hörsten, Stephan; Prickaerts, Jos; Temel, Yasin

2012-02-01

67

FUS Transgenic Rats Develop the Phenotypes of Amyotrophic Lateral Sclerosis and Frontotemporal Lobar Degeneration  

OpenAIRE

Fused in Sarcoma (FUS) proteinopathy is a feature of frontotemporal lobar dementia (FTLD), and mutation of the fus gene segregates with FTLD and amyotrophic lateral sclerosis (ALS). To study the consequences of mutation in the fus gene, we created transgenic rats expressing the human fus gene with or without mutation. Overexpression of a mutant (R521C substitution), but not normal, human FUS induced progressive paralysis resembling ALS. Mutant FUS transgenic rats developed progressive paralys...

Huang, Cao; Zhou, Hongxia; Tong, Jianbin; Chen, Han; Liu, Yong-jian; Wang, Dian; Wei, Xiaotao; Xia, Xu-gang

2011-01-01

68

A transgenic rat expressing human APP with the Swedish Alzheimer's disease mutation  

DEFF Research Database (Denmark)

In recent years, transgenic mice have become valuable tools for studying mechanisms of Alzheimer's disease (AD). With the aim of developing an animal model better for memory and neurobehavioural testing, we have generated a transgenic rat model of AD. These animals express human amyloid precursor protein (APP) containing the Swedish AD mutation. The highest level of expression in the brain is found in the cortex, hippocampus, and cerebellum. Starting after the age of 15 months, the rats show increased tau phosphorylation and extracellular Abeta staining. The Abeta is found predominantly in cerebrovascular blood vessels with very rare diffuse plaques. We believe that crossing these animals with mutant PS1 transgenic rats will result in accelerated plaque formation similar to that seen in transgenic mice.

Folkesson, Ronnie; Malkiewicz, Katarzyna

2007-01-01

69

Transgenic rats with green, red, and blue fluorescence: powerful tools for bioimaging, cell trafficking, and differentiation  

Science.gov (United States)

The rat represents a perfect animal for broadening medical experiments, because its physiology has been well understood in the history of experimental animals. In addition, its larger body size takes enough advantage for surgical manipulation, compared to the mouse. Many rat models mimicking human diseases, therefore, have been used in a variety of biomedical studies including physiology, pharmacology, transplantation, and immunology. In an effort to create the specifically designed rats for biomedical research and regenerative medicine, we have developed the engineered rat system on the basis of transgenic technology and succeeded in establishing various transgenic rat strains. The transgenic rats with green fluorescent protein (GFP) were generated in the two different strains (Wistar and Lewis), in which GFP is driven under the chicken beta-actin promoter and cytomegalovirus enhancer (CAG promoter). Their GFP expression levels were different in each organ, but the Lewis line expressed GFP strongly and ubiquitously in most of the organs compared with that of Wistar. For red fluorescence, DsRed2 was transduced to the Wistar rats: one line specifically expresses DsRed2 in the liver under the mouse albumin promoter, another is designed for the Cre/LoxP system as the double reporter rat (the initial DsRed2 expression turns on GFP in the presence of Cre recombinase). LacZ-transgenic rats represent blue color, and LacZ is driven the CAG (DA) or ROSA26 promoter (Lewis). Our unique transgenic rats" system highlights the powerful performance for the elucidation of many cellular processes in regenerative medicine, leading to innovative medical treatments.

Murakami, Takashi; Kobayashi, Eiji

2005-04-01

70

CD46 expression does not overcome the intracellular block of measles virus replication in transgenic rats.  

OpenAIRE

The study of measles pathogenesis and the testing of improved vaccine candidates is hampered by the lack of a small animal model which is susceptible to infection by the intranasal route. With the identification of CD46 as a measles virus (MV) receptor, it was feasible to generate transgenic rats to overcome this problem. Although there was widespread expression of CD46 in the transgenic Sprague-Dawley rats, no measles-like disease could be induced after various routes of infection. The expre...

Niewiesk, S.; Schneider-schaulies, J.; Ohnimus, H.; Jassoy, C.; Schneider-schaulies, S.; Diamond, L.; Logan, J. S.; Ter Meulen, V.

1997-01-01

71

Can organic and transgenic soy be used as a substitute for animal protein by rats?  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in english We evaluated the protein quality of organic and transgenic soy fed to rats throughout life. Thirty female Wistar rats were divided into three groups (N = 10): organic soy group (OSG) receiving organic soy-based diet, genetically modified soy group (GMSG) receiving transgenic soy-based diet, and a co [...] ntrol group (CG) receiving casein-based diet. All animals received water and isocaloric diet (10% protein), ad libitum for 291 days. After this, the weight of GMSG animals (290.9 ± 9.1 g) was significantly lower (P

L.L., Soares; A.M.M., Lucas; G.T., Boaventura.

2005-04-01

72

Development of transgenic rats producing human ?-amyloid precursor protein as a model for Alzheimer's disease: Transgene and endogenous APP genes are regulated tissue-specifically  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background Alzheimer's disease (AD is a devastating neurodegenerative disorder that affects a large and growing number of elderly individuals. In addition to idiopathic disease, AD is also associated with autosomal dominant inheritance, which causes a familial form of AD (FAD. Some instances of FAD have been linked to mutations in the ?-amyloid protein precursor (APP. Although there are numerous mouse AD models available, few rat AD models, which have several advantages over mice, have been generated. Results Fischer 344 rats expressing human APP driven by the ubiquitin-C promoter were generated via lentiviral vector infection of Fischer 344 zygotes. We generated two separate APP-transgenic rat lines, APP21 and APP31. Serum levels of human amyloid-beta (A?40 were 298 pg/ml for hemizygous and 486 pg/ml for homozygous APP21 animals. Serum A?42 levels in APP21 homozygous rats were 135 pg/ml. Immunohistochemistry in brain showed that the human APP transgene was expressed in neurons, but not in glial cells. These findings were consistent with independent examination of enhanced green fluorescent protein (eGFP in the brains of eGFP-transgenic rats. APP21 and APP31 rats expressed 7.5- and 3-times more APP mRNA, respectively, than did wild-type rats. Northern blots showed that the human APP transgene, driven by the ubiquitin-C promoter, is expressed significantly more in brain, kidney and lung compared to heart and liver. A similar expression pattern was also seen for the endogenous rat APP. The unexpected similarity in the tissue-specific expression patterns of endogenous rat APP and transgenic human APP mRNAs suggests regulatory elements within the cDNA sequence of APP. Conclusion This manuscript describes the generation of APP-transgenic inbred Fischer 344 rats. These are the first human AD model rat lines generated by lentiviral infection. The APP21 rat line expresses high levels of human APP and could be a useful model for AD. Tissue-specific expression in the two transgenic rat lines and in wild-type rats contradicts our current understanding of APP gene regulation. Determination of the elements that are responsible for tissue-specific expression of APP may enable new treatment options for AD.

Chan Anthony WS

2008-02-01

73

Modified impact of emotion on temporal discrimination in a transgenic rat model of Huntington disease  

OpenAIRE

Huntington's disease (HD) is characterized by triad of motor, cognitive, and emotional symptoms along with neuropathology in fronto-striatal circuit and limbic system including amygdala. Emotional alterations, which have a negative impact on patient well-being, represent some of the earliest symptoms of HD and might be related to the onset of the neurodegenerative process. In the transgenic rat model (tgHD rats), evidence suggest emotional alterations at the symptomatic stage along with neuro...

AlexisFaure; BruceLBrown; HoaPNguyen; StephanVon Horsten

2013-01-01

74

Tauroursodeoxycholic acid prevents retinal degeneration in transgenic P23H rats  

OpenAIRE

Purpose. To evaluate the preventive effect of tauroursodeoxycholic acid (TUDCA) on photoreceptor degeneration, synaptic connectivity and functional activity of the retina in the transgenic P23H rat, an animal model of autosomal dominant retinitis pigmentosa (RP). Methods. P23H line-3 rats were injected with TUDCA once a week from postnatal day (P)21 to P120, in parallel with vehicle-administered controls. At P120, functional activity of the retina was evaluated by electroretinographic (ERG) r...

Ferna?ndez Sa?nchez, Laura; Lax Zapata, Pedro; Pinilla Lozano, Isabel; Marti?n Nieto, Jose?; Cuenca Navarro, Nicola?s

2011-01-01

75

Expression of the Mu Opioid Receptor in the Human Immunodeficiency Virus Type 1 Transgenic Rat Model?  

OpenAIRE

Opioids, via the mu opioid receptor (MOR), can exacerbate bacterial infections and the immunopathogenesis of human immunodeficiency virus type 1 (HIV-1) infection. Recently, an HIV-1 transgenic (HIV-1Tg) rat model containing circulating HIV-1 gp120 was created. Using real-time reverse transcription-PCR, we found that MOR mRNA levels were significantly higher in the peritoneal macrophages of the HIV-1Tg rat than those in control animals. Lipopolysaccharide, a bacterial endotoxin, induced secre...

Chang, Sulie L.; Beltran, Jose A.; Swarup, Shilpa

2007-01-01

76

Cognitive impairment in the Tg6590 transgenic rat model of Alzheimer's disease  

DEFF Research Database (Denmark)

Recently, interest in the rat as an animal model of Alzheimer's disease (AD) has been growing. We have previously described the Tg6590 transgenic rat line expressing the amyloid precursor protein containing the Swedish AD mutation (K670M/N671L) that shows early stages of Abeta deposition, predominantly in cerebrovascular blood vessels, after 15 months of age. Here we show that by the age of 9 months, that is long before the appearance of Abeta deposits, the Tg6590 rats exhibit deficits in the Morris water maze spatial navigation task and altered spontaneous behaviour in the open-field test. The levels of soluble Abeta were elevated both in the hippocampus and cortex of transgenic animals. Magnetic resonance imaging showed no major changes in the brains of transgenic animals, although they tended to have enlarged lateral ventricles when compared to control animals. The Tg6590 transgenic rat line should prove a suitable model of early AD for advanced studies including serial cerebrospinal fluid sampling, electrophysiology, neuroimaging or complex behavioural testing.

Kloskowska, Ewa; Pham, Therese M

2010-01-01

77

A two-generation reproduction study with transgenic Bt rice TT51 in Wistar rats.  

Science.gov (United States)

TT51 is a transgenic Bt rice created by fusion a synthetic CryAb/CryAc gene into rice MingHui63. A significant number of animal feeding studies with transgenic crops have been carried out with the rapid development of transgenic crops. However, the evidence is far from identifying whether certain novel transgenic crops possess potential danger for human or animal health after long-term consumption. Rice-based diets, containing 60% ordinary grocery rice, MingHui63 rice or TT51 rice by weight, were fed to two generations of male and female rats in order to determine the potential reproductive effects of TT51. In this study, both clinical performance variables and histopathological responses were examined and compared between groups. There were no significant differences between groups on body weights, food consumption, reproductive data and relative organ/body weights. There were some statistically significant differences in hematology and serum chemistry parameters, but no histological abnormalities were seen in the brain, heart, liver, spleen, kidneys, stomach, small intestine, thymus, ovaries, uterus, testes and epididymides. Based on the results, under the circumstance of this study TT51 show no significant differences on reproduction performance of rats compared with MingHui63 and the control. PMID:24309144

Wang, Er Hui; Yu, Zhou; Hu, Jing; Jia, Xu Dong; Xu, Hai Bin

2014-03-01

78

Neurobehavioral Alterations in HIV-1 Transgenic Rats: Evidence for Dopaminergic Dysfunction  

OpenAIRE

Clinical studies have provided evidence that the progression of HIV-1-associated neurocognitive disorders (HAND) involves alterations in dopamine (DA) systems. Drugs of abuse that act on the brain DA system, such as cocaine (Coc), may exacerbate HIV-1 infection and consequent behavioral and neurological manifestations. In the present study, we used the HIV-1 transgenic (Tg) rat, which constitutively expresses 7 of the 9 HIV-1 genes, to assess potential DA system alterations in three behaviora...

Moran, L. M.; Booze, R. M.; Webb, K. M.; Mactutus, C. F.

2012-01-01

79

A transgenic Alzheimer rat with plaques, tau pathology, behavioral impairment, oligomeric A? and frank neuronal loss  

OpenAIRE

Alzheimer’s disease (AD) is hallmarked by amyloid plaques, neurofibrillary tangles, and widespread cortical neuronal loss (Selkoe, 2001). The ‘amyloid cascade hypothesis’ posits that cerebral amyloid sets neurotoxic events into motion that precipitate Alzheimer dementia (Hardy and Allsop, 1991). Yet, faithful recapitulation of all AD features in widely used transgenic (Tg) mice engineered to overproduce A? peptides has been elusive. We have developed a Tg rat model (line TgF344-AD) exp...

Cohen, Robert M.; Rezai-zadeh, Kavon; Weitz, Tara M.; Rentsendorj, Altan; Gate, David; Spivak, Inna; Bholat, Yasmin; Vasilevko, Vitaly; Glabe, Charles G.; Breunig, Joshua J.; Rakic, Pasko; Davtyan, Hayk; Agadjanyan, Michael G.; Kepe, Vladimir; Barrio, Jorge

2013-01-01

80

Chromosome assignment of Cd36 transgenes in two rat SHR lines by FISH and linkage mapping of transgenic insert in the SHR-TG19 line.  

Czech Academy of Sciences Publication Activity Database

Ro?. 48, ?. 4 (2002), s. 139-144. ISSN 0015-5500 R&D Projects: GA ?R GV204/98/K015 Institutional research plan: CEZ:AV0Z5011922 Keywords : spontaneously hypertensive rat * Cd36 * transgenic lines Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 0.615, year: 2002

Liška, F.; Levan, G.; Helou, K.; Sladká, M.; Pravenec, Michal; Zídek, Václav; Landa, Vladimír; K?en, Vladimír

2002-01-01

81

Chronic alcohol ingestion exacerbates skeletal muscle myopathy in HIV-1 transgenic rats  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background Separately, chronic alcohol ingestion and HIV-1 infection are associated with severe skeletal muscle derangements, including atrophy and wasting, weakness, and fatigue. One prospective cohort study reported that 41% of HIV-infected patients met the criteria for alcoholism, however; few reports exist on the co-morbid effects of these two disease processes on skeletal muscle homeostasis. Thus, we analyzed the atrophic effects of chronic alcohol ingestion in HIV-1 transgenic rats and identified alterations to several catabolic and anabolic factors. Findings Relative plantaris mass, total protein content, and fiber cross-sectional area were reduced in each experimental group compared to healthy, control-fed rats. Alcohol abuse further reduced plantaris fiber area in HIV-1 transgenic rats. Consistent with previous reports, gene levels of myostatin and its receptor activin IIB were not increased in HIV-1 transgenic rat muscle. However, myostatin and activin IIB were induced in healthy and HIV-1 transgenic rats fed alcohol for 12 weeks. Catabolic signaling factors such as TGF?1, TNF?, and phospho-p38/total-p38 were increased in all groups compared to controls. There was no effect on IL-6, leukemia inhibitory factor (LIF, cardiotrophin-1 (CT-1, or ciliary neurotrophic factor (CNTF in control-fed, transgenic rats. However, the co-morbidity of chronic alcohol abuse and HIV-1-related protein expression decreased expression of the two anabolic factors, CT-1 and CNTF. Conclusions Consistent with previous reports, alcohol abuse accentuated skeletal muscle atrophy in an animal model of HIV/AIDS. While some catabolic pathways known to drive alcoholic or HIV-1-associated myopathies were also elevated in this co-morbid model (e.g., TGF?1, consistent expression patterns were not apparent. Thus, specific alterations to signaling mechanisms such as the induction of the myostatin/activin IIB system or reductions in growth factor signaling via CT-1- and CNTF-dependent mechanisms may play larger roles in the regulation of muscle mass in alcoholic, HIV-1 models.

Bratina Margaux A

2011-08-01

82

Nonmammalian gonadotropin-releasing hormone molecules in the brain of promoter transgenic rats  

OpenAIRE

Mammalian gonadotropin-releasing hormone (GnRH1) and nonmammalian immunoreactive GnRH subtypes were examined in transgenic rats carrying an enhanced GFP (EGFP) reporter gene driven by a rat GnRH1 promoter. Double-label immunocytochemistry was performed on EGFP+/GnRH1 brain sections by using antisera against GnRH1, GnRH2 (chicken II), GnRH3 (salmon), or seabream GnRH. EGFP+/GnRH1 neurons were in the septal–preoptic hypothalamus but not in the midbrain, consistent with GnRH1-immunopositive ne...

Parhar, Ishwar S.; Soga, Tomoko; Ogawa, Satoshi; Ogawa, Sonoko; Pfaff, Donald W.; Sakuma, Yasuo

2005-01-01

83

In vivo cell kinetics of the bone marrow transplantation using dual colored transgenic rat system  

Science.gov (United States)

Because bone marrow is an adequate site for bone marrow stem cells, intra-bone marrow - bone marrow transplantation (IBM-BMT) is an efficient strategy for bone marrow transplantation (BMT). However, the fate of the transplanted cells remains unclear. Herein, we established a dual-colored transgenic rat system utilizing green fluorescent protein (GFP) and a luciferase (luc) marker. We then utilized this system to investigate the in vivo kinetics of transplanted bone marrow cells (BMCs) after authentic intravenous (IV)-BMT or IBM-BMT. The in vivo fate of the transplanted cells was tracked using an in vivo luminescent imaging technique; alterations in peripheral blood chimerism were also followed using flow cytometry. IBM-BMT and IV-BMT were performed using syngeneic and allogeneic rat combinations. While no difference in the proliferation pattern was observed between the two treatment groups at 7 days after BMT, different distribution patterns were clearly observed during the early phase. In the IBM-BMT-treated rats, the transplanted BMCs were engrafted immediately at the site of the injected bone marrow and expanded more rapidly than in the IV-BMT-treated rats during this phase. Graft-versus-host disease was also visualized. Our bio-imaging system using dual-colored transgenic rats is a powerful tool for performing quantitative and morphological assessments in vivo.

Kai, Kotaro; Teraoka, Satoshi; Adachi, Yasushi; Ikehara, Susumu; Murakami, Takashi; Kobayashi, Eiji

2008-02-01

84

In vivo gene delivery to the pulmonary circulation in rats: transgene distribution and vascular inflammatory response.  

Science.gov (United States)

Although gene delivery to the pulmonary circulation has both experimental and therapeutic potential, the delivery methods, distribution of transgene, and subsequent inflammatory response have been poorly characterized to date. To address these issues, we utilized a 0.76-mm OD (outside diameter) end hole catheter inserted into the internal jugular vein of adult Sprague-Dawley rats, directing the tip into a pulmonary capillary wedge position. We then compared infusion of polycationic lipid:DNA complexes to replication-defective adenovirus with respect to magnitude and distribution of transgene expression using either chloramphenicol acetyltransferase (CAT) or human placental alkaline phosphatase (hpAP) reporter genes. Both lipid:DNA and adenovirus resulted in detectable transgene expression, though maximum lung CAT activity using lipid (gamma AP-DLRIE/DOPE) was approximately 2% of maximum activity using adenovirus (Ad-CAT). Further characterization of expression after transfection with 10(8) pfu (plaque forming units) of Ad-CAT demonstrated persistence of transgene for at least 14 days (lung CAT activity 27% of maximum). Alkaline phosphatase staining demonstrated that both large and small pulmonary arteries as well as the alveolar wall expressed transgene. Although little inflammatory response was detected in conduit arteries, a predominantly mononuclear cell infiltrate surrounded small pulmonary arteries as well as the alveolar spaces in transfected areas of lung. We conclude that percutaneous catheter-mediated gene delivery to the pulmonary circulation in rats using non-viral and viral vectors is feasible. Although an inflammatory response to first generation replication-defective adenovirus was detected, it appeared to be largely restricted to the distal pulmonary circulation and airspace. This technique should prove useful for investigations requiring overexpression of novel genes in the pulmonary artery wall, and could ultimately be used to develop gene-based therapies for pulmonary vascular diseases. PMID:9191465

Rodman, D M; San, H; Simari, R; Stephan, D; Tanner, F; Yang, Z; Nabel, G J; Nabel, E G

1997-06-01

85

Inducible Gene Manipulations in Brain Serotonergic Neurons of Transgenic Rats  

OpenAIRE

The serotonergic (5-HT) system has been implicated in various physiological processes and neuropsychiatric disorders, but in many aspects its role in normal and pathologic brain function is still unclear. One reason for this might be the lack of appropriate animal models which can address the complexity of physiological and pathophysiological 5-HT functioning. In this respect, rats offer many advantages over mice as they have been the animal of choice for sophisticated neurophysiological and ...

Weber, Tillmann; Scho?nig, Kai; Tews, Bjo?rn; Bartsch, Dusan

2011-01-01

86

Megaesophagus in a line of transgenic rats: a model of achalasia.  

Science.gov (United States)

Megaesophagus is defined as the abnormal enlargement or dilatation of the esophagus, characterized by a lack of normal contraction of the esophageal walls. This is called achalasia when associated with reduced or no relaxation of the lower esophageal sphincter (LES). To date, there are few naturally occurring models for this disease. A colony of transgenic (Pvrl3-Cre) rats presented with megaesophagus at 3 to 4 months of age; further breeding studies revealed a prevalence of 90% of transgene-positive animals having megaesophagus. Affected rats could be maintained on a total liquid diet long term and were shown to display the classic features of dilated esophagus, closed lower esophageal sphincter, and abnormal contractions on contrast radiography and fluoroscopy. Histologically, the findings of muscle degeneration, inflammation, and a reduced number of myenteric ganglia in the esophagus combined with ultrastructural lesions of muscle fiber disarray and mitochondrial changes in the striated muscle of these animals closely mimic that seen in the human condition. Muscle contractile studies looking at the response of the lower esophageal sphincter and fundus to electrical field stimulation, sodium nitroprusside, and L-nitro-L-arginine methyl ester also demonstrate the similarity between megaesophagus in the transgenic rats and patients with achalasia. No primary cause for megaesophagus was found, but the close parallel to the human form of the disease, as well as ease of care and manipulation of these rats, makes this a suitable model to better understand the etiology of achalasia as well as study new management and treatment options for this incurable condition. PMID:24457157

Pang, J; Borjeson, T M; Muthupalani, S; Ducore, R M; Carr, C A; Feng, Y; Sullivan, M P; Cristofaro, V; Luo, J; Lindstrom, J M; Fox, J G

2014-11-01

87

Polyethylenimine-mediated expression of transgenes in the acinar cells of rats salivary glands in vivo  

Science.gov (United States)

Non viral-mediated transfection of plasmid DNA provides a fast and reliable way to express various transgenes in selected cell populations in live animals. Here, we show an improvement of a previously published method that is based on injecting plasmid DNA into the ductal system of the salivary glands in live rats. Specifically, using complexes between plasmid DNA and polyethyleneimine (PEI) we show that the expression of the transgenes is directed selectively to the salivary acinar cells. PEI does not affect the ability of cells to undergo regulated exocytosis, which was one of the main drawbacks of the previous methods. Moreover PEI does not affect the proper localization and targeting of transfected proteins, as shown for the apical plasma membrane water channel aquaporin 5 (AQP5). Overall, this approach, coupled with the use of intravital microscopy, permits to conduct localization and functional studies under physiological conditions, in a rapid, reliable, and affordable fashion. PMID:25621283

Sramkova, Monika; Parente, Laura; Wigand, Timothy; Aye, Myo-Pale'; Shitara, Akiko; Weigert, Roberto

2015-01-01

88

Light-evoked Somatosensory Perception of Transgenic Rats That Express Channelrhodopsin-2 in Dorsal Root Ganglion Cells  

OpenAIRE

In vertebrate somatosensory systems, each mode of touch-pressure, temperature or pain is sensed by sensory endings of different dorsal root ganglion (DRG) neurons, which conducted to the specific cortical loci as nerve impulses. Therefore, direct electrical stimulation of the peripheral nerve endings causes an erroneous sensation to be conducted by the nerve. We have recently generated several transgenic lines of rat in which channelrhodopsin-2 (ChR2) transgene is driven by the Thy-1.2 promot...

Ji, Zhi-gang; Ito, Shin; Honjoh, Tatsuya; Ohta, Hiroyuki; Ishizuka, Toru; Fukazawa, Yugo; Yawo, Hiromu

2012-01-01

89

Dominant dwarfism in transgenic rats by targeting human growth hormone (GH) expression to hypothalamic GH-releasing factor neurons.  

OpenAIRE

Expression of human growth hormone (hGH) was targeted to growth hormone-releasing (GRF) neurons in the hypothalamus of transgenic rats. This induced dominant dwarfism by local feedback inhibition of GRF. One line, bearing a single copy of a GRF-hGH transgene, has been characterized in detail, and has been termed Tgr (for Transgenic growth-retarded). hGH was detected by immunocytochemistry in the brain, restricted to the median eminence of the hypothalamus. Low levels were also detected in the...

Flavell, D. M.; Wells, T.; Wells, S. E.; Carmignac, D. F.; Thomas, G. B.; Robinson, I. C.

1996-01-01

90

HIV-1 transgene expression in rats causes oxidant stress and alveolar epithelial barrier dysfunction  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background HIV-infected individuals are at increased risk for acute and chronic airway disease even though there is no evidence that the virus can infect the lung epithelium. Although HIV-related proteins including gp120 and Tat can directly cause oxidant stress and cellular dysfunction, their effects in the lung are unknown. The goal of this study was to determine the effects of HIV-1 transgene expression in rats on alveolar epithelial barrier function. Alveolar epithelial barrier function was assessed by determining lung liquid clearance in vivo and alveolar epithelial monolayer permeability in vitro. Oxidant stress in the alveolar space was determined by measuring the glutathione redox couple by high performance liquid chromatography, and the expression and membrane localization of key tight junction proteins were assessed. Finally, the direct effects of the HIV-related proteins gp120 and Tat on alveolar epithelial barrier formation and tight junction protein expression were determined. Results HIV-1 transgene expression caused oxidant stress within the alveolar space and impaired epithelial barrier function even though there was no evidence of overt inflammation within the airways. The expression and membrane localization of the tight junction proteins zonula occludens-1 and occludin were decreased in alveolar epithelial cells from HIV-1 transgenic rats. Further, treating alveolar epithelial monolayers from wild type rats in vitro with recombinant gp120 or Tat for 24 hours reproduced many of the effects on zonula occludens-1 and occludin expression and membrane localization. Conclusion Taken together, these data indicate that HIV-related proteins cause oxidant stress and alter the expression of critical tight junction proteins in the alveolar epithelium, resulting in barrier dysfunction.

Jacob Barbara A

2009-02-01

91

Angiotensin II induced inflammation in the kidney and in the heart of double transgenic rats  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background We are investigating a double transgenic rat (dTGR model, in which rats transgenic for the human angiotensinogen and renin genes are crossed. These rats develop moderately severe hypertension but die of end-organ cardiac and renal damage by week 7. The heart shows necrosis and fibrosis, whereas the kidneys resemble the hemolytic-uremic syndrome vasculopathy. Surface adhesion molecules (ICAM-1 and VCAM-1 are expressed early on the endothelium, while the corresponding ligands are found on circulating leukocytes. Leukocyte infiltration in the vascular wall accompanies PAI-1, MCP-1, iNOS and Tissue Factor expression. Furthermore we show evidence that Ang II causes the upregulation of NF-kB in our model. Methods We started PDTC-treatment on four weeks old dTGR (200 mg/kg sc and age-matched SD rats.. Blood-pressure- and albuminuria- measurements were monitored during the treatement period (four weeks. The seven weeks old animals were killed, hearts and kidneys were isolated and used for immunohistochemical-and electromobility shift assay analsis. Results Chronic treatment with the antioxidant PDTC decreased blood pressure (162 ± 8 vs. 190 ± 7 mm Hg, p = 0.02. Cardiac hypertrophy index was significantly reduced (4.90 ± 0.1 vs. 5.77 ± 0.1 mg/g, p Conclusion Our data show that inhibition of NF-?B by PDTC markedly reduces inflammation, iNOS expression in the dTGR most likely leading to decreased cytotoxicity, and cell proliferation. Thus, NF-?B activation plays an important role in ANG II-induced end-organ damage.

Haller Hermann

2002-01-01

92

Altered emotional and motivational processing in the transgenic rat model for Huntington's disease.  

Science.gov (United States)

Huntington disease (HD) is caused by an expansion of CAG repeat in the Huntingtin gene. Patients demonstrate a triad of motor, cognitive and psychiatric symptoms. A transgenic rat model (tgHD rats) carrying 51 CAG repeats demonstrate progressive striatal degeneration and polyglutamine aggregates in limbic structures. In this model, emotional function has only been investigated through anxiety studies. Our aim was to extend knowledge on emotional and motivational function in symptomatic tgHD rats. We subjected tgHD and wild-type rats to behavioral protocols testing motor, emotional, and motivational abilities. From 11 to 15 months of age, animals were tested in emotional perception of sucrose using taste reactivity, acquisition, extinction, and re-acquisition of discriminative Pavlovian fear conditioning as well as reactivity to changes in reinforcement values in a runway Pavlovian approach task. Motor tests detected the symptomatic status of tgHD animals from 11 months of age. In comparison to wild types, transgenic animals exhibited emotional blunting of hedonic perception for intermediate sucrose concentration. Moreover, we found emotional alterations with better learning and re-acquisition of discriminative fear conditioning due to a higher level of conditioned fear to aversive stimuli, and hyper-reactivity to a negative hedonic shift in reinforcement value interpreted in term of greater frustration. Neuropathological assessment in the same animals showed a selective shrinkage of the central nucleus of the amygdala. Our results showing emotional blunting and hypersensitivity to negative emotional situations in symptomatic tgHD animals extend the face validity of this model regarding neuropsychiatric symptoms as seen in manifest HD patients, and suggest that some of these symptoms may be related to amygdala dysfunction. PMID:21111837

Faure, A; Höhn, S; Von Hörsten, S; Delatour, B; Raber, K; Le Blanc, P; Desvignes, N; Doyère, V; El Massioui, N

2011-01-01

93

A progressive dopaminergic phenotype associated with neurotoxic conversion of ?-synuclein in BAC-transgenic rats  

DEFF Research Database (Denmark)

Conversion of soluble ?-synuclein into insoluble and fibrillar inclusions is a hallmark of Parkinson's disease and other synucleinopathies. Accumulating evidence points towards a relationship between its generation at nerve terminals and structural synaptic pathology. Little is known about the pathogenic impact of ?-synuclein conversion and deposition at nigrostriatal dopaminergic synapses in transgenic mice, mainly owing to expression limitations of the ?-synuclein construct. Here, we explore whether both the rat as a model and expression of the bacterial artificial chromosome construct consisting of human full-length wild-type ?-synuclein could exert dopaminergic neuropathological effects. We found that the human promoter induced a pan-neuronal expression, matching the rodent ?-synuclein expression pattern, however, with prominent C-terminally truncated fragments. Ageing promoted conversion of both full-length and C-terminally truncated ?-synuclein species into insolube and proteinase K-resistant fibres, with strongest accumulation in the striatum, resembling biochemical changes seen in human Parkinson's disease. Transgenic rats develop early changes in novelty-seeking, avoidance and smell before the progressive motor deficit. Importantly, the observed pathological changes were associated with severe loss of the dopaminergic integrity, thus resembling more closely the human pathology.

Nuber, Silke; Harmuth, Florian

2013-01-01

94

Inducible and reversible gene silencing by stable integration of an shRNA-encoding lentivirus in transgenic rats  

OpenAIRE

Currently, tools to generate loss-of-function mutations in rats are limited. Therefore, we have developed a lentiviral single-vector system for the temporal control of ubiquitous shRNA expression. Here, we report transgenic rats carrying an insulin receptor-specific shRNA transcribed from a regulatable promoter and identified by concomitant EGFP expression. In the absence of the inducer doxycycline (Dox), we observed no siRNA expression. However, Dox treatment at very low concentrations led t...

Herold, Marco J.; Den Brandt, Jens; Seibler, Jost; Reichardt, Holger M.

2008-01-01

95

Enhancement of metabolizing herbicides in young tubers of transgenic potato plants with the rat CYP1A1 gene.  

Science.gov (United States)

A rat P450 monooxygenase gene ( CYP1A1) was introduced into potato plants to enhance the metabolism of the environmental contaminants in subterranean organs. The CYP1A1 gene was kept under the control of the potato patatin promoter to enhance tuber-specific expression. A total of 106 transgenic plants (PAT1A1 plants) were obtained following selection by a resistance test to kanamycin and PCR analysis. PAT1A1 plants treated with 10% exogenous sucrose showed a higher activity of monooxgenase in the leaves than the non-transgenic plants. This indicated that the activity enhanced by 10% sucrose was due to the patatin promoter containing the sucrose-inducted elements. One representative transgenic plant, Ag2197, was selected on the basis of monooxgenase activity in the leaves and Western blot analysis. Ag2197 was found to accumulate a large amount of CYP1A1 mRNA and protein in the developing tuber but not in the mature tuber. The residual herbicides, atrazine and chlortoluron, were analyzed in the micro-tubers of Ag2197 and non-transgenic plants. The amount of residual herbicides in Ag2197 was much lower than that in the non-transgenic plant, indicating that the transgenic plant metabolized the herbicides to a detoxified form. The transgenic plants produced in this study might be useful for the phytoremediation of chemical pollution in the soil. PMID:12582499

Yamada, T.; Ishige, T.; Shiota, N.; Inui, H.; Ohkawa, H.; Ohkawa, Y.

2002-09-01

96

Modified impact of emotion on temporal discrimination in a transgenic rat model of Huntington disease.  

Science.gov (United States)

Huntington's disease (HD) is characterized by triad of motor, cognitive, and emotional symptoms along with neuropathology in fronto-striatal circuit and limbic system including amygdala. Emotional alterations, which have a negative impact on patient well-being, represent some of the earliest symptoms of HD and might be related to the onset of the neurodegenerative process. In the transgenic rat model (tgHD rats), evidence suggest emotional alterations at the symptomatic stage along with neuropathology of the central nucleus of amygdala (CE). Studies in humans and animals demonstrate that emotion can modulate time perception. The impact of emotion on time perception has never been tested in HD, nor is it known if that impact could be part of the presymptomatic emotional phenotype of the pathology. The aim of this paper was to characterize the effect of emotion on temporal discrimination in presymptomatic tgHD animals. In the first experiment, we characterized the acute effect of an emotion (fear) conditioned stimulus on temporal discrimination using a bisection procedure, and tested its dependency upon an intact central amygdala. The second experiment was aimed at comparing presymptomatic homozygous transgenic animals at 7-months of age and their wild-type littermates (WT) in their performance on the modulation of temporal discrimination by emotion. Our principal findings show that (1) a fear cue produces a short-lived decrease of temporal precision after its termination, and (2) animals with medial CE lesion and presymptomatic tgHD animals demonstrate an alteration of this emotion-evoked temporal distortion. The results contribute to our knowledge about the presymptomatic phenotype of this HD rat model, showing susceptibility to emotion that may be related to dysfunction of the central nucleus of amygdala. PMID:24133419

Faure, Alexis; Es-Seddiqi, Mouna; Brown, Bruce L; Nguyen, Hoa P; Riess, Olaf; von Hörsten, Stephan; Le Blanc, Pascale; Desvignes, Nathalie; Bozon, Bruno; El Massioui, Nicole; Doyère, Valérie

2013-01-01

97

Nonmammalian gonadotropin-releasing hormone molecules in the brain of promoter transgenic rats.  

Science.gov (United States)

Mammalian gonadotropin-releasing hormone (GnRH1) and nonmammalian immunoreactive GnRH subtypes were examined in transgenic rats carrying an enhanced GFP (EGFP) reporter gene driven by a rat GnRH1 promoter. Double-label immunocytochemistry was performed on EGFP(+)/GnRH1 brain sections by using antisera against GnRH1, GnRH2 (chicken II), GnRH3 (salmon), or seabream GnRH. EGFP(+)/GnRH1 neurons were in the septal-preoptic hypothalamus but not in the midbrain, consistent with GnRH1-immunopositive neurons in WT rats. Apparent coexpression of EGFP(+)/GnRH1 with other GnRH subtypes was observed. All EGFP(+) neurons in the septal-preoptic hypothalamus were GnRH1-immunopositive. However, only approximately 80% of GnRH1-immunopositive neurons were EGFP(+), which awaits further elucidation. GnRH subtypes-immunopositive fibers and EGFP(+)/GnRH1 fibers were conspicuous in the organum vasculosum of the lamina terminalis, median eminence, and surrounding the ependymal walls of the third ventricle and the aqueduct in the midbrain. These results demonstrate that the expression of the EGFP-GnRH1 transgene is restricted to the bona fide GnRH1 population and provide clear morphological evidence supporting the existence of GnRH1 neuronal subpopulations in the septal-preoptic hypothalamus, which might be driven by different segments of the GnRH promoter. This genetic construct permits analyses of promoter usage in GnRH neurons, and our histochemical approaches open questions about functional relations among isoforms of this peptide, which regulates reproductive physiology in its behavioral and endocrine aspects. PMID:15824321

Parhar, Ishwar S; Soga, Tomoko; Ogawa, Satoshi; Ogawa, Sonoko; Pfaff, Donald W; Sakuma, Yasuo

2005-04-19

98

L-NAME protects against acute light damage in albino rats, but not against retinal degeneration in P23H and S334ter transgenic rats.  

Science.gov (United States)

Two previous studies have shown that N(G)-nitro-L-arginine methyl ester (L-NAME), an inhibitor of neuronal nitric oxide synthase, protects retinas of albino rats and mice from damaging levels of light. The aims of the present study were two-fold: (1) to confirm the protective effect of L-NAME on wild type albino rats and (2) to determine if L-NAME protects the retinas of transgenic rats with P23H and S334ter rhodopsin mutations. In the first study, albino rats born and raised in 5-10 lux cyclic light were injected intraperitoneally with either L-NAME or its inactive isomer D-NAME 30 min before being placed in bright light (2700 lux) for 24hr. Electroretinograms (ERGs) were recorded before light treatment and 2 days after cessation of exposure, and eyes were enucleated for morphologic evaluation. L-NAME, but not D-NAME provided structural protection of photoreceptor cells from light damage. The functional rescue was not statistically significant between the drug treated groups. In the second study, albino WT, P23H transgenic, and S334ter transgenic rats were born and raised in 400 lux cyclic light. Three week old animals received daily intraperitoneal injections of L-NAME or D-NAME for 4 weeks, and the same drugs were added to their drinking water. At 7 weeks of age, the ERG sensitivity curves and the outer nuclear layer thickness of both transgenic groups were significantly reduced compared to WT controls. However, administration of L-NAME did not protect against retinal degeneration caused by the rhodopsin mutation in either strain of transgenic (P23H and S334ter) rats. Thus, although photoreceptor cell death in light damage and inherited retinal degenerations share a common apoptotic mechanism, there must be significant 'up-stream' differences that allow selective neuroprotection by L-NAME. PMID:12634110

Káldi, Ildikó; Dittmar, Mark; Pierce, Paula; Anderson, Robert E

2003-04-01

99

Expression of ATP-binding cassette membrane transporters in a HIV-1 transgenic rat model.  

Science.gov (United States)

P-glycoprotein (P-gp, product of Mdr1a and Mdr1b genes), multidrug resistance associated proteins (Mrps), and breast cancer resistance protein (Bcrp), all members of the ATP-binding cassette (ABC) membrane-associated drug transporters superfamily, can significantly restrict the entry of antiretroviral drugs (ARVs) into organs which exhibit a barrier function such as the central nervous system (CNS) and the male genital tract (MGT). In vitro, HIV-1 viral proteins such as glycoprotein-120 (gp120) and transcriptional transactivator (tat) have been shown to alter the expression of these transporters and ARVs permeability. The objective of this study was to compare mRNA expression of these transporters, in vivo, in several tissues obtained from HIV-1 transgenic rats (Tg-rat) (8 and 24 weeks) with those of age-matched wild-type rats. At 24 weeks, significant changes in several drug transporter mRNA expressions were observed, in particular, in brain, kidney, liver and testes. These findings suggest that HIV-1 viral proteins can alter the expression of ABC drug transporters, in vivo, in the context of HIV-1 and further regulate ARVs permeability in several organs including the CNS and MGT, two sites which have been reported to display very low ARVs permeability in the clinic. PMID:24472536

Robillard, Kevin R; Hoque, Md Tozammel; Bendayan, Reina

2014-02-21

100

Overexpression of the rat sarcoplasmic reticulum Ca2+ ATPase gene in the heart of transgenic mice accelerates calcium transients and cardiac relaxation.  

OpenAIRE

The Ca2+ ATPase of the sarcoplasmic reticulum (SERCA2) plays a dominant role in lowering cytoplasmic calcium levels during cardiac relaxation and reduction of its activity has been linked to delayed diastolic relaxation in hypothyroid and failing hearts. To determine the contractile alterations resulting from increased SERCA2 expression, we generated transgenic mice overexpressing a rat SERCA2 transgene. Characterization of a heterozygous transgenic mouse line (CJ5) showed that the amount of ...

He, H.; Giordano, F. J.; Hilal-dandan, R.; Choi, D. J.; Rockman, H. A.; Mcdonough, P. M.; Bluhm, W. F.; Meyer, M.; Sayen, M. R.; Swanson, E.; Dillmann, W. H.

1997-01-01

101

Photoreceptor differentiation and integration of retinal progenitor cells transplanted into transgenic rats.  

Science.gov (United States)

Previous studies evaluating neural stem cells transplanted into the mature retina have demonstrated limited levels of graft-host integration and photoreceptor differentiation. The purpose of this investigation is to enhance photoreceptor cell differentiation and integration of retinal progenitor cells (RPC) following subretinal transplantation into retinal degenerate rats by optimization of isolation, expansion, and transplantation procedures. RPCs were isolated from human placental alkaline phosphatase (hPAP)-positive embryonic day 17 (E17) rat retina and expanded in serum-free defined media. RPCs at passage 2 underwent in vitro induction with all trans retinoic acid or were transplanted into the subretinal space of post-natal day (P) 17 S334ter-3 and S334ter-5 transgenic rats. Animals were examined post-operatively by ophthalmoscopy and optical coherence tomography (OCT) at weeks 1 and 4. Differentiation profiles of RPCs, both in vitro and in vivo were analysed microscopically by immunohistochemistry for various retinal cell specific markers. Our results demonstrated that the majority of passage 2 RPCs differentiated into retina-specific neurons expressing rhodopsin after in vitro induction. Following subretinal transplantation, grafted cells formed a multi-layer cellular sheet in the subretinal space in both S334ter-3 and S334ter-5 rats. Prominent retina-specific neuronal differentiation was observed in both rat lines as evidenced by recoverin or rhodopsin staining in 80% of grafted cells. Less than 5% of the grafted cells expressed glial fibrillary acidic protein. Synapsin-1 (label for nerve terminals) positive neural processes were present at the graft-host interface. Expression profiles of the grafted RPCs were similar to those of RPCs induced to differentiate in vitro using all-trans retinoic acid. In contrast to our previous study, grafted RPCs can demonstrate extensive rhodopsin expression, organize into layers, and show some features of apparent integration with the host retina following subretinal transplantation in slow and fast retinal degenerate rats. The similarity of the in vitro and in vivo RPC differentiation profiles suggests that intrinsic signals may have a significant contribution to RPC cell fate determination. PMID:15781279

Qiu, Guanting; Seiler, Magdalene J; Mui, Cathy; Arai, Shinichi; Aramant, Robert B; de Juan, Eugene; Sadda, SriniVas

2005-04-01

102

Meloxicam Blocks Neuroinflammation, but Not Depressive-Like Behaviors, in HIV-1 Transgenic Female Rats  

Science.gov (United States)

Adolescents living with human immunodeficiency virus (HIV) comprise approximately 12% of the HIV-positive population worldwide. HIV-positive adolescents experience a higher rate of clinical depression, a greater risk of sexual and drug abuse behaviors, and a decreased adherence to highly active antiretroviral therapies (HAART). Using adolescent HIV-1 transgenic rats (HIV-1 tg) that display related immune response alterations and pathologies, this study tested the hypothesis that developmental expression of HIV-1-related proteins induces a depressive-like phenotype that parallels a decrease in hippocampal cell proliferation and an increase in pro-inflammatory cytokine expression in the hippocampus. Consistent with this hypothesis, adolescent HIV-1 tg rats demonstrated a depressive-like behavioral phenotype, had decreased levels of cell proliferation, and exhibited elevated expression of monocyte chemotactic protein-1 (Mcp-1) in the hippocampus relative to controls. Subsequently, we tested the ability of meloxicam, a selective COX-2 inhibitor, to attenuate behavioral deficits via inflammatory mechanisms. Daily meloxicam treatments did not alter the behavioral profile despite effectively reducing hippocampal inflammatory gene expression. Together, these data support a biological basis for the co-morbid manifestation of depression in HIV-positive patients as early as in adolescence and suggest that modifications in behavior manifest independent of inflammatory activity in the hippocampus. PMID:25271421

Nemeth, Christina L.; Glasper, Erica R.; Harrell, Constance S.; Malviya, Sanjana A.; Otis, Jeffrey S.; Neigh, Gretchen N.

2014-01-01

103

Progress Toward a Human CD4/CCR5 Transgenic Rat Model for De Novo Infection by Human Immunodeficiency Virus Type 1  

OpenAIRE

The development of a permissive small animal model for the study of human immunodeficiency virus type (HIV)-1 pathogenesis and the testing of antiviral strategies has been hampered by the inability of HIV-1 to infect primary rodent cells productively. In this study, we explored transgenic rats expressing the HIV-1 receptor complex as a susceptible host. Rats transgenic for human CD4 (hCD4) and the human chemokine receptor CCR5 (hCCR5) were generated that express the transgenes in CD4+ T lymph...

Keppler, Oliver T.; Welte, Frank J.; Ngo, Tuan A.; Chin, Peggy S.; Patton, Kathryn S.; Tsou, Chia-lin; Abbey, Nancy W.; Sharkey, Mark E.; Grant, Robert M.; You, Yun; Scarborough, John D.; Ellmeier, Wilfried; Littman, Dan R.; Stevenson, Mario; Charo, Israel F.

2002-01-01

104

Fat-specific transgenic expression of resistin in the spontaneously hypertensive rat impairs fatty acid re-esterification.  

Czech Academy of Sciences Publication Activity Database

Ro?. 30, ?. 7 (2006), s. 1157-1159. ISSN 0307-0565 R&D Projects: GA ?R(CZ) GA301/03/0751; GA MZd(CZ) NB7403; GA MŠk(CZ) 1M0520 Grant ostatní: HHMI(US) 55005624 Institutional research plan: CEZ:AV0Z50110509 Keywords : spontaneously hypertensive rat * transgenic resistin * fatty acid reesterification Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 4.055, year: 2006

Pravenec, Michal; Kazdová, L.; Cahová, M.; Landa, Vladimír; Zídek, Václav; Mlejnek, Petr; Šimáková, Miroslava; Wang, J.; Qi, N.; Kurtz, T. W.

2006-01-01

105

Neuronal driven pre-plaque inflammation in a transgenic rat model of Alzheimer's disease.  

Science.gov (United States)

Chronic brain inflammation is associated with Alzheimer's disease (AD) and is classically attributed to amyloid plaque deposition. However, whether the amyloid pathology can trigger early inflammatory processes before plaque deposition remains a matter of debate. To address the possibility that a pre-plaque inflammatory process occurs, we investigated the status of neuronal, astrocytic, and microglial markers in pre- and post-amyloid plaque stages in a novel transgenic rat model of an AD-like amyloid pathology (McGill-R-Thy1-APP). In this model, we found a marked upregulation of several classical inflammatory markers such as COX-2, IL-1?, TNF-?, and fractalkine (CX3CL1) in the cerebral cortex and hippocampus. Interestingly, many of these markers were highly expressed in amyloid beta-burdened neurons. Activated astrocytes and microglia were associated with these A?-burdened neurons. These findings confirm the occurrence of a proinflammatory process preceding amyloid plaque deposition and suggest that A?-burdened neurons play a crucial role in initiating inflammation in AD. PMID:24831823

Hanzel, Cecilia E; Pichet-Binette, Alexa; Pimentel, Luisa S B; Iulita, M Florencia; Allard, Simon; Ducatenzeiler, Adriana; Do Carmo, Sonia; Cuello, A Claudio

2014-10-01

106

Longitudinal analysis of the behavioral phenotype in a novel transgenic rat model of early stages of Alzheimer's disease.  

Science.gov (United States)

Intraneuronal accumulation of amyloid ? (iA?) has been linked to mild cognitive impairment that may precede Alzheimer's disease (AD) onset. This neuropathological trait was recently mimicked in a novel animal model of AD, the hemizygous transgenic McGill-R-Thy1-APP (Tg(+/-)) rat. The characterization of the behavioral phenotypes in this animal model could provide a baseline of efficacy for earlier therapeutic interventions. The aim of the present study was to undertake a longitudinal study of A? accumulation and a comprehensive behavioral evaluation of this transgenic rat model. We assessed exploratory activity, anxiety-related behaviors, recognition memory, working memory, spatial learning and reference memory at 3, 6, and 12 months of age. In parallel, we measured A? by ELISA, Western blots and semiquantitative immunohistochemistry in hippocampal samples. SDS-soluble A? peptide accumulated at low levels (~9 pg/mg) without differences among ages. However, Western blots showed SDS-resistant A? oligomers (~30 kDa) at 6 and 12 months, but not at 3 months. When compared to wild-type (WT), male Tg(+/-) rats exhibited a spatial reference memory deficit in the Morris Water Maze (MWM) as early as 3 months of age, which persisted at 6 and 12 months. In addition, Tg(+/-) rats displayed a working memory impairment in the Y-maze and higher anxiety levels in the Open Field (OF) at 6 and 12 months of age, but not at 3 months. Exploratory activity in the OF was similar to that of WT at all-time points. Spatial learning in the MWM and the recognition memory, as assessed by the Novel Object Recognition Test, were unimpaired at any time point. The data from the present study demonstrate that the hemizygous transgenic McGill-R-Thy1-APP rat has a wide array of behavioral and cognitive impairments from young adulthood to middle-age. The low A? burden and early emotional and cognitive deficits in this transgenic rat model supports its potential use for drug discovery purposes in early AD. PMID:25278855

Galeano, Pablo; Martino Adami, Pamela V; Do Carmo, Sonia; Blanco, Eduardo; Rotondaro, Cecilia; Capani, Francisco; Castaño, Eduardo M; Cuello, A Claudio; Morelli, Laura

2014-01-01

107

Antihypertensive action of soluble epoxide hydrolase inhibition in Ren-2 transgenic rats is mediated by suppression of the intrarenal renin-angiotensin system  

OpenAIRE

The present study was undertaken to evaluate the hypothesis that the antihypertensive action of soluble epoxide hydrolase (sEH) inhibition is mediated by an increased availability in intrarenal epoxyeicosatrienoic acids (EETs) with consequent improvement of renal haemodynamic autoregulatory efficiency and of the pressure-natriuresis relationship.Ren-2 transgenic rats (TGR), a model of angiotensin II (ANG II)-dependent hypertension, and normotensive transgene-negative Hannover Sprague-Dawley (...

Varcabova?, S?a?rka; Huskova?, Zuzana; Kramer, Herbert J.; Hwang, Sung Hee; Hammock, Bruce D.; Imig, John D.; Kitada, Kento; C?ervenka, Lude?k

2013-01-01

108

Age-related autocrine diabetogenic effects of transgenic resistin in spontaneously hypertensive rats: gene expression profile analysis.  

Czech Academy of Sciences Publication Activity Database

Ro?. 43, ?. 7 (2011), s. 372-379. ISSN 1094-8341 R&D Projects: GA MŠk(CZ) ME08006; GA MŠk(CZ) 1M0510; GA AV ?R(CZ) IAA500110805; GA MZd(CZ) NS9759 Grant ostatní: Fondation Leducq(FR) 06CVD03 Institutional research plan: CEZ:AV0Z50110509 Keywords : transgenic rat * adipose tissue * insulin resistance * autocrine effects Subject RIV: FB - Endocrinology, Diabetology, Metabolism , Nutrition Impact factor: 2.735, year: 2011

Pravenec, Michal; Zídek, Václav; Landa, Vladimír; Šimáková, Miroslava; Mlejnek, Petr; Šilhavý, J.; Maxová, M.; Kazdová, L.; Seidman, J. G.; Seidman, Ch. E.; Eminaga, S.; Gorham, J.; Wang, J.; Kurtz, T. W.

2011-01-01

109

The Relationship of Photoreceptor Degeneration to Retinal Vascular Development and Loss in Mutant Rhodopsin Transgenic and RCS Rats  

OpenAIRE

The early loss of photoreceptors in some retinal degenerations in mice has been shown to have a profound effect on vascular development of the retina. To better characterize this relationship, we have examined the formation of retinal blood vessels during the first month of life in 8 lines of transgenic rats with different ages of onset and rates of photoreceptor cell loss mediated by the expression of mutant rhodopsin (P23H and S334ter). The number of capillary profiles in the superficial pl...

Pennesi, Mark E.; Nishikawa, Shimpei; Matthes, Michael T.; Yasumura, Douglas; Lavail, Matthew M.

2008-01-01

110

HIV-susceptible transgenic rats allow rapid preclinical testing of antiviral compounds targeting virus entry or reverse transcription  

OpenAIRE

The current testing of anti-HIV drugs is hampered by the lack of a small animal that is readily available and easy to handle; can be infected systemically with HIV type 1 (HIV-1); harbors the major HIV-1 target cells in a physiological frequency, organ distribution, and activation state; and is established as a pharmacological model. Here, we explored the potential of outbred Sprague–Dawley rats that transgenically express the HIV-1 receptor complex on CD4 T cells and macrophages as a model...

Goffinet, Christine; Allespach, Ina; Keppler, Oliver T.

2007-01-01

111

Visual Properties of Transgenic Rats Harboring the Channelrhodopsin-2 Gene Regulated by the Thy-1.2 Promoter  

OpenAIRE

Channelrhodopsin-2 (ChR2), one of the archea-type rhodopsins from green algae, is a potentially useful optogenetic tool for restoring vision in patients with photoreceptor degeneration, such as retinitis pigmentosa. If the ChR2 gene is transferred to retinal ganglion cells (RGCs), which send visual information to the brain, the RGCs may be repurposed to act as photoreceptors. In this study, by using a transgenic rat expressing ChR2 specifically in the RGCs under the regulation of a Thy-1.2 pr...

Tomita, Hiroshi; Sugano, Eriko; Fukazawa, Yugo; Isago, Hitomi; Sugiyama, Yuka; Hiroi, Teru; Ishizuka, Toru; Mushiake, Hajime; Kato, Megumi; Hirabayashi, Masumi; Shigemoto, Ryuichi; Yawo, Hiromu; Tamai, Makoto

2009-01-01

112

Impact of Age at Administration, Lysosomal Storage, and Transgene Regulatory Elements on AAV2/8-Mediated Rat Liver Transduction  

Science.gov (United States)

Liver-directed gene transfer is being investigated for the treatment of systemic or liver-specific diseases. Recombinant vectors based on adeno-associated virus serotype 8 (AAV2/8) efficiently transduce liver cells allowing long term transgene expression after a single administration in animal models and in patients. We evaluated the impact on AAV2/8-mediated rat liver transduction of the following variables: i) age at vector administration, ii) presence of lysosomal storage in liver cells, and iii) regulatory elements included in the transgene expression cassette. We found that systemic administration of AAV2/8 to newborn rats results in vector genome dilution and reduced transduction efficacy when compared to adult injected animals, presumably due to hepatocyte proliferation. Accumulation of glycosaminoglycans in lysosomes does not impact on levels and distribution of AAV2/8-mediated liver transduction. Transgene expression occurs in hepatocytes but not in Kupffer or liver endothelial cells when the liver-specific thyroxine-binding-globulin promoter is used. However, extra-hepatic transduction is observed in the spleen and kidney of animals injected at birth. The use of target sequences for the hematopoietic-specific microRNA miR142-3p does not improve liver transduction efficacy neither reduce immune responses to the lysosomal enzyme arylsulfatase B. The inclusion of a variant of the Woodchuck hepatitis virus post-transcriptional regulatory element (WPRE-m) decreases AAV2/8-mediated liver transduction levels. As AAV2/8-mediated liver gene transfer is entering in the clinical arena, these data will provide relevant information to the design of efficient AAV2/8-based therapeutic strategies. PMID:22428010

Cotugno, Gabriella; Annunziata, Patrizia; Barone, Maria Vittoria; Karali, Marianthi; Banfi, Sandro; Auricchio, Alberto

2012-01-01

113

Impact of age at administration, lysosomal storage, and transgene regulatory elements on AAV2/8-mediated rat liver transduction.  

Science.gov (United States)

Liver-directed gene transfer is being investigated for the treatment of systemic or liver-specific diseases. Recombinant vectors based on adeno-associated virus serotype 8 (AAV2/8) efficiently transduce liver cells allowing long term transgene expression after a single administration in animal models and in patients.We evaluated the impact on AAV2/8-mediated rat liver transduction of the following variables: i) age at vector administration, ii) presence of lysosomal storage in liver cells, and iii) regulatory elements included in the transgene expression cassette. We found that systemic administration of AAV2/8 to newborn rats results in vector genome dilution and reduced transduction efficacy when compared to adult injected animals, presumably due to hepatocyte proliferation. Accumulation of glycosaminoglycans in lysosomes does not impact on levels and distribution of AAV2/8-mediated liver transduction. Transgene expression occurs in hepatocytes but not in Kupffer or liver endothelial cells when the liver-specific thyroxine-binding-globulin promoter is used. However, extra-hepatic transduction is observed in the spleen and kidney of animals injected at birth. The use of target sequences for the hematopoietic-specific microRNA miR142-3p does not improve liver transduction efficacy neither reduce immune responses to the lysosomal enzyme arylsulfatase B. The inclusion of a variant of the Woodchuck hepatitis virus post-transcriptional regulatory element (WPRE-m) decreases AAV2/8-mediated liver transduction levels.As AAV2/8-mediated liver gene transfer is entering in the clinical arena, these data will provide relevant information to the design of efficient AAV2/8-based therapeutic strategies. PMID:22428010

Cotugno, Gabriella; Annunziata, Patrizia; Barone, Maria Vittoria; Karali, Marianthi; Banfi, Sandro; Auricchio, Alberto

2012-01-01

114

Characterization of dsRed2-positive cells in the doublecortin-dsRed2 transgenic adult rat retina.  

Science.gov (United States)

Doublecortin (DCX) is predominantly expressed in neuronal precursor cells and young immature neurons of the developing and adult brain, where it is involved in neuronal differentiation, migration and plasticity. Moreover, its expression pattern reflects neurogenesis, and transgenic DCX promoter-driven reporter models have been previously used to investigate adult neurogenesis. In this study, we characterize dsRed2 reporter protein-expressing cells in the adult retina of the transgenic DCX promoter-dsRed2 rat model, with the aim to identify cells with putative neurogenic activity. Additionally, we confirmed the expression of the dsRed2 protein in DCX-expressing cells in the adult hippocampal dentate gyrus. Adult DCX-dsRed2 rat retinas were analyzed by immunohistochemistry for expression of DCX, NF200, Brn3a, Sox2, NeuN, calbindin, calretinin, PKC-a, Otx2, ChAT, PSA-NCAM and the glial markers GFAP and CRALBP, followed by confocal laser-scanning microscopy. In addition, brain sections of transgenic rats were analyzed for dsRed2 expression and co-localization with DCX, NeuN, GFAP and Sox2 in the cortex and dentate gyrus. Endogenous DCX expression in the adult retina was confined to horizontal cells, and these cells co-expressed the DCX promoter-driven dsRed2 reporter protein. In addition, we encountered dsRed2 expression in various other cell types in the retina: retinal ganglion cells (RGCs), a subpopulation of amacrine cells, a minority of bipolar cells and in perivascular cells. Since also RGCs expressed dsRed2, the DCX-dsRed2 rat model might offer a useful tool to study RGCs in vivo under various conditions. Müller glial cells, which have previously been identified as cells with stem cell features and with neurogenic potential, did express neither endogenous DCX nor the dsRed2 reporter. However, and surprisingly, we identified a perivascular glial cell type expressing the dsRed2 reporter, enmeshed with the glia/stem cell marker GFAP and colocalizing with the neural stem cell marker Sox2. These findings suggest the so far undiscovered existence of perivascular associated cell with neural stem cell-like properties in the adult retina. PMID:25138677

Trost, A; Schroedl, F; Marschallinger, J; Rivera, F J; Bogner, B; Runge, C; Couillard-Despres, S; Aigner, L; Reitsamer, H A

2014-12-01

115

A Vitamin A Deficient Diet Enhances Proinflammatory Cytokine, Mu Opioid Receptor, and HIV-1 Expression in the HIV-1 Transgenic Rat  

OpenAIRE

The HIV-1 (HIV) transgenic (Tg) rat develops several immune abnormalities in association with clinical impairments that are similar to what are seen with HIV infection in humans. In HIV infection, retinoids and opioids can have separate and potentially combined effects on the clinical course of HIV disease. In these studies, the effects of a vitamin A deficient diet on T cell proinflammatory cytokine and mu opioid receptor (MOR) expression were examined in the Tg and in wild-type (WT) rats. T...

Royal, Walter; III; Wang, Huiyun; Jones, Odell; Tran, Hieu; Bryant, Joseph L.

2007-01-01

116

The relationship of photoreceptor degeneration to retinal vascular development and loss in mutant rhodopsin transgenic and RCS rats.  

Science.gov (United States)

The early loss of photoreceptors in some retinal degenerations in mice has been shown to have a profound effect on vascular development of the retina. To better characterize this relationship, we have examined the formation of retinal blood vessels during the first month of life in 8 lines of transgenic rats with different ages of onset and rates of photoreceptor cell loss mediated by the expression of mutant rhodopsin (P23H and S334ter). The number of capillary profiles in the superficial plexus (SP) and deep capillary plexus (DCP) of the retina were quantified in retinal sections taken at postnatal day (P) 8, 10, 12, 15 and 30. In normal wild-type rats, the SP and DCP had mostly established mature, adult patterns by P15, as previously shown. In the transgenic rats, the loss of photoreceptors had relatively little effect on the SP. By contrast, the loss of photoreceptors during vascular development had a major impact on the DCP. In the two lines with early and most rapid photoreceptor loss, S334ter-7 and S334ter-3, where about 90% and 65%, respectively, of the photoreceptors were already lost by P15, the DCP either failed to form (S334ter-7) or the number of capillary profiles was less than 7% of controls (S334ter-3). In lines where almost all photoreceptors were still present at P15 (S334ter-4, S334ter-9, P23H-2 and P23H-3), the number of profiles in the DCP were the same as in wild-type controls at P30. In two lines with an intermediate rate of degeneration (S334ter-5 and P23H-1), where only about 25% of the photoreceptors were lost by P15, there was an intermediate number of vascular profiles in the DCP at P30. Thus, a very close relationship between the number of photoreceptors and vessel profiles in the DCP during its development exists in the transgenic rats, and the loss of photoreceptors results in the failure or inhibition of the DCP to develop. Several mechanisms may explain this relationship including changes in the level of physiological oxygen tension or alteration in the release of angiogenic factors that normally drive vessel development. Analysis of older transgenic retinas up to 1 year of age revealed that (1) vascular profiles are lost from the DCP in essentially all lines once fewer than about 30-33% of photoreceptors remain; (2) in those lines where the DCP essentially did not develop (S334ter-7 and S334ter-3), the effect of photoreceptor absence was permanent, and there was no late vascularization of the DCP; (3) the number of capillary profiles in the SP remained no different from controls in any of the lines, despite long-standing loss of photoreceptors; and (4) neovascularization of the RPE by retinal capillaries occurred with a latency of 60-180 days after the loss of photoreceptors, except in S334ter-7 rats, where neovascularization essentially did not occur. Analysis of RCS rats was carried out for comparison. PMID:18848932

Pennesi, Mark E; Nishikawa, Shimpei; Matthes, Michael T; Yasumura, Douglas; LaVail, Matthew M

2008-12-01

117

Sterol regulatory element binding protein 2 overexpression is associated with reduced adipogenesis and ectopic fat accumulation in transgenic spontaneously hypertensive rats.  

Czech Academy of Sciences Publication Activity Database

Ro?. 63, ?. 5 (2014), s. 587-590. ISSN 0862-8408 R&D Projects: GA MŠk(CZ) LH12061 Institutional support: RVO:67985823 Keywords : sterol regulatory element binding protein 2 * transgenic * spontaneously hypertensive rat * lipid metabolism Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 1.487, year: 2013

Landa, Vladimír; Zídek, Václav; Mlejnek, Petr; Šimáková, Miroslava; Šilhavý, Jan; Trnovská, J.; Kazdová, L.; Pravenec, Michal

2014-01-01

118

Benzo[a]pyrene-enhanced mutagenesis by man-made mineral fibres in the lung of gama-lacI transgenic rats.  

Czech Academy of Sciences Publication Activity Database

Ro?. 595, - (2006), s. 167-173. ISSN 0027-5107 Institutional research plan: CEZ:AV0Z50390512 Keywords : transgenic rats * mineral fibres * mutations Subject RIV: DN - Health Impact of the Environment Quality Impact factor: 4.111, year: 2006

Topinka, Jan; Loli, P.; Hurbánková, M.; Ková?iková, Z.; Volkovová, K.; Wolff, T.; Oesterle, D.; Kyrtopoulos, S.A.; Georgiadis, P.

2006-01-01

119

Dual, HLA-B27 Subtype-dependent Conformation of a Self-peptide  

OpenAIRE

The products of the human leukocyte antigen subtypes HLA-B*2705 and HLA-B*2709 differ only in residue 116 (Asp vs. His) within the peptide binding groove but are differentially associated with the autoimmune disease ankylosing spondylitis (AS); HLA-B*2705 occurs in AS-patients, whereas HLA-B*2709 does not. The subtypes also generate differential T cell repertoires as exemplified by distinct T cell responses against the self-peptide pVIPR (RRKWRRWHL). The crystal structures described here show...

Hu?lsmeyer, Martin; Fiorillo, Maria Teresa; Bettosini, Francesca; Sorrentino, Rosa; Saenger, Wolfram; Ziegler, Andreas; Uchanska-ziegler, Barbara

2004-01-01

120

Is There an HLA-B27 Associated Peptide Responsible for Ankylosin Spondylitis.  

Czech Academy of Sciences Publication Activity Database

Praha : Verlag, 2006, s. 96-96. [International Mass Spectrometry Conference /17./. Prague (CZ), 27.08.2006-01.09.2006] R&D Projects: GA MZd 1A8631 Institutional research plan: CEZ:AV0Z50200510 Keywords : biomarkers * fourier transform * mass spectrometry Subject RIV: EE - Microbiology, Virology

Novák, Petr; Man, Petr; Stod?lková, Eva; Ivašková, E.; Flieger, Miroslav

121

A progressive dopaminergic phenotype associated with neurotoxic conversion of ?-synuclein in BAC-transgenic rats  

OpenAIRE

Conversion of soluble ?-synuclein into insoluble and fibrillar inclusions is a hallmark of Parkinson’s disease and other synucleinopathies. Accumulating evidence points towards a relationship between its generation at nerve terminals and structural synaptic pathology. Little is known about the pathogenic impact of ?-synuclein conversion and deposition at nigrostriatal dopaminergic synapses in transgenic mice, mainly owing to expression limitations of the ?-synuclein constr...

Nuber, Silke; Harmuth, Florian; Kohl, Zacharias; Adame, Anthony; Trejo, Margaritha; Scho?nig, Kai; Zimmermann, Frank; Bauer, Claudia; Casadei, Nicolas; Giel, Christiane; Calaminus, Carsten; Pichler, Bernd J.; Jensen, Poul H.; Mu?ller, Christian P.; Amato, Davide

2013-01-01

122

Role of HIV-1 Infection in Addictive Behavior: A Study of a HIV-1 Transgenic Rat Model  

Directory of Open Access Journals (Sweden)

Full Text Available Epidemiological research indicates that drug abuse is prevalent among individuals infected with HIV-1. Evidence from preclinical research also suggests that drugs of abuse exacerbate the progression of neuropathological changes in the HIV-1 infected brain probably through common mechanisms of neuronal injury. The effects of HIV-1 on the efficacy and abuse potential of controlled drugs such as morphine, however, has not been explored. The current study reports that the noninfectious HIV-1 transgenic (HIV-1 Tg rat shows up-regulated expression of the mu opioid receptor (MOR at the transcriptional level and functional supersensitivity to morphine, a MOR agonist. Compared to nontransgenic control rats, the HIV-1 Tg rats also show greater motivation to run in a wheel, a behavior that is known to be associated with increased drug self-administration. These results suggest the potential role of HIV-1 infection in enhancing vulnerability to addiction and this possibility warrants further investigation to better understand the link between HIV-1 infection and the abuse of drugs including opioids.

Sulie L. Chang

2006-01-01

123

Inducible and reversible gene silencing by stable integration of an shRNA-encoding lentivirus in transgenic rats.  

Science.gov (United States)

Currently, tools to generate loss-of-function mutations in rats are limited. Therefore, we have developed a lentiviral single-vector system for the temporal control of ubiquitous shRNA expression. Here, we report transgenic rats carrying an insulin receptor-specific shRNA transcribed from a regulatable promoter and identified by concomitant EGFP expression. In the absence of the inducer doxycycline (Dox), we observed no siRNA expression. However, Dox treatment at very low concentrations led to a rapid induction of the siRNA and ablation of INSR protein expression. As anticipated, blood glucose levels increased, whereas insulin signaling and glucose regulation were impaired. Importantly, this phenotype was reversible (i.e., discontinuation of Dox treatment led to INSR re-expression and remission of diabetes symptoms). The lentiviral system offers a simple tool for reversible gene ablation in the rat and can be used for other species that cannot be manipulated by conventional recombination techniques. PMID:19017805

Herold, Marco J; van den Brandt, Jens; Seibler, Jost; Reichardt, Holger M

2008-11-25

124

In Vivo Expression of Angiotensin-(1-7) Lowers Blood Pressure and Improves Baroreflex Function in Transgenic (mRen2)27 Rats  

OpenAIRE

Transgenic (mRen2)27 rats are hypertensive with impaired baroreflex sensitivity for control of heart rate compared to Hannover Sprague-Dawley rats. We assessed blood pressure and baroreflex function in male hemizygous (mRen2)27 rats (30-40 wks of age) instrumented for arterial pressure recordings and receiving into the cisterna magna either an Ang-(1-7) fusion protein or a control fusion protein (CTL-FP). The maximum reduction in mean arterial pressure achieved was -38 ± 7 mm Hg on day 3, ac...

Garcia-espinosa, Maria A.; Shaltout, Hossam A.; Gallagher, Patricia E.; Chappell, Mark C.; Diz, Debra I.

2012-01-01

125

Mineralocorticoid receptor antagonism attenuates glomerular filtration barrier remodeling in the transgenic Ren2 rat  

OpenAIRE

Recent evidence suggests that mineralocorticoid receptor (MR) antagonism has beneficial effects on tissue oxidative stress and insulin metabolic signaling as well as reducing proteinuria. However, the mechanisms by which MR antagonism corrects both renin-angiotensin-aldosterone system (RAAS) impairments in renal insulin metabolic signaling and filtration barrier/podocyte injury remain unknown. To explore this potential beneficial interactive effect of MR antagonism we used young transgenic (m...

Whaley-connell, Adam; Habibi, Javad; Wei, Yongzhong; Gutweiler, Alex; Jellison, Jessica; Wiedmeyer, Charles E.; Ferrario, Carlos M.; Sowers, James R.

2009-01-01

126

Primary T-cells from human CD4/CCR5-transgenic rats support all early steps of HIV-1 replication including integration, but display impaired viral gene expression  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background In vivo studies on HIV-1 pathogenesis and testing of antiviral strategies have been hampered by the lack of an immunocompetent small animal model that is highly susceptible to HIV-1 infection. Since native rodents are non-permissive, we developed transgenic rats that selectively express the HIV-1 receptor complex, hCD4 and hCCR5, on relevant target cells. These animals display a transient low-level plasma viremia after HIV-1YU-2 infection, demonstrating HIV-1 susceptibility in vivo. However, unlike macrophages, primary CD4 T-cells from double-transgenic animals fail to support viral spread ex vivo. To identify quantitative limitations or absolute blocks in this rodent species, we quantitatively assessed the efficiency of key steps in the early phase of the viral replication cycle in a side-by-side comparison in infected cell lines and primary T-cells from hCD4/hCCR5-transgenic rats and human donors. Results Levels of virus entry, HIV-1 cDNA synthesis, nuclear import, and integration into the host genome were shown to be remarkably similar in cell lines and, where technically accessible, in primary T-cells from both species. In contrast, a profound impairment at the level of early HIV gene expression was disclosed at the single-cell level in primary rat T-cells and most other rat-derived cells. Macrophages were a notable exception, possibly reflecting the unique transcriptional milieu in this evolutionarily conserved target cell of all lentiviruses. Importantly, transient trans-complementation by ex vivo nucleofection with the Tat-interacting protein Cyclin T1 of human origin markedly elevated HIV gene expression in primary rat T-cells. Conclusion This is the first study that has quantitatively determined the efficiency of consecutive steps in the HIV-1 replication cycle in infected primary HIV target cells from a candidate transgenic small animal and compared it to human cells. Unlike cells derived from mice or rabbits, rat cells complete all of the early steps in the HIV-1 replication cycle, including provirus integration in vivo, with high efficiency. A deficiency in gene expression was disclosed at the single cell level and could be counteracted by the human pTEFb transcription complex factor Cyclin T1. Collectively, these results provide the basis for the advancement of this transgenic rat model through strategies aimed at boosting HIV-1 gene expression in primary rat CD4 T-cells, including human Cyclin T1 transgenesis.

Hermann Volker

2007-07-01

127

Remyelination of spinal cord axons by olfactory ensheathing cells and Schwann cells derived from a transgenic rat expressing alkaline phosphatase marker gene  

OpenAIRE

Transplantation of cell suspensions containing olfactory ensheathing cells (OECs) has been reported to remyelinate demyelinated axons in the spinal cord with a Schwann cell (SC)-like pattern of myelination. However, questions have been raised recently as to whether OECs can form SC-like myelin. To address this issue we prepared SCs and OECs from transgenic rats in which a marker gene, human placental alkaline phosphatase (hPAP), is linked to the ubiquitously active promoter of the R26 gene. S...

Akiyama, Yukinori; Lankford, Karen; Radtke, Christine; Greer, Charles A.; Kocsis, Jeffery D.

2004-01-01

128

Combined inhibition of 20-hydroxyeicosatetraenoic acid formation and of epoxyeicosatrienoic acids degradation attenuates hypertension and hypertension-induced end-organ damage in Ren-2 transgenic rats  

OpenAIRE

Abstract Recent studies have shown that the renal cytochrome P-450 metabolites of arachidonic acid: the vasoconstrictor 20-hydroxyeicosatetraenoic acid (20-HETE), and the vasodilator epoxyeicosatrienoic acids (EETs) play an important role in the pathophysiology of angiotensin II (ANG II)-dependent forms of hypertension and the associated target organ damage. The present studies were performed in Ren-2 renin transgenic rats (TGR) to evaluate the effects of chronic selective inhibiti...

C?erti?kova? Cha?bova?, Ve?ra; Walkowska, Agnieszka; Kompanowska-jezierska, Elzbieta; Sadowski, Janusz; Kujal, Peter; Vernerova?, Zdena; Van?ourkova?, Zdenka; Kopkan, Libor; Kramer, Herbert J.; Falck, John R.; Imig, John D.; Hammock, Bruce D.; Vane?c?kova?, Ivana; C?ervenka, Lude?k

2010-01-01

129

Fumaric Acid Esters Can Block Pro-Inflammatory Actions of Human CRP and Ameliorate Metabolic Disturbances in Transgenic Spontaneously Hypertensive Rats  

OpenAIRE

Inflammation and oxidative stress have been implicated in the pathogenesis of metabolic disturbances. Esters of fumaric acid, mainly dimethyl fumarate, exhibit immunomodulatory, anti-inflammatory, and anti-oxidative effects. In the current study, we tested the hypothesis that fumaric acid ester (FAE) treatment of an animal model of inflammation and metabolic syndrome, the spontaneously hypertensive rat transgenically expressing human C-reactive protein (SHR-CRP), will ameliorate inflammation,...

S?ilhavy?, Jan; Zi?dek, Va?clav; Mlejnek, Petr; Landa, Vladimi?r; S?ima?kova?, Miroslava; Strnad, Hynek; Oliyarnyk, Olena; S?kop, Vojte?ch; Kazdova?, Ludmila; Kurtz, Theodore; Pravenec, Michal

2014-01-01

130

Fumaric Acid Esters Can Block Pro-Inflammatory Actions of Human CRP and Ameliorate Metabolic Disturbances in Transgenic Spontaneously Hypertensive Rats.  

Czech Academy of Sciences Publication Activity Database

Ro?. 9, ?. 7 (2014), e101906. ISSN 1932-6203 R&D Projects: GA MZd(CZ) NT14325; GA MŠk(CZ) LH12061; GA MŠk(CZ) LL1204 Institutional support: RVO:67985823 ; RVO:68378050 Keywords : fumaric acid esters * C-reactive protein * transgenic * spontaneously hypertensive rat Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.534, year: 2013

Šilhavý, Jan; Zídek, Václav; Mlejnek, Petr; Landa, Vladimír; Šimáková, Miroslava; Strnad, Hynek; Oliyarnyk, O.; Škop, V.; Kazdová, L.; Kurtz, T.; Pravenec, Michal

2014-01-01

131

Comparative effect of direct renin inhibition and AT1R blockade on glomerular filtration barrier injury in the transgenic Ren2 rat  

OpenAIRE

Renin-angiotensin system (RAS) activation contributes to kidney injury through oxidative stress. Renin is the rate-limiting step in angiotensin (ANG II) generation. Recent work suggests renin inhibition improves proteinuria comparable to ANG type 1 receptor (AT1R) blockade (ARB). Thereby, we investigated the relative impact of treatment with a renin inhibitor vs. an ARB on renal oxidative stress and associated glomerular structural and functional changes in the transgenic Ren2 rat, which mani...

Whaley-connell, Adam; Nistala, Ravi; Habibi, Javad; Hayden, Melvin R.; Schneider, Rebecca I.; Johnson, Megan S.; Tilmon, Roger; Rehmer, Nathan; Ferrario, Carlos M.; Sowers, James R.

2009-01-01

132

Plasmid-based genetic modification of human bone marrow-derived stromal cells: analysis of cell survival and transgene expression after transplantation in rat spinal cord  

OpenAIRE

Abstract Background Bone marrow-derived stromal cells (MSC) are attractive targets for ex vivo cell and gene therapy. In this context, we investigated the feasibility of a plasmid-based strategy for genetic modification of human (h)MSC with enhanced green fluorescent protein (EGFP) and neurotrophin (NT)3. Three genetically modified hMSC lines (EGFP, NT3, NT3-EGFP) were established and used to study cell survival and transgene expression following transplantation in rat spinal cord. Results Fi...

Fi, Tendeloo Viggo; Haese Patrick, D.; Vermeulen Katrien; Chatterjee Shyama; Spaepen Gie; Daans Jasmijn; Ronsyn Mark W; Van Marck Eric; Ysebaert Dirk; Berneman Zwi N; Jorens Philippe G; Ponsaerts Peter

2007-01-01

133

Imatinib ameliorates renal morphological changes in Cyp1a1-Ren2 transgenic rats with inducible ANG II-dependent malignant hypertension  

OpenAIRE

The present study was performed to assess the effects of the platelet-derived growth factor (PDGF) receptor kinase inhibitor imatinib mesylate on the renal morphological changes occurring during the development of malignant hypertension in transgenic rats with inducible expression of the Ren2 gene [TGR(Cyp1a1Ren2)]. Arterial blood pressure was measured by radiotelemetry in male Cyp1a1-Ren2 rats during control conditions and during dietary administration of indole-3-carbinol (I3C; 0.3%) for 14...

Graciano, Miguel L.; Mitchell, Kenneth D.

2011-01-01

134

Cardiac remodeling during and after renin-angiotensin system stimulation in Cyp1a1-Ren2 transgenic rats  

DEFF Research Database (Denmark)

This study investigated renin-angiotensin system (RAS)-induced cardiac remodeling and its reversibility in the presence and absence of high blood pressure (BP) in Cyp1a1-Ren2 transgenic inducible hypertensive rats (IHR). In IHR (pro)renin levels and BP can be dose-dependently titrated by oral administration of indole-3-carbinol (I3C). Young (four-weeks old) and adult (30-weeks old) IHR were fed I3C for four weeks (leading to systolic BP >200 mmHg). RAS-stimulation was stopped and animals were followed-up for a consecutive period. Cardiac function and geometry was determined echocardiographically and the hearts were excised for molecular and immunohistochemical analyses. Echocardiographic studies revealed that four weeks of RAS-stimulation incited a cardiac remodeling process characterized by increased left ventricular (LV) wall thickness, decreased LV volumes, and shortening of the left ventricle. Hypertrophic genes were highly upregulated, whereas in substantial activation a fibrotic response was absent. Four weeks after withdrawal of I3C, (pro)renin levels were normalized in all IHR. While in adult IHR BP returned to normal, hypertension was sustained in young IHR. Despite the latter, myocardial hypertrophy was fully regressed in both young and adult IHR. We conclude that (pro)renin-induced severe hypertension in IHR causes an age-independent fully reversible myocardial concentric hypertrophic remodeling, despite a continued elevated BP in young IHR.

Heijnen, Bart Fj; Pelkmans, Leonie Pj

2013-01-01

135

TRANSGENE-MEDIATED EXPRESSION OF TUMOR NECROSIS FACTOR SOLUBLE RECEPTOR ATTENUATES MORPHINE TOLERANCE IN RATS  

OpenAIRE

Opiate/narcotic analgesics are the most effective treatments for chronic severe pain, but their clinical utility is often hampered by the development of analgesic tolerance. Recent evidence suggests chronic morphine may activate glial cells to release proinflammatory cytokines. In this study, we used herpes simplex virus (HSV) vectors-based gene transfer to dorsal root ganglion to produce a local release of p55 TNF soluble receptor in the spinal cord in rats with morphine tolerance. Subcutane...

Sun, Jingxia; Liu, Shue; Mata, Marina; Fink, David J.; Hao, Shuanglin

2011-01-01

136

Transgene-mediated expression of tumor necrosis factor soluble receptor attenuates morphine tolerance in rats.  

Science.gov (United States)

Opiate/narcotic analgesics are the most effective treatments for chronic severe pain, but their clinical utility is often hampered by the development of analgesic tolerance. Recent evidence suggests chronic morphine may activate glial cells to release proinflammatory cytokines. In this study, we used herpes simplex virus (HSV) vector-based gene transfer to dorsal root ganglion to produce a local release of p55 tumor necrosis factor (TNF) soluble receptor in the spinal cord in rats with morphine tolerance. Subcutaneous inoculation of HSV vectors expressing p55 TNF soluble receptor into the plantar surface of the hindpaws enhanced the antinociceptive effect of acute morphine in rats. Subcutaneous inoculation of those vectors into hindpaws also delayed the development of chronic morphine tolerance in rats. TNF soluble receptor expressed by HSV vector reduced gene transcription of spinal TNF? and interleukin-1? (IL-1?) induced by repeated morphine. Furthermore, we found that TNF soluble receptor mediated by HSV reversed the upregulation of protein level of TNF? and IL-1? and phosphorylation of p38 mitogen-activated protein kinase induced by repeated morphine. These results support the concept that proinflammatory cytokines may have an important role in the pathogenesis induced by morphine. This study provides a novel approach to treating morphine tolerance. PMID:21614028

Sun, J; Liu, S; Mata, M; Fink, D J; Hao, S

2012-01-01

137

Pulsatile luteinizing hormone and follicle-stimulating hormone secretion and gonadotropin subunit mRNA levels in the ovariectomized GPR-4 transgenic rat.  

Science.gov (United States)

Genetic targeting of the cAMP-specific phosphodiesterase 4D1 (PDE4D1) to gonadotropin-releasing hormone (GnRH) neurons in the GPR-4 transgenic rat resulted in decreased luteinizing hormone (LH) pulse frequency in castrated female and male rats. A similar decrease in the intrinsic GnRH pulse frequency was observed in GT1 GnRH cells expressing the PDE4D1 phosphodiesterase. We have extended these findings in ovariectomized (OVX) GPR-4 rats by asking what effect transgene expression had on pulsatile LH and follicle-stimulating hormone (FSH) secretion, plasma and pituitary levels of LH and FSH, and levels of the alpha-glycoprotein hormone subunit (alpha-GSU), LH-beta and FSH-beta subunit mRNAs. In OVX GPR-4 rats the LH pulse frequency but not pulse amplitude was decreased by 50% compared to wild-type littermate controls. Assaying the same samples for FSH, the FSH pulse frequency and amplitude were unchanged. The plasma and anterior pituitary levels of LH in the GPR-4 rats were significantly decreased by approximately 45%, while the plasma but not anterior pituitary level of FSH was significantly decreased by 25%. As measured by real-time RT-PCR, the mRNA levels for the alpha-GSU in the GPR-4 rats were significantly decreased by 41%, the LH-beta subunit by 38% and the FSH-beta subunit by 28%. We conclude that in the castrated female GPR-4 rats the decreased GnRH pulse frequency results in decreased levels of LH and FSH and in the alpha- and beta-subunit mRNA levels. PMID:14688441

El Majdoubi, Mohammed; Paruthiyil, Sreenivasan; Weiner, Richard I

2003-12-01

138

Different mechanisms of acute versus long-term antihypertensive effects of soluble epoxide hydrolase inhibition: studies in Cyp1a1-Ren-2 transgenic rats.  

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Recent studies have shown that the long-term antihypertensive action of soluble epoxide hydrolase inhibition (sEH) in angiotensin-II (AngII)-dependent hypertension might be mediated by the suppression of intrarenal AngII levels. To test this hypothesis, we examined the effects of acute (2 days) and chronic (14 days) sEH inhibition on blood pressure (BP) in transgenic rats with inducible AngII-dependent hypertension. AngII-dependent malignant hypertension was induced by 10 days' dietary administration of indole-3-carbinol (I3C), a natural xenobiotic that activates the mouse renin gene in Cyp1a1-Ren-2 transgenic rats. BP was monitored by radiotelemetry. Acute and chronic sEH inhibition was achieved using cis-4-(4-(3-adamantan-1-yl-ureido)cyclohexyloxy) benzoic acid, given at doses of 0.3, 3, 13, 26, 60 and 130 mg/L in drinking water. At the end of experiments, renal concentrations of epoxyeicosatrienoic acids, their inactive metabolites dihydroxyeicosatrienoic acids and AngII were measured. Acute BP-lowering effects of sEH inhibition in I3C-induced rats was associated with a marked increase in renal epoxyeicosatrienoic acids to dihydroxyeicosatrienoic acids ratio and acute natriuresis. Chronic treatment with cis-4-(4-(3-adamantan-1-yl-ureido)cyclohexyloxy) benzoic acid in I3C-induced rats elicited dose-dependent persistent BP lowering associated with a significant reduction of plasma and kidney AngII levels. Our findings show that the acute BP-lowering effect of sEH inhibition in I3C-induced Cyp1a1-Ren-2 transgenic rats is mediated by a substantial increase in intrarenal epoxyeicosatrienoic acids and their natriuretic action without altering intrarenal renin-angiotensin system activity. Long-term antihypertensive action of cis-4-(4-(3-adamantan-1-yl-ureido)cyclohexyloxy) benzoic acid in I3C-induced Cyp1a1-Ren-2 transgenic rats is mediated mostly by suppression of intrarenal AngII concentration. PMID:25224811

Sporková, Alexandra; Jíchová, Sárka; Husková, Zuzana; Kopkan, Libor; Nishiyama, Akira; Hwang, Sung H; Hammock, Bruce D; Imig, John D; Kompanowska-Jezierska, Elzbieta; Sadowski, Janusz; Kramer, Herbert J; Cervenka, Lud?k

2014-12-01

139

Stable expression of the alkaline phosphatase marker gene by neural cells in culture and after transplantation into the CNS using cells derived from a transgenic rat.  

Science.gov (United States)

Multipotent stem cells and more developmentally restricted precursors have previously been isolated from the developing nervous system and their properties analyzed by culture assays in vitro and by transplantation in vivo. However, the variety of labeling techniques that have been used to identify grafted cells in vivo have been unsatisfactory. In this article we describe the characteristics of cells isolated from a transgenic rat in which the marker gene human placental alkaline phosphatase (hPAP) is linked to the ubiquitously active R26 gene promoter. We show that hPAP is readily detected in embryonic neuroepithelial stem cells, neuronal-restricted precursor cells, and glial-restricted precursor cells. Transgene expression is robust and can be detected by both immunocytochemistry and histochemistry. Furthermore, the levels of hPAP on the cell surface are sufficient for live cell labeling and fluorescence-activated cell sorting. Expression of hPAP is stable in isolated cells in culture and in cells transplanted into the spinal cord for at least 1 month. We submit that cells isolated from this transgenic rat will be valuable for studies of neural development and regeneration. PMID:11869033

Mujtaba, Tahmina; Han, Steve S W; Fischer, Itzhak; Sandgren, Eric P; Rao, Mahendra S

2002-03-01

140

Plasmid-based genetic modification of human bone marrow-derived stromal cells: analysis of cell survival and transgene expression after transplantation in rat spinal cord  

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Full Text Available Abstract Background Bone marrow-derived stromal cells (MSC are attractive targets for ex vivo cell and gene therapy. In this context, we investigated the feasibility of a plasmid-based strategy for genetic modification of human (hMSC with enhanced green fluorescent protein (EGFP and neurotrophin (NT3. Three genetically modified hMSC lines (EGFP, NT3, NT3-EGFP were established and used to study cell survival and transgene expression following transplantation in rat spinal cord. Results First, we demonstrate long-term survival of transplanted hMSC-EGFP cells in rat spinal cord under, but not without, appropriate immune suppression. Next, we examined the stability of EGFP or NT3 transgene expression following transplantation of hMSC-EGFP, hMSC-NT3 and hMSC-NT3-EGFP in rat spinal cord. While in vivo EGFP mRNA and protein expression by transplanted hMSC-EGFP cells was readily detectable at different time points post-transplantation, in vivo NT3 mRNA expression by hMSC-NT3 cells and in vivo EGFP protein expression by hMSC-NT3-EGFP cells was, respectively, undetectable or declined rapidly between day 1 and 7 post-transplantation. Further investigation revealed that the observed in vivo decline of EGFP protein expression by hMSC-NT3-EGFP cells: (i was associated with a decrease in transgenic NT3-EGFP mRNA expression as suggested following laser capture micro-dissection analysis of hMSC-NT3-EGFP cell transplants at day 1 and day 7 post-transplantation, (ii did not occur when hMSC-NT3-EGFP cells were transplanted subcutaneously, and (iii was reversed upon re-establishment of hMSC-NT3-EGFP cell cultures at 2 weeks post-transplantation. Finally, because we observed a slowly progressing tumour growth following transplantation of all our hMSC cell transplants, we here demonstrate that omitting immune suppressive therapy is sufficient to prevent further tumour growth and to eradicate malignant xenogeneic cell transplants. Conclusion In this study, we demonstrate that genetically modified hMSC lines can survive in healthy rat spinal cord over at least 3 weeks by using adequate immune suppression and can serve as vehicles for transgene expression. However, before genetically modified hMSC can potentially be used in a clinical setting to treat spinal cord injuries, more research on standardisation of hMSC culture and genetic modification needs to be done in order to prevent tumour formation and transgene silencing in vivo.

Van Tendeloo Viggo FI

2007-12-01

141

Increased neuronal activity in the OVLT of Cyp1a1-Ren2 transgenic rats with inducible Ang II-dependent malignant hypertension.  

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The contribution of angiotensin II (Ang II) to the pathophysiology of hypertension is established based on facts that high levels of circulating Ang II increase vasoconstriction of peripheral arteries causing a rise in blood pressure (BP). In addition, circulating Ang II has various effects on the central nervous system, including the osmosensitive neurons in the organum vasculosum of the lamina terminalis (OVLT). Osmosensitive neurons in the OVLT transduce hypertonicity via the activation of the nonselective cation channel known as transient receptor potential vanilloid 1 (TRPV1), causing membrane depolarization, followed by increased action potential discharge. This effect is absent in mice lacking expression of the TRPV1 gene. Most observations related to the importance of the OVLT in cardiovascular control are mainly based on models of lesion of the entire preoptic periventricular tissue. However, it remains unclear whether neuronal activity and TRPV1 protein expression levels alter in the OVLT of Cyp1a1-Ren2 transgenic rats with inducible Ang II-dependent malignant hypertension. C-fos was used as a marker of neuronal activity. Immunostaining was used to demonstrate distribution of c-fos positive neurons in the OVLT of Cyp1a1Ren2 transgenic rats. Western blot analysis showed increased c-fos and TRPV1 total protein expression levels in the OVLT of hypertensive rats. The present findings demonstrate increased c-fos and TRPV1 expression levels in the OVLT of Cyp1a1-Ren2 transgenic rats with Ang II-dependent malignant hypertension. PMID:22579820

Issa, Alexandra T; Miyata, Kayoko; Heng, Vibol; Mitchell, Kenneth D; Derbenev, Andrei V

2012-06-21

142

Chronic bilateral renal denervation attenuates renal injury in a transgenic rat model of diabetic nephropathy.  

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Bilateral renal denervation (BRD) has been shown to reduce hypertension and improve renal function in both human and experimental studies. We hypothesized that chronic intervention with BRD may also attenuate renal injury and fibrosis in diabetic nephropathy. This hypothesis was examined in a female streptozotocin-induced diabetic (mRen-2)27 rat (TGR) shown to capture the cardinal features of human diabetic nephropathy. Following diabetic induction, BRD/sham surgeries were conducted repeatedly (at the week 3, 6, and 9 following induction) in both diabetic and normoglycemic animals. Renal denervation resulted in a progressive decrease in systolic blood pressure from first denervation to termination (at 12 wk post-diabetic induction) in both normoglycemic and diabetic rats. Renal norepinephrine content was significantly raised following diabetic induction and ablated in denervated normoglycemic and diabetic groups. A significant increase in glomerular basement membrane thickening and mesangial expansion was seen in the diabetic kidneys; this morphological appearance was markedly reduced by BRD. Immunohistochemistry and protein densitometric analysis of diabetic innervated kidneys confirmed the presence of significantly increased levels of collagens I and IV, ?-smooth muscle actin, the ANG II type 1 receptor, and transforming growth factor-?. Renal denervation significantly reduced protein expression of these fibrotic markers. Furthermore, BRD attenuated albuminuria and prevented the loss of glomerular podocin expression in these diabetic animals. In conclusion, BRD decreases systolic blood pressure and reduces the development of renal fibrosis, glomerulosclerosis, and albuminuria in this model of diabetic nephropathy. The evidence presented strongly suggests that renal denervation may serve as a therapeutic intervention to attenuate the progression of renal injury in diabetic nephropathy. PMID:24899056

Yao, Yimin; Fomison-Nurse, Ingrid C; Harrison, Joanne C; Walker, Robert J; Davis, Gerard; Sammut, Ivan A

2014-08-01

143

Enhanced cardiovascular alteration and Fos expression induced by central salt loading in a conscious rat transgenic for the metallothionein-vasopressin fusion gene.  

Science.gov (United States)

The present study is an investigation of the responses of the cardiovascular system and Fos expression to intracerebroventricular (i.c.v.) administration of hypertonic saline (HS) in conscious arginine vasopressin (AVP)-overexpressing transgenic (Tg) and control rats. Central HS (0.3, 0.67, or 1.0M NaCl, 1 microl/min for 20 min) significantly increased the mean arterial blood pressure (MABP) and Fos-like immunoreactivity (FLI) in the paraventricular nucleus (PVN) and supraoptic nucleus (SON) of the hypothalamus, the area postrema (AP), the median preoptic nucleus (MnPO), and the organum vasculosum laminae terminalis (OVLT) in both Tg and control rats. The changes in MABP and FLI were significantly larger in Tg rats than in control rats. i.c.v. pretreatment with the AVP V1 receptor antagonist, OPC-21268, blocked the increase in MABP and significantly decreased the Fos expression in the PVN (posterior magnocellular (pm) component) induced by 0.3 M HS in the Tg rats. The present study demonstrates an increased responsiveness to i.c.v. administration of HS in AVP Tg rats, suggesting the relationship between the vasopressinergic drive and central cardiovascular response via, at least in part, the V1 receptor in the PVN magnocellular neurons. PMID:16039738

Chu, Chun-Ping; Kato, Kazuo; Jin, Qing-Hua; Qiu, De-Lai; Yu, Nan-Shou; Oiso, Yutaka; Kannan, Hiroshi

2005-10-01

144

Antihypertensive action of soluble epoxide hydrolase inhibition in Ren-2 transgenic rats is mediated by suppression of the intrarenal renin-angiotensin system.  

Science.gov (United States)

The aim of the present study was to evaluate the hypothesis that the antihypertensive effects of inhibition of soluble epoxide hydrolase (sEH) are mediated by increased intrarenal availability of epoxyeicosatrienoic acids (EETs), with consequent improvement in renal haemodynamic autoregulatory efficiency and the pressure-natriuresis relationship. Ren-2 transgenic rats (TGR), a model of angiotensin (Ang) II-dependent hypertension, and normotensive transgene-negative Hannover Sprague-Dawley (HanSD) rats were treated with the sEH inhibitor cis-4-(4-(3-adamantan-1-yl-ureido)cyclohexyloxy)benzoic acid (c-AUCB; 26 mg/L) for 48 h. Then, the effects on blood pressure (BP), autoregulation of renal blood flow (RBF) and glomerular filtration rate (GFR), and on the pressure-natriuresis relationship in response to stepwise reductions in renal arterial pressure (RAP) were determined. Treatment with c-AUCB did not significantly change BP, renal autoregulation or pressure-natriuresis in normotensive HanSD rats. In contrast, c-AUCB treatment significantly reduced BP, increased intrarenal bioavailability of EETs and significantly suppressed AngII levels in TGR. However, treatment with c-AUCB did not significantly improve the autoregulatory efficiency of RBF and GFR in response to reductions of RAP and to restore the blunted pressure-natriuresis relationship in TGR. Together, the data indicate that the antihypertensive actions of sEH inhibition in TGR are predominantly mediated via significant suppression of intrarenal renin-angiotensin system activity. PMID:23039246

Varcabova, Sarka; Huskova, Zuzana; Kramer, Herbert J; Hwang, Sung Hee; Hammock, Bruce D; Imig, John D; Kitada, Kento; Cervenka, Ludek

2013-04-01

145

Transgene therapy for rat anti-Thy1.1 glomerulonephritis via mesangial cell vector with a polyethylenimine/decorin nanocomplex  

OpenAIRE

Polyethylenimine (PEI), a cationic polymer, is one of the most efficient non-viral vectors for transgene therapy. Decorin (DCN), a leucine-rich proteoglycan secreted by glomerular mesangial cells (MC), is a promising anti-fibrotic agent for the treatment of glomerulonephritis. In this study, we used PEI–DCN nanocomplexes with different N/P ratios to transfect MC in vitro and deliver the MC vector with PEI–DCN expressing into rat anti-Thy1.1 nephritis kidney tissue via injection into the l...

Sun, Jian-yong; Sun, Yu; Wu, Hui-juan; Zhang, Hong-xia; Zhao, Zhong-hua; Chen, Qi; Zhang, Zhi-gang

2012-01-01

146

Progressive changes in microglia and macrophages in spinal cord and peripheral nerve in the transgenic rat model of amyotrophic lateral sclerosis  

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Full Text Available Abstract Background The role of neuroinflammation in motor neuron death of amyotrophic lateral sclerosis (ALS is unclear. The human mutant superoxide dismutase-1 (hmSOD1-expressing murine transgenic model of ALS has provided some insight into changes in microglia activity during disease progression. The purpose of this study was to gain further knowledge by characterizing the immunological changes during disease progression in the spinal cord and peripheral nerve using the more recently developed hmSOD1 rat transgenic model of ALS. Methods Using immunohistochemistry, the extent and intensity of tissue CD11b expression in spinal cord, lumbar nerve roots, and sciatic nerve were evaluated in hmSOD1 rats that were pre-clinical, at clinical onset, and near disease end-stage. Changes in CD11b expression were compared to the detection of MHC class II and CD68 microglial activation markers in the ventral horn of the spinal cord, as well as to the changes in astrocytic GFAP expression. Results Our study reveals an accumulation of microglia/macrophages both in the spinal cord and peripheral nerve prior to clinical onset based on CD11b tissue expression. The microglia formed focal aggregates in the ventral horn and became more widespread as the disease progressed. Hypertrophic astrocytes were not prominent in the ventral horn until after clinical onset, and the enhancement of GFAP did not have a strong correlation to increased CD11b expression. Detection of MHC class II and CD68 expression was found in the ventral horn only after clinical onset. The macrophages in the ventral nerve root and sciatic nerve of hmSOD1 rats were observed encircling axons. Conclusions These findings describe for the first time in the hmSOD1 rat transgenic model of ALS that enhancement of microglia/macrophage activity occurs pre-clinically both in the peripheral nerve and in the spinal cord. CD11b expression is shown to be a superior indicator for early immunological changes compared to other microglia activation markers and astrogliosis. Furthermore, we suggest that the early activity of microglia/macrophages is involved in the early phase of motor neuron degeneration and propose that studies involving immunomodulation in hmSOD1transgenic models need to consider effects on macrophages in peripheral nerves as well as to microglia in the spinal cord.

Hickey William F

2010-01-01

147

Primary T-cells from human CD4/CCR5-transgenic rats support all early steps of HIV-1 replication including integration, but display impaired viral gene expression  

OpenAIRE

Abstract Background In vivo studies on HIV-1 pathogenesis and testing of antiviral strategies have been hampered by the lack of an immunocompetent small animal model that is highly susceptible to HIV-1 infection. Since native rodents are non-permissive, we developed transgenic rats that selectively express the HIV-1 receptor complex, hCD4 and hCCR5, on relevant target cells. These animals display a transient low-level plasma viremia after HIV-1YU-2 infection, demonstrating HIV-1 susceptibilit...

Hermann Volker; Tervo Hanna-Mari; Allespach Ina; Michel Nico; Goffinet Christine; Kräusslich Hans-Georg; Greene Warner C; Keppler Oliver T

2007-01-01

148

Human cyclin T1 expression ameliorates a T-cell-specific transcriptional limitation for HIV in transgenic rats, but is not sufficient for a spreading infection of prototypic R5 HIV-1 strains ex vivo  

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Full Text Available Abstract Background Cells derived from native rodents have limits at distinct steps of HIV replication. Rat primary CD4 T-cells, but not macrophages, display a profound transcriptional deficit that is ameliorated by transient trans-complementation with the human Tat-interacting protein Cyclin T1 (hCycT1. Results Here, we generated transgenic rats that selectively express hCycT1 in CD4 T-cells and macrophages. hCycT1 expression in rat T-cells boosted early HIV gene expression to levels approaching those in infected primary human T-cells. hCycT1 expression was necessary, but not sufficient, to enhance HIV transcription in T-cells from individual transgenic animals, indicating that endogenous cellular factors are critical co-regulators of HIV gene expression in rats. T-cells from hCD4/hCCR5/hCycT1-transgenic rats did not support productive infection of prototypic wild-type R5 HIV-1 strains ex vivo, suggesting one or more significant limitation in the late phase of the replication cycle in this primary rodent cell type. Remarkably, we identify a replication-competent HIV-1 GFP reporter strain (R7/3 YU-2 Env that displays characteristics of a spreading, primarily cell-to-cell-mediated infection in primary T-cells from hCD4/hCCR5-transgenic rats. Moreover, the replication of this recombinant HIV-1 strain was significantly enhanced by hCycT1 transgenesis. The viral determinants of this so far unique replicative ability are currently unknown. Conclusion Thus, hCycT1 expression is beneficial to de novo HIV infection in a transgenic rat model, but additional genetic manipulations of the host or virus are required to achieve full permissivity.

Littman Dan R

2009-01-01

149

Establishment and characterization of conditionally immortalized endothelial cell lines from the thoracic duct and inferior vena cava of tsA58/EGFP double-transgenic rats.  

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The basic biology of blood vascular endothelial cells has been well documented. However, little is known about that of lymphatic endothelial cells, despite their importance under normal and pathological conditions. The lack of a lymphatic endothelial cell line has hampered progress in this field. The objective of this study has been to establish and characterize lymphatic and venous endothelial cell lines derived from newly developed tsA58/EGFP transgenic rats harboring the temperature-sensitive simian virus 40 (SV40) large T-antigen and enhanced green fluorescent protein (EGFP). Endothelial cells were isolated from the transgenic rats by intraluminal enzymatic digestion. The cloned cell lines were named TR-LE (temperature-sensitive rat lymphatic endothelial cells from thoracic duct) and TR-BE (temperature-sensitive rat blood-vessel endothelial cells from inferior vena cava), respectively, and cultured on fibronectin-coated dishes in HuMedia-EG2 supplemented with 20% fetal bovine serum and Endothelial Mitogen at a permissive temperature, 33 degrees C. A temperature shift to 37 degrees C resulted in a decrease in proliferation with degradation of the large T-antigen and cleavage of poly (ADP-ribose) polymerase. TR-LE cells expressed lymphatic endothelial markers VEGFR-3 (vascular endothelial growth factor receptor), LYVE-1 (a lymphatic endothelial receptor), Prox-1 (a homeobox gene product), and podoplanin (a glomerular podocyte membrane mucoprotein), together with endothelial markers CD31, Tie-2, and VEGFR-2, whereas TR-BE cells expressed CD31, Tie-2, and VEGFR-2, but no lymphatic endothelial markers. Thus, these conditionally immortalized and EGFP-expressing lymphatic and vascular endothelial cell lines might represent an important tool for the study of endothelial cell functions in vitro. PMID:16773315

Matsuo, Mitsuhiro; Koizumi, Keiichi; Yamada, Sanae; Tomi, Masatoshi; Takahashi, Ri-ichi; Ueda, Masatsugu; Terasaki, Tetsuya; Obinata, Masuo; Hosoya, Ken-ichi; Ohtani, Osamu; Saiki, Ikuo

2006-12-01

150

Transgenic Fish  

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Fish into which foreign DNA is artificially introduced and integrated into their genome are called transgenic fish. Since the development of the first transgenic fish in 1985, techniques to produce transgenic fish have improved tremendously, resulting in the production of genetically modified (GM) ...

151

In vivo expression of angiotensin-(1-7) lowers blood pressure and improves baroreflex function in transgenic (mRen2)27 rats.  

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Transgenic (mRen2)27 rats are hypertensive with impaired baroreflex sensitivity for control of heart rate compared with Hannover Sprague-Dawley rats. We assessed blood pressure and baroreflex function in male hemizygous (mRen2)27 rats (30-40 weeks of age) instrumented for arterial pressure recordings and receiving into the cisterna magna either an Ang-(1-7) fusion protein or a control fusion protein (CTL-FP). The maximum reduction in mean arterial pressure achieved was -38 ± 7 mm Hg on day 3, accompanied by a 55% enhancement in baroreflex sensitivity in Ang-(1-7) fusion protein-treated rats. Both the high-frequency alpha index (HF-?) and heart rate variability increased, suggesting increased parasympathetic tone for cardiac control. The mRNA levels of several components of the renin-angiotensin system in the dorsal medulla were markedly reduced including renin (-80%), neprilysin (-40%), and the AT1a receptor (-40%). However, there was a 2-fold to 3-fold increase in the mRNA levels of the phosphatases PTP-1b and dual-specificity phosphatase 1 in the medulla of Ang-(1-7) fusion protein-treated rats. Our finding that replacement of Ang-(1-7) in the brain of (mRen2)27 rats reverses in part the hypertension and baroreflex impairment is consistent with a functional deficit of Ang-(1-7) in this hypertensive strain. We conclude that the increased mRNA expression of phosphatases known to counteract the phosphoinositol 3 kinase and mitogen-activated protein kinases, and the reduction of renin and AT1a receptor mRNA levels may contribute to the reduction in arterial pressure and improvement in baroreflex sensitivity in response to Ang-(1-7). PMID:22526299

Garcia-Espinosa, Maria A; Shaltout, Hossam A; Gallagher, Patricia E; Chappell, Mark C; Diz, Debra I

2012-08-01

152

Adeno-associated viral vector serotypes 1 and 5 targeted to the neonatal rat and pig striatum induce widespread transgene expression in the forebrain  

DEFF Research Database (Denmark)

Viral vector-mediated gene transfer has emerged as a powerful means to target transgene expression in the central nervous system. Here we characterized the efficacy of serotypes 1 and 5 recombinant adeno-associated virus (rAAV) vectors encoding green fluorescent protein (GFP) after stereotaxic delivery to the neonatal rat and minipig striatum. The efficiency of GFP expression and the phenotype of GFP-positive cells were assessed within the forebrain at different time points up to 12 months after surgery. Both rAAV1-GFP and rAAV5-GFP delivery resulted in transduction of the striatum as well as striatal input and output areas, including large parts of the cortex. In both species, rAAV5 resulted in a more widespread transgene expression compared to rAAV1. In neonatal rats, rAAV5 also transduced several other areas such as the olfactory bulbs, hippocampus, and septum. Phenotypic analysis of the GFP-positive cells, performed using immunohistochemistry and confocal microscopy, showed that most of the GFP-positive cells by either serotype were NeuN-positive neuronal profiles. The rAAV5 vector further displayed the ability to transduce non-neuronal cell types in both rats and pigs, albeit at a low frequency. Our results show that striatal delivery of rAAV5 vectors in the neonatal brain represents a useful tool to express genes of interest both in the basal ganglia and the neocortex. Furthermore, we apply, for the first time, viral vector-mediated gene transfer to the pig brain providing the opportunity to study effects of genetic manipulation in this non-primate large animal species. Finally, we generated an atlas of the Göttingen minipig brain for guiding future studies in this large animal species.

Kornum, Birgitte R; Stott, Simon R W

2010-01-01

153

Parallel age-associated changes in brain and plasma neuronal pentraxin receptor levels in a transgenic APP/PS1 rat model of Alzheimer's disease.  

Science.gov (United States)

Neuronal pentraxin receptor (NPR) is a synaptic protein implicated in AMPA receptor trafficking at excitatory synapses. Since glutamate neurotransmission is disrupted in Alzheimer's disease (AD), NPR levels measured from plasma represent a potential biomarker for synaptic dysfunction associated with AD. We sought to determine the relationship between AD pathology and brain and plasma NPR levels by examining age-associated NPR levels in these compartments in a transgenic APP/PS1 rat model of AD. NPR levels in cortical homogenate were similar in wild-type (Wt) and APP/PS1 rats at 3months of age (prior to A? plaque deposition), but significantly increased in APP/PS1 rats by 9 and 18-20months of age (after the onset of plaque deposition). These age-dependent differences were driven by proportional increases in NPR in membrane-associated cortical fractions. Genotype-related differences in NPR expression were also seen in the hippocampus, which exhibits significant A? pathology, but not in the cerebellum, which does not. Plasma analyses revealed increased levels of a 26kDa NPR fragment in APP/PS1 rats relative to Wt rats by 18-20months of age, which correlated with the levels of full-length NPR in cortex. Our findings indicate that cerebral accumulation of NPR and A? occurs with similar temporal and regional patterns in the APP/PS1 model, and suggest that a 26kDa plasma NPR fragment may represent a peripheral biomarker of this process. PMID:25449907

Bilousova, Tina; Taylor, Karen; Emirzian, Ana; Gylys, Raymond; Frautschy, Sally A; Cole, Gregory M; Teng, Edmond

2015-02-01

154

Effect of visible light on normal and P23H-3 transgenic rat retinas: characterization of a novel retinoic acid derivative present in the P23H-3 retina.  

Science.gov (United States)

Transgenic rats with the P23H mutation in rhodopsin exhibit increased susceptibility to light damage, compared with normal animals. It is known that light-induced retinal damage requires repetitive bleaching of rhodopsin and that photoreceptor cell loss is by apoptosis; however, the underlying molecular mechanism(s) leading to photoreceptor cell death are still unknown. Photoproducts, such as all-trans retinal or other retinoid metabolites, released by the extensive bleaching of rhodopsin could lead to activation of degenerative processes, especially in animals genetically predisposed to retinal degenerations. Using wild-type and transgenic rats carrying the P23H opsin mutation, we evaluated the effects of acute intense visible light on retinoid content, type and distribution in ocular tissues. Rats were exposed to green light (480-590 nm) for 0, 5, 10, 30 and 120 min. Following light treatment, rats were sacrificed and neural retinas were dissected free of the retinal pigment epithelium. Retinoids were extracted from retinal tissues and then subjected to HPLC and mass spectral analysis. We found that the light exposure affected relative levels of retinoids in the neural retina and retinal pigment epithelium of wild-type and P23H rat eyes similarly. In the P23H rat retina but not the wild-type rat retina, we found a retinoic acid-like compound with an absorbance maximum of 357 nm and a mass of 304 daltons. Production of this retinoic acid-like compound in transgenic rats is influenced by the age of the animals and the duration of light exposure. It is possible that this unique retinoid may be involved in the process of light-induced retinal degeneration. PMID:16336041

Duncan, Todd; Wiggert, Barbara; Whittaker, Noel; Darrow, Ruth; Organisciak, Daniel T

2006-01-01

155

Peptides from HLA-B27 molecules of patients with ankylosing spondylitis reveal increased glutamic acid/glutamine ratio.  

Czech Academy of Sciences Publication Activity Database

Oslo : Verlag, 2006, s. 12-12. [European Immunogenetics and Histocompatibility conference /20./. Oslo (NO), 08.06.2006-11.06.2006] R&D Projects: GA MZd 1A8631 Institutional research plan: CEZ:AV0Z50200510; CEZ:AV0Z50520514 Keywords : sped * peptide Subject RIV: EE - Microbiology, Virology

Stod?lková, Eva; Ivašková, E.; Sedlá?ková, M.; Pohl, J.; Votruba, Jaroslav; Man, Petr; ?apková, Jana; Ivanyi, J.; Flieger, Miroslav

156

Free radical trap phenyl-N-tert-butylnitrone protects against light damage but does not rescue P23H and S334ter rhodopsin transgenic rats from inherited retinal degeneration.  

Science.gov (United States)

Phenyl-N-tert-butylnitrone (PBN) protects rat retinas against light damage. Because the degenerative process involved in light damage and inherited retinal degeneration both lead to a common final cell death, apoptosis, we used transgenic rats with a P23H or S334ter rhodopsin mutation to test the effects of PBN on retinal degeneration and light damage and the susceptibility of the transgenic rats to light damage. In the first study, 3-week-old mutant and wild-type rats were given no drug, 0.25% PBN in drinking water, or 0.25% PBN in drinking water plus three daily intraperitoneal injections of PBN (100 mg/kg, i.p., every 8 hr). Electroretinograms were recorded at postnatal day 49, after which the rats were killed for morphometric analysis. There was no photoreceptor rescue by PBN in P23H or S334ter rats, as evidenced by equivalent loss of function and photoreceptor cells in the three treatment groups. In the second study, P23H, S334ter, and wild-type rats were exposed for 24 hr to 2700 lux light. The rats were untreated or treated with PBN (50 mg/kg per injection, every 6 hr, starting before exposure). ERGs were recorded before and 1 d after exposure. Animals were killed 6 d later for morphometric analysis. PBN protected wild-type and P23H but not S334ter retinas from light damage. S334ter retinas were relatively less susceptible to light damage than P23H and wild-type rats. The results suggest that the initiating event(s) that causes photoreceptor cell death in the mutated rats is different from that which occurs in light damage, although both ultimately undergo an apoptotic cell death. PMID:12853423

Ranchon, Isabelle; LaVail, Matthew M; Kotake, Yashige; Anderson, Robert E

2003-07-01

157

Overexpression of HIF-1? Transgene in the Renal Medulla Attenuated Salt Sensitive Hypertension in Dahl S Rats  

OpenAIRE

Hypoxia inducible factor (HIF)-1?-mediated gene activation in the renal medulla in response to high salt intake plays an important role in the control of salt sensitivity of blood pressure. High salt-induced activation of HIF-1? in the renal medulla is blunted in Dahl S rats. The present study determined whether the impairment of the renal medullary HIF-1? pathway was responsible for salt sensitive hypertension in Dahl S rats. Renal medullary HIF-1? levels were induced by either transfect...

Zhu, Qing; Wang, Zhengchao; Xia, Min; Li, Pin-lan; Zhang, Fan; Li, Ningjun

2012-01-01

158

Transgenic tea  

OpenAIRE

Like most of the important crop plants of the world, transgenic technology has also been extended to tea. Both biolistic and Agrobacterium mediated transformation methods have been employed to transform explants like leaves and somatic embryos. While gusand nptil genes were used to optimize parameters and develop protocols for transgenic production, plants expressing stress tolerance genes (osmotin) have also been produced. These methods have opened a whole new era for developing tea plants a...

Bhattacharya, Amita; Saini, U.; Ahuja, P. S.

2006-01-01

159

Regressive and reactive changes in the connectivity patterns of rod and cone pathways of P23H transgenic rat retina  

OpenAIRE

We have used the P23H line 1 homozygous albino rat to study how progressive photoreceptor degeneration affects rod and cone relay pathways. We examined P23H retinas at different stages of degeneration by confocal microscopy of immunostained sections and electroretinogram (ERG) recordings. By 21 days of age in the P23H rat retina, there is already substantial loss of rods and reduction in rod bipolar dendrites along with reduction of metabotropic glutamate receptor 6 (mGluR6) and rod-assoc...

Cuenca Navarro, Nicola?s; Pinilla Lozano, Isabel; Sauve?, Yves; Lu, B.; Wang, Shaomei; Lund, Raymond D.

2004-01-01

160

Sperm parameters and epididymis function in transgenic rats overexpressing the Ca2+-binding protein regucalcin: a hidden role for Ca2+ in sperm maturation?  

Science.gov (United States)

Sperm undergo maturation acquiring progressive motility and the ability to fertilize oocytes through exposure to the components of the epididymal fluid (EF). Although the establishment of a calcium (Ca(2+)) gradient along the epididymis has been described, its direct effects on epididymal function remain poorly explored. Regucalcin (RGN) is a Ca(2+)-binding protein, regulating the activity of Ca(2+)-channels and Ca(2+)-ATPase, for which a role in male reproductive function has been suggested. This study aimed at comparing the morphology, assessed by histological analysis, and function of epididymis, by analysis of sperm parameters, antioxidant potential and Ca(2+) fluxes, between transgenic rats overexpressing RGN (Tg-RGN) and their wild-type littermates. Tg-RGN animals displayed an altered morphology of epididymis and lower sperm counts and motility. Tissue incubation with (45)Ca(2+) showed also that epididymis of Tg-RGN displayed a diminished rate of Ca(2+)-influx, indicating unbalanced Ca(2+) concentrations in the epididymal lumen. Sperm viability and the frequency of normal sperm, determined by the one-step eosin-nigrosin staining technique and the Diff-Quik staining method, respectively, were higher in Tg-RGN. Moreover, sperm of Tg-RGN rats showed a diminished incidence of tail defects. Western blot analysis demonstrated the presence of RGN in EF as well as its higher expression in the corpus region. The results presented herein demonstrated the importance of maintaining Ca(2+)-levels in the epididymal lumen and suggest a role for RGN in sperm maturation. Overall, a new insight into the molecular mechanisms driving epididymal sperm maturation was obtained, which could be relevant to development of better approaches in male infertility treatment and contraception. PMID:23615721

Correia, S; Oliveira, P F; Guerreiro, P M; Lopes, G; Alves, M G; Canário, A V M; Cavaco, J E; Socorro, Sílvia

2013-09-01

161

Remyelination of spinal cord axons by olfactory ensheathing cells and Schwann cells derived from a transgenic rat expressing alkaline phosphatase marker gene.  

Science.gov (United States)

Transplantation of cell suspensions containing olfactory ensheathing cells (OECs) has been reported to remyelinate demyelinated axons in the spinal cord with a Schwann cell (SC)-like pattern of myelination. However, questions have been raised recently as to whether OECs can form SC-like myelin. To address this issue we prepared SCs and OECs from transgenic rats in which a marker gene, human placental alkaline phosphatase (hPAP), is linked to the ubiquitously active promoter of the R26 gene. SCs were prepared from the sciatic nerve and OECs from the outer nerve-fiber layer of the olfactory bulb. Positive S100 and p75 immunostaining indicated that >95% of cells in culture displayed either SC or OEC phenotypes. Suspensions of either SCs or OECs were transplanted into an X-irradiation/ethidium bromide demyelinating lesion in the spinal cord. We observed extensive SC-like remyelination following either SC or OEC transplantation 3 weeks after injection of the cells. Alkaline phosphatase (ALP) chromagen reaction product was associated clearly with the myelin-forming cells. Thus, cell suspensions that are enriched in either SCs or OECs result in peripheral-like myelin when transplanted in vivo. PMID:16799702

Akiyama, Yukinori; Lankford, Karen; Radtke, Christine; Greer, Charles A; Kocsis, Jeffery D

2004-02-01

162

Disease-Inducible Transgene Expression from a Recombinant Adeno-Associated Virus Vector in a Rat Arthritis Model  

OpenAIRE

Rheumatoid arthritis (RA) is a systemic autoimmune disease affecting 1% of the world’s population, with significant morbidity and mortality. In this study, we investigated a recombinant adeno-associated virus (rAAV) vector for its potential application in RA gene therapy. rAAV encoding Escherichia coli ?-galactosidase was injected into rat joints which had already been induced into acute arthritis after local lipopolysaccharide (LPS) administration, and the efficiency of in vivo transducti...

Pan, Ru-yu; Xiao, Xiao; Chen, Show-li; Li, Juan; Lin, Leou-chyr; Wang, Hsian-jenn; Tsao, Yeou-ping

1999-01-01

163

Retinal ganglion cell survival and axon regeneration in WldS transgenic rats after optic nerve crush and lens injury  

OpenAIRE

Abstract Background We have previously shown that the slow Wallerian degeneration mutation, whilst delaying axonal degeneration after optic nerve crush, does not protect retinal ganglion cell (RGC) bodies in adult rats. To test the effects of a combination approach protecting both axons and cell bodies we performed combined optic nerve crush and lens injury, which results in both enhanced RGC survival as well as axon regeneration past the lesion site in wildtype animals. Results As previously...

Lorber, Barbara; Tassoni, Alessia; Bull, Natalie D.; Moschos, Marilita M.; Martin, Keith R.

2012-01-01

164

Transgene-mediated enkephalin expression attenuates signs of naloxone-precipitated morphine withdrawal in rats with neuropathic pain.  

Science.gov (United States)

Chronic morphine exposure induces physical dependence and tolerance. Previous studies have shown that there is a decrease in met-enkephalin levels in states of morphine physical dependence, and that increasing enkephalin during opiate physical withdrawal ameliorates the severity of the morphine withdrawal syndrome. In order to investigate the role of spinal opioid peptide in the phenomenon of naloxone-precipitated withdrawal we examined the effect of herpes simplex virus vector-mediated overexpression of proenkephalin in lumbar dorsal root ganglia in rats with neuropathic pain treated with morphine. The morphine physical dependence was induced by chronic administration of intraperitoneal (IP) morphine for 2 weeks. Rats with neuropathic pain inoculated subcutaneously with the vector-mediated overexpression of proenkephalin showed a significant reduction in jumps, 'wet-dog' shakes, diarrhea and ptosis precipitated by naloxone after 2 weeks of morphine treatment. The global withdrawal score was also reduced significantly by vector-mediated overexpression of proenkephalin. These studies demonstrate a role for opioid peptide in the spinal cord in mediating some of the withdrawal response. PMID:18761380

Hao, Shuanglin; Hu, Jian; Fink, David J

2009-01-30

165

Regressive and reactive changes in the connectivity patterns of rod and cone pathways of P23H transgenic rat retina.  

Science.gov (United States)

We have used the P23H line 1 homozygous albino rat to study how progressive photoreceptor degeneration affects rod and cone relay pathways. We examined P23H retinas at different stages of degeneration by confocal microscopy of immunostained sections and electroretinogram (ERG) recordings. By 21 days of age in the P23H rat retina, there is already substantial loss of rods and reduction in rod bipolar dendrites along with reduction of metabotropic glutamate receptor 6 (mGluR6) and rod-associated bassoon staining. The cone pathway is relatively unaffected. By 150 days, when rods are absent from much of the retina, some rod bipolars remain and dendrites of rod and cone bipolar cells form synaptic complexes associated with cones and horizontal cell processes. These complexes include foci of mGluR6 and bassoon staining; they develop further by 270 days of age. Over the course of degeneration, beginning at 21 days, bipolar axon terminals atrophy and the inner retina undergoes further changes including a reduced and disorganized AII amacrine cell population and thinning of the inner plexiform layer. Electroretinogram (ERG) results at 23 days show reductions in a-wave amplitude, in rod and cone-associated b-waves (using a double flash paradigm) and in the amplitude of oscillatory potentials (OPs). By 38 days, rod scotopic a-wave responses and OPs are lost. B-wave amplitudes decline until 150 days, at which point they are purely cone-driven and remain stable up to 250 days. The results show that during the course of photoreceptor loss in the P23H rat, there are progressive degenerative changes, particularly in the rod relay pathway, and these are reflected in the changing ERG response patterns. Later reactive changes involving condensation of cone terminals and neurotransmitter receptors associated with rod and cone bipolar dendrites and with horizontal cell processes suggest that at this stage, there are likely to be complex changes in the relay of sensory information through the retina. PMID:15262321

Cuenca, N; Pinilla, I; Sauvé, Y; Lu, B; Wang, S; Lund, R D

2004-01-01

166

The vestibulo- and preposito-cerebellar cholinergic neurons of a ChAT-tdTomato transgenic rat exhibit heterogeneous firing properties and the expression of various neurotransmitter receptors.  

Science.gov (United States)

Cerebellar function is regulated by cholinergic mossy fiber inputs that are primarily derived from the medial vestibular nucleus (MVN) and prepositus hypoglossi nucleus (PHN). In contrast to the growing evidence surrounding cholinergic transmission and its functional significance in the cerebellum, the intrinsic and synaptic properties of cholinergic projection neurons (ChPNs) have not been clarified. In this study, we generated choline acetyltransferase (ChAT)-tdTomato transgenic rats, which specifically express the fluorescent protein tdTomato in cholinergic neurons, and used them to investigate the response properties of ChPNs identified via retrograde labeling using whole-cell recordings in brainstem slices. In response to current pulses, ChPNs exhibited two afterhyperpolarisation (AHP) profiles and three firing patterns; the predominant AHP and firing properties differed between the MVN and PHN. Morphologically, the ChPNs were separated into two types based on their soma size and dendritic extensions. Analyses of the firing responses to time-varying sinusoidal current stimuli revealed that ChPNs exhibited different firing modes depending on the input frequencies. The maximum frequencies in which each firing mode was observed were different between the neurons that exhibited distinct firing patterns. Analyses of the current responses to the application of neurotransmitter receptor agonists revealed that the ChPNs expressed (i) AMPA- and NMDA-type glutamate receptors, (ii) GABAA and glycine receptors, and (iii) muscarinic and nicotinic acetylcholine receptors. The current responses mediated by these receptors of MVN ChPNs were not different from those of PHN ChPNs. These findings suggest that ChPNs receive various synaptic inputs and encode those inputs appropriately across different frequencies. PMID:24593297

Zhang, Yue; Kaneko, Ryosuke; Yanagawa, Yuchio; Saito, Yasuhiko

2014-04-01

167

Angiotensin-converting enzyme inhibition, but not AT1 receptor blockade, in the solitary tract nucleus improves baroreflex sensitivity in anesthetized transgenic hypertensive (mRen2)27 rats  

OpenAIRE

Transgenic hypertensive (mRen2)27 rats overexpress the murine Ren2 gene and have impaired baroreflex sensitivity (BRS) for control of the heart rate. Removal of endogenous angiotensin (Ang)-(1–7) tone using a receptor blocker does not further lower BRS. Therefore, we assessed whether blockade of Ang II with a receptor antagonist or combined reduction in Ang II and restoration of endogenous Ang-(1-7) levels with Ang-converting enzyme (ACE) inhibition will improve BRS in these animals. Bilate...

Isa, Katsunori; Arnold, Amy C.; Westwood, Brian M.; Chappell, Mark C.; Diz, Debra I.

2011-01-01

168

Prostate cancer in a transgenic mouse.  

OpenAIRE

Progress toward understanding the biology of prostate cancer has been slow due to the few animal research models available to study the spectrum of this uniquely human disease. To develop an animal model for prostate cancer, several lines of transgenic mice were generated by using the prostate-specific rat probasin promoter to derive expression of the simian virus 40 large tumor antigen-coding region. Mice expressing high levels of the transgene display progressive forms of prostatic disease ...

Greenberg, N. M.; Demayo, F.; Finegold, M. J.; Medina, D.; Tilley, W. D.; Aspinall, J. O.; Cunha, G. R.; Donjacour, A. A.; Matusik, R. J.; Rosen, J. M.

1995-01-01

169

Inhibition of soluble epoxide hydrolase improves the impaired pressure–natriuresis relationship and attenuates the development of hypertension and hypertension-associated end-organ damage in Cyp1a1-Ren-2 transgenic rats  

Science.gov (United States)

Objective In the present study, we compared the effects of treatment with the novel soluble epoxide hydrolase (sEH) inhibitor (c-AUCB) with those of the AT1 receptor antagonist losartan on blood pressure (BP), autoregulation of renal blood flow (RBF) and on glomerular filtration rate (GFR) and the pressure–natriuresis relationship in response to stepwise reduction in renal arterial pressure (RAP) in Cyp1a1-Ren-2 transgenic rats. Methods Hypertension was induced in Cyp1a1-Ren-2 rats through dietary administration for 11 days of the natural xenobiotic indole-3-carbinol (I3C) which activates the renin gene. Treatment with c-AUCB and losartan was started 48 h before initiating administration of the diet containing I3C. Rats were prepared for renal functional studies to evaluate in-vivo renal autoregulatory efficiency when RAP was gradually decreased by an aortic clamp. Results I3C administration resulted in the development of severe hypertension which was associated with markedly lower basal RBF and GFR and substantially impaired autoregulatory efficiency as well as a suppression of the pressure–natriuresis relationship when compared with noninduced rats. Treatment with c-AUCB significantly decreased BP, improved autoregulatory efficiency of RBF and GFR and the slope of pressure–natriuresis relationship. Treatment with losartan completely prevented the impaired autoregulation and pressure–natriuresis relationship as well as the development of hypertension in I3C-induced rats. Conclusion Our present findings indicate that chronic treatment with the sEH inhibitor c-AUCB substantially attenuates the development of malignant hypertension in I3C-induced rats likely via improvement of the renal autoregulatory efficiency and the pressure–natriuresis relationship. PMID:21720266

Honetschlägerová, Zuzana; Sporková, Alexandra; Kopkan, Libor; Husková, Zuzana; Hwang, Sung H.; Hammock, Bruce D.; Imig, John D.; Kramer, Herbert J.; Kujal, Petr; Vernerová, Zdenka; Chábová, V?ra ?.; Tesa?, Vladimír; ?ervenka, Lud?k

2013-01-01

170

Peptides Eluted from HLA-B27 of Human Splenocytes and blood Cells Reveal a Partially Different Motif Compared to In Vitro Grown Cell Lines.  

Czech Academy of Sciences Publication Activity Database

Montreal, 2004, s. 22. [International Congress of Immunology and Annual Conference of FOCIS /12./. Montreal (CA), 18.07.2004-23.07.2004] R&D Projects: GA MZd NI7327 Institutional research plan: CEZ:AV0Z5020903 Keywords : hla * hla -b Subject RIV: EE - Microbiology, Virology

Stod?lková, Eva; Man, Petr; Pohl, J.; Van Nguyen, D.; Vaingátová, Silvie; Ivašková, E.; Pla, M.; ?apková, J.; Sedlá?ková, M.; Ivanyi, P.; Flieger, Miroslav

171

HLA-B73: An atypical HLA-B molecule carrying a Bw6-epitope motif variant and a B pocket identical to HLA-B27  

Energy Technology Data Exchange (ETDEWEB)

HLA-B73, first described by Mayr and Kirnbauer (1981), is a poorly characterized allospecificity, serologically related to the B7-CREG. We polymerase chain reaction-amplified, cloned and sequenced the HLA-B alleles of the B-LCL LE023, established from a Spanish Caucasoid individual expressing HLA-B73. 5 refs., 2 figs.

Vilches, C.; Pablo, R. de; Herrero, M.J.; Moreno, M.E.; Kreisler, M. [Hospital Puerta de Hierro, Madrid (Spain)

1994-12-31

172

Peptides eluted from HLA-B27 of human splenocytes and blood cells reveal a similar but partially different profile compared to in vitro grown cell lines.  

Czech Academy of Sciences Publication Activity Database

Ro?. 94, - (2004), s. 261-265. ISSN 0165-2478 R&D Projects: GA MZd NI7327 Keywords : hla -b*2705 * peptide profile * lc-ms/ms analysis Subject RIV: EE - Microbiology, Virology Impact factor: 2.136, year: 2004

Stod?lková, Eva; Man, Petr; Pohl, J.; Nguyen, V. D.; Vaingátová, Silvie; Ivašková, E.; Pla, M.; ?apková, Jana; Sedlá?ková, M.; Ivanyi, P.; Flieger, Miroslav

2004-01-01

173

Heterologous introns can enhance expression of transgenes in mice.  

OpenAIRE

In a previous study we showed that genomic constructs were expressed more efficiently in transgenic mice than constructs that were identical except for the lack of introns. Using the mouse metallothionein promoter-rat growth hormone gene construct as a model, we show that the first intron of the rat growth hormone gene is essential for high-level expression, whereas the other three introns are less effective. Several heterologous introns placed 3' of the coding region of an intronless rat gro...

Palmiter, R. D.; Sandgren, E. P.; Avarbock, M. R.; Allen, D. D.; Brinster, R. L.

1991-01-01

174

Tet-Transgenic Rodents: a comprehensive, up-to date database  

OpenAIRE

Here we introduce the "Tet-Transgenic Rodents" database, documenting most of the published Tet-transgenic mouse lines generated in the past 2 decades. Aside from the >500 mouse lines listed, it also includes the first of the recently reported Tet-transgenic rat models. Since the Tet technology comprises two essential components, a cis-acting promoter (P(tet)) and a trans-acting transactivator, the database has been organized accordingly. One section of the database summarizes the different tr...

Schoenig, K.; Freundlieb, S.; Gossen, M.

2013-01-01

175

[Inheritance and expression stability of transgene in transgenic animals].  

Science.gov (United States)

Transgenic technology is one of the most hotspots in biology. In the past decade, the progress in animal cloning has provided an alternative method to improve transgenic efficiency. Many kinds of transgenic animals have been successfully produced via the combination of transfection and nuclear transfer. However, the ultimate aim of transgenesis is not to produce several transgenic animals, but to service for the needs of human. In animal production, transgenic technology has been used to breed new livestock, which has received a lot of attention in China. It has been evidenced that inheritance and expression instability of transgene in transgenic animals is still the major limitation, which is attributed to position effect, epigenetic modification, and hereditary efficiency of transgene. In this review, we discussed the three points for promoting the industrialization of animal transgenic breeding. PMID:21586397

Kong, Qing-Ran; Liu, Zhong-Hua

2011-05-01

176

Transgenic animal models for the functional analysis of vasoactive peptides  

Directory of Open Access Journals (Sweden)

Full Text Available The interplay of vasoactive peptide systems is an essential determinant of blood pressure regulation in mammals. While the endothelin and the renin-angiotensin systems raise blood pressure by inducing vasoconstriction and sodium retention, the kallikrein-kinin and the natriuretic-peptide systems reduce arterial pressure by eliciting vasodilatation and natriuresis. Transgenic technology has proven to be very useful for the functional analysis of vasoactive peptide systems. As an outstanding example, transgenic rats overexpressing the mouse Ren-2 renin gene in several tissues become extremely hypertensive. Several other transgenic rat and mouse strains with genetic modifications of components of the renin-angiotensin system have been developed in the past decade. Moreover, in recent years gene-targeting technology was employed to produce mouse strains lacking these proteins. The established animal models as well as the main insights gained by their analysis are summarized in this review.

Bader M.

1998-01-01

177

Transgenic animal models for the functional analysis of vasoactive peptides  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in english The interplay of vasoactive peptide systems is an essential determinant of blood pressure regulation in mammals. While the endothelin and the renin-angiotensin systems raise blood pressure by inducing vasoconstriction and sodium retention, the kallikrein-kinin and the natriuretic-peptide systems red [...] uce arterial pressure by eliciting vasodilatation and natriuresis. Transgenic technology has proven to be very useful for the functional analysis of vasoactive peptide systems. As an outstanding example, transgenic rats overexpressing the mouse Ren-2 renin gene in several tissues become extremely hypertensive. Several other transgenic rat and mouse strains with genetic modifications of components of the renin-angiotensin system have been developed in the past decade. Moreover, in recent years gene-targeting technology was employed to produce mouse strains lacking these proteins. The established animal models as well as the main insights gained by their analysis are summarized in this review.

M., Bader.

1998-09-01

178

Phytoremediation with transgenic trees  

Energy Technology Data Exchange (ETDEWEB)

In the present paper actual trends in the use of transgenic trees for phytoremediation of contaminated soils are reviewed. In this context a current field trial in which transgenic poplars with enhanced GSH synthesis and hence elevated capacity for phytochelatin production are compared with wildtype plants for the removal of heavy metals at different levels of contamination and under different climatic conditions. The studies are carried out with grey poplar (Populus tremula x P. alba), wildtype plants and plants overexpressing the gene for {gamma}-glutamylcysteine synthetase (gshI) from E. coli in the cytosol. The expression of this gene in poplar leads to two- to four-fold enhanced GSH concentrations in the leaves. In greenhouse experiments under controlled conditions these transgenic poplars showed a high potential for uptake and detoxification of heavy metals and pesticides. This capacity is evaluated in field experiments. Further aims of the project are to elucidate (a) the stability of the transgene under field conditions and (b) the possibility of horizontal gene transfer to microorganisms in the rhizosphere. The results will help to assess the biosafety risk of the use of transgenic poplar for phytoremediation of soils. (orig.)

Peuke, A.D.; Rennenberg, H. [Inst. fuer Forstbotanik und Baumphysiologie, Professur fuer Baumphysiologie, Freiburg im Breisgau (Germany)

2005-04-01

179

Inducible transgenic mouse models.  

Science.gov (United States)

Inducible transgenic mouse models allow for the activation of genes in specific cells and tissues at specific times. Expression levels are dependent on the dose of the agent administered. Effective experimental models are characterized by low background levels of the regulated gene and induction to high levels with sub-physiological levels of inducing agents. The most commonly used methods to control gene expression in mouse models are based on the tet-operon/repressor bi-transgenic system and the estrogen receptor (ER) ligand-binding domain. Less commonly used systems to control gene expression in transgenic mice take advantage of the ligand-binding domain of the progesterone receptor, and the lac and GAL4 inducible systems. The tetracycline-regulated transgenic models are typically designed to activate the expression of the gene of interest in a specific cell type at a specific point in time. The ER is most commonly fused with Cre recombinase, although it can be used with transcription factors, kinases, etc., that are active in the nucleus. Cre-ER transgenes allow for the induction of recombinase activity in specific cells at defined time points. Cre recombinase is most often found in combination with conditional alleles to inactivate gene expression. When used for gene activation, Cre removes stop cassettes from transgenes and thus allows the expression of reporter or other molecules. Thus, the tetracycline-regulated and Cre-ER systems are complementary in mouse models, with utility in the cell-specific activation and inactivation of gene expression. PMID:21080277

Saunders, Thomas L

2011-01-01

180

Calcium electrotransfer for termination of transgene expression in muscle  

DEFF Research Database (Denmark)

Gene electrotransfer is expanding in clinical use, thus we have searched for an emergency procedure to stop transgene expression in case of serious adverse events. Calcium is cytotoxic at high intracellular levels, so we tested effects of calcium electrotransfer on transgene expression in muscle. A clinical grade calcium solution (20 ?l, 168 mM) was injected into transfected mouse or rat tibialis cranialis muscle. Ca(2+) uptake was quantified using calcium 45 ((45)Ca), and voltage and time between injection and pulsation were varied. Extinction of transgene expression was investigated by using both in vivo imaging of infrared fluorescent "Katushka" and erythropoietin evaluated by ELISA and hemoglobin. Histology was performed. Electrotransfer of Katushka and erythropoietin yielded significant expression. Maximal calcium uptake occurred after injection of Ca(2+) before electropulsing using eight high voltage pulses of 1000 V/cm. Using these parameters, in vivo imaging showed that transgene expression significantly decreased 4 hr after Ca(2+) electrotransfer and was eliminated within 24 hr. Similarly, serum erythropoietin was reduced by 46% at 4 hr and to control levels at 2 days. Histological analyses showed muscle damage and subsequent regeneration. Electrotransfer of isotonic CaCl(2) terminates transgenic protein expression in muscles and may be used for contingency elimination of transgene expression.

Hojman, Pernille; Spanggaard, Iben

2011-01-01

181

Hypothalamic vasopressin response to stress and various physiological stimuli: visualization in transgenic animal models.  

Science.gov (United States)

Arginine vasopressin (AVP) is involved in the homeostatic responses numerous life-threatening conditions, for example, the promotion of water conservation during periods of dehydration, and the activation of the hypothalamo-pituitary adrenal axis by emotional stress. Recently, we generated new transgenic animals that faithfully express an AVP-enhanced green fluorescent protein (eGFP) fusion gene in the paraventricular nucleus (PVN), the supraoptic nucleus (SON) and the suprachiasmatic nucleus (SCN) of the hypothalamus. In these transgenic rats, marked increases in eGFP fluorescence and fusion gene expression were observed in the magnocellular division of the PVN and the SON, but not the SCN, after osmotic challenges, such as dehydration and salt loading, and both acute and chronic nociceptive stimuli. In the parvocellular division of the PVN, eGFP expression was increased after acute and chronic pain, bilateral adrenalectomy, endotoxin shock and restraint stress. In the extra-hypothalamic areas of the brain, eGFP expression was induced in the locus coeruleus after the intracerebroventricular administration of colchicine. Next, we generated another transgenic rat that expresses a fusion gene comprised of c-fos promoter-enhancer sequences driving the expression of monomeric red fluorescent protein 1 (mRFP1). In these transgenic rats, abundant nuclear fluorescence of mRFP1 was observed in the PVN, the SON and other osmosensitive areas after acute osmotic stimulation. Finally, we generated a double transgenic rat that expresses both the AVP-eGFP and c-fos-mRFP1 fusion genes. In this double transgenic rat, we have observed nuclear mRFP1 fluorescence in eGFP-positive neurons after acute osmotic stimulation. These unique transgenic rats provide an exciting new tool to examine neuroendocrine responses to physiological and stressful stimuli in both in vivo and in vitro preparations. PMID:21185297

Ueta, Yoichi; Dayanithi, Govindin; Fujihara, Hiroaki

2011-02-01

182

Pharmacogenetic heterogeneity of transgene expression in muscle and tumours  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background Recombinant adenoviruses are employed to deliver a therapeutic transgene in the liver, muscle or tumour tissue. However, to rationalise this delivery approach, the factors of variation between individuals need to be identified. It is assumed that differences between inbred strains of laboratory animals are considered to reflect differences between patients. Previously we showed that transgene expression in the liver of different rat strains was dependent on the transcription efficiency of the transgene. In the present paper we investigated if transfection of muscle and tumour tissue were also subject to such variations. Methods Variation, in transgene expression, after intramuscular gene delivery was determined in different rodent strains and gene expression in tumours was investigated in different human and rodent cell lines as well as in subcutaneously implanted rodent tumours. The molecular mechanisms involved in transgene expression were dissected using an adenovirus encoding luciferase. The luciferase activity, the viral DNA copies and the luciferase transcripts were assessed in cultured cells as well as in the tissues. Results Large differences of luciferase activity, up to 2 logs, were observed between different rodent strains after intramuscular injection of Ad Luciferase. This inter-strain variation of transgene expression was due to a difference in transcription efficiency. The transgene expression level in tumour cell lines of different tissue origin could be explained largely by the difference of infectibility to the adenovirus. In contrast, the main step responsible for luciferase activity variation, between six human breast cancer cell lines with similar phenotype, was at the transcriptional level. Conclusion Difference in transcriptional efficiency in muscles as observed between different inbred strains and between human breast cancer cell lines may be expected to occur between individual patients. This might have important consequences for clinical gene therapy. The variation between tumour types and tissues within a species are mainly at the levels of infectivity.

Attema Joline

2003-08-01

183

Transgenic A1 adenosine receptor overexpression increases myocardial resistance to?ischemia  

OpenAIRE

Activation of myocardial A1 adenosine receptors (A1AR) protects the heart from ischemic injury. In this study transgenic mice were created using the cardiac-specific ?-myosin heavy chain promoter and rat A1AR cDNA. Heart membranes from two transgene positive lines displayed ?1,000-fold overexpression of A1AR (6,574 ± 965 and 10,691 ± 1,002 fmol per mg of protein vs. 8 ± 5 fmol per mg of protein in control hearts). Compared with control hearts, transgenic Langendorff-perfused hearts had ...

Matherne, G. Paul; Linden, Joel; Byford, Anne M.; Gauthier, Naomi S.; Headrick, John P.

1997-01-01

184

Regeneration of transgenic tamarillo plants.  

Science.gov (United States)

Media were developed to regenerate shoots from leaf pieces of tamarillo (Cyphomandra betacea (Cav.) Sendtner). Shoots were derived via organogenesis and could be easily rooted and transferred to the growth chamber. Transgenic tamarillo plants were produced using the binary vector pKIWI110 in the avirulent Agrobacterium strain LBA4404. All transgenic plants were kanamycin resistant and some plants expressed the ? D-glucuronidase (gusA) reporter gene and were chlorsulfuron resistant. Molecular evidence for transformation was obtained using PCR (polymerase chain reaction) and Southern hybridization. Inheritance of the transgenic phenotypes was demonstrated in seedling progeny. PMID:24197262

Atkinson, R G; Gardner, R C

1993-04-01

185

Extra-prostatic Transgene-associated Neoplastic Lesions in Transgenic Adenocarcinoma of the Mouse Prostate (TRAMP) Mice.  

Science.gov (United States)

Male transgenic adenocarcinoma of the mouse prostate (TRAMP) mice are frequently used in prostate cancer research because their prostates consistently develop a series of preneoplastic and neoplastic lesions. Disease progression in TRAMP mouse prostates culminates in metastatic, poorly differentiated carcinomas with neuroendocrine features. The androgen dependence of the rat probasin promoter largely limits transgene expression to the prostatic epithelium. However, extra-prostatic transgene-positive lesions have been described in TRAMP mice, including renal tubuloacinar carcinomas, neuroendocrine carcinomas of the urethra, and phyllodes-like tumors of the seminal vesicle. Here, we describe the histologic and immunohistochemical features of 2 novel extra-prostatic lesions in TRAMP mice: primary anaplastic tumors of uncertain cell origin in the midbrain and poorly differentiated adenocarcinomas of the submandibular salivary gland. These newly characterized tumors apparently result from transgene expression in extra-prostatic locations rather than representing metastatic prostate neoplasms because lesions were identified in both male and female mice and in male TRAMP mice without histologically apparent prostate tumors. In this article, we also calculate the incidences of the urethral carcinomas and renal tubuloacinar carcinomas, further elucidate the biological behavior of the urethral carcinomas, and demonstrate the critical importance of complete necropsies even when evaluating presumably well characterized phenotypes in genetically engineered mice. PMID:24742627

Berman-Booty, Lisa D; Thomas-Ahner, Jennifer M; Bolon, Brad; Oglesbee, Michael J; Clinton, Steven K; Kulp, Samuel K; Chen, Ching-Shih; La Perle, Krista M D

2015-02-01

186

ALIMENTOS TRANSGÉNICOS TRANSGENIC FOODS  

Directory of Open Access Journals (Sweden)

Full Text Available Gracias al gran avance de la tecnología, la ingeniería genética y la biología molecular, se han desarrollado los productos transgénicos. En sus inicios, los productos modificados genéticamente tenían como objeto obtener ventajas en las áreas de la agricultura y ganadería. Posteriormente esta técnica se comenzó a aplicar en el ámbito de la producción de alimentos para el consumo humano. Se ha generado mucha controversia en relación a su utilización. Esta revisión tiene por objeto revisar la información científica disponible en relación a las aplicaciones, ventajas y potenciales riesgos para la salud humana y el medio ambiente asociados al consumo de los alimentos transgénicosDue to the advancements in technology, genetic engineering and molecular biology, have develop transgenic foods. Initially, genetically modified plants were produced to confer advantages in agriculture and animal husbandry. Later this technique was applied to the production of food for human consumption, generating a great deal of controversy. This review discusses the available scientific evidence in relation to the advantages and potential risks of genetically modified foods

María Soledad Reyes S.

2003-04-01

187

ALIMENTOS TRANSGÉNICOS / TRANSGENIC FOODS  

Scientific Electronic Library Online (English)

Full Text Available SciELO Chile | Language: Spanish Abstract in spanish Gracias al gran avance de la tecnología, la ingeniería genética y la biología molecular, se han desarrollado los productos transgénicos. En sus inicios, los productos modificados genéticamente tenían como objeto obtener ventajas en las áreas de la agricultura y ganadería. Posteriormente esta técnica [...] se comenzó a aplicar en el ámbito de la producción de alimentos para el consumo humano. Se ha generado mucha controversia en relación a su utilización. Esta revisión tiene por objeto revisar la información científica disponible en relación a las aplicaciones, ventajas y potenciales riesgos para la salud humana y el medio ambiente asociados al consumo de los alimentos transgénicos Abstract in english Due to the advancements in technology, genetic engineering and molecular biology, have develop transgenic foods. Initially, genetically modified plants were produced to confer advantages in agriculture and animal husbandry. Later this technique was applied to the production of food for human consump [...] tion, generating a great deal of controversy. This review discusses the available scientific evidence in relation to the advantages and potential risks of genetically modified foods

María Soledad, Reyes S.; Jaime, Rozowski N.

2003-04-01

188

Temporal Expression of Mutant LRRK2 in Adult Rats Impairs Dopamine Reuptake  

Directory of Open Access Journals (Sweden)

Full Text Available Parkinson's disease (PD results from progressive degeneration of dopaminergic neurons. Most PD cases are sporadic, but some have pathogenic mutation in the individual genes. Mutation of the leucine-rich repeat kinase-2 (LRRK2 gene is associated with familial and sporadic PD, as exemplified by G2019S substitution. While constitutive expression of mutant LRRK2 in transgenic mice fails to induce neuron death, transient expression of the disease gene by viral delivery causes a substantial loss of dopaminergic neurons in mice. To further assess LRRK2 pathogenesis, we created inducible transgenic rats expressing human LRRK2 with G2019S substitution. Temporal overexpression of LRRK2G2019S in adult rats impaired dopamine reuptake by dopamine transporter (DAT and thus enhanced locomotor activity, the phenotypes that were not observed in transgenic rats constitutively expressing the gene throughout life time. Reduced DAT binding activity is an early sign of dopaminergic dysfunction in asymptomatic subjects carrying pathogenic mutation in LRRK2. Our transgenic rats recapitulated the initiation process of dopaminergic dysfunction caused by pathogenic mutation in LRRK2. Inducible transgenic approach uncovered phenotypes that may be obscured by developmental compensation in constitutive transgenic rats. Finding in inducible LRRK2 transgenic rats would guide developing effective strategy in transgenic studies: Inducible expression of transgene may induce greater phenotypes than constitutive gene expression, particularly in rodents with short life time.

Hongxia Zhou, Cao Huang, Jianbin Tong, Weimin C Hong, Yong-Jian Liu, Xu-Gang Xia

2011-01-01

189

Changes in function of antigen-specific lymphocytes correlating with progression towards diabetes in a transgenic model.  

OpenAIRE

Mice that express influenza hemagglutinin under control of the rat insulin promoter (INS-HA) as well as a class II major histocompatibility complex (MHC)-restricted HA-specific transgenic TCR (TCR-HA), develop early insulitis with huge infiltrates, but progress late and irregularly to diabetes. Initially, in these mice, INS-HA modulates the reactivity of antigen-specific lymphocytes, such that outside the pancreas they do not cause lethal shock like their naive counterparts in single transgen...

Sarukhan, A.; Lanoue, A.; Franzke, A.; Brousse, N.; Buer, J.; Von Boehmer, H.

1998-01-01

190

Pancreatic expression and secretion of human islet amyloid polypeptide in a transgenic mouse.  

Science.gov (United States)

Islet amyloid polypeptide (IAPP) is a secretory product of the pancreatic beta-cell, which is the primary constituent of the islet amyloid that develops in type II diabetes. To study the role the inherent amyloidogenicity of human IAPP (hIAPP) plays in the formation of islet amyloid deposits and to investigate a possible hormonal role for IAPP, transgenic mice expressing hIAPP were developed. The transgene was composed of a fragment of an hIAPP cDNA linked to the rat insulin II promoter. One line of transgenic mice expressed the transgene and synthesized hIAPP in their pancreatic islets. IAPP-like immunoreactivity in pancreatic extracts and plasma were two- to threefold greater in the transgenic mice compared with nontransgenic control mice. Although plasma concentrations of immunoreactive insulin (IRI) and glucose were equal in transgenic and control mice, the pancreatic content of IRI was nearly twofold greater in the transgenic animals, and proinsulin mRNA was significantly elevated, suggesting increased rates of insulin biosynthesis. Pancreatic samples obtained from transgenic mice up to 19 months of age had no evidence of islet amyloid. These results indicate that an increased level of synthesis of the amyloidogenic hIAPP is not sufficient to cause islet amyloid deposition. However, the increased synthesis and storage of insulin in the islets of the transgenic mice are consistent with either a direct regulatory effect of IAPP on the beta-cell or indirect stimulation of insulin production through IAPP-induced insulin resistance. PMID:7958499

D'Alessio, D A; Verchere, C B; Kahn, S E; Hoagland, V; Baskin, D G; Palmiter, R D; Ensinck, J W

1994-12-01

191

Human antibody production in transgenic animals.  

Science.gov (United States)

Fully human antibodies from transgenic animals account for an increasing number of new therapeutics. After immunization, diverse human monoclonal antibodies of high affinity can be obtained from transgenic rodents, while large animals, such as transchromosomic cattle, have produced respectable amounts of specific human immunoglobulin (Ig) in serum. Several strategies to derive animals expressing human antibody repertoires have been successful. In rodents, gene loci on bacterial artificial chromosomes or yeast artificial chromosomes were integrated by oocyte microinjection or transfection of embryonic stem (ES) cells, while ruminants were derived from manipulated fibroblasts with integrated human chromosome fragments or human artificial chromosomes. In all strains, the endogenous Ig loci have been silenced by gene targeting, either in ES or fibroblast cells, or by zinc finger technology via DNA microinjection; this was essential for optimal production. However, comparisons showed that fully human antibodies were not as efficiently produced as wild-type Ig. This suboptimal performance, with respect to immune response and antibody yield, was attributed to imperfect interaction of the human constant region with endogenous signaling components such as the Ig?/? in mouse, rat or cattle. Significant improvements were obtained when the human V-region genes were linked to the endogenous CH-region, either on large constructs or, separately, by site-specific integration, which could also silence the endogenous Ig locus by gene replacement or inversion. In animals with knocked-out endogenous Ig loci and integrated large IgH loci, containing many human Vs, all D and all J segments linked to endogenous C genes, highly diverse human antibody production similar to normal animals was obtained. PMID:25467949

Brüggemann, Marianne; Osborn, Michael J; Ma, Biao; Hayre, Jasvinder; Avis, Suzanne; Lundstrom, Brian; Buelow, Roland

2015-04-01

192

Transgenic approaches to crop improvement.  

Science.gov (United States)

Transgenic crops are now grown commercially on several million hectares, principally in North America. To date, the predominant crops are maize (corn), soybean, cotton, and potatoes. In addition, there have been field trials of transgenics from at least 52 species including all the major field crops, vegetables, and several herbaceous and woody species. This review summarizes recent data relating to such trials, particularly in terms of the trends away from simple, single gene traits such as herbicide and insect resistance towards more complex agronomic traits such as growth rate and increased photosynthetic efficiency. Much of the recent information is derived from inspection of patent databases, a useful source of information on commercial priorities. The review also discusses the time scale for the introduction of these transgenes into breeding populations and their eventual release as new varieties. PMID:10938856

Dunwell, J M

2000-02-01

193

Progress on researches of transgenic alfalfa  

International Nuclear Information System (INIS)

In this paper, the progress on the researches of transgenic alfalfa in the past two decades had been reviewed in the aspects of regeneration system, transformation, improvement of the important traits and so on. Moreover, such problems as variation of transgene expression and safety of transgenic plant had also been discussed and propose had been given for the future research work. (authors)

194

TRANSGENIC FISH: In Genomics and Genetics  

Science.gov (United States)

Fish into which foreign DNA is artificially introduced and integrated into their genome are called transgenic fish. Since the development of the first transgenic fish in 1985, techniques to produce transgenic fish have improved tremendously, resulting in the production of genetically modified (GM)...

195

Transcription-dependent silencing of inducible convergent transgenes in transgenic mice  

OpenAIRE

Abstract Background Silencing of transgenes in mice is a common phenomenon typically associated with short multi-copy transgenes. We have investigated the regulation of the highly inducible human granulocyte-macrophage colony-stimulating-factor gene (Csf2) in transgenic mice. Results In the absence of any previous history of transcriptional activation, this transgene was expressed in T lineage cells at the correct inducible level in all lines of mice tested. In contrast, the transgene was sil...

Calero-Nieto Fernando J; Bert Andrew G; Cockerill Peter N

2010-01-01

196

Identifying and genotyping transgene integration loci.  

Science.gov (United States)

The random germline integration of genetically engineered transgenes has been a powerful technique to study the role of particular genes in variety of biological processes. Although the identification of the transgene insertion site is often not essential for functional analysis of the transgene, identifying the site can have practical benefit. Enabling one to distinguish between animals that are homozygous or hemizygous for the transgene locus could facilitate breeding strategies to produce animals with a large number of genetic markers. Furthermore, founder lines generated with the same transgene construct may exhibit different phenotypes and levels of transgene expression depending on the site of integration. The goal of this report was to develop a rapid protocol for the identification and verification of transgene insertion sites. To identify host genomic sequences at the coagulation Factor X transgene integration site, DNA from a tail snip of the transgenic mouse was digested with NcoI and circularized using T4 DNA ligase. Using appropriately positioned PCR primers annealing to a transgene fragment distal to a terminal transgene restriction site (NcoI), one could amplify a fragment containing the transgene terminal region and extending into the flanking genomic sequence at the insertion site. DNA sequence determination of the amplicon permitted identification of the insertion site using a BLASTN search. FISH analysis of a metaphase spread of primary fibroblasts derived from the transgenic mouse was consistent with the identification of insertion site near the end of mouse chromosome 14. Identification of transgene insertion sites will facilitate genotyping strategies useful for the construction of mice with multiple engineered genetic markers and to distinguish among different founder lines generated by the same transgene. Furthermore, identification of the insertion site is necessary to analyze unexpected phenotypes that might be caused by insertional inactivation of an endogenous gene. PMID:18612840

Liang, Zhong; Breman, Amy Marie; Grimes, Brenda R; Rosen, Elliot D

2008-10-01

197

Prolonged survival of pancreatic islet allografts mediated by adenovirus immunoregulatory transgenes.  

OpenAIRE

The adenovirus (Ad) early region 3 (E3) genes code for at least four proteins that inhibit the host immune responses mediated by cytotoxic T lymphocytes and tumor necrosis factor alpha. To evaluate the potential use of these immunoregulatory viral functions in facilitating allogeneic cell transplantation, the Ad E3 genes were expressed in pancreatic beta cells in transgenic mice under control of the rat insulin II promoter. Transgenic H-2b/d (C57BL/6 x BALB/c) islets, expressing the Ad E3 gen...

Efrat, S.; Fejer, G.; Brownlee, M.; Horwitz, M. S.

1995-01-01

198

Production and analysis of transgenic mice with ectopic expression of parvalbumin.  

Science.gov (United States)

Transgenic mice expressing rat parvalbumin under the control of the human metallothionein IIA (MTII A), SV-40 early, and neuron-specific enolase (NSE) promoters were produced. Ectopic expression was analyzed by RNA polymerase chain reaction and RNase protection in combination with immunohistochemistry. From a total of 25 transgenic lines 18 were found to express the transgene. Expression strength and tissue specificity were dependent upon the promoter used and varied considerably among animal lines produced with the same construct. Highest constitutive MT IIA-driven expression was found in lung, liver, heart, and kidney, as well as in brain, and lower amounts of transgene expression were found in spleen, testis, and muscle. Immunohistochemistry of tissue sections of metallothionein-parvalbumin transgenic strain 29 in the non-induced state revealed that ectopic PV mRNA is translated into protein. Short-term induction of the MT IIA promoter by CdSO4 or CdCl2 leads to a shift in tissue specificity and does not increase ectopic expression in tissues where the transgene is active in the noninduced state. As expected the NSE promoter showed highest activity in brain. However, NSE-driven expression could also be detected to various degrees in all investigated tissues. SV-40-dependent PV expression showed no tissue preference and varied considerably among different strains. Except for the observation that the SV-40-PV construct showed lower yields in transgenic production and reduced numbers of positive offspring no obvious impairment of growth or behavior as a consequence of transgenic PV expression could be detected. PMID:7532934

Castillo, M B; Celio, M R; Andressen, C; Gotzos, V; Rülicke, T; Berger, M C; Weber, J; Berchtold, M W

1995-02-20

199

Malignant melanoma in transgenic mice.  

OpenAIRE

Ocular and cutaneous melanomas arose in new inbred lines of transgenic mice having an integrated recombinant gene comprised of the tyrosinase promoter, expressed in pigment cells, and the simian virus 40 early-region transforming sequences. The tumors were hypomelanotic and were histopathologically similar to corresponding human melanomas. Eye melanomas often originated at a young age, chiefly from the retinal pigment epithelium, also from the choroid, and rarely from the ciliary body. The ey...

Bradl, M.; Klein-szanto, A.; Porter, S.; Mintz, B.

1991-01-01

200

Transgenic trees and forestry biosafety  

Scientific Electronic Library Online (English)

Full Text Available SciELO Chile | Language: English Abstract in english The benefits from the development of transgenic trees are expected from the improvement of traits as growth and form, wood quality, industrial processes, disease and insect resistance, herbicide tolerance, ecological restoration, rooting ability, etc. One of the first reported field trials with gene [...] tically modified forest trees was established in Belgium in 1988 and the characteristic evaluated was herbicide tolerance in poplars. Since then, there have been more than 200 reported trials, involving at least 15 forest species. The majority of the field trials have been carried out in the USA (64%). More than 50% of the field trials are done with Populus species and the main target traits are herbicide tolerance (31%), followed by marker genes (23%) and insect resistance (14%). Until today, there is only one report on commercial-scale production of transgenic forest trees which is Populus nigra with the Bt gene release in China in 2002 and established on commercial plantations in 2003. Operational application of GMO's in forestry depends on technical, economical, political and public aspects, but the development of adequate regulatory frameworks and public acceptance of transgenic trees will define the future of this technology in forestry.

Sofía, Valenzuela; Claudio, Balocchi; Jaime, Rodríguez.

2006-06-01

201

Improving expression of reporter transgene in stem cell by construction of different lentiviral vectors  

International Nuclear Information System (INIS)

For stem cell trafficking applications, it is imperative to express transgenes at desired and stable levels. In recent years, lentivirus-mediated gene transfer was shown to be an efficient method to stably introduce genetic modifications in target cells, even if these are in proliferative or nonproliferative states. Moreover, transgene expression levels can be controlled by using different promoters. The present study was designed to compare the potency of various promoters regulating expression of imaging reporter genes in embryonic H9c2 cardiomyoblasts derived from rat heart. Lentiviral vector was produced by the transient transfection of plasmids carrying required genes and those encoding for virus coating proteins into 293T cells. Harvested viral constructs were incubated with Hela and H9c2 cells, respectively. Transgene expressions were detected by several imaging modalities and evaluated by enzymatic assays. Results - We observed that the level of stable transgene expression in lentivirus-transduced myoblasts could be modulated over several orders of magnitude, with the Ubiquitin (Ub) promoter exhibiting the highest activity, intermediate expression was observed with the CAG promoter, whereas expression observed with the CMV promoter was very weak. We observed that the level of stable transgene expression in lentivirus-transduced myoblasts could be modulated over several orders of magnitude, with the Ubiquitin (Ub) promoter exhibiting the highest activity, interter exhibiting the highest activity, intermediate expression was observed with the CAG promoter, whereas expression observed with the CMV promoter was very weak. Here we show that lentivirus-mediated gene transfer allows efficient and stable transgene expression in embryonic cardiomyoblasts in vitro and that transgene expression levels can be varied by using different well-characterized gene promoters. In vivo trials about gene expression will probably further determine the potential of long-term trafficking stem cells using lentivirus

202

Improving expression of reporter transgene in stem cell by construction of different lentiviral vectors  

Energy Technology Data Exchange (ETDEWEB)

For stem cell trafficking applications, it is imperative to express transgenes at desired and stable levels. In recent years, lentivirus-mediated gene transfer was shown to be an efficient method to stably introduce genetic modifications in target cells, even if these are in proliferative or nonproliferative states. Moreover, transgene expression levels can be controlled by using different promoters. The present study was designed to compare the potency of various promoters regulating expression of imaging reporter genes in embryonic H9c2 cardiomyoblasts derived from rat heart. Lentiviral vector was produced by the transient transfection of plasmids carrying required genes and those encoding for virus coating proteins into 293T cells. Harvested viral constructs were incubated with Hela and H9c2 cells, respectively. Transgene expressions were detected by several imaging modalities and evaluated by enzymatic assays. Results - We observed that the level of stable transgene expression in lentivirus-transduced myoblasts could be modulated over several orders of magnitude, with the Ubiquitin (Ub) promoter exhibiting the highest activity, intermediate expression was observed with the CAG promoter, whereas expression observed with the CMV promoter was very weak. We observed that the level of stable transgene expression in lentivirus-transduced myoblasts could be modulated over several orders of magnitude, with the Ubiquitin (Ub) promoter exhibiting the highest activity, intermediate expression was observed with the CAG promoter, whereas expression observed with the CMV promoter was very weak. Here we show that lentivirus-mediated gene transfer allows efficient and stable transgene expression in embryonic cardiomyoblasts in vitro and that transgene expression levels can be varied by using different well-characterized gene promoters. In vivo trials about gene expression will probably further determine the potential of long-term trafficking stem cells using lentivirus.

Tae, Seong Ho; Min, Jung Joon [Chonnam National University Medical School, Gwangju (Korea, Republic of); Le, Uyenchi N.; Padmanabhan, Parasuraman [Singapore Bio-Imaging Imaging Consortium, Singapore (Singapore)

2007-07-01

203

Adenovirus-mediated gene transfer to rat testis in vivo.  

Science.gov (United States)

To study transgene expression in the adult rat testis in vivo, an adenovirus vector carrying a lacZ transgene with a nuclear localization signal was used as a marker. The adenovirus vector was first tested on rat Sertoli cell-germ cell cocultures in vitro; it efficiently mediated transgene expression in Sertoli cells but not germ cells. This vector was then delivered to the interstitial compartment of adult rat testes by intratesticular injection, resulting in Leydig cells expressing the transgene. Alternatively, delivering the vector to the intratubular compartment by rete testis injection resulted in expression of the transgene by Sertoli cells of the seminiferous epithelium and principal cells of the epididymis. In vivo, each cell type expressed the transgene by 2 days postinfection, and expression persisted for at least 10 days; however, later time points were associated with a loss of transgene expression and focal interstitial inflammation. This study documents the ability of adenovirus to mediate gene transfer to specific testicular cells, providing a powerful tool to study the short-term effects of specific genes on spermatogenesis in vivo. PMID:9116152

Blanchard, K T; Boekelheide, K

1997-02-01

204

Myosin light chain enhancer activates muscle-specific, developmentally regulated gene expression in transgenic mice.  

OpenAIRE

The rat myosin light chain (MLC)1/3 gene locus contains a potent muscle-specific enhancer, located downstream of the coding region, greater than 24 kilobases away from the MLC1 transcription start site. To assess the role of this enhancer in the activation of MLC expression during development, transgenic mice were generated carrying multiple copies of a MLC1 promoter-chloramphenicol acetyltransferase (CAT) transcription unit linked to a genomic fragment including the enhancer. CAT expression ...

Rosenthal, N.; Kornhauser, J. M.; Donoghue, M.; Rosen, K. M.; Merlie, J. P.

1989-01-01

205

Inducible Nephrin Transgene Expression in Podocytes Rescues Nephrin-Deficient Mice from Perinatal Death  

OpenAIRE

Mutations leading to nephrin loss result in massive proteinuria both in humans and mice. Early perinatal lethality of conventional nephrin knockout mice makes it impossible to determine the role of nephrin protein in the adult kidney and in extra-renal tissues. Herein, we studied whether podocyte-specific, doxycycline-inducible, rat nephrin expression can rescue nephrin-deficient mice from perinatal lethality. Fourteen littermates out of 72 lacked endogenous nephrin and expressed transgenic r...

Juhila, Juuso; Lassila, Markus; Roozendaal, Ramon; Lehtonen, Eero; Messing, Marcel; Langer, Brigitte; Kerjaschki, Dontscho; Verbeek, J. Sjef; Holthofer, Harry

2010-01-01

206

Transcription-dependent silencing of inducible convergent transgenes in transgenic mice  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background Silencing of transgenes in mice is a common phenomenon typically associated with short multi-copy transgenes. We have investigated the regulation of the highly inducible human granulocyte-macrophage colony-stimulating-factor gene (Csf2 in transgenic mice. Results In the absence of any previous history of transcriptional activation, this transgene was expressed in T lineage cells at the correct inducible level in all lines of mice tested. In contrast, the transgene was silenced in a specific subset of lines in T cells that had encountered a previous episode of activation. Transgene silencing appeared to be both transcription-dependent and mediated by epigenetic mechanisms. Silencing was accompanied by loss of DNase I hypersensitive sites and inability to recruit RNA polymerase II upon stimulation. This pattern of silencing was reflected by increased methylation and decreased acetylation of histone H3 K9 in the transgene. We found that silenced lines were specifically associated with a single pair of tail-to-tail inverted repeated copies of the transgene embedded within a multi-copy array. Conclusions Our study suggests that epigenetic transgene silencing can result from convergent transcription of inverted repeats which can lead to silencing of an entire multi-copy transgene array. This mechanism may account for a significant proportion of the reported cases of transgene inactivation in mice.

Calero-Nieto Fernando J

2010-01-01

207

2008 FHB Analysis of Transgenic Barley Lines  

Science.gov (United States)

Transgenic lines have been developed with the goal of reducing FHB and DON in barley. Replicated field trials for FHB reaction of 48 Conlon transgenic lines were conducted in 2008 in Langdon, ND and Rosemount, MN. The Langdon trials consisted of three replicates in hill plots in an inoculated misted...

208

[Progress in transgenic fish techniques and application].  

Science.gov (United States)

Transgenic technique provides a new way for fish breeding. Stable lines of growth hormone gene transfer carps, salmon and tilapia, as well as fluorescence protein gene transfer zebra fish and white cloud mountain minnow have been produced. The fast growth characteristic of GH gene transgenic fish will be of great importance to promote aquaculture production and economic efficiency. This paper summarized the progress in transgenic fish research and ecological assessments. Microinjection is still the most common used method, but often resulted in multi-site and multi-copies integration. Co-injection of transposon or meganuclease will greatly improve the efficiency of gene transfer and integration. "All fish" gene or "auto gene" should be considered to produce transgenic fish in order to eliminate misgiving on food safety and to benefit expression of the transferred gene. Environmental risk is the biggest obstacle for transgenic fish to be commercially applied. Data indicates that transgenic fish have inferior fitness compared with the traditional domestic fish. However, be-cause of the genotype-by-environment effects, it is difficult to extrapolate simple phenotypes to the complex ecological interactions that occur in nature based on the ecological consequences of the transgenic fish determined in the laboratory. It is critical to establish highly naturalized environments for acquiring reliable data that can be used to evaluate the environ-mental risk. Efficacious physical and biological containment strategies remain to be crucial approaches to ensure the safe application of transgenic fish technology. PMID:21586396

Ye, Xing; Tian, Yuan-Yuan; Gao, Feng-Ying

2011-05-01

209

Transposon-mediated chromosomal integration of transgenes in the parasitic nematode Strongyloides ratti and establishment of stable transgenic lines.  

Science.gov (United States)

Genetic transformation is a potential tool for analyzing gene function and thereby identifying new drug and vaccine targets in parasitic nematodes, which adversely affect more than one billion people. We have previously developed a robust system for transgenesis in Strongyloides spp. using gonadal microinjection for gene transfer. In this system, transgenes are expressed in promoter-regulated fashion in the F1 but are silenced in subsequent generations, presumably because of their location in repetitive episomal arrays. To counteract this silencing, we explored transposon-mediated chromosomal integration of transgenes in S. ratti. To this end, we constructed a donor vector encoding green fluorescent protein (GFP) under the control of the Ss-act-2 promoter with flanking inverted tandem repeats specific for the piggyBac transposon. In three experiments, free-living Strongyloides ratti females were transformed with this donor vector and a helper plasmid encoding the piggyBac transposase. A mean of 7.9% of F1 larvae were GFP-positive. We inoculated rats with GFP-positive F1 infective larvae, and 0.5% of 6014 F2 individuals resulting from this host passage were GFP-positive. We cultured GFP-positive F2 individuals to produce GFP-positive F3 L3i for additional rounds of host and culture passage. Mean GFP expression frequencies in subsequent generations were 15.6% in the F3, 99.0% in the F4, 82.4% in the F5 and 98.7% in the F6. The resulting transgenic lines now have virtually uniform GFP expression among all progeny after at least 10 generations of passage. Chromosomal integration of the reporter transgenes was confirmed by Southern blotting and splinkerette PCR, which revealed the transgene flanked by S. ratti genomic sequences corresponding to five discrete integration sites. BLAST searches of flanking sequences against the S. ratti genome revealed integrations in five contigs. This result provides the basis for two powerful functional genomic tools in S. ratti: heritable transgenesis and insertional mutagenesis. PMID:22912584

Shao, Hongguang; Li, Xinshe; Nolan, Thomas J; Massey, Holman C; Pearce, Edward J; Lok, James B

2012-01-01

210

Disease-associated polymorphisms in ERAP1 do not alter endoplasmic reticulum stress in patients with ankylosing spondylitis.  

Science.gov (United States)

The mechanism by which human leukocyte antigen B27 (HLA-B27) contributes to ankylosing spondylitis (AS) remains unclear. Genetic studies demonstrate that association with and interaction between polymorphisms of endoplasmic reticulum aminopeptidase 1 (ERAP1) and HLA-B27 influence the risk of AS. It has been hypothesised that ERAP1-mediated HLA-B27 misfolding increases endoplasmic reticulum (ER) stress, driving an interleukin (IL) 23-dependent, pro-inflammatory immune response. We tested the hypothesis that AS-risk ERAP1 variants increase ER-stress and concomitant pro-inflammatory cytokine production in HLA-B27(+) but not HLA-B27(-) AS patients or controls. Forty-nine AS cases and 22 healthy controls were grouped according to HLA-B27 status and AS-associated ERAP1 rs30187 genotypes: HLA-B27(+)ERAP1(risk), HLA-B27(+)ERAP1(protective), HLA-B27(-)ERAP1(risk) and HLA-B27(-)ERAP1(protective). Expression levels of ER-stress markers GRP78 (8?kDa glucose-regulated protein), CHOP (C/EBP-homologous protein) and inflammatory cytokines were determined in peripheral blood mononuclear cell and ileal biopsies. We found no differences in ER-stress gene expression between HLA-B27(+) and HLA-B27(-) cases or healthy controls, or between cases or controls stratified by carriage of ERAP1 risk or protective alleles in the presence or absence of HLA-B27. No differences were observed between expression of IL17A or TNF (tumour necrosis factor) in HLA-B27(+)ERAP1(risk), HLA-B27(+)ERAP1(protective) and HLA-B27(-)ERAP1(protective) cases. These data demonstrate that aberrant ERAP1 activity and HLA-B27 carriage does not alter ER-stress levels in AS, suggesting that ERAP1 and HLA-B27 may influence disease susceptibility through other mechanisms. PMID:25354578

Kenna, T J; Lau, M C; Keith, P; Ciccia, F; Costello, M-E; Bradbury, L; Low, P-L; Agrawal, N; Triolo, G; Alessandro, R; Robinson, P C; Thomas, G P; Brown, M A

2015-01-01

211

Transgene-induced gene silencing in plants.  

Science.gov (United States)

RNAi is the most important reverse genetics tool to trigger transgenic gene silencing, which is now applied widely to investigate gene function and also practically applied to enhance resistance to biotic and abiotic stress. Recently, the most effective way to induce transgenic gene silencing is to introduce inverted repeat (IR) double-stranded RNA (dsRNA) or artificial microRNA (amiRNA) instead of a transgenic sense or antisense strand of genes. The stable transgenic plants can be acquired through Agrobacterium tumefaciens-mediated transformation of binary vectors containing an RNAi hairpin construct or amiRNA precursor backbone sequence. Here we primarily describe these two methods' vector construction, plant transformation, and transgenic line verification. PMID:25740359

Jin, Yun; Guo, Hui-Shan

2015-01-01

212

Transgenic expression of human cytoxic T-lymphocyte associated antigen4-Immunoglobulin (hCTLA4Ig) by porcine skin for xenogeneic skin grafting.  

Science.gov (United States)

Porcine skin is frequently used as a substitute of human skin to cover large wounds in clinic practice of wound care. In our previous work, we found that transgenic expression of human cytoxicT-lymphocyte associated antigen4-immunoglobulin (hCTLA4Ig) in murine skin graft remarkably prolonged its survival in xenogeneic wounds without extensive immunosuppression in recipients, suggesting that transgenic hCTLA4Ig expression in skin graft may be an effective and safe method to prolong xenogeneic skin graft survival. In this work, using a transgene construct containing hCTLA4Ig coding sequence under the drive of human Keratine 14 (k14) promoter, hCTLA4Ig transgenic pigs were generated by somatic nuclear transfer. The derived transgenic pigs were healthy and exhibited no signs of susceptibility to infection. The hCTLA4Ig transgene was stably transmitted through germline over generations, and thereby a transgenic pig colony was established. In the derived transgenic pigs, hCTLA4Ig expression in skin was shown to be genetically stable over generations, and detected in heart, kidney and corneal as well as in skin. Transgenic hCTLA4Ig protein in pigs exhibited expected biological activity as it suppressed human lymphocyte proliferation in human mixed lymphocyte culture to extents comparable to those of commercially purchased purified hCTLA4Ig protein. In skin grafting from pigs to rats, transgenic porcine skin grafts exhibited remarkably prolonged survival compared to the wild-type skin grafts derived from the same pig strain (13.33 ± 3.64 vs. 6.25 ± 2.49 days, P work can be used as a reproducible resource to provide porcine skin grafts with extended survival for wound coverage, and also as donors to investigate the impacts of hCTLA4Ig on xenotransplantation of other organs (heart, kidney and corneal) due to the ectopic transgenic hCTLA4Ig expression. PMID:25236862

Wang, Yong; Yang, Hua-Qiang; Jiang, Wen; Fan, Na-Na; Zhao, Ben-Tian; Ou-Yang, Zhen; Liu, Zhao-Ming; Zhao, Yu; Yang, Dong-Shan; Zhou, Xiao-Yang; Shang, Hai-Tao; Wang, Lu-Lu; Xiang, Peng-Ying; Ge, Liang-Peng; Wei, Hong; Lai, Liang-Xue

2015-04-01

213

Glyphostate-drift but not herbivory alters the rate of transgene flow from single and stacked trait transgenic canola (Brassica napus L.) to non-transgenic B. napus and B. rapa  

Science.gov (United States)

While transgenic plants can offer agricultural benefits, the escape of transgenes out of crop fields is a major environmental concern. Escape of transgenic herbicide resistance has occurred between transgenic Brassica napus (canola) and weedy species in numerous locations. In t...

214

HLA class I associations of ankylosing spondylitis in the white population in the United Kingdom.  

OpenAIRE

OBJECTIVE: To investigate the HLA class I associations of ankylosing spondylitis (AS) in the white population, with particular reference to HLA-B27 subtypes. METHODS: HLA-B27 and -B60 typing was performed in 284 white patients with AS. Allele frequencies of HLA-B27 and HLA-B60 from 5926 white bone marrow donors were used for comparison. HLA-B27 subtyping was performed by single strand conformation polymorphism (SSCP) in all HLA-B27 positive AS patients, and 154 HLA-B27 positive ethnically mat...

Brown, Ma; Pile, Kd; Kennedy, Lg; Calin, A.; Darke, C.; Bell, J.; Wordsworth, Bp; Corne?lis, F.

1996-01-01

215

Comparison of the antibody responses to the 77 Klebsiella capsular types in ankylosing spondylitis and various rheumatic diseases.  

OpenAIRE

The production of antibodies to Klebsiella capsular polysaccharides was measured in sera from either HLA-B27-positive (HLA-B27+) or HLA-B27-negative (HLA-B27-) patients with classical ankylosing spondylitis (n = 54). These sera were compared with sera from patients with various rheumatic diseases (n = 82) and HLA-B27+ or HLA-B27- healthy individuals (n = 85). All sera were analyzed by means of an enzyme-linked immunosorbent assay specific to each of the 77 Klebsiella serotypes. The sera from ...

Sahly, H.; Kekow, J.; Podschun, R.; Schaff, M.; Gross, W. L.; Ullmann, U.

1994-01-01

216

Leukemia Inhibitory Factor Induces Neurotransmitter Switching in Transgenic Mice  

Science.gov (United States)

Leukemia inhibitory factor (LIF) is a cytokine growth factor that induces rat sympathetic neurons to switch their neurotransmitter phenotype from noradrenergic to cholinergic in vitro. To test whether LIF can influence neuronal differentiation in vivo, we generated transgenic mice that expressed LIF in pancreatic islets under the control of the insulin promoter and evaluated the neurotransmitter phenotype of the pancreatic sympathetic innervation. We also used the insulin promoter to coexpress nerve growth factor in the islets, which greatly increased the density of sympathetic innervation and facilitated analysis of the effects of LIF. Our data demonstrate that tyrosine hydroxylase and catecholamines declined and choline acetyltransferase increased in response to LIF. We conclude that LIF can induce neurotransmitter switching of sympathetic neurons in vivo.

Bamber, Bruce A.; Masters, Brian A.; Hoyle, Gary W.; Brinster, Ralph L.; Palmiter, Richard D.

1994-08-01

217

AN APPROACH TO TRANSGENIC CROP MONITORING  

Science.gov (United States)

Remote sensing by aerial or satellite images may provide a method of identifying transgenic pesticidal crop distribution in the landscape. Genetically engineered crops containing bacterial gene(s) that express an insecticidal protein from Bacillus thuringiensis (Bt) are regulated...

218

Reversing insect adaptation to transgenic insecticidal plants.  

OpenAIRE

The refuge-high-dose strategy for delaying insect adaptation to transgenic plants produces non-transgenic plants that enable survival of susceptible individuals. Previous theoretical work has suggested three requirements for success of the refuge-high-dose strategy: a low initial frequency of the resistance allele, extensive mating between resistant and susceptible adults and recessive inheritance of resistance. In order to understand an observed decrease in resistance frequency and improve t...

Carrie?re, Y.; Tabashnik, B. E.

2001-01-01

219

Transgenic Wheat, Barley and Oats: Future Prospects  

Science.gov (United States)

Following the success of transgenic maize and rice, methods have now been developed for the efficient introduction of genes into wheat, barley and oats. This review summarizes the present position in relation to these three species, and also uses information from field trial databases and the patent literature to assess the future trends in the exploitation of transgenic material. This analysis includes agronomic traits and also discusses opportunities in expanding areas such as biofuels and biopharming.

Dunwell, Jim M.

220

Strategies for antiviral resistance in transgenic plants  

OpenAIRE

Genetic engineering offers a means of incorporating new virus resistance traits into existing desirable plant cultivars. The initial attempts to create transgenes conferring virus resistance were based on the pathogen-derived resistance concept. The expression of the viral coat protein gene in transgenic plants was shown to induce protective effects similar to classical cross protection, and was therefore distinguished as 'coat-protein-mediated' protection. Since then, a large variety of vira...

Prins, M. W.; Laimer, M.; Noris, E.; Schubert, J.; Wassenegger, M.; Tepfer, M.

2008-01-01

221

TRANSGENIC FISH MODEL IN ENVIRONMENTAL TOXICOLOGY  

Directory of Open Access Journals (Sweden)

Full Text Available A number of experiments and the use of drugs have been performed in fish. The fish may be used as model organism in various biological experiments, including environmental toxicology. Aquatic animals are being engineered to increase aquaculture production, for medical and industrial research, and for ornamental reasons. Fish have been found to play an important role in assessing potential risks associated with exposure to toxic substances in aquatic environment. Hence, it has been thought that the development of transgenic fish can enhance the use of fish in environmental toxicology. India has developed experimental transgenics of rohu fish, zebra fish, cat fish and singhi fish. Genes, promoters and vectors of indigenous origin are now available for only two species namely rohu and singhi for engineering growth. Development of fish model carrying identical transgenes to those found in rodents is beneficial and has shown that several aspects of in vivo mutagenesis are similar between the two classes of vertebrates. Fish shows the frequencies of spontaneous mutations similar to rodents and respond to mutagen exposure consistent with known mutagenic mechanisms. The feasibility of in vivo mutation analysis using transgenic fish has been demonstrated and the potential value of transgenic fish as a comparative animal model has been illustrated. Therefore, the transgenic fish can give the significant contribution to study the environmental toxicity in animals as a whole.

Madhuri Sharma

2012-05-01

222

Transgenic animals and their application in medicine  

Directory of Open Access Journals (Sweden)

Full Text Available Transgenic animals are animals that are genetically altered to have traits that mimic symptoms of specific human pathologies. They provide genetic models of various human diseases which are important in understanding disease and developing new targets. In early 1980 Gordon and co-workers described the first gene addition experiment using the microinjection technology and since then the impact of transgenic technology on basic research has been significant. Within 20 years of its inception, ATryn the first drug approved by USFDA from transgenic animals was developed and it has opened door to drugs from transgenic animals. In addition, they are looked upon as potential future donors for xenotransplantation. With increasing knowledge about the genetics and improvements in the transgenetic technology numerous useful applications like biologically safe new-generation drugs based on human regulatory proteins are being developed.Various aspects of concern in the coming years are the regulatory guidelines, ethical issues and patents related to the use of transgenic animals. This modern medicine is on the threshold of a pharmacological revolution. Use of transgenic animals will provide solutions for drug research, xenotransplantation, clinical trials and will prove to be a new insight in drug development.

Bagle TR, Kunkulol RR, Baig MS, More SY

2013-01-01

223

Soil persistence of DNA from transgenic poplar.  

Science.gov (United States)

The presence of recombinant DNA in soil cultivated with white poplars (Populus alba L.) expressing either the bar transgene for herbicide tolerance or the StSy transgene for resveratrol production, respectively, was investigated in a greenhouse over a 20-month period. The bar trial included the transgenic lines 5P56 and 6EA22P56 and the untransformed line, while the StSy trial was established with the transgenic lines 5EAC1 and 12EAC1 and with the untransformed line. All the transgenic poplars harbored the nptII marker gene. Plantlets were cultivated in pots, and soil samples were mixed in order to obtain composite pools which were used for molecular analyses. The 35SCaMV-bar (1504 bp), 35SCaMV-StSy (1403 bp) and NosP-nptII (1188 bp) sequences were detected in total DNA extracted from soil samples taken at different times after planting, using PCR/Southern blot hybridization. Microcosm experiments, carried out to assess the effects of temperature and DNA purity on transgene persistence, revealed only a partial correlation between the intensity of hybridization signals and the parameters tested. PMID:19833075

Bonadei, Martina; Balestrazzi, Alma; Frigerio, Barbara; Carbonera, Daniela

2009-01-01

224

Transgenic technologies to induce sterility  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract The last few years have witnessed a considerable expansion in the number of tools available to perform molecular and genetic studies on the genome of Anopheles mosquitoes, the vectors of human malaria. As a consequence, knowledge of aspects of the biology of mosquitoes, such as immunity, reproduction and behaviour, that are relevant to their ability to transmit disease is rapidly increasing, and could be translated into concrete benefits for malaria control strategies. Amongst the most important scientific advances, the development of transgenic technologies for Anopheles mosquitoes provides a crucial opportunity to improve current vector control measures or design novel ones. In particular, the use of genetic modification of the mosquito genome could provide for a more effective deployment of the sterile insect technique (SIT against vector populations in the field. Currently, SIT relies on the release of radiation sterilized males, which compete with wild males for mating with wild females. The induction of sterility in males through the genetic manipulation of the mosquito genome, already achieved in a number of other insect species, could eliminate the need for radiation and increase the efficiency of SIT-based strategies. This paper provides an overview of the mechanisms already in use for inducing sterility by transgenesis in Drosophila and other insects, and speculates on possible ways to apply similar approaches to Anopheles mosquitoes.

Wimmer Ernst A

2009-11-01

225

Increased Expression of the Na,K-ATPase alpha4 Isoform Enhances Sperm Motility in Transgenic Mice1  

Science.gov (United States)

The Na,K-ATPase alpha4 (ATP1A4) isoform is specifically expressed in male germ cells and is highly prevalent in spermatozoa. Although selective inhibition of alpha4 activity with ouabain has been shown to affect sperm motility, a more direct analysis of the role of this isoform in sperm movement has not yet been demonstrated. To establish this, we engineered transgenic mice that express the rat alpha4 isoform fused to green fluorescent protein in male germ cells, under the control of the mouse protamine 1 promoter. We showed that the rat Atp1a4 transgene is expressed in mouse spermatozoa and that it is localized to the sperm flagellum. In agreement with increased expression of the alpha4 isoform, sperm from transgenic mice displayed higher alpha4-specific Na,K-ATPase activity and binding of fluorescently labeled ouabain than wild-type mice. In contrast, expression and activity of ATP1A1 (alpha1), the other Na,K-ATPase alpha isoform present in sperm, remained unchanged. Similar to wild-type mice, mice expressing the alpha4 transgene exhibited normal testis and sperm morphology and no differences in fertility. However, compared to wild-type mice, sperm from transgenic mice displayed plasma membrane hyperpolarization and higher total and progressive motility. Other parameters of motility also increased, including straight-line, curvilinear, and average path velocities and amplitude of lateral head displacement. In addition, sperm from the transgenic mice showed enhanced sperm hyperactive motility, but no changes in progesterone-induced acrosome reaction. Altogether, these results provide new genetic evidence for the role of the ATP1A4 isoform in sperm motility, under both noncapacitating and capacitating conditions. PMID:20826726

Jimenez, Tamara; Sanchez, Gladis; McDermott, Jeffrey P.; Nguyen, Anh-Nguyet; Kumar, T. Rajendra; Blanco, Gustavo

2010-01-01

226

Formation of multinucleated giant cells and microglial degeneration in rats expressing a mutant Cu/Zn superoxide dismutase gene  

OpenAIRE

Abstract Background Microglial neuroinflammation is thought to play a role in the pathogenesis of amyotrophic lateral sclerosis (ALS). The purpose of this study was to provide a histopathological evaluation of the microglial neuroinflammatory response in a rodent model of ALS, the SOD1G93A transgenic rat. Methods Multiple levels of the CNS from spinal cord to cerebral cortex were studied in SOD1G93A transgenic rats during three stages of natural disease progression, including presymptomatic, ...

Streit Wolfgang J; Xue Qing-Shan; Fendrick Sarah E

2007-01-01

227

Synthesis of minus-strand copies of a viral transgene during viral infections of transgenic plants.  

Science.gov (United States)

Viral transgenes designed to provide resistance to specific plant viruses frequently consist of the coat protein gene and a contiguous 3' untranslated region (3'UTR) of viral origin. In many RNA viruses the viral 3'UTR establishes a recognition and initiation site for viral RNA replication. Thus the transgenic transcript may contain a functional virus replication site. Experiments were designed to determine if a challenging virus would recognize this replication site on a nuclear derived transcript and synthesize the complementary RNA. These data demonstrate that upon infection by a virus that recognizes the viral replication site, a full-length complement of the transgenic transcript is produced. In these experiments the replication complex of Brome Mosaic bromovirus recognized the transgenic transcript derived from a Cowpea Chlorotic Mottle bromovirus transgene. The resulting RNA may contribute to RNA recombination events. PMID:16965831

Deng, Min; Schneider, William L; Allison, Richard F

2006-12-01

228

Transgenic mouse models for lung cancer.  

Science.gov (United States)

Transgenic technology allows the ability to target regulatory genes to the lungs in a cell-specific fashion. Using this technology, we have generated a model to investigate the phenotypic consequences of targeting oncogenes to particular cell types in the lungs and are developing a second model for the regulated expression of oncogenes in the lung. The transgenic model involves the constitutive expression of simian virus 40 large T antigen in the Clara cells of mouse lungs. This model has been used to investigate changes in expression of cell cycle regulatory genes in the Clara cells during the transformation process, as well as the expression of the transcription factors regulating the expression of Clara cell differentiation markers. The second model we are developing investigates the regulated expression of the genes in the lungs. This system is based on the establishment of two types of transgenic lines. The regulator line consists of a chimeric transcriptional factor placed under the control of a lung-specific SPC (surfactant protein C) promoter. This chimeric regulator is composed of a transcription activation domain, the GAL4 DNA-binding domain, and a truncated progesterone receptor that is responsive to RU 486, but not to endogenous progesterone. The second transgenic mouse line contains the silent target transgene under the control of a minimal promoter with upstream activating sequences (UAS) that are recognized by the regulator transgene. Upon breeding these two lines, the resulting bitransgenic mice can then be induced to express the target transgene only with the administration of RU 486. Two generations of regulators have been evaluated on their ability to regulate the expression of a growth hormone reporter gene. This system demonstrated the inducible expression of the reporter genes in the distal airways of the lungs. PMID:11195456

Zhao, B; Magdaleno, S; Chua, S; Wang, Y L; Burcin, M; Elberg, D; Finegold, M; Tsai, S; DeMayo, F J

2000-12-01

229

Development of a BAC vector for integration-independent and tight regulation of transgenes in rodents via the Tet system.  

Science.gov (United States)

The establishment of functional transgenic mouse lines is often limited by problems caused by integration site effects on the expression construct. Similarly, tetracycline (Tet) controlled transcription units most commonly used for conditional transgene expression in mice are strongly influenced by their genomic surrounding. Using bacterial artificial chromosome (BAC) technology in constitutive expression systems, it has been shown that integration site effects resulting in unwanted expression patterns can be largely eliminated. Here we describe a strategy to minimize unfavourable integration effects on conditional expression constructs based on a 75 kb genomic BAC fragment. This fragment was derived from a transgenic mouse line, termed LC-1, which carries the Tet-inducible genes luciferase and cre (Schönig et al. 2002). Animals of this mouse line have previously been shown to exhibit optimal expression properties in terms of tightness in the off state and the absolute level of induction, when mated to appropriate transactivator expressing mice. Here we report the cloning and identification of the transgenic LC-1 integration site which was subsequently inserted into a bacterial artificial chromosome. We demonstrate that this vector facilitates the efficient generation of transgenic mouse and rat lines, where the Tet-controlled expression unit is shielded from perturbations caused by the integration site. PMID:20640885

Schönig, Kai; Kentner, David; Gossen, Manfred; Baldinger, Tina; Miao, Jun; Welzel, Katrin; Vente, Andreas; Bartsch, Dusan; Bujard, Hermann

2011-06-01

230

The ecological risks of transgenic plants.  

Science.gov (United States)

Biotechnologies have been utilized "ante litteram" for thousands of years to produce food and drink and genetic engineering techniques have been widely applied to produce many compounds for human use, from insulin to other medicines. The debate on genetically modified (GM) organisms broke out all over the world only when GM crops were released into the field. Plant ecologists, microbiologists and population geneticists carried out experiments aimed at evaluating the environmental impact of GM crops. The most significant findings concern: the spread of transgenes through GM pollen diffusion and its environmental impact after hybridisation with closely related wild species or subspecies; horizontal gene transfer from transgenic plants to soil microbes; the impact of insecticide proteins released into the soil by transformed plants on non-target microbial soil communities. Recent developments in genetic engineering produced a technology, dubbed "Terminator", which protects patented genes introduced in transgenic plants by killing the seeds in the second generation. This genetic construct, which interferes so heavily with fundamental life processes, is considered dangerous and should be ex-ante evaluated taking into account the data on "unexpected events", as here discussed, instead of relying on the "safe until proven otherwise" claim. Awareness that scientists, biotechnologists and genetic engineers cannot answer the fundamental question "how likely is that transgenes will be transferred from cultivated plants into the natural environment?" should foster long-term studies on the ecological risks and benefits of transgenic crops. PMID:14595899

Giovannetti, Manuela

2003-01-01

231

Rosuvastatin Can Block Pro-Inflammatory Actions of Transgenic Human C-Reactive Protein Without Reducing its Circulating Levels.  

Czech Academy of Sciences Publication Activity Database

Ro?. 32, ?. 2 (2014), s. 59-65. ISSN 1755-5914 R&D Projects: GA MŠk(CZ) LL1204; GA ?R(CZ) GA13-04420S; GA MZd(CZ) NT14325; GA MŠk(CZ) 7E10067 Grant ostatní: MH CZ - DRO(CZ) IN0002301 Institutional support: RVO:67985823 Keywords : C-reactive protein * transgenic * spontaneously hypertensive rat * rosuvastatin Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.536, year: 2013

Šilhavý, Jan; Zídek, Václav; Landa, Vladimír; Šimáková, Miroslava; Mlejnek, Petr; Škop, V.; Oliyarnyk, O.; Kazdová, L.; Mancini, M.; Saar, K.; Schulz, H.; Hübner, N.; Kurtz, T. W.; Pravenec, Michal

2014-01-01

232

Functional conservation between rodents and chicken of regulatory sequences driving skeletal muscle gene expression in transgenic chickens  

OpenAIRE

Abstract Background Regulatory elements that control expression of specific genes during development have been shown in many cases to contain functionally-conserved modules that can be transferred between species and direct gene expression in a comparable developmental pattern. An example of such a module has been identified at the rat myosin light chain (MLC) 1/3 locus, which has been well characterised in transgenic mouse studies. This locus contains two promoters encoding two alternatively...

Taylor Lorna; Lillico Simon G; Sherman Adrian; McGrew Michael J; Sang Helen

2010-01-01

233

Detection of membrane-bound HLA-G translated products with a specific monoclonal antibody.  

OpenAIRE

A monomorphic anti-HLA-G monoclonal antibody (mAb) was obtained by immunization of HLA-B27/human beta 2-microglobulin double-transgenic mice with transfected murine L cells expressing both HLA-G and human beta 2-microglobulin. This mAb, designated BFL.1, specifically recognizes, by flow cytometry analysis, the immunizing HLA-G-expressing cells, whereas it does not bind to parental untransfected or to HLA-B7- and HLA-A3-transfected L cells, suggesting that it distinguishes between classical HL...

Bensussan, A.; Mansur, I. G.; Mallet, V.; Rodriguez, A. M.; Girr, M.; Weiss, E. H.; Brem, G.; Boumsell, L.; Gluckman, E.; Dausset, J.

1995-01-01

234

Lectin cDNA and transgenic plants derived therefrom  

Science.gov (United States)

Transgenic plants containing cDNA encoding Gramineae lectin are described. The plants preferably contain cDNA coding for barley lectin and store the lectin in the leaves. The transgenic plants, particularly the leaves exhibit insecticidal and fungicidal properties.

Raikhel, Natasha V. (Okemos, MI)

2000-10-03

235

Age-related changes in the dorsal skin histology in Mini and Wistar rats  

OpenAIRE

Mini rats (Jcl: WistarTGN(ARGHGEN)1Nts (MRs) are Wistar rat (WR)-derived transgenic rats in which the expression of growth hormone (GH) gene is suppressed by the presence of antisense transgene. The plasma GH level of MRs is reduced to 40 to 60% of that of WRs. In this study, to evaluate the influence of GH deficiency on the skin nature, age-related changes in the dorsal skin histology were compared between male MRs and WRs. Although there were no essential dif...

Ikawa, A.; Ishii, Y.; Suzuki, K.; Yasoshima, A.; Suzuki, N.; Nakayama, Hiroyuki; Takahashi, S.; Doi, K.

2002-01-01

236

Detection of potential transgenic plant DNA recipients among soil bacteria  

OpenAIRE

The likelihood of gene transfer from transgenic plants to bacteria is dependent on gene number and the presence of homologous sequences. The large number of transgene copies in transplastomic (transgenes contained in the chloroplast genome) plant cells as well as the prokaryotic origin of the transgene, may thus significantly increase the likelihood of gene transfer to bacteria that colonize plant tissues. In order to assess the probability of such transfer, the length of homologous DNA seque...

Monier, Jean-michel; Bernillon, Dominique; Kay, Elizabeth; Faugier, Aure?lie; Rybalka, Oleksandra; Dessaux, Yves; Simonet, Pascal; Vogel, Timothy

2007-01-01

237

New doxycycline-inducible transgenic lines in Xenopus  

OpenAIRE

We have characterized two new transgenic Xenopus lines enabling transgene expression using the Tet-On inducible system. An inducer line expresses the doxycycline- (Dox-) activated transcription factor rtTA under control of the ubiquitous promoter CMV. A responder line enables Dox-inducible expression of a dominant positive thyroid hormone receptor via a tetracycline responsive transgenic promoter (TRE). Dox-induced expression of transgenic GFP mRNA was detectable after 3 hours and increased u...

Rankin, Scott A.; Zorn, Aaron M.; Buchholz, Daniel R.

2011-01-01

238

Transgenic cultures: from the economic viewpoint  

Directory of Open Access Journals (Sweden)

Full Text Available The introduction of transgenic seeds for agricultural purposes poses modification to their production, due to the potential for reaching desired characteristics such as greater yield, this being fundamental in an economic environment characterised by open market conditions. However, acceptance of products resulting from genetic engineering is far from becoming a simple process; discussion relating to the predominance of private sector interests, the monopoly of knowledge and the safety of such seeds/food is currently in the spotlight. This article presents the main points of debate regarding adoption of transgenic cultures, contributing to discussion about this topic for Colombia.

Mauricio Mosquera

2011-12-01

239

Stress-induced activation of the sympathoadrenal system is determined by genetic background in rat models of tauopathy.  

Science.gov (United States)

Stress may accelerate onset of neurodegenerative diseases in vulnerable subjects and, vice versa, neurodegeneration affects the responsiveness to stressors. We investigated the neuroendocrine response to immobilization stress in normotensive Wistar-Kyoto rats (WKY), spontaneously hypertensive rats (SHR), and transgenic rats of respective WKY and SHR strains overexpressing human truncated tau protein. Plasma levels of epinephrine, norepinephrine, and corticosterone were determined. An immobilization-induced elevation of epinephrine and norepinephrine was significantly reduced in WKY transgenic rats compared to WKY wild-type rats, while no differences were seen between SHR transgenic and SHR wild-type animals. Our data have shown that sympathoadrenal system response to stress strongly depends on both tau protein-induced neurodegeneration and genetic background of experimental animals. PMID:25147110

Lejavova, Katarina; Ondicova, Katarina; Horvathova, Lubica; Hegedusova, Noemi; Cubinkova, Veronika; Vargovic, Peter; Manz, Georg; Filipcik, Peter; Mravec, Boris; Novak, Michal; Kvetnansky, Richard

2015-01-01

240

Cooperation between Stat3 and Akt Signaling Leads to Prostate Tumor Development in Transgenic Mice12  

OpenAIRE

In this report, we describe the development of a transgenic mouse in which a rat probasin promoter (ARR2Pb) was used to direct prostate specific expression of a constitutively active form of signal transducer and activator of transcription 3 (i.e., Stat3C). ARR2Pb.Stat3C mice exhibited hyperplasia and prostate intraepithelial neoplasia (PIN) lesions in both ventral and dorsolateral prostate lobes at 6 and 12 months; however, no adenocarcinomas were detected. The effect of combined loss of PTE...

Blando, Jorge M.; Carbajal, Steve; Abel, Erika; Beltran, Linda; Conti, Claudio; Fischer, Susan; Digiovanni, John

2011-01-01

241

The substantive equivalence of transgenic (Bt and Chi) and non-transgenic cotton based on metabolite profiles.  

Science.gov (United States)

Compositional studies comparing transgenic with non-transgenic counterpart plants are almost universally required by governmental regulatory bodies. In the present study, two T(2) transgenic cotton lines containing chitinase (Line 11/57) and Bt lines (Line 61) were compared with non-transgenic counterpart. To do this, biochemical characteristics of leaves and seeds, including amino acids, fatty acids, carbohydrates, anions, and cations contents of the studied lines were analyzed using GC/MS, high-performance liquid chromatography (HPLC), and ion chromatography (IC) analyzers, respectively. polymerase chain reaction (PCR) and Western blot analyses confirmed the presence and expression of Chi and Bt genes in the studied transgenic lines. Although, compositional analysis of leaves contents confirmed no significant differences between transgenic and non-transgenic counterpart lines, but it was shown that glucose content of chitinase lines, fructose content of transgenic lines (Bt and chitinase) and asparagine and glutamine of chitinase lines were significantly higher than the non-transgenic counterpart plants. Both the transgenic lines (Bt and chitinase) showed significant decrease in the amounts of sodium in comparison to the non-transgenic counterpart plants. The experiments on the seeds showed that histidine, isoleucine, leucine, and phenylalanine contents of all transgenic and non-transgenic lines were the same, whereas other amino acids were significantly increased in the transgenic lines. Surprisingly, it was observed that the concentrations of stearic acid, myristic acid, oleic acid, and linoleic acid in the chitinase line were significantly different than those of non-transgenic counterpart plants, but these components were the same in both Bt line and its non-transgenic counterpart. It seems that more changes observed in the seed contents than leaves is via this point that seeds are known as metabolites storage organs, so they show greater changes in the metabolites contents comparing to the leaves. PMID:24374853

Modirroosta, Bentol Hoda; Tohidfar, Masoud; Saba, Jalal; Moradi, Foad

2014-03-01

242

Production of recombinant proteins in milk of transgenic and non-transgenic goats  

Directory of Open Access Journals (Sweden)

Full Text Available Among all the transgenic mammalians produced so far, goats have represented an excellent model of transgenesis when considering the factors such as the market demand for protein, volume of milk produced per lactation and reproductive rate. Various recombinant proteins have been obtained from the transgenic and non-transgenic goats, and among these, human antithrombin, produced by the transgenic goats, was the first recombinant protein of animal origin to be released as a drug for the clinical use in humans. This review reports the aspects inherent to the production of recombinant proteins in the goats, from the production of the animal bioreactors up to the expression of these proteins in their milk.

Raylene Ramos Moura

2011-10-01

243

Improved production of genetically modified fetuses with homogeneous transgene expression after transgene integration site analysis and recloning in cattle.  

Science.gov (United States)

Animal cloning by nuclear transfer (NT) has made the production of transgenic animals using genetically modified donor cells possible and ensures the presence of the gene construct in the offspring. The identification of transgene insertion sites in donor cells before cloning may avoid the production of animals that carry undesirable characteristics due to positional effects. This article compares blastocyst development and competence to establish pregnancies of bovine cloned embryos reconstructed with lentivirus-mediated transgenic fibroblasts containing either random integration of a transgene (random integration group) or nuclear transfer derived transgenic fibroblasts with known transgene insertion sites submitted to recloning (recloned group). In the random integration group, eGFP-expressing bovine fetal fibroblasts were selected by fluorescence activated cell sorting (FACS) and used as nuclei donor cells for NT. In the recloned group, a fibroblast cell line derived from a transgenic cloned fetus was characterized regarding transgene insertion and submitted to recloning. The recloned group had higher blastocyst production (25.38 vs. 14.42%) and higher percentage of 30-day pregnancies (14.29 vs. 2.56%) when compared to the random integration group. Relative eGFP expression analysis in fibroblasts derived from each cloned embryo revealed more homogeneous expression in the recloned group. In conclusion, the use of cell lines recovered from transgenic fetuses after identification of the transgene integration site allowed for the production of cells and fetuses with stable transgene expression, and recloning may improve transgenic animal yields. PMID:21241190

Bressan, Fabiana Fernandes; Dos Santos Miranda, Moyses; Perecin, Felipe; De Bem, Tiago Henrique; Pereira, Flavia Thomaz Verechia; Russo-Carbolante, Elisa Maria; Alves, Daiani; Strauss, Bryan; Bajgelman, Marcio; Krieger, José Eduardo; Binelli, Mario; Meirelles, Flavio Vieira

2011-02-01

244

Progress in Xenotransplantation Research Employing Transgenic Pigs  

Directory of Open Access Journals (Sweden)

Full Text Available Microinjection of foreign DNA into pronuclei of a fertilized oocyte has predominantly been used for the generation of transgenic livestock. This technology works reliably, but is inefficient and results in random integration and variable expression patterns in the transgenic offspring. Nevertheless, remarkable achievements have been made with this technology with regard to xenotransplantation. Transgenic pigs that express human complement regulating proteins have been tested in their ability to serve as donors in human organ transplantation (i.e. xenotransplantation. In vitro and in vivo data convincingly show that the hyperacute rejection response can be overcome in a clinically acceptable manner by successfully employing this strategy. The recent developments in nuclear transfer and its merger with the growing genomic data allow targeted and regulatable transgenesis. Systems for efficient homologous recombination in somatic cells are being developed and the first knock-out pigs, carrying a deletion in the a-galactosyltransferase gene, were recently generated. It is anticipated that poly-transgenic pigs will be available as donors for functional xenografts within a few years. Similarly, pigs may serve as donors for a variety of xenogenic cells and tissues. The availability of these technologies is essential to maintain "genetic security" and to ensure absence of unwanted side effects.

H. Niemann

2003-06-01

245

Ethics and Transgenic Crops: a Review  

Scientific Electronic Library Online (English)

Full Text Available SciELO Chile | Language: English Abstract in english This article represents a review of some of the ethical dilemmas that have arisen as a result of the development and deployment of transgenic crop plants. The potential for transgenic crops to alleviate human hunger and the possible effects on human health are discussed. Risks and benefits to the en [...] vironment resulting from genetic engineering of crops for resistance to biotic and abiotic stresses are considered, in addition to effects on biodiversity. The socio-economic impacts and distribution of benefits from transgenic technologies are reviewed. Fundamental issues of man’s relationship with nature and the environment, and theological matters are also addressed. An almost unprecedented amount of discussion has been stimulated on the merits and demerits of genetic engineering of crop plants, and has divided both the public and scientific communities. The arguments for and against transgenics are invariably based on visions of the new technology from widely different ethical perspectives.

Jonathan, Robinson.

1999-08-15

246

Transgenic plants with increased calcium stores  

Science.gov (United States)

The present invention provides transgenic plants over-expressing a transgene encoding a calcium-binding protein or peptide (CaBP). Preferably, the CaBP is a calcium storage protein and over-expression thereof does not have undue adverse effects on calcium homeostasis or biochemical pathways that are regulated by calcium. In preferred embodiments, the CaBP is calreticulin (CRT) or calsequestrin. In more preferred embodiments, the CaBP is the C-domain of CRT, a fragment of the C-domain, or multimers of the foregoing. In other preferred embodiments, the CaBP is localized to the endoplasmic reticulum by operatively associating the transgene encoding the CaBP with an endoplasmic reticulum localization peptide. Alternatively, the CaBP is targeted to any other sub-cellular compartment that permits the calcium to be stored in a form that is biologically available to the plant. Also provided are methods of producing plants with desirable phenotypic traits by transformation of the plant with a transgene encoding a CaBP. Such phenotypic traits include increased calcium storage, enhanced resistance to calcium-limiting conditions, enhanced growth and viability, increased disease and stress resistance, enhanced flower and fruit production, reduced senescence, and a decreased need for fertilizer production. Further provided are plants with enhanced nutritional value as human food or animal feed.

Wyatt, Sarah (Inventor); Tsou, Pei-Lan (Inventor); Robertson, Dominique (Inventor); Boss, Wendy (Inventor)

2004-01-01

247

Transgenic Mouse Model of Chronic Beryllium Disease  

Energy Technology Data Exchange (ETDEWEB)

Animal models provide powerful tools for dissecting dose-response relationships and pathogenic mechanisms and for testing new treatment paradigms. Mechanistic research on beryllium exposure-disease relationships is severely limited by a general inability to develop a sufficient chronic beryllium disease animal model. Discovery of the Human Leukocyte Antigen (HLA) - DPB1Glu69 genetic susceptibility component of chronic beryllium disease permitted the addition of this human beryllium antigen presentation molecule to an animal genome which may permit development of a better animal model for chronic beryllium disease. Using FVB/N inbred mice, Drs. Rubin and Zhu, successfully produced three strains of HLA-DPB1 Glu 69 transgenic mice. Each mouse strain contains a haplotype of the HLA-DPB1 Glu 69 gene that confers a different magnitude of odds ratio (OR) of risk for chronic beryllium disease: HLA-DPB1*0401 (OR = 0.2), HLA-DPB1*0201 (OR = 15), HLA-DPB1*1701 (OR = 240). In addition, Drs. Rubin and Zhu developed transgenic mice with the human CD4 gene to permit better transmission of signals between T cells and antigen presenting cells. This project has maintained the colonies of these transgenic mice and tested the functionality of the human transgenes.

Gordon, Terry

2009-05-26

248

Viable transgenic goats derived from skin cells.  

Science.gov (United States)

The current study was undertaken to evaluate the possibility of expanding transgenic goat herds by means of somatic cell nuclear transfer (NT) using transgenic goat cells as nucleus donors. Skin cells from adult, transgenic goats were first synchronized at quiescent stage (G0) by serum starvation and then induced to exit G0 and proceed into G1. Oocytes collected from superovulated donors were enucleated, karyoplast-cytoplast couplets were constructed, and then fused and activated simultaneously by a single electrical pulse. Fused couplets were either co-cultured with oviductal cells in TCM-199 medium (in vitro culture) or transferred to intermediate recipient goat oviducts (in vivo culture) until final transfer. The resulting morulae and blastocysts were transferred to the final recipients. Pregnancies were confirmed by ultrasonography 25-30 days after embryo transfer. In vitro cultured NT embryos developed to morulae and blastocyst stages but did not produce any pregnancies while 30% (6/20) of the in vivo derived morulae and blastocysts produced pregnancies. Two of these pregnancies were resorbed early in gestation. Of the four recipients that maintained pregnancies to term, two delivered dead fetuses 2-3 days after their due dates, and two recipients gave birth to healthy kids at term. Fluorescence in situ hybridization (FISH) analysis confirmed that both kids were transgenic and had integration sites consistent with those observed in the adult cell line. PMID:15359599

Behboodi, Esmail; Memili, Erdogan; Melican, David T; Destrempes, Margaret M; Overton, Susan A; Williams, Jennifer L; Flanagan, Peter A; Butler, Robin E; Liem, Hetty; Chen, Li How; Meade, Harry M; Gavin, William G; Echelard, Yann

2004-06-01

249

Making BAC transgene constructs with lambda-red recombineering system for transgenic animals or cell lines.  

Science.gov (United States)

The genomic DNA libraries based on Bacteria Artificial Chromosomes (BAC) are the foundation of whole genomic mapping, sequencing, and annotation for many species like mice and humans. With their large insert size, BACs harbor the gene-of-interest and nearby transcriptional regulatory elements necessary to direct the expression of the gene-of-interest in a temporal and cell-type specific manner. When replacing a gene-of-interest with a transgene in vivo, the transgene can be expressed with the same patterns and machinery as that of the endogenous gene. This chapter describes in detail a method of using lambda-red recombineering to make BAC transgene constructs with the integration of a transgene into a designated location within a BAC. As the final BAC construct will be used for transfection in cell lines or making transgenic animals, specific considerations with BAC transgenes such as genotyping, BAC coverage and integrity as well as quality of BAC DNA will be addressed. Not only does this approach provide a practical and effective way to modify large DNA constructs, the same recombineering principles can apply to smaller high copy plasmids as well as to chromosome engineering. PMID:25239742

Holmes, Scott; Lyman, Suzanne; Hsu, Jen-Kang; Cheng, JrGang

2015-01-01

250

Can transgenic maize affect soil microbial communities?  

Science.gov (United States)

The aim of the experiment was to determine if temporal variations of belowground activity reflect the influence of the Cry1Ab protein from transgenic maize on soil bacteria and, hence, on a regulatory change of the microbial community (ability to metabolize sources belonging to different chemical guilds) and/or a change in numerical abundance of their cells. Litter placement is known for its strong influence on the soil decomposer communities. The effects of the addition of crop residues on respiration and catabolic activities of the bacterial community were examined in microcosm experiments. Four cultivars of Zea mays L. of two different isolines (each one including the conventional crop and its Bacillus thuringiensis cultivar) and one control of bulk soil were included in the experimental design. The growth models suggest a dichotomy between soils amended with either conventional or transgenic maize residues. The Cry1Ab protein appeared to influence the composition of the microbial community. The highly enhanced soil respiration observed during the first 72 h after the addition of Bt-maize residues can be interpreted as being related to the presence of the transgenic crop residues. This result was confirmed by agar plate counting, as the averages of the colony-forming units of soils in conventional treatments were about one-third of those treated with transgenic straw. Furthermore, the addition of Bt-maize appeared to induce increased microbial consumption of carbohydrates in BIOLOG EcoPlates. Three weeks after the addition of maize residues to the soils, no differences between the consumption rate of specific chemical guilds by bacteria in soils amended with transgenic maize and bacteria in soils amended with conventional maize were detectable. Reaped crop residues, comparable to post-harvest maize straw (a common practice in current agriculture), rapidly influence the soil bacterial cells at a functional level. Overall, these data support the existence of short Bt-induced ecological shifts in the microbial communities of croplands' soils. PMID:17009863

Mulder, Christian; Wouterse, Marja; Raubuch, Markus; Roelofs, Willem; Rutgers, Michiel

2006-09-29

251

Transgenic expression of COL1A1-chloramphenicol acetyltransferase fusion genes in bone: differential utilization of promoter elements in vivo and in cultured cells.  

OpenAIRE

To directly compare the patterns of collagen promoter expression in cells and tissues, the activity of COL1A1 fusion genes in calvariae of neonatal transgenic mice and in primary bone cell cultures derived by sequential digestion of transgenic calvariae was measured. ColCAT3.6 contains 3.6 kb (positions -3521 to +115) of the rat COL1A1 gene ligated to the chloramphenicol acetyltransferase (CAT) reporter gene. ColCAT2.3 and ColCAT1.7 are 5' deletion mutants which contain 2,296 and 1,672 bp, re...

Krebsbach, P. H.; Harrison, J. R.; Lichtler, A. C.; Woody, C. O.; Rowe, D. W.; Kream, B. E.

1993-01-01

252

Glucose metabolic gene expression in growth hormone transgenic coho salmon.  

Science.gov (United States)

Salmonids are generally known to be glucose intolerant. However, previous studies have shown that growth hormone (GH) transgenic coho salmon display modified nutritional regulation of glycolysis and lipogenesis compared to non-transgenic fish, suggesting the potential for better use of glucose in GH transgenic fish. To examine this in detail, GH transgenic and non-transgenic coho salmon were subjected to glucose tolerance test and subsequent metabolic assessments. After intra-peritoneal injection of 250mg/kg glucose, we analysed post-injection kinetics of glycaemia and expression of several key target genes highly involved in glucose homeostasis in muscle and liver tissues. Our data show no significant differences in plasma glucose levels during peak hyperglycaemia (3-6h after injection), demonstrating a similar glucose tolerance between transgenic and non transgenic. However, and unrelated to the hyperglycaemic episode, GH transgenic fish return to a slightly lower basal glycaemia values 24h after injection. Correspondingly, GH transgenic fish exhibited higher mRNA levels of glucokinase (GK) and glucose-6-phosphate dehydrogenase (G6PDH) in liver, and glucose transporter (GLUT4) in muscle. These data suggest that these metabolic actors may be involved in different glucose use in GH transgenic fish, which would be expected to influence the glucose challenge response. Overall, our data demonstrate that GH transgenic coho salmon may be a pertinent animal model for further study of glucose metabolism in carnivorous fish. PMID:24486143

Panserat, Stéphane; Kamalam, Biju Sam; Fournier, Jeanne; Plagnes-Juan, Elisabeth; Woodward, Krista; Devlin, Robert H

2014-04-01

253

Compensation of the AKT signaling by ERK signaling in transgenic mice hearts overexpressing TRIM72  

International Nuclear Information System (INIS)

The AKT and ERK signaling pathways are known to be involved in cell hypertrophy, proliferation, survival and differentiation. Although there is evidence for crosstalk between these two signaling pathways in cellulo, there is less evidence for cross talk in vivo. Here, we show that crosstalk between AKT and ERK signaling in the hearts of TRIM72-overexpressing transgenic mice (TRIM72-Tg) with alpha-MHC promoter regulates and maintains their heart size. TRIM72, a heart- and skeletal muscle-specific protein, downregulates AKT-mTOR signaling via IRS-1 degradation and reduces the size of rat cardiomyocytes and the size of postnatal TRIM72-Tg hearts. TRIM72 expression was upregulated by hypertrophic inducers in cardiomyocytes, while IRS-1 was downregulated by IGF-1. TRIM72 specifically regulated IGF-1-dependent AKT-mTOR signaling, resulting in a reduction of the size of cardiomyocytes. Postnatal TRIM72-Tg hearts were smaller than control-treated hearts with inhibition of AKT-mTOR signaling. However, adult TRIM72-Tg hearts were larger than of control despite the suppression of AKT-mTOR signaling. Activation of ERK, PKC-?, and JNK were observed to be elevated in adult TRIM72-Tg, and these signals were mediated by ET-1 via the ET receptors A and B. Altogether, these results suggest that AKT signaling regulates cardiac hypertrophy in physiological conditions, and ERK signaling compensates for the absence of AKT signaling during TRIM72 overexpression, leading to pathological hypertrophy. -- Highlights: • TRIM72 inhibits AKT signaling through ubiquitination of IRS-1 in cardiac cells. • TRIM72 regulates the size of cardiac cells. • TRIM72 regulates size of postnatal TRIM72-overexpressing transgenic mice hearts. • Adult TRIM72-overexpressing transgenic mice hearts showed cardiac dysfunction. • Adult TRIM72 transgenic mice hearts showed higher expression of endothelin receptors

254

Compensation of the AKT signaling by ERK signaling in transgenic mice hearts overexpressing TRIM72  

Energy Technology Data Exchange (ETDEWEB)

The AKT and ERK signaling pathways are known to be involved in cell hypertrophy, proliferation, survival and differentiation. Although there is evidence for crosstalk between these two signaling pathways in cellulo, there is less evidence for cross talk in vivo. Here, we show that crosstalk between AKT and ERK signaling in the hearts of TRIM72-overexpressing transgenic mice (TRIM72-Tg) with alpha-MHC promoter regulates and maintains their heart size. TRIM72, a heart- and skeletal muscle-specific protein, downregulates AKT-mTOR signaling via IRS-1 degradation and reduces the size of rat cardiomyocytes and the size of postnatal TRIM72-Tg hearts. TRIM72 expression was upregulated by hypertrophic inducers in cardiomyocytes, while IRS-1 was downregulated by IGF-1. TRIM72 specifically regulated IGF-1-dependent AKT-mTOR signaling, resulting in a reduction of the size of cardiomyocytes. Postnatal TRIM72-Tg hearts were smaller than control-treated hearts with inhibition of AKT-mTOR signaling. However, adult TRIM72-Tg hearts were larger than of control despite the suppression of AKT-mTOR signaling. Activation of ERK, PKC-?, and JNK were observed to be elevated in adult TRIM72-Tg, and these signals were mediated by ET-1 via the ET receptors A and B. Altogether, these results suggest that AKT signaling regulates cardiac hypertrophy in physiological conditions, and ERK signaling compensates for the absence of AKT signaling during TRIM72 overexpression, leading to pathological hypertrophy. -- Highlights: • TRIM72 inhibits AKT signaling through ubiquitination of IRS-1 in cardiac cells. • TRIM72 regulates the size of cardiac cells. • TRIM72 regulates size of postnatal TRIM72-overexpressing transgenic mice hearts. • Adult TRIM72-overexpressing transgenic mice hearts showed cardiac dysfunction. • Adult TRIM72 transgenic mice hearts showed higher expression of endothelin receptors.

Ham, Young-Mi, E-mail: youngmi_ham@hms.harvard.edu [College of Life Science and Biotechnology, Korea University, Seoul (Korea, Republic of); Department of Cell Biology, Harvard Medical School, Boston, MA 02115 (United States); Mahoney, Sarah Jane [Department of Cell Biology, Harvard Medical School, Boston, MA 02115 (United States)

2013-06-10

255

Studies of an expanded trinucleotide repeat in transgenic mice  

Energy Technology Data Exchange (ETDEWEB)

Spinal and bulbar muscular atrophy (SBMA) is a progressive motor neuron disease caused by expansion of a trinucleotide repeat in the androgen receptor gene (AR{sup exp}). AR{sup exp} repeats expand further or contract in approximately 25% of transmissions. Analogous {open_quotes}dynamic mutations{close_quotes} have been reported in other expanded trinucleotide repeat disorders. We have been developing a mouse model of this disease using a transgenic approach. Expression of the SBMA AR was documented in transgenic mice with an inducible promoter. No phenotypic effects of transgene expression were observed. We have extended our previous results on stability of the expanded trinucleotide repeat in transgenic mice in two lines carrying AR{sup exp}. Tail DNA was amplified by PCR using primers spanning the repeat on 60 AR{sup exp} transgenic mice from four different transgenic lines. Migration of the PCR product through an acrylamide gel showed no change of the 45 CAG repeat length in any progeny. Similarly, PCR products from 23 normal repeat transgenics showed no change from the repeat length of the original construct. Unlike the disease allele in humans, the expanded repeat AR cDNA in transgenic mice showed no change in repeat length with transmission. The relative stability of CAG repeats seen in the transgenic mice may indicate either differences in the fidelity of replicative enzymes, or differences in error identification and repair between mice and humans. Integration site or structural properties of the transgene itself might also play a role.

Bingham, P.; Wang, S.; Merry, D. [Univ. of Pennsylvania, Philadelphia, PA (United States)

1994-09-01

256

Effect of transgene number of spontaneous and radiation-induced micronuclei in lacl transgenic mice  

International Nuclear Information System (INIS)

Lacl transgenic mice are widely used for the measurement of mutations in specific target issues. The lacl transgene is present in mice as 40 tandem repeats; this sequence is homozygous (contained in both copies of chromosome 5) in C57Bl/6 mice, and is hemizygous in B6C3F1 mice. Previous reports have indicated that tandem repeats can produce chromosome instability, fragile sites, and other effects. To determine whether the presence of the transgene effects micronucleus induction we compared the response of nontransgenic (NTR) to hemizygous (HEMI) transgenic B6C3F1 mice and to hemizygous and homozygous (HOMO) transgenic C57Bl/6 mice. Five mice/group were irradiated with 500 cGy from a 137Cs source. Bone marrow was harvested 24 hr after treatment and 2000 polychromatic erythrocytes (PCE) were analyzed per animal. The presence or absence of the lacl transgene had no effect in unirradiated mice on the percent of micronucleated PCE (MN) or on the ratio of PCE to total red blood cells for either strain: B6C3F1 mice had MN frequencies of 0.26% and 0.20% for NTR and HEMI mice, respectively; C57Bl/6 mice had MN frequencies of 0.34%, 0.32%, and 0.38% for NTR, HEMI, and HOMO mice, respectively. Radiation-induced micronucleus frequencies were significantly higher in HEMI lacl B6C3F1 mice (2.85%) than in NTR litter mates (1.59%); the converse was true in C57Bl/6 mice: NTR were 2.45%, HEMI were 1.25%, HOMO were 1.65%. These data suggest that the lacl transgene does not cauggest that the lacl transgene does not cause chromosome instability as measured by spontaneous micronucleus levels. However, the response of these transgenic mice to a variety of clastogenic agents needs to be investigated before they are integrated into standard in vivo assays for chromosome damage

257

A multidisciplinary approach involving comparative 'OMICS' of transgenic and non-transgenic soybeam seeds  

International Nuclear Information System (INIS)

Complete text of publication follows. Soybean culture has an expressive impact in the economy of many countries, being the commercialization of its by-products, which presents many benefits in terms of health and nutritional aspects, and which also includes a fuel alternative (biodiesel), the main factor for the large soybean production. Part of this impact is due to the transgenic modification of soybean, conferring enhanced characteristics to the culture, such as tolerance to fungicides (Y. Kim et al., J. Microbiol. Biotechnol., 16 (2006), 25-31.). Due to the insertion of hexogen genes, some proteome modification is possible (S. Natarajan et al., Anal. Biochem., 342 (2005), 214-220), and recently some metallome modification was reported by our research group (A. Sussulini et al., J. Anal. At. Spectrom., 22 (2007), 1501-1506). Then, the aim of this work is to enlarge the results in terms of 'omics' when considering transgenic and non-transgenic soybean seeds. For this task, the identification of more than 140 soybean proteins using MALDI-QTOF-MS after 2D-PAGE protein separation (369±46 and 376±42 protein spots in the 4-7 pI range for transgenic and non-transgenic soybean seeds, respectively), the analysis of the protein expression using image program, the analysis of some enzymes (SOD, GR, APX, CAT) involved in the ROS production, the mapping of 80 protein spots using SR-XRF, and the metal identification of more than 30 spots using ICP-MS was carried out. In terms using ICP-MS was carried out. In terms of metal distribution when considering some proteins, the results displayed a great ability of proteins bind different metal ions. High iron (sucrose binding protein homolog S-64 - 57,922 kDa), chromium (protein not identified), lead (seed maturation protein PM 41 - 15,103 kDa), copper and tin (trypsin inhibitor (kunitz), chain A - 20,417 kDa) contents were achieved in the non-transgenic soybean, while high magnesium (actin - 50,281 kDa), barium (protein not identified) and ruthenium (protein not identified) contents were achieved in the transgenic soybean. The results put in evidence the possibility to find a biomarker candidate for differentiating transgenic and non-transgenic organisms. Financial support from Fundacao de Amparo a Pesquisa do Estado de Sao Paulo - FAPESP, Conselho Nacional de Desenvolvimento Cientifico e Tecnologico - CNPq, Fianciadora de Estudos e Projetos - FINEP, and Proteome Network of the Sao Paulo state - Brazilian National Laboratory of Synchrotron Radiation are highly acknowledged.

258

Transgene mobilization and regulatory uncertainty for non-GE fruit products of transgenic rootstocks.  

Science.gov (United States)

Genetically engineered (GE) rootstocks may offer some advantages for biotechnology applications especially in woody perennial crops such as grape or walnut. Transgrafting combines horticultural grafting practices with modern GE methods for crop improvement. Here, a non-GE conventional scion (upper stem portion) is grafted onto a transgenic GE rootstock. Thus, the scion does not contain the genetic modification present in the rootstock genome. We examined transgene presence in walnut and tomato GE rootstocks and non-GE fruit-bearing scions. Mobilization of transgene DNA, protein, and mRNA across the graft was not detected. Though transgenic siRNA mobilization was not observed in grafted tomatoes or walnut scions, transgenic siRNA signal was detected in walnut kernels. Prospective benefits from transgrafted plants include minimized risk of GE pollen flow (Lev-Yadun and Sederoff, 2001), possible use of more than one scion per approved GE rootstock which could help curb the estimated US$136 million (CropLife International, 2011) cost to bring a GE crop to international markets, as well as potential for improved consumer and market acceptance since the consumable product is not itself GE. Thus, transgrafting provides an alternative option for agricultural industries wishing to expand their biotechnology portfolio. PMID:22749907

Haroldsen, Victor M; Chi-Ham, Cecilia L; Bennett, Alan B

2012-10-31

259

Caudal dysgenesis in islet-1 transgenic mice  

OpenAIRE

Maternal diabetes during pregnancy is responsible for the occurrence of diabetic embryopathy, a spectrum of birth defects that includes heart abnormalities, neural tube defects, and caudal dysgenesis syndromes. Here, we report that mice transgenic for the homeodomain transcription factor Isl-1 develop profound caudal growth defects that resemble human sacral/caudal agenesis. Isl-1 is normally expressed in the pancreas and is required for pancreas development and endocrine cell differentiation...

Muller, Yunhua Li; Yueh, Yir Gloria; Yaworsky, Paul J.; Salbaum, J. Michael; Kappen, Claudia

2003-01-01

260

Transgenic nonhuman primates for neurodegenerative diseases  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Animal models that represent human diseases constitute an important tool in understanding the pathogenesis of the diseases, and in developing effective therapies. Neurodegenerative diseases are complex disorders involving neuropathologic and psychiatric alterations. Although transgenic and knock-in mouse models of Alzheimer's disease, (AD, Parkinson's disease (PD and Huntington's disease (HD have been created, limited representation in clinical aspects has been recognized and the rodent models lack true neurodegeneration. Chemical induction of HD and PD in nonhuman primates (NHP has been reported, however, the role of intrinsic genetic factors in the development of the diseases is indeterminable. Nonhuman primates closely parallel humans with regard to genetic, neuroanatomic, and cognitive/behavioral characteristics. Accordingly, the development of NHP models for neurodegenerative diseases holds greater promise for success in the discovery of diagnoses, treatments, and cures than approaches using other animal species. Therefore, a transgenic NHP carrying a mutant gene similar to that of patients will help to clarify our understanding of disease onset and progression. Additionally, monitoring disease onset and development in the transgenic NHP by high resolution brain imaging technology such as MRI, and behavioral and cognitive testing can all be carried out simultaneously in the NHP but not in other animal models. Moreover, because of the similarity in motor repertoire between NHPs and humans, it will also be possible to compare the neurologic syndrome observed in the NHP model to that in patients. Understanding the correlation between genetic defects and physiologic changes (e.g. oxidative damage will lead to a better understanding of disease progression and the development of patient treatments, medications and preventive approaches for high risk individuals. The impact of the transgenic NHP model in understanding the role which genetic disorders play in the development of efficacious interventions and medications is foreseeable.

Chan Anthony WS

2004-06-01

261

Transgenic cultures: from the economic viewpoint  

OpenAIRE

The introduction of transgenic seeds for agricultural purposes poses modification to their production, due to the potential for reaching desired characteristics such as greater yield, this being fundamental in an economic environment characterised by open market conditions. However, acceptance of products resulting from genetic engineering is far from becoming a simple process; discussion relating to the predominance of private sector interests, the monopoly of knowledge and the safety of suc...

Mauricio Mosquera

2011-01-01

262

Manipulating transgenes using a chromosome vector  

OpenAIRE

Recent technological advances have enabled us to visualize the organization and dynamics of local chromatin structures; however, the comprehensive mechanisms by which chromatin organization modulates gene regulation are poorly understood. We designed a human artificial chromosome vector that allowed manipulation of transgenes using a method for delivering chromatin architectures into different cell lines from human to fish. This methodology enabled analysis of de novo construction, epigenetic...

Ikeno, Masashi; Suzuki, Nobutaka; Hasegawa, Yoshinori; Okazaki, Tsuneko

2009-01-01

263

Phytoremediation of polychlorinated biphenyls by transgenic tobacco.  

Czech Academy of Sciences Publication Activity Database

Chania : Technical University of Crete, 2008 - (Kalogerakis, N.; Fava, F.; Banwart, S.). s. 295-295 ISBN 978-960-8475-12-0. [European Bioremediation Conference /4./. 03.09.2008-06.09.2008, Chania] R&D Projects: GA MŠk 1M06030 Institutional research plan: CEZ:AV0Z40550506 Keywords : phytoremediation * PCB * transgenic plants * bphC Subject RIV: EI - Biotechnology ; Bionics

Chrastilová, Z.; Macková, M.; Nováková, M.; Szekeres, M.; Macek, Tomáš

264

Transgene Detection by Digital Droplet PCR  

OpenAIRE

Somatic gene therapy is a promising tool for the treatment of severe diseases. Because of its abuse potential for performance enhancement in sports, the World Anti-Doping Agency (WADA) included the term ‘gene doping’ in the official list of banned substances and methods in 2004. Several nested PCR or qPCR-based strategies have been proposed that aim at detecting long-term presence of transgene in blood, but these strategies are hampered by technical limitations. We developed a digital dro...

Moser, Dirk A.; Braga, Luca; Raso, Andrea; Zacchigna, Serena; Giacca, Mauro; Simon, Perikles

2014-01-01

265

Zebrafish transgenic Enhancer TRAP line database (ZETRAP)  

OpenAIRE

Abstract Background The zebrafish, Danio rerio, is used as a model organism to study vertebrate genetics and development. An effective enhancer trap (ET) in zebrafish using the Tol2 transposon has been demonstrated. This approach could be used to study embryogenesis of a vertebrate species in real time and with high resolution. Description The information gathered during the course of systematic investigation of many ET transgenic lines have been collected and compiled in the form of an onlin...

Emelyanov Alexander; Parinov Sergey; Kondrichin Igor; Gh, Choo Benjamin; Go William; Toh Wei-chang; Korzh Vladimir

2006-01-01

266

Toxins for Transgenic Resistance to Hemipteran Pests  

OpenAIRE

The sap sucking insects (Hemiptera), which include aphids, whiteflies, plant bugs and stink bugs, have emerged as major agricultural pests. The Hemiptera cause direct damage by feeding on crops, and in some cases indirect damage by transmission of plant viruses. Current management relies almost exclusively on application of classical chemical insecticides. While the development of transgenic crops expressing toxins derived from the bacterium Bacillus thuringiensis (Bt) ha...

Bonning, Bryony C.; Chougule, Nanasaheb P.

2012-01-01

267

Can Transgenic Maize Affect Soil Microbial Communities?  

OpenAIRE

The aim of the experiment was to determine if temporal variations of belowground activity reflect the influence of the Cry1Ab protein from transgenic maize on soil bacteria and, hence, on a regulatory change of the microbial community (ability to metabolize sources belonging to different chemical guilds) and/or a change in numerical abundance of their cells. Litter placement is known for its strong influence on the soil decomposer communities. The effects of the addition of crop residues on r...

Mulder, Christian; Wouterse, Marja; Raubuch, Markus; Roelofs, Willem; Rutgers, Michiel

2006-01-01

268

Transgenic approaches to western corn rootworm control.  

Science.gov (United States)

The western corn rootworm, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae) is a significant corn pest throughout the United States corn belt. Rootworm larvae feed on corn roots causing yield losses and control expenditures that are estimated to exceed US$1 billion annually. Traditional management practices to control rootworms such as chemical insecticides or crop rotation have suffered reduced effectiveness due to the development of physiological and behavioral resistance. Transgenic maize expressing insecticidal proteins are very successful in protecting against rootworm damage and preserving corn yield potential. However, the high rate of grower adoption and early reliance on hybrids expressing a single mode of action and low-dose traits threatens the durability of commercialized transgenic rootworm technology for rootworm control. A summary of current transgenic approaches for rootworm control and the corresponding insect resistance management practices is included. An overview of potential new modes of action based on insecticidal proteins, and especially RNAi targeting mRNA coding for essential insect proteins is provided. PMID:23604211

Narva, Kenneth E; Siegfried, Blair D; Storer, Nicholas P

2013-01-01

269

Arsenic biotransformation and volatilization in transgenic rice.  

Science.gov (United States)

• Biotransformation of arsenic includes oxidation, reduction, methylation, and conversion to more complex organic arsenicals. Members of the class of arsenite (As(III)) S-adenosylmethyltransferase enzymes catalyze As(III) methylation to a variety of mono-, di-, and trimethylated species, some of which are less toxic than As(III) itself. However, no methyltransferase gene has been identified in plants. • Here, an arsM gene from the soil bacterium Rhodopseudomonas palustris was expressed in Japonica rice (Oryza sativa) cv Nipponbare, and the transgenic rice produced methylated arsenic species, which were measured by inductively coupled plasma mass spectrometry (ICP-MS) and high-performance liquid chromatography-inductively coupled plasma mass spectrometry (HPLC-ICP-MS). • Both monomethylarsenate (MAs(V)) and dimethylarsenate (DMAs(V)) were detected in the roots and shoots of transgenic rice. After 12 d exposure to As(III), the transgenic rice gave off 10-fold greater volatile arsenicals. • The present study demonstrates that expression of an arsM gene in rice induces arsenic methylation and volatilization, theoretically providing a potential stratagem for phytoremediation. PMID:21517874

Meng, Xiang-Yan; Qin, Jie; Wang, Li-Hong; Duan, Gui-Lan; Sun, Guo-Xin; Wu, Hui-Lan; Chu, Cheng-Cai; Ling, Hong-Qing; Rosen, Barry P; Zhu, Yong-Guan

2011-07-01

270

Using metabolomics to estimate unintended effects in transgenic crop plants: problems, promises and opportunities  

Science.gov (United States)

Transgenic crops are widespread in some countries and sectors of the agro-economy, but are also highly contentious. Proponents of transgenic crop improvement often cite the “substantial equivalence” of transgenic crops to their non-transgenic parents and sibling varieties. Opponents of transgenic cr...

271

Production of recombinant proteins in milk of transgenic and non-transgenic goats  

Scientific Electronic Library Online (English)

Full Text Available Among all the transgenic mammalians produced so far, goats have represented an excellent model of transgenesis when considering the factors such as the market demand for protein, volume of milk produced per lactation and reproductive rate. Various recombinant proteins have been obtained from the tra [...] nsgenic and non-transgenic goats, and among these, human antithrombin, produced by the transgenic goats, was the first recombinant protein of animal origin to be released as a drug for the clinical use in humans. This review reports the aspects inherent to the production of recombinant proteins in the goats, from the production of the animal bioreactors up to the expression of these proteins in their milk.

Raylene Ramos, Moura; Luciana Magalhães, Melo; Vicente José de Figueirêdo, Freitas.

2011-10-01

272

New doxycycline-inducible transgenic lines in Xenopus.  

Science.gov (United States)

We have characterized two new transgenic Xenopus lines enabling transgene expression using the Tet-On inducible system. An inducer line expresses the doxycycline- (Dox-) activated transcription factor rtTA under control of the ubiquitous promoter CMV. A responder line enables Dox-inducible expression of a dominant positive thyroid hormone receptor via a tetracycline responsive transgenic promoter (TRE). Dox-induced expression of transgenic GFP mRNA was detectable after 3 hr and increased up to 10- to 50-fold by 2 days depending on dose of Dox. Induced GFP mRNA expression returned to uninduced levels within 3 days upon Dox removal. Treatment of rtTA inducer and TRE responder double transgenic animals with Dox caused acceleration of metamorphic changes in thyroid hormone-response gene expression and morphology. These transgenic lines will be made available through the new Xenopus Stock Center and will serve as valuable tools for genetic analysis of development and metamorphosis. PMID:21491543

Rankin, Scott A; Zorn, Aaron M; Buchholz, Daniel R

2011-06-01

273

Rheumatic manifestations of inflammatory bowel disease  

Directory of Open Access Journals (Sweden)

Full Text Available This article reviews the literature concerning rheumatic manifestations of inflammatory bowel disease (IBD, including common immune-mediated pathways, frequency, clinical course and therapy. Musculoskeletal complications are frequent and well-recognized manifestations in IBD, and affect up to 33% of patients with IBD. The strong link between the bowel and the osteo-articular system is suggested by many clinical and experimental observations, notably in HLA-B27 transgenic rats. The autoimmune pathogenic mechanisms shared by IBD and spondyloarthropathies include genetic susceptibility to abnormal antigen presentation, aberrant recognition of self, the presence of autoantibodies against specific antigens shared by the colon and other extra-colonic tissues, and increased intestinal permeability. The response against microorganisms may have an important role through molecular mimicry and other mechanisms. Rheumatic manifestations of IBD have been divided into peripheral arthritis, and axial involvement, including sacroiliitis, with or without spondylitis, similar to idiopathic ankylosing spondylitis. Other periarticular features can occur, including enthesopathy, tendonitis, clubbing, periostitis, and granulomatous lesions of joints and bones. Osteoporosis and osteomalacia secondary to IBD and iatrogenic complications can also occur. The management of the rheumatic manifestations of IBD consists of physical therapy in combination with local injection of corticosteroids and nonsteroidal anti-inflammatory drugs; caution is in order however, because of their possible harmful effects on intestinal integrity, permeability, and even on gut inflammation. Sulfasalazine, methotrexate, azathioprine, cyclosporine and leflunomide should be used for selected indications. In some cases, tumor necrosis factor-? blocking agents should be considered as first-line therapy.

Tatiana Sofía Rodríguez-Reyna, Cynthia Martínez-Reyes, Jesús Kazúo Yamamoto-Furusho

2009-11-01

274

Pronuclear Microinjection and Oviduct Transfer Procedures for Transgenic Mouse Production  

OpenAIRE

Transgenic mouse technology is a powerful method for studying gene function and creating animal models of human diseases. Currently, the most widely used method for generating transgenic mice is the pronuclear microinjection method. In this method, a transgenic DNA construct is physically microinjected into the pronucleus of a fertilized egg. The injected embryos are subsequently transferred into the oviducts of pseudopregnant surrogate mothers. A portion of the mice born to these surrogate m...

Liu, Chengyu; Xie, Wen; Gui, Changyun; Du, Yubin

2013-01-01

275

The Applications of Transgenic Rabbits in Agriculture and Biomedicine  

OpenAIRE

During the last decades transgenic rabbits have provided suitable biological model for regulating and manipulating of interest genes. Several studies showed transgenic rabbits can be produced by different methods. In the last two decades pronucleotide microinjection was conventional and common method for produce transgenic rabbits but the current studies demonstrated that SMGT method provides a simple and straightforward technology to introduce DNA into rabbits which has many advantages in co...

Kh. Hosseini; Tahmoorespur, M.; Zabetian, M.

2011-01-01

276

Illegitimate Cre-dependent chromosome rearrangements in transgenic mouse spermatids  

OpenAIRE

The bacteriophage P1 Cre/loxP system has become a powerful tool for in vivo manipulation of the genomes of transgenic mice. Although in vitro studies have shown that Cre can catalyze recombination between cryptic “pseudo-loxP” sites in mammalian genomes, to date there have been no reports of loxP-site infidelity in transgenic animals. We produced lines of transgenic mice that use the mouse Protamine 1 (Prm1) gene promoter to express Cre recombinase in postm...

Schmidt, Edward E.; Taylor, Deborah S.; Prigge, Justin R.; Barnett, Sheila; Capecchi, Mario R.

2000-01-01

277

Variable patterns of expression of luciferase in transgenic tobacco leaves.  

OpenAIRE

A carboxyl-terminally modified firefly luciferase, encoded as a gene fusion to the neomycin phosphotransferase gene (which confers kanamycin resistance), was found to be enzymatically active for both enzymes when expressed in bacteria and in transgenic plants. A military-type starlight vision system was used to conveniently analyze the pattern of gene expression in transgenic tobacco plant leaves. Transgenic tobacco plants which expressed luciferase uniformly in all areas of the leaf, and ass...

Barnes, W. M.

1990-01-01

278

Hepatic steatosis in transgenic mice overexpressing human histone deacetylase 1  

International Nuclear Information System (INIS)

It is generally thought that histone deacetylases (HDACs) play important roles in the transcriptional regulation of genes. However, little information is available concerning the specific functions of individual HDACs in disease states. In this study, two transgenic mice lines were established which harbored the human HDAC1 gene. Overexpressed HDAC1 was detected in the nuclei of transgenic liver cells, and HDAC1 enzymatic activity was significantly higher in the transgenic mice than in control littermates. The HDAC1 transgenic mice exhibited a high incidence of hepatic steatosis and nuclear pleomorphism. Molecular studies showed that HDAC1 may contribute to nuclear pleomorphism through the p53/p21 signaling pathway

279

Transgenes are dispensable for the RNA degradation step of cosuppression  

OpenAIRE

Cosuppression results in the degradation of RNA from host genes and homologous transgenes after transcription in the nucleus. By using grafting experiments, we have shown previously that a systemic signal mediates the propagation of cosuppression of Nia host genes and 35S-Nia2 transgenes from silenced 35S-Nia2 transgenic stocks to nonsilenced 35S-Nia2 transgenic scions but not to wild-type scions. Here, we examined the requirements for triggering and maintenance of cosuppression in various ty...

Palauqui, Jean-christophe; Vaucheret, Herve?

1998-01-01

280

Comparative metallomics of transgenic and non-transgenic soybeans using HPLC-ICP-MS  

International Nuclear Information System (INIS)

Complete text of publication follows. In the last years, many soybean varieties have been developed, and due to these modifications, the proteins composition and profile can be affected, causing changes in the species proteome (S. Natarajan et al., Anal. Biochem., 342 (2005), 214-220.). With the proteome modifications, the metallome of this specie, defined as the total content of metals and metalloids in a cell or tissue (J. Spuznar, Analyst, 130 (2005), 442-465.), can also be affected (A. Sussulini et al., J. Anal. At. Spectrom., 22 (2007), 1501-1506.). So, the aim of this work is to amplify the information about the transgenic and non-transgenic soybeans metallome, and doing that we expect to find biomarkers that can differentiate the transgenic and non-transgenic soybeans physiologically. For that purpose a SEC column (GE Healthcare, model Superdex 200) was employed for the separation of the proteins, which were extracted using the mobile phase of the chromatographic system (90 mmol.L-1 phosphate buffer - pH 7.2). After the chromatographic separation, the eluate was passed through a DAD Series 200 detector (PerkinElmer), the fractions were collected and latter introduced into the ICP-MS (PerkinElmer, model ELANDRC-e) for the element-selective detection. The calibration of the column using purified proteins of known molecular weight allowed the calculation of the approximate masses of the eight fractions (1800-800 kDa; 800-420 kDa; 420-120 kDa; 100-23 kDa kDa; 800-420 kDa; 420-120 kDa; 100-23 kDa; 23-7 kDa; 7-2 kDa; 2-0.4 kDa and 0.4-0.2 kDa, respectively) identified in the transgenic and non-transgenic soybeans after 95 min of separation using a flow rate of 0.25 mL.min-1. A wide range of elements could be identified in all the fractions, including: Cu, Zn, Mn, Mg, Ni, Cr, Hg, Fe and Pb. Differences in the detectability of elements in the transgenic and non-transgenic soybeans were found, specially for Hg where the counts were two times higher in the transgenic soybean. Elements were found in the two samples that were not common for both of them, such as Sr identified only in fraction 2 of the non-transgenic soybean and Th in fraction 4 of the transgenic soybean. Financial support from Fundacao de Amparo a Pesquisa do Estado de Sao Paulo - FAPESP and Conselho Nacional de Desenvolvimento Cientifico e Tecnologico - CNPq are highly acknowledged.

281

Functional conservation between rodents and chicken of regulatory sequences driving skeletal muscle gene expression in transgenic chickens  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background Regulatory elements that control expression of specific genes during development have been shown in many cases to contain functionally-conserved modules that can be transferred between species and direct gene expression in a comparable developmental pattern. An example of such a module has been identified at the rat myosin light chain (MLC 1/3 locus, which has been well characterised in transgenic mouse studies. This locus contains two promoters encoding two alternatively spliced isoforms of alkali myosin light chain. These promoters are differentially regulated during development through the activity of two enhancer elements. The MLC3 promoter alone has been shown to confer expression of a reporter gene in skeletal and cardiac muscle in transgenic mice and the addition of the downstream MLC enhancer increased expression levels in skeletal muscle. We asked whether this regulatory module, sufficient for striated muscle gene expression in the mouse, would drive expression in similar domains in the chicken. Results We have observed that a conserved downstream MLC enhancer is present in the chicken MLC locus. We found that the rat MLC1/3 regulatory elements were transcriptionally active in chick skeletal muscle primary cultures. We observed that a single copy lentiviral insert containing this regulatory cassette was able to drive expression of a lacZ reporter gene in the fast-fibres of skeletal muscle in chicken in three independent transgenic chicken lines in a pattern similar to the endogenous MLC locus. Reporter gene expression in cardiac muscle tissues was not observed for any of these lines. Conclusions From these results we conclude that skeletal expression from this regulatory module is conserved in a genomic context between rodents and chickens. This transgenic module will be useful in future investigations of muscle development in avian species.

Taylor Lorna

2010-02-01

282

Genetic approaches for studying transgene inheritance and genetic recombination in three successive generations of transformed tobacco  

OpenAIRE

Transgene integration into plant genomes is a complex process accompanied by molecular rearrangements. Classic methods that are normally used to study transgenic population genetics are generally inadequate for assessing such integration. Two major characteristics of transgenic populations are that a transgenic genome may harbor many copies of the transgene and that molecular rearrangements can create an unstable transgenic locus. In this work, we examined the segregation of T1, T2 and T3 tra...

Kalthoum Tizaoui; Mohamed Elyes Kchouk

2012-01-01

283

Characterization of a Maize Wip1 Promoter in Transgenic Plants  

Directory of Open Access Journals (Sweden)

Full Text Available The Maize Wip1 gene encodes a wound-induced Bowman-Birk inhibitor (BBI protein which is a type of serine protease inhibitor, and its expression is induced by wounding or infection, conferring resistance against pathogens and pests. In this study, the maize Wip1 promoter was isolated and its function was analyzed. Different truncated Wip1 promoters were fused upstream of the GUS reporter gene and transformed into Arabidopsis, tobacco and rice plants. We found that (1 several truncated maize Wip1 promoters led to strong GUS activities in both transgenic Arabidopsis and tobacco leaves, whereas low GUS activity was detected in transgenic rice leaves; (2 the Wip1 promoter was not wound-induced in transgenic tobacco leaves, but was induced by wounding in transgenic rice leaves; (3 the truncated Wip1 promoter had different activity in different organs of transgenic tobacco plants; (4 the transgenic plant leaves containing different truncated Wip1 promoters had low GUS transcripts, even though high GUS protein level and GUS activities were observed; (5 there was one transcription start site of Wip1 gene in maize and two transcription start sites of GUS in Wip1::GUS transgenic lines; (6 the adjacent 35S promoter which is present in the transformation vectors enhanced the activity of the truncated Wip1 promoters in transgenic tobacco leaves, but did not influence the disability of truncated Wip1231 promoter to respond to wounding signals. We speculate that an ACAAAA hexamer, several CAA trimers and several elements similar to ACAATTAC octamer in the 5'-untranslated region might contribute to the strong GUS activity in Wip1231 transgenic lines, meanwhile, compared to the 5'-untranslated region from Wip1231 transgenic lines, the additional upstream open reading frames (uORFs in the 5'-untranslated region from Wip1737 transgenic lines might contribute to the lower level of GUS transcript and GUS activity.

Shengxue Zhang

2013-12-01

284

Transgene detection by digital droplet PCR.  

Science.gov (United States)

Somatic gene therapy is a promising tool for the treatment of severe diseases. Because of its abuse potential for performance enhancement in sports, the World Anti-Doping Agency (WADA) included the term 'gene doping' in the official list of banned substances and methods in 2004. Several nested PCR or qPCR-based strategies have been proposed that aim at detecting long-term presence of transgene in blood, but these strategies are hampered by technical limitations. We developed a digital droplet PCR (ddPCR) protocol for Insulin-Like Growth Factor 1 (IGF1) detection and demonstrated its applicability monitoring 6 mice injected into skeletal muscle with AAV9-IGF1 elements and 2 controls over a 33-day period. A duplex ddPCR protocol for simultaneous detection of Insulin-Like Growth Factor 1 (IGF1) and Erythropoietin (EPO) transgenic elements was created. A new DNA extraction procedure with target-orientated usage of restriction enzymes including on-column DNA-digestion was established. In vivo data revealed that IGF1 transgenic elements could be reliably detected for a 33-day period in DNA extracted from whole blood. In vitro data indicated feasibility of IGF1 and EPO detection by duplex ddPCR with high reliability and sensitivity. On-column DNA-digestion allowed for significantly improved target detection in downstream PCR-based approaches. As ddPCR provides absolute quantification, it ensures excellent day-to-day reproducibility. Therefore, we expect this technique to be used in diagnosing and monitoring of viral and bacterial infection, in detecting mutated DNA sequences as well as profiling for the presence of foreign genetic material in elite athletes in the future. PMID:25375130

Moser, Dirk A; Braga, Luca; Raso, Andrea; Zacchigna, Serena; Giacca, Mauro; Simon, Perikles

2014-01-01

285

Human prion strain selection in transgenic mice  

OpenAIRE

Transgenic (Tg) mice expressing chimeras of mouse and human prion proteins (PrP) have shorter incubation periods for Creutzfeldt-Jakob disease (CJD) prions than mice expressing full-length human PrP. Increasing the sequence similarity of the chimeric PrP to mouse PrP, by reverting human residues to mouse, resulted in a Tg line, denoted Tg22372, which was susceptible to sporadic (s) CJD prions in ~110 days 1. Reversion of one additional residue (M111V) resulted in a new Tg line, termed Tg1014,...

Giles, Kurt; Glidden, David V.; Patel, Smita; Korth, Carsten; Groth, Darlene; Lemus, Azucena; Dearmond, Stephen J.; Prusiner, Stanley B.

2010-01-01

286

Overexpression of full-length centrobin rescues limb malformation but not male fertility of the hypodactylous (hd) rats  

OpenAIRE

Rat hypodactyly (hd) mutation is characterized by abnormal spermatogenesis and sperm decapitation, limb malformation (missing digits II and III) and growth retardation. We have previously reported centrobin (centrosome BRCA2-interacting protein) truncation at the C-terminus in the hd mutant. Here, we report data from a transgenic rescue experiment carried out to determine a role of centrobin in pathogenesis of hd. The transgenic construct, consisting of full-length-coding cDNA linked to a ubi...

Liska, F.; Gosele, C.; Popova, E.; Chylikova, B.; Krenova, D.; Kren, V.; Bader, M.; Tres, L. L.; Hubner, N.; Kierszenbaum, A. L.

2013-01-01

287

Biodiversity (Communications arising): maize transgene results in Mexico are artefacts.  

OpenAIRE

Quist and Chapela's conclusion that the transgenes they claim to have detected in native maize in Oaxaca, Mexico, are predominantly reassorted and inserted into a "diversity of genomic contexts" seems to be based on an artefact arising from the inverse polymerase chain reaction (i-PCR) they used to amplify sequences flanking 35S transgenes from cauliflower mosaic virus (CaMV).

Kaplinsky, N.; Braun, D.; Lisch, D.; Hay, A.; Hake, S.; Freeling, M.

2002-01-01

288

Bacterial Diversity in Rhizospheres of Nontransgenic and Transgenic Corn  

OpenAIRE

Bacterial diversity in transgenic and nontransgenic corn rhizospheres was determined. In greenhouse and field studies, metabolic profiling and molecular analysis of 16S rRNAs differentiated bacterial communities among soil textures but not between corn varieties. We conclude that bacteria in corn rhizospheres are affected more by soil texture than by cultivation of transgenic varieties.

Fang, Min; Kremer, Robert J.; Motavalli, Peter P.; Davis, Georgia

2005-01-01

289

[Transgene complete silencing may associate with rearrangement of retroviral vector].  

Science.gov (United States)

Transgene silencing is one of two major obstacles in both basic biomedical research for transgene and clinical practice of gene therapy. Based on the model of HT1080 cell clones, which transduced single copy of retroviral vector MGPN2, the mechanism of transgene silencing was explored in this investigation by a serial molecular techniques. In the HT1080 cell clone that absence of GFP protein synthesized, no significant aberration of epigenetic modification was detected, but the transcript size and the sequence changed that resulted in the reading frame shift. In addition to chromosomal position effect leading to transgene silencing, the transcript reading frame shift associated with retroviral vector rearrangements could induce complete silencing of transgene. PMID:21604499

Wang, Dan; Xiao, Lejia; Ma, Qingxin; Zhai, Fen; Ren, Sichong; Li, Changlong

2011-04-01

290

MRI of Transgene Expression: Correlation to Therapeutic Gene Expression  

Directory of Open Access Journals (Sweden)

Full Text Available Magnetic resonance imaging (MRI can provide highresolution 3D maps of structural and functional information, yet its use of mapping in vivo gene expression has only recently been explored. A potential application for this technology is to noninvasively image transgene expression. The current study explores the latter using a nonregulatable internalizing engineered transferrin receptor (ETR whose expression can be probed for with a superparamagnetic Tf-CLIO probe. Using an HSV-based amplicon vector system for transgene delivery, we demonstrate that: 1 ETR is a sensitive MR marker gene; 2 several transgenes can be efficiently expressed from a single amplicon; 3 expression of each transgene results in functional gene product; and 4 ETR gene expression correlates with expression of therapeutic genes when the latter are contained within the same amplicon. These data, taken together, suggest that MRI of ETR expression can serve as a surrogate for measuring therapeutic transgene expression.

Tomotsugu Ichikawa

2002-01-01

291

Advancing environmental risk assessment for transgenic biofeedstock crops  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Transgenic modification of plants is a key enabling technology for developing sustainable biofeedstocks for biofuels production. Regulatory decisions and the wider acceptance and development of transgenic biofeedstock crops are considered from the context of science-based risk assessment. The risk assessment paradigm for transgenic biofeedstock crops is fundamentally no different from that of current generation transgenic crops, except that the focus of the assessment must consider the unique attributes of a given biofeedstock crop and its environmental release. For currently envisioned biofeedstock crops, particular emphasis in risk assessment will be given to characterization of altered metabolic profiles and their implications relative to non-target environmental effects and food safety; weediness and invasiveness when plants are modified for abiotic stress tolerance or are domesticated; and aggregate risk when plants are platforms for multi-product production. Robust risk assessments for transgenic biofeedstock crops are case-specific, initiated through problem formulation, and use tiered approaches for risk characterization.

Wolt Jeffrey D

2009-11-01

292

Transgenic crops. Processes, products and problems  

International Nuclear Information System (INIS)

Transgenic crops are a natural extension of plant breeding technologies, offering new opportunities for increasing the productivity of agriculture and reducing the cost of food production, for increasing the appeal, nutritional content and quality of fresh and processed foods, and for reducing the environmental damage of agricultural practices. These new transgenic traits will be combined with continuing improvements in the latest varieties developed via breeding technologies, spurring investment in both. These technologies are inherently compatible with and necessary for meeting the challenges now facing the world, namely, economic growth and development, environmental protection and remediation, and human needs for food, shelter and a decent quality of life. The first products from genetically engineered crops are beginning to enter commerce. This is a critical time for issues that will shape public acceptance and for adoption of regulatory and trade policies that encourage rather than discourage investment in and use of this technology. Further investment in the tools for transforming crops and in the basic and applied sciences that will provide a pipeline of new genes is also needed. (author). 24 refs, 1 tab

293

[Biofuels, food security and transgenic crops].  

Science.gov (United States)

Soaring global food prices are threatening to push more poor people back below the poverty line; this will probably become aggravated by the serious challenge that increasing population and climate changes are posing for food security. There is growing evidence that human activities involving fossil fuel consumption and land use are contributing to greenhouse gas emissions and consequently changing the climate worldwide. The finite nature of fossil fuel reserves is causing concern about energy security and there is a growing interest in the use of renewable energy sources such as biofuels. There is growing concern regarding the fact that biofuels are currently produced from food crops, thereby leading to an undesirable competition for their use as food and feed. Nevertheless, biofuels can be produced from other feedstocks such as lingo-cellulose from perennial grasses, forestry and vegetable waste. Biofuel energy content should not be exceeded by that of the fossil fuel invested in its production to ensure that it is energetically sustainable; however, biofuels must also be economically competitive and environmentally acceptable. Climate change and biofuels are challenging FAO efforts aimed at eradicating hunger worldwide by the next decade. Given that current crops used in biofuel production have not been domesticated for this purpose, transgenic technology can offer an enormous contribution towards improving biofuel crops' environmental and economic performance. The present paper critically presents some relevant relationships between biofuels, food security and transgenic plant technology. PMID:19722000

Acosta, Orlando; Chaparro-Giraldo, Alejandro

2009-01-01

294

Investigations into the hypothesis of transgenic cannabis.  

Science.gov (United States)

The unusual concentration of cannabinoids recently found in marijuana samples submitted to the forensic laboratory for chemical analysis prompted an investigation into whether genetic modifications have been made to the DNA of Cannabis sativa L. to increase its potency. Traditional methods for the detection of genetically modified organisms (GMO) were used to analyze herbal cannabis preparations. Our analyses support the hypothesis that marijuana samples submitted to forensic laboratories and characterized by an abnormal level of ?(9)-THC are the product of breeding selection rather than of transgenic modifications. Further, this research has shown a risk of false positive results associated with the poor quality of the seized samples and probably due to the contamination by other transgenic vegetable products. On the other hand, based on these data, a conclusive distinction between the hypothesis of GMO plant contamination and the other of genetic modification of cannabis cannot be made requiring further studies on comparative chemical and genetic analyses to find out an explanation for the recently detected increased potency of cannabis. PMID:22211569

Cascini, Fidelia

2012-05-01

295

Germline competent embryonic stem cells derived from rat blastocysts.  

Science.gov (United States)

Rats have important advantages over mice as an experimental system for physiological and pharmacological investigations. The lack of rat embryonic stem (ES) cells has restricted the availability of transgenic technologies to create genetic models in this species. Here, we show that rat ES cells can be efficiently derived, propagated, and genetically manipulated in the presence of small molecules that specifically inhibit GSK3, MEK, and FGF receptor tyrosine kinases. These rat ES cells express pluripotency markers and retain the capacity to differentiate into derivatives of all three germ layers. Most importantly, they can produce high rates of chimerism when reintroduced into early stage embryos and can transmit through the germline. Establishment of authentic rat ES cells will make possible sophisticated genetic manipulation to create models for the study of human diseases. PMID:19109898

Li, Ping; Tong, Chang; Mehrian-Shai, Ruty; Jia, Li; Wu, Nancy; Yan, Youzhen; Maxson, Robert E; Schulze, Eric N; Song, Houyan; Hsieh, Chih-Lin; Pera, Martin F; Ying, Qi-Long

2008-12-26

296

Increased Expression of the Na,K-ATPase alpha4 Isoform Enhances Sperm Motility in Transgenic Mice1  

OpenAIRE

The Na,K-ATPase alpha4 (ATP1A4) isoform is specifically expressed in male germ cells and is highly prevalent in spermatozoa. Although selective inhibition of alpha4 activity with ouabain has been shown to affect sperm motility, a more direct analysis of the role of this isoform in sperm movement has not yet been demonstrated. To establish this, we engineered transgenic mice that express the rat alpha4 isoform fused to green fluorescent protein in male germ cells, under the control of the mous...

Jimenez, Tamara; Sanchez, Gladis; Mcdermott, Jeffrey P.; Nguyen, Anh-nguyet; Kumar, T. Rajendra; Blanco, Gustavo

2010-01-01

297

Inhibition of mammary carcinoma development in HER-2/neu transgenic mice through induction of autoimmunity by xenogeneic DNA vaccination.  

OpenAIRE

Plasmid DNA vectors encoding the full-length (VR1012/HER-2-FL) or only the extracellular and transmembrane domains (VR1012/HER-2-ECD-TM) of human (h) HER-2/neu proto-oncogene were used to vaccinate HER-2/neu transgenic mice (N202) engineered to overexpress the rat (r) neu proto-oncogene product (r-p185(neu)). Both the full-length and the deleted vaccines were significantly (P = 0.0001 and P = 0.06, respectively) more active than the empty vector (VR1012/EV) in preventing and delaying HER-2/ne...

Cavallo, Federica

2005-01-01

298

Ectopic growth of hippocampal mossy fibers in a mutated GAP-43 transgenic mouse with impaired spatial memory retention  

OpenAIRE

In a previous study, it was shown that transgenic mice, designated G-NonP, forget the location of a water maze hidden platform when tested 7 days after the last training day (Holahan and Routtenberg, 2008). The memory loss in G-NonP mice might be related to altered hippocampal architecture suggested by the fact that in the rat, 7 days after water maze training, there is discernible mossy fiber (MF) growth (Holahan et al., 2006; Rekart et al., 2007). In the present report, we studied the distr...

Holahan, Matthew R.; Honegger, Kyle S.; Routtenberg, Aryeh

2010-01-01

299

A set of highly informative rat simple sequence length polymorphism (SSLP markers and genetically defined rat strains  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background The National Bio Resource Project for the Rat in Japan (NBRP-Rat is focusing on collecting, preserving and distributing various rat strains, including spontaneous mutant, transgenic, congenic, and recombinant inbred (RI strains. To evaluate their value as models of human diseases, we are characterizing them using 109 phenotypic parameters, such as clinical measurements, internal anatomy, metabolic parameters, and behavioral tests, as part of the Rat Phenome Project. Here, we report on a set of 357 simple sequence length polymorphism (SSLP markers and 122 rat strains, which were genotyped by the marker set. Results The SSLP markers were selected according to their distribution patterns throughout the whole rat genome with an average spacing of 7.59 Mb. The average number of informative markers between all possible pairs of strains was 259 (72.5% of 357 markers, showing their high degree of polymorphism. From the genetic profile of these rat inbred strains, we constructed a rat family tree to clarify their genetic background. Conclusion These highly informative SSLP markers as well as genetically and phenotypically defined rat strains are useful for designing experiments for quantitative trait loci (QTL analysis and to choose strategies for developing new genetic resources. The data and resources are freely available at the NBRP-Rat web site 1.

Yamasaki Ken-ichi

2006-04-01

300

Suppressed phosphorylation of collapsin response mediator protein-2 in the hippocampus of HCNP precursor transgenic mice.  

Science.gov (United States)

We previously reported a novel peptide, Hippocampal Cholinergic Neurostimulating Peptide (HCNP), which induces acetylcholine synthesis by increasing the amount of choline acetyltransferase (ChAT) in medial septal nuclei. The HCNP precursor protein (HCNP-pp), composed of 186 amino acids, is an inhibitory factor of the c-Raf/MEK cascade and may be involved in fetal rat brain development via the inhibition of phosphorylation of Erk. To clarify the involvement of HCNP in hippocampal cholinergic circuitry, we previously generated HCNP-pp transgenic (HCNP-pp Tg) mice using the promoter of the ? subunit of Ca(2+) calmodulin-dependent protein kinase II (CaMKII?). These mice showed increased levels of ChAT in medial septal nuclei at 12 weeks of age, and the phenotype of depressive mood at 30 weeks of age. Here, through proteomic analysis we investigated the alteration of protein expression in the hippocampus of HCNP-pp Tg mice compared with wild-type littermate mice. We demonstrate that the activation of collapsin response mediator protein-2 (CRMP-2) is increased in the transgenic mice at 12 weeks of age when compared with wild-type littermate mice. PMID:20682295

Kanamori, Tetsuko; Matsukawa, Noriyuki; Kobayashi, Hatasu; Uematsu, Norihiko; Sagisaka, Takafumi; Toyoda, Takanari; Kato, Daisuke; Oikawa, Shinji; Ojika, Kosei

2010-10-01

301

Welfare assessment in transgenic pigs expressing green fluorescent protein (GFP)  

DEFF Research Database (Denmark)

Since large animal transgenesis has been successfully attempted for the first time about 25 years ago, the technology has been applied in various lines of transgenic pigs. Nevertheless one of the concerns with the technology—animal welfare—has not been approached through systematic assessment and statements regarding the welfare of transgenic pigs have been based on anecdotal observations during early stages of transgenic programs. The main aim of the present study was therefore to perform an extensive welfare assessment comparing heterozygous transgenic animals expressing GFP with wildtype animals along various stages of post natal development. The protocol used covered reproductory performance and behaviour in GFP and wildtype sows and general health and development, social behaviour, exploratory behaviour and emotionality in GFP and wildtype littermates from birth until an age of roughly 4 months. The absence of significant differences between GFP and wildtype animals in the parameters observed suggests that the transgenic animals in question are unlikely to suffer from deleterious effects of transgene expression on their welfare and thus support existing anecdotal observations of pigs expressing GFP as healthy. Although the results are not surprising in the light of previous experience, they give a more solid fundament to the evaluation of GFP expression as being relatively non-invasive in pigs. The present study may furthermore serve as starting point for researchers aiming at a systematic characterization of welfare relevant effects in the line of transgenic pigs they are working with.

Huber, Reinhard C.; Remuge, Liliana

2012-01-01

302

Design and Management of Field Trials of Transgenic Cereals  

Science.gov (United States)

The development of gene transformation systems has allowed the introgression of alien genes into plant genomes, thus providing a mechanism for broadening the genetic resources available to plant breeders. The design and the management of field trials vary according to the purpose for which transgenic cereals are developed. Breeders study the phenotypic and genotypic stability of transgenic plants, monitor the increase in homozygosity of transgenic genotypes under field conditions, and develop backcross generations to transfer the introduced genes into secondary transgenic cereal genotypes. For practical purposes, they may also multiply seed of the transgenic lines to produce sufficient amounts of grain for the detailed analysis of trait(s) of interest, to determine the field performance of transgenic lines, and to compare them with the non-transformed parental genotypes. Prior to variety registration, the Distinctness, Uniformity and Stability (DUS) tests and Value for Cultivation and Use (VCU) experiments are carried out in field trials. Field testing includes specific requirements for transgenic cereals to assess potential environmental risks. The capacity of the pollen to survive, establish and disseminate in the field test environment, the potential for gene transfer, the effects of products expressed by the introduced sequences and phenotypic and genotypic instability that might cause deleterious effects must all be specifically monitored, as required by EU Directives 2003/701/EC (1) on the release of genetically modified higher plants in the environment.

Bed?, Zoltán; Rakszegi, Mariann; Láng, László

303

Evaluating potential risks of transgenic arthropods for pest management programmes  

International Nuclear Information System (INIS)

Genetic modification using recombinant DNA methods can now be used, almost routinely, to transform pest and beneficial arthropods and such genetically engineered insects and mites could be used to improve pest management programs. Genetic manipulation with recombinant DNA techniques may generate concerns about risk, requiring additional time and resources to resolve. Risk assessments must be conducted prior to releasing transgenic arthropods into the environment for either short term experiments or permanent establishment. Potential risk issues to be resolved include whether: the inserted gene(s) (trait) is stable; the traits can be horizontally transferred to other populations or species; released arthropods will perform as expected (especially with regard to their geographic distribution, host or prey specificity; released arthropods will have unintended environmental effects; and, in the case of short term releases, the released arthropods can be recovered from field sites. If the transgenic arthropods strain(s) perform well in preliminary, short term releases and risk assessments are completed satisfactorily, permanent releases into the environment may follow. Many pest management programs, especially those involving replacement of pest populations by the transgenic population, will require permanent establishment in the environment and the use of 'drive mechanisms', have been proposed to achieve this. Because efficacy can be severely compromised by 'transgene sil be severely compromised by 'transgene silencing', plant molecular biologists are now attempting to stabilize gene expression by building in 'insulators'. Transgene silencing occurs in Drosophila and will no doubt be a factor in other transgenic arthropods. (author)

304

Changes in physiological properties of rat ganglion cells during retinal degeneration  

OpenAIRE

Retinitis pigmentosa (RP) is a leading cause of degenerative vision loss, yet its progressive effects on visual signals transmitted from the retina to the brain are not well understood. The transgenic P23H rat is a valuable model of human autosomal dominant RP, exhibiting extensive similarities to the human disease pathology, time course, and electrophysiology. In this study, we examined the physiological effects of degeneration in retinal ganglion cells (RGCs) of P23H rats aged between P37 a...

Sekirnjak, Chris; Jepson, Lauren H.; Hottowy, Pawel; Sher, Alexander; Dabrowski, Wladyslaw; Litke, A. M.; Chichilnisky, E. J.

2011-01-01

305

Faster Recovery of Cerebral Perfusion in SOD1-Overexpressed Rats after Cardiac Arrest and Resuscitation  

OpenAIRE

Protracted hypoperfusion is one of the hallmarks of secondary cerebral derangement after cardiac arrest and resuscitation (CAR), and reactive oxygen species have been implicated in reperfusion abnormalities. Using transgenic (Tg) rats overexpressing copper zinc superoxide dismutase (SOD1), we investigated the role of this intrinsic antioxidant in the restoration of cerebral blood flow (CBF) after CAR. Nine Tg and 11 wild-type (WT) rats were subjected to 14-min cardiac arrest, and CBF was meas...

Xu, Y.; Liachenko, S. M.; Tang, P.; Chan, P. H.

2009-01-01

306

Inheritance of transgenes in transgenic Bt lines resistance to Helicoerpa armigera in upland cotton.  

Science.gov (United States)

Six transgenic Bt cotton cultivars (lines) including GKsu12, GK19, MR1, GK5, 109B, and SGK1 are highly resistant to bollworm from the seedling to boll-setting stages in bioassays with detached cotton leaves, though there are differences in resistant level and Bt toxin content in these transgenic cottons. Genetics analysis reveals that the resistance to Helicoverpa armigera in these six transgenic Bt cotton cultivars (lines) are controlled by one pair of dominant genes. Allelic tests further demonstrate some populations are in Mendel segregation for two nonallelic genes, i.e., the inserted Bt gene in GKsu12 is nonallelic to that of SGK1, GK5, 109B, and GK19 and Bt genes in GK19 and SGK1 are likely inserted in the same or in close proximity (genetically closely linked), while some F(2) produce abnormal segregation patterns, with a segregation of resistance to Helicoerpa armigera vary between 15:1 and 3:1, though their Bt segregation fit into 15:1 by PCR analysis, suggesting Bt gene silence in these populations. Two genes silence may occur in these populations due to the homologous sequence by crossing since the silenced individuals accounted for 1/16 of the F(2) populations for allelic test. To those silenced populations, one of their parents all showed high resistance to bollworm. PMID:23143492

Zhang, Baolong; Guo, Wangzhen; Zhang, Tianzhen

2013-01-01

307

Transgenic fish systems and their application in ecotoxicology.  

Science.gov (United States)

Abstract The use of transgenics in fish is a relatively recent development for advancing understanding of genetic mechanisms and developmental processes, improving aquaculture, and for pharmaceutical discovery. Transgenic fish have also been applied in ecotoxicology where they have the potential to provide more advanced and integrated systems for assessing health impacts of chemicals. The zebrafish (Daniorerio) is the most popular fish for transgenic models, for reasons including their high fecundity, transparency of their embryos, rapid organogenesis and availability of extensive genetic resources. The most commonly used technique for producing transgenic zebrafish is via microinjection of transgenes into fertilized eggs. Transposon and meganuclease have become the most reliable methods for insertion of the genetic construct in the production of stable transgenic fish lines. The GAL4-UAS system, where GAL4 is placed under the control of a desired promoter and UAS is fused with a fluorescent marker, has greatly enhanced model development for studies in ecotoxicology. Transgenic fish have been developed to study for the effects of heavy metal toxicity (via heat-shock protein genes), oxidative stress (via an electrophile-responsive element), for various organic chemicals acting through the aryl hydrocarbon receptor, thyroid and glucocorticoid response pathways, and estrogenicity. These models vary in their sensitivity with only very few able to detect responses for environmentally relevant exposures. Nevertheless, the potential of these systems for analyses of chemical effects in real time and across multiple targets in intact organisms is considerable. Here we illustrate the techniques used for generating transgenic zebrafish and assess progress in the development and application of transgenic fish (principally zebrafish) for studies in environmental toxicology. We further provide a viewpoint on future development opportunities. PMID:25394772

Lee, Okhyun; Green, Jon M; Tyler, Charles R

2015-02-01

308

Generation and characterization of human heme oxygenase-1 transgenic pigs.  

Science.gov (United States)

Xenotransplantation using transgenic pigs as an organ source is a promising strategy to overcome shortage of human organ for transplantation. Various genetic modifications have been tried to ameliorate xenograft rejection. In the present study we assessed effect of transgenic expression of human heme oxygenase-1 (hHO-1), an inducible protein capable of cytoprotection by scavenging reactive oxygen species and preventing apoptosis caused by cellular stress during inflammatory processes, in neonatal porcine islet-like cluster cells (NPCCs). Transduction of NPCCs with adenovirus containing hHO-1 gene significantly reduced apoptosis compared with the GFP-expressing adenovirus control after treatment with either hydrogen peroxide or hTNF-? and cycloheximide. These protective effects were diminished by co-treatment of hHO-1 antagonist, Zinc protoporphyrin IX. We also generated transgenic pigs expressing hHO-1 and analyzed expression and function of the transgene. Human HO-1 was expressed in most tissues, including the heart, kidney, lung, pancreas, spleen and skin, however, expression levels and patterns of the hHO-1 gene are not consistent in each organ. We isolate fibroblast from transgenic pigs to analyze protective effect of the hHO-1. As expected, fibroblasts derived from the hHO-1 transgenic pigs were significantly resistant to both hydrogen peroxide damage and hTNF-? and cycloheximide-mediated apoptosis when compared with wild-type fibroblasts. Furthermore, induction of RANTES in response to hTNF-? or LPS was significantly decreased in fibroblasts obtained from the hHO-1 transgenic pigs. These findings suggest that transgenic expression of hHO-1 can protect xenografts when exposed to oxidative stresses, especially from ischemia/reperfusion injury, and/or acute rejection mediated by cytokines. Accordingly, hHO-1 could be an important candidate molecule in a multi-transgenic pig strategy for xenotransplantation. PMID:23071605

Yeom, Hye-Jung; Koo, Ok Jae; Yang, Jaeseok; Cho, Bumrae; Hwang, Jong-Ik; Park, Sol Ji; Hurh, Sunghoon; Kim, Hwajung; Lee, Eun Mi; Ro, Han; Kang, Jung Taek; Kim, Su Jin; Won, Jae-Kyung; O'Connell, Philip J; Kim, Hyunil; Surh, Charles D; Lee, Byeong-Chun; Ahn, Curie

2012-01-01

309

WP1: transgenic opto-animals  

Science.gov (United States)

We aim to create a set of genetic tools where permanent opsin expression (ChR or NpHR) is precisely limited to the population of neurons that express immediate early gene c-fos during a specific temporal window of behavioral training. Since the c-fos gene is only expressed in neurons that form experience-dependent ensemble, this approach will result in specific labeling of a small subset of cells that create memory trace for the learned behavior. To this end we employ two alternative inducible gene expression systems: Tet Expression System and Cre/lox System. In both cases, the temporal window for opsin induction is controlled pharmacologically, by doxycycline or tamoxifen, respectively. Both systems will be used for creating lines of transgenic animals.

U?arowska, E.; Czajkowski, Rafa?; Konopka, W.

2014-11-01

310

Novel Transgenic Rice-Based Vaccines.  

Science.gov (United States)

Oral vaccination can induce both systemic and mucosal antigen-specific immune responses. To control rampant mucosal infectious diseases, the development of new effective oral vaccines is needed. Plant-based vaccines are new candidates for oral vaccines, and have some advantages over the traditional vaccines in cost, safety, and scalability. Rice seeds are attractive for vaccine production because of their stability and resistance to digestion in the stomach. The efficacy of some rice-based vaccines for infectious, autoimmune, and other diseases has been already demonstrated in animal models. We reported the efficacy in mice, safety, and stability of a rice-based cholera toxin B subunit vaccine called MucoRice-CTB. To advance MucoRice-CTB for use in humans, we also examined its efficacy and safety in primates. The potential of transgenic rice production as a new mucosal vaccine delivery system is reviewed from the perspective of future development of effective oral vaccines. PMID:25027548

Azegami, Tatsuhiko; Itoh, Hiroshi; Kiyono, Hiroshi; Yuki, Yoshikazu

2014-07-16

311

Overexpression of Glyoxalase-I Reduces Hyperglycemia-induced Levels of Advanced Glycation End Products and Oxidative Stress in Diabetic Rats*  

OpenAIRE

The reactive advanced glycation end product (AGE) precursor methylglyoxal (MGO) and MGO-derived AGEs are associated with diabetic vascular complications and also with an increase in oxidative stress. Glyoxalase-I (GLO-I) transgenic rats were used to explore whether overexpression of this MGO detoxifying enzyme reduces levels of AGEs and oxidative stress in a rat model of diabetes. Rats were made diabetic with streptozotocin, and after 12 weeks, plasma and multiple tissues were isolated for an...

Brouwers, O.; Niessen, P. M.; Ferreira, I.; Miyata, T.; Scheffer, P. G.; Teerlink, T.; Schrauwen, P.; Brownlee, M.; Stehouwer, C. D. A.; Schalkwijk, C. G.

2010-01-01

312

Unstable expression of transgene is associated with the methylation of CAG promoter in the offspring from the same litter of homozygous transgenic mice.  

Science.gov (United States)

Transgenic animals have been established for studying gene function, improving animals' production traits, and providing organ models for the exploration of human diseases. However, the stability of inheritance and transgene expression in transgenic animals has gained extensive attention. The unstable expression of transgene through DNA methyltransferase (DNMT) targeting to the methylation of transgenic DNA such as CAG promoter and Egfp coding region in homozygous transgenic animals is still unknown. In the present study, the offspring from the same litter of homozygous transgenic mice carrying ubiquitously expressed enhanced green fluorescence protein driven by CMV early enhancer/chicken ?-actin (CAG) promoter was observed to have unstable expression of transgene Egfp, quantitative PCR, western blot and bisulfite sequencing were conducted to quantify the expressional characteristics and methylation levels in various tissues. The correlation between transgene expression and methylation was analyzed. We have found that transgene expression is dependent on the methylation of CAG promoter, but not Egfp coding region. We have also characterized the correlation between the methylation of CAG promoter and DNMT, and found that only Dnmt3b expression is correlated with the methylation of CAG promoter. In conclusion, Dnmt3b-related methylation of CAG promoter can inhibit the transgene expression and may result in the unstable expression of transgene in the offspring from the same litter of homozygous transgenic mice. PMID:24804614

Zhou, Yang; Zhang, Teng; Zhang, Qin-Kai; Jiang, Ying; Xu, Deng-Gao; Zhang, Min; Shen, Wei; Pan, Qing-Jie

2014-08-01

313

Mutagenesis by man-made mineral fibres in the lung of rats.  

Czech Academy of Sciences Publication Activity Database

Ro?. 595, - (2006), s. 174-183. ISSN 0027-5107 Institutional research plan: CEZ:AV0Z50390512 Keywords : transgenic rats * mineral fibres Subject RIV: DN - Health Impact of the Environment Quality Impact factor: 4.111, year: 2006

Topinka, Jan; Loli, P.; Dušinská, M.; Hurbánková, M.; Ková?iková, Z.; Volkovová, K.; Kažimírová, A.; Baran?oková, M.; Tatrai, E.; Wolff, T.; Oesterle, D.; Kyrtopoulos, S.A.; Georgiadis, P.

2006-01-01

314

Transgenic approaches to a non-transgenic release of sterile male Lepidoptera  

International Nuclear Information System (INIS)

Successful implementation of the Sterile Insect Technique (SIT) against codling moth, Cydia pomonella (L.) (Tortricidae), in British Columbia, Canada, resulted in demands for the expansion of codling moth SIT and a related suppression strategy, radiation-induced inherited sterility (IS), in other countries. In the current SIT programme, both sterile males and females are released to control the pest population. There are compelling reasons to believe that both codling moth SIT and IS would benefit if efficient ways could be found to produce and release only males. Recently, a new scheme for genetic sexing in Lepidoptera has been proposed. The scheme is based on the construction of transgenic females carrying a dominant conditional lethal gene in the female-determining chromosome W. Following this scheme we intend to develop transgenic sexing strains in the codling moth. This requires basic knowledge of codling moth genome and appropriate molecular tools for codling moth transgenesis. We performed a detailed analysis of codling moth karyotype with a particular focus on the identification and characterization of sex chromosomes. Here we summarize our data on codling moth cytogenetics and discuss the potential of codling moth sex chromosomes for their use in developing transgenic sexing strains. The karyotype of codling moth consists of 2n=56 chromosomes, which can be classified into five groups according to their sizes: extra large (3 pairs), large (3 pairs), medium (15rge (3 pairs), large (3 pairs), medium (15 pairs), small (5 pairs), and dot-like (2 pairs). Females are heterogametic with a W-Z sex chromosome pair, males are homogametic with two Z chromosomes. The W and Z chromosomes represent the two largest elements in female chromosome complements. While the Z is composed of euchromatin and resembles to autosomes, the W consists largely of heterochromatin. For successful development of transgenic sexing strains in the codling moth, it is required to insert a conditional dominant lethal mutation (a transgene) into the W chromosome. Theoretically, the transgene insertion is a matter of probability, which is dependent on the size of the W relative to the rest of the genome. In the codling moth, the W is one of two largest chromosomes, comprising about 4% of the female genome, which should make it a good target for transgenesis with the probability of insertion of 1 in 25 (if only females are included) or with the overall probability of 1 in 50 (since both female and male embryos are exposed). However, since the W chromosome is mainly composed of heterochromatin, silencing of the transgene expression might be a serious problem. Different ways how to overcome this problem are discussed. For further characterisation of the codling moth W chromosome we employed advanced methods of molecular cytogenetics, genomic in situ hybridisation (GISH) and comparative genomic hybridisation (CGH). GISH detected the W chromosome by strong binding of the Cy3-labelled, female-derived DNA probe. With CGH, both the Cy3-labelled female-derived probe and Fluor-X labelled male-derived probe evenly bound to the W. This suggested that the W is composed predominantly of repetitive DNA sequences occurring scattered in other chromosomes but accumulated in the W. Finally, we prepared W-specific probes by laser microdissection of the W chromatin followed by DOP-PCR and PCR labelling. The probes stained the W with a high specificity in a chromosome-painting manner. DNA fragments of the microdissected W chromatin were cloned and sequenced. The W-sequence analysis revealed no homology to any DNA sequenced so far. Several cloned sequences were found to originate exclusively from the W chromosome. These unique sequences can be very useful as molecular markers of the W chromosome in codling moth transgenesis. The demonstrated ways of W chromosome identification will facilitate the development of genetic sexing strains in the codling moth

315

[Effects of agricultural activities and transgenic crops on agricultural biodiversity].  

Science.gov (United States)

Agricultural biodiversity is a key part of the ecosystem biodiversity, but it receives little concern. The monoculture, environmental pollution and habitat fragmentation caused by agricultural activities have threatened agricultural biodiversity over the past 50 years. To optimize agricultural management measures for crop production and environmental protection, we reviewed the effects of agricultural activities, including cultivation patterns, plastic mulching, chemical additions and the cultivation of transgenic crops, on agricultural biodiversity. The results showed that chemical pesticides and fertilizers had the most serious influence and the effects of transgenic crops varied with other factors like the specific transgene inserted in crops. The environmental risk of transgenic crops should be assessed widely through case-by-case methods, particularly its potential impacts on agricultural biodiversity. It is important to consider the protection of agricultural biodiversity before taking certain agricultural practices, which could improve agricultural production and simultaneously reduce the environmental impacts. PMID:25757330

Zhang, Xi-Tao; Luo, Hong-Bing; Li, Jun-Sheng; Huang, Hai; Liu, Yong-Bo

2014-09-01

316

Microinjection of A. aegypti Embryos to Obtain Transgenic Mosquitoes  

OpenAIRE

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.

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

2007-01-01

317

Simplifying Transgene Locus Structure Through Cre-lox Recombination.  

Science.gov (United States)

Transgene silencing is often associated with multicopy integrations, which occur frequently during plant transformation. Transgene expression can be restored in a number of multicopy loci by converting them to single copy. This chapter describes a plant transformation protocol based on use of the Cre-lox system, which allows conversion of a multicopy transgene locus into single copy. The strategy is based on designing a transformation vector with lox sites, developing transgenic lines, and introducing Cre activity to initiate Cre-lox recombination, which leads to the simplification of a multicopy locus to a single- or low-copy state. This method is compatible with both gene gun and Agrobacterium-mediated gene delivery and should be particularly useful for crops that are difficult to transform. PMID:25740358

Srivastava, Vibha; Ow, David W

2015-01-01

318

Designer proton-channel transgenic algae for photobiological hydrogen production  

Energy Technology Data Exchange (ETDEWEB)

A designer proton-channel transgenic alga for photobiological hydrogen production that is specifically designed for production of molecular hydrogen (H.sub.2) through photosynthetic water splitting. The designer transgenic alga includes proton-conductive channels that are expressed to produce such uncoupler proteins in an amount sufficient to increase the algal H.sub.2 productivity. In one embodiment the designer proton-channel transgene is a nucleic acid construct (300) including a PCR forward primer (302), an externally inducible promoter (304), a transit targeting sequence (306), a designer proton-channel encoding sequence (308), a transcription and translation terminator (310), and a PCR reverse primer (312). In various embodiments, the designer proton-channel transgenic algae are used with a gas-separation system (500) and a gas-products-separation and utilization system (600) for photobiological H.sub.2 production.

Lee, James Weifu (Knoxville, TN)

2011-04-26

319

Hybridization between transgenic and wild plants: environmental risk  

Directory of Open Access Journals (Sweden)

Full Text Available Genetically modified products are widely commercialized in agricultural production. These include resistant plants to diseases, insects or herbicides, plants with capacity for longer storing times or better nutritional quality. However, there are some concerns and critics from environmental organizations on the risk associated to transgenic plants or organisms genetically modified (OGM. This review discusses the vertical gene transfer (plant/Plant within the OGM context. Although transgenic hybrids have been reported between transgenic plants and their wild relatives, the extent of the environmental risk has not been evaluated per se. The risk depends on the plant species involved, the transgenes, and the ecosystem where the plants are located. Studies on biosafety assessment must be evaluated case by case. Biotechnology and conventional methods allow to control gen flow and decrease the risk of gene transfer among species.

Alejandro Chaparro Giraldo

2011-12-01

320

Preferential silencing of a common dominant rhodopsin mutation does not inhibit retinal degeneration in a transgenic model.  

Science.gov (United States)

Autosomal dominant retinitis pigmentosa caused by the frequent rhodopsin P23H mutation is characterized by progressive photoreceptor cell death eventually leading to blindness and for which no therapies are available. Considering the gain-of-function effect exerted by the P23H mutation, strategies aimed at silencing the expression of the mutated allele, like RNA interference, are desirable. We have designed small interfering RNAs (siRNA) to silence specifically the P23H rhodopsin allele expressed by a transgenic rat model of the disease. We have selected in vitro one siRNA and generated an adeno-associated viral (AAV) vector expressing the short hairpin RNA (shRNA) based on the selected siRNA. In vitro the shRNA significantly inhibits the expression of the P23H but not the wild-type rhodopsin allele. Subretinal administration of the AAV2/5 vector encoding the shRNA in P23H transgenic rats results in inhibition of rhodopsin P23H expression that is not able to prevent or block photoreceptor degeneration. Since rhodopsin is the most abundant rod photoreceptor protein, systems resulting in more robust shRNA expression in the retina may be required to achieve therapeutic efficacy in vivo. PMID:16979943

Tessitore, Alessandra; Parisi, Fabiana; Denti, Michela Alessandra; Allocca, Mariacarmela; Di Vicino, Umberto; Domenici, Luciano; Bozzoni, Irene; Auricchio, Alberto

2006-11-01

321

Devitalization of Transgenic Seed That Preserves DNA and Protein Integrity  

OpenAIRE

Agricultural biotechnology companies have been asked to provide intact transgenic seed to regulatory agencies as reference materials for evaluating transgene and protein detection methods (PCR and immunoassay). Due to intellectual-property and product-stewardship considerations, submission of devitalized seed prior to regulatory approval is preferable in any given country. Commonly used devitalization procedures, such as heating or autoclaving, degrade the protein and/or DNA rendering the see...

Schafer, Barry W.; Cai, Charles Q.; Embrey, Shawna K.; Herman, Rod A.; Song, Ping

2008-01-01

322

A regulatable transgene expression system for cultured Plasmodium falciparum parasites  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background The ability to transfect and create transgenic cultured malaria parasites has transformed the study of Plasmodium falciparum over the last decade. With the completion of the annotated genome sequence, the process of gene discovery now routinely includes gene knockouts, over-expression and complementation analysis. However, while this technology has proven extremely valuable, significant limitations exist. In particular, P. falciparum DNA is often unstable and difficult to clone because of its AT-rich, repetitive nature. As a result, transgene expression constructs can be difficult to assemble due to the need to include two expression cassettes on a single plasmid, one to drive expression of the transgene of interest and a second for expression of the selectable marker. In addition, transgene expression levels are usually not regulatable, making it difficult to assess phenotypes that are sensitive to the amount of protein expressed. Results A plasmid based system for transgene expression is described that uses a single, bidirectional promoter to drive expression of both the transgene and the selectable marker, thus greatly reducing the size of the construct and enhancing stability. Further, by altering the concentration of drug used for selection, it is possible to modulate the copy number of the concatameric episomes and thereby regulate the expression level of the transgene through a range greater than 10 fold. Conclusion The transgene expression system described here should prove useful for both routine protein over-expression and complementation experiments as well as for experiments in which precisely manipulating the expression level of candidate proteins is desirable. This should provide an additional level of precision to the tools used to study the molecular biology of malaria parasites.

Raskolnikov Dima

2008-05-01

323

Purification of Plasmid and BAC Transgenic DNA Constructs  

OpenAIRE

Pronuclear microinjection is the most used method for generating transgenic mice. The quality of DNA to be microinjected is a key determinant of the success rate of this method. DNA purity is a critical factor because trace amounts of many substances, when microinjected into the pronucleus of the fertilized egg, can kill or prevent the further development of the embryo. Avoiding all contaminants is not a trivial issue, because most transgenic fragments need to be purified from agarose gels. S...

Liu, Chengyu; Du, Yubin; Xie, Wen; Gui, Changyun

2013-01-01

324

Detection of potential transgenic plant DNA recipients among soil bacteria.  

Science.gov (United States)

The likelihood of gene transfer from transgenic plants to bacteria is dependent on gene number and the presence of homologous sequences. The large number of transgene copies in transplastomic (transgenes contained in the chloroplast genome) plant cells as well as the prokaryotic origin of the transgene, may thus significantly increase the likelihood of gene transfer to bacteria that colonize plant tissues. In order to assess the probability of such transfer, the length of homologous DNA sequences required between the transgene and the genome of the bacterial host was assessed. In addition, the probability that bacteria, which co-infect diseased plants, are transformable and have sequences similar to the flanking regions of the transgene was evaluated. Using Acinetobacter baylyi strain BD143 and transplastomic tobacco plants harboring the aadA gene (streptomycin and spectinomycin resistance), we found that sequences identical to the flanking regions containing as few as 55 nucleotides were sufficient for recombination to occur. Consequently, a collection of bacterial isolates able to colonize tobacco plant tissue infected by Ralstonia solanacearum strain K60 was obtained, screened for DNA sequence similarity with the chloroplastic genes accD and rbcL flanking the transgene, and tested for their ability to uptake extracellular DNA (broad host-range pBBR1MCS plasmids) by natural or electro-transformation. Results showed that among the 288 bacterial isolates tested, 8% presented DNA sequence similarity with one or both chloroplastic regions flanking the transgene. Two isolates, identified as Pseudomonas sp. and Acinetobacter sp., were able to integrate exogenous plasmid DNA by electro-transformation and natural transformation, respectively. Our data suggest that transplastomic plant DNA recipients might be present in soil bacterial communities. PMID:17961481

Monier, Jean-Michel; Bernillon, Dominique; Kay, Elizabeth; Faugier, Aurélie; Rybalka, Oleksandra; Dessaux, Yves; Simonet, Pascal; Vogel, Timothy M

2007-01-01

325

Enhancing shoot recovery from transgenic avocado somatic embryos  

OpenAIRE

Use of biotechnological tools in avocado (Persea americana Mill.) is hampered by difficulties in obtaining mature somatic embryos with an acceptable germination capacity. Use of semi-permeable cellulose acetate membranes on top of maturation medium has improved the quality of obtained embryos and their germination rate; however, in the case of transgenic embryos the conversion rate is still rather low. In this investigation, a protocol for recovery of transgenic plants has been developed. Mat...

Palomo-ri?os, Elena; Vidoy, Isabel; Barcelo?-mun?oz, Araceli; Mercado, Jose Angel; Pliego-alfaro, Fernando

2013-01-01

326

Transgene-induced mutation of the murine steel locus.  

OpenAIRE

The product of the steel locus is essential for normal development of three distinct populations of stem cells--the neural crest-derived melanoblasts, germ cells, and blood cell precursors. Many mutant alleles at steel are lethal in homozygotes and produce coat color dilution in heterozygotes. We have identified a transgenic mouse with diluted pigmentation that closely resembles that of steel heterozygotes. We have demonstrated that the site of transgene insertion is genetically linked to the...

Keller, S. A.; Liptay, S.; Hajra, A.; Meisler, M. H.

1990-01-01

327

Developments in transgenic fish in the People's Republic of China.  

Science.gov (United States)

In the People's Republic of China, genetically modified (GM) fish are being developed primarily to produce desirable alterations to growth rates or feed-conversion efficiency. Up to the present, no transgenic fish have been commercially approved for human consumption. This review introduces advances in the People's Republic of China in transgenic fish studies, biosafety studies of fast-growth GM fish, and the regulation of GM fish. PMID:16110897

Fu, C; Hu, W; Wang, Y; Zhu, Z

2005-04-01

328

Enhanced phytoremediation of volatile environmental pollutants with transgenic trees  

OpenAIRE

Small, volatile hydrocarbons, including trichloroethylene, vinyl chloride, carbon tetrachloride, benzene, and chloroform, are common environmental pollutants that pose serious health effects. We have developed transgenic poplar (Populus tremula × Populus alba) plants with greatly increased rates of metabolism and removal of these pollutants through the overexpression of cytochrome P450 2E1, a key enzyme in the metabolism of a variety of halogenated compounds. The transgenic poplar plants exh...

Doty, Sharon L.; James, C. Andrew; Moore, Allison L.; Vajzovic, Azra; Singleton, Glenda L.; Ma, Caiping; Khan, Zareen; Xin, Gang; Kang, Jun Won; Park, Jin Young; Meilan, Richard; Strauss, Steven H.; Wilkerson, Jasmine; Farin, Federico; Strand, Stuart E.

2007-01-01

329

Time-Course Expression Profiles of Hair Cycle-Associated Genes in Male Mini Rats after Depilation of Telogen-Phase Hairs  

OpenAIRE

Jcl:WistarTGN(ARGHGEN)1Nts rat (Mini rat) is a growth hormone (GH)-deficient transgenic rat. The hair cycle in the dorsal skin of male Mini rats enters a long-lasting telogen phase after eights weeks of age, but depilation can induce a transient hair cycle again. In this study, a time-course profiling of genes expression was done on the dorsal skin of male Mini rats along the progression of depilation-induced hair cycle using DNA microarray analysis. As a result, 1,215 probe sets including 1,...

Aya Umeda-Ikawa; Isao Shimokawa; Kunio Doi

2009-01-01

330

A pre-breeding screening program for transgenic boars based on fluorescence in situ hybridization assay.  

Science.gov (United States)

For efficient transgenic herd expansion, only the transgenic animals that possess the ability to transmit transgene into next generation are considered for breeding. However, for transgenic pig, practically lacking a pre-breeding screening program, time, labor and money is always wasted to maintain non-transgenic pigs, low or null transgenic transmission pigs and the related fruitless gestations. Developing a pre-breeding screening program would make the transgenic herd expansion more economical and efficient. In this technical report, we proposed a three-step pre-breeding screening program for transgenic boars simply through combining the fluorescence in situ hybridization (FISH) assay with the common pre-breeding screening workflow. In the first step of screening, combined with general transgenic phenotype analysis, FISH is used to identify transgenic boars. In the second step of screening, combined with conventional semen test, FISH is used to detect transgenic sperm, thus to identify the individuals producing high quality semen and transgenic sperm. In the third step of screening, FISH is used to assess the in vitro fertilization embryos, thus finally to identify the individuals with the ability to produce transgenic embryos. By this three-step screening, the non-transgenic boars and boars with no ability to produce transgenic sperm or transgenic embryos would be eliminated; therefore only those boars could produce transgenic offspring are maintained and used for breeding and herd expansion. It is the first time a systematic pre-breeding screening program is proposed for transgenic pigs. This program might also be applied in other transgenic large animals, and provide an economical and efficient strategy for herd expansion. PMID:24788205

Bou, Gerelchimeg; Sun, Mingju; Lv, Ming; Zhu, Jiang; Li, Hui; Wang, Juan; Li, Lu; Liu, Zhongfeng; Zheng, Zhong; He, Wenteng; Kong, Qingran; Liu, Zhonghua

2014-08-01

331

Transgene rescue in the mammary gland is associated with transcription but does not require translation of BLG transgenes.  

Science.gov (United States)

Many transgenes, particularly those comprising cDNA sequences fail to be expressed when they are introduced into transgenic mice. We have previously shown that this problem can be overcome in the mammary gland by co-integrating a poorly expressed cDNA transgene, comprising the sheep beta-lactoglobulin promoter, with the efficiently expressed, unmodified beta-lactoglobulin gene. In this report we demonstrate that the transcription of the beta-lactoglobulin gene is associated with this effect because co-integration with a non-transcribed beta-lactoglobulin gene fails to rescue expression. By contrast, co-integration with a translationally inactivated beta-lactoglobulin transgene does rescue the expression of the second gene, but without the co-production of beta-lactoglobulin protein. PMID:9032973

Yull, F; Binas, B; Harold, G; Wallace, R; Clark, A J

1997-01-01

332

Comparative study of transgenic and non-transgenic maize (Zea mays) flours commercialized in Brazil, focussing on proteomic analyses.  

Science.gov (United States)

Genetically modified foods are a major concern around the world due to the lack of information concerning their safety and health effects. This work evaluates differences, at the proteomic level, between two types of crop samples: transgenic (MON810 event with the Cry1Ab gene, which confers resistance to insects) and non-transgenic maize flour commercialized in Brazil. The 2-D DIGE technique revealed 99 differentially expressed spots, which were collected in 2-D PAGE gels and identified via mass spectrometry (nESI-QTOF MS/MS). The abundance of protein differences between the transgenic and non-transgenic samples could arise from genetic modification or as a result of an environmental influence pertaining to the commercial sample. The major functional category of proteins identified was related to disease/defense and, although differences were observed between samples, no toxins or allergenic proteins were found. PMID:25766830

Vidal, Nádia; Barbosa, Herbert; Jacob, Silvana; Arruda, Marco

2015-08-01

333

Effect of 60Co ?-rays irradiation on antioxidant enzymes activities in transgenic and non-transgenic tobacco seedlings  

International Nuclear Information System (INIS)

Changes of activities of antioxidant enzymes in pprI-transgenic tobacco seedlings and non-transgenic tobacco seedlings after different doses 60Co ?-rays irradiation were studied. The results showed that the activity of superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) in pprI-transgenic tobacco seedlings and non-transgenic tobacco seedlings were gradually increased after different doses 60Co ?-rays irradiation. The activity of SOD was to the maximum at 100 Gy treatment, but the activity of POD and CAT at 300 Gy treatment, and then these three antioxidant enzymes gradually decreased with the increase of irradiation dose. Quantitative real-time PCR analysis also revealed that the over-express of these antioxidant enzymes were induced after different doses 60Co ?-rays irradiation and were consistent with the variance of their enzymic activities, which enhanced the tolerance of tobacco against irradiation. (authors)

334

Composite potato plants with transgenic roots on non-transgenic shoots: a model system for studying gene silencing in roots  

DEFF Research Database (Denmark)

Composite plants, with transgenic roots on a non-transgenic shoot, can be obtained by shoot explant transformation with Agrobacterium rhizogenes. The aim of this study was to generate composite potato plants (Solanum tuberosum) to be used as a model system in future studies on root-pathogen interactions and gene silencing in the roots. The proportion of transgenic roots among the roots induced was high (80-100 %) in the four potato cultivars tested (Albatros, Desirée, Sabina and Saturna). No wild-type adventitious roots were formed at mock inoculation site. All strains of A. rhizogenes tested induced phenotypically normal roots which, however, showed a reduced response to cytokinin as compared with non-transgenic roots. Nevertheless, both types of roots were infected to a similar high rate with the zoospores of Spongospora subterranea, a soilborne potato pathogen. The transgenic roots of composite potato plants expressed significantly higher amounts of ?-glucuronidase (GUS) than the roots of a GUS-transgenic potato line event. Silencing of the uidA transgene (GUS) was tested by inducing roots on the GUS-transgenic cv. Albatros event with strains of A. rhizogenes over-expressing either the uidA sense or antisense transcripts, or inverted-repeat or hairpin uidA RNA. The three last mentioned constructs caused 2.5-4.0 fold reduction in the uidA mRNA expression. In contrast, over-expression of uidA resulted in over 3-fold increase in the uidA mRNA and GUS expression, indicating that sense-mediated silencing (co-suppression) was not functional in roots. The results suggest that composite plants offer a useful experimental system for potato research, which has gained little previous attention.

Horn, Patricia; Santala, Johanna

2014-01-01

335

Non-Homologous End Joining Plays a Key Role in Transgene Concatemer Formation in Transgenic Zebrafish Embryos  

OpenAIRE

This study focused on concatemer formation and integration pattern of transgenes in zebrafish embryos. A reporter plasmid based on enhanced green fluorescent protein (eGFP) driven by Cytomegalovirus (CMV) promoter, pCMV-pax6in-eGFP, was constructed to reflect transgene behavior in the host environment. After removal of the insertion fragment by double digestion with various combinations of restriction enzymes, linearized pCMV-pax6in-eGFP vectors were generated with different combinations of 5...

Jun Dai, Xiaojuan Cui

2010-01-01

336

Positive and negative selection of T cells in T-cell receptor transgenic mice expressing a bcl-2 transgene.  

OpenAIRE

To explore the role of bcl-2 in T-cell development, a bcl-2 transgene was introduced into mice expressing a T-cell receptor (TCR) transgene encoding reactivity for the mouse male antigen HY presented by the H-2Db class I antigen of the major histocompatibility complex (MHC). Normal thymic development is contingent on the ability of immature thymocytes to interact with self-MHC molecules presented by thymic stroma (positive selection). Thus, thymocyte numbers are low in femal...

Strasser, A.; Harris, A. W.; Von Boehmer, H.; Cory, S.

1994-01-01

337

RNAi-mediated knockdown of IKK1 in transgenic mice using a transgenic construct containing the human H1 promoter.  

Science.gov (United States)

Inhibition of gene expression through siRNAs is a tool increasingly used for the study of gene function in model systems, including transgenic mice. To achieve perdurable effects, the stable expression of siRNAs by an integrated transgenic construct is necessary. For transgenic siRNA expression, promoters transcribed by either RNApol II or III (such as U6 or H1 promoters) can be used. Relatively large amounts of small RNAs synthesis are achieved when using RNApol III promoters, which can be advantageous in knockdown experiments. To study the feasibility of H1 promoter-driven RNAi-expressing constructs for protein knockdown in transgenic mice, we chose IKK1 as the target gene. Our results indicate that constructs containing the H1 promoter are sensitive to the presence of prokaryotic sequences and to transgene position effects, similar to RNApol II promoters-driven constructs. We observed variable expression levels of transgenic siRNA among different tissues and animals and a reduction of up to 80% in IKK1 expression. Furthermore, IKK1 knockdown led to hair follicle alterations. In summary, we show that constructs directed by the H1 promoter can be used for knockdown of genes of interest in different organs and for the generation of animal models complementary to knockout and overexpression models. PMID:24523631

Moreno-Maldonado, Rodolfo; Murillas, Rodolfo; Navarro, Manuel; Page, Angustias; Suarez-Cabrera, Cristian; Alameda, Josefa P; Bravo, Ana; Casanova, M Llanos; Ramirez, Angel

2014-01-01

338

[Determination of nutrient elements in transgenic insect-resistant cotton tissues by three different spectroscopical methods].  

Science.gov (United States)

In order to find out the effects of exogenous genes, such as Bt and Bt coupled with CpTI, on nutrition metabolism in transgenic plants, totally eleven types of nutrient elements in transgenic Bt (Z30) and Bt-CpTI (CCRI41 and SGK321) cotton were determined using methods of flame atomic absorption spectroscopy, flame atomic emission spectroscopy and spectrophotometry at flowering stage and boll-opening stage. The results showed that the chemical composition of plant nutrition in transgenic insect-resistant cotton differed in comparison with non-transgenic cotton counterparts related to varieties, tissues and stages. The content of total N in transgenic cotton changed most significantly. Especially, it increased by 21% for transgenic Bt cotton Z30 compared to non-transgenic cotton Z16. These changes in total N content were probably caused by both transgenes expression in transgenic cotton and other processes not studied in this experiment. The content of Mg, Na and Cu in transgenic cotton varied significantly only in some certain varieties or tissues. It was unobvious how the incorporation of transgenes impacted on the content of organic C, total P, total S, K, Ca, Fe and Zn in transgenic cotton. The authors speculated that there were no significant changes in utilization and accumulation of these nutrient elements between transgenic insect-resistant cotton and their non-transgenic cotton counterparts (Z16, CCRI23 and SY321, respectively). PMID:20101981

Sun, Cai-Xia; Zhang, Yu-Lan; Sun, Yu-Quan; Yang, Lei; Wang, Jie; Cui, Zhen-Bo

2009-11-01

339

In utero recombinant adeno-associated virus gene transfer in mice, rats, and primates  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background Gene transfer into the amniotic fluid using recombinant adenovirus vectors was shown previously to result in high efficiency transfer of transgenes into the lungs and intestines. Adenovirus mediated in utero gene therapy, however, resulted in expression of the transgene for less than 30 days. Recombinant adenovirus associated viruses (rAAV have the advantage of maintaining the viral genome in daughter cells thus providing for long-term expression of transgenes. Methods Recombinant AAV2 carrying green fluorescent protein (GFP was introduced into the amniotic sac of fetal rodents and nonhuman primates. Transgene maintenance and expression was monitor. Results Gene transfer resulted in rapid uptake and long-term gene expression in mice, rats, and non-human primates. Expression and secretion of the reporter gene, GFP, was readily demonstrated within 72 hours post-therapy. In long-term studies in rats and nonhuman primates, maintenance of GFP DNA, protein expression, and reporter gene secretion was documented for over one year. Conclusions Because only multipotential stem cells are present at the time of therapy, these data demonstrated that in utero gene transfer with AAV2 into stem cells resulted in long-term systemic expression of active transgene roducts. Thus, in utero gene transfer via the amniotic fluid may be useful in treatment of gene disorders.

Marrero Luis

2003-09-01

340

Containment and competition: transgenic animals in the One Health agenda.  

Science.gov (United States)

The development of the One World, One Health agenda coincides in time with the appearance of a different model for the management of human-animal relations: the genetic manipulation of animal species in order to curtail their ability as carriers of human pathogens. In this paper we examine two examples of this emergent transgenic approach to disease control: the development of transgenic chickens incapable of shedding avian flu viruses, and the creation of transgenic mosquitoes refractory to dengue or malaria infection. Our analysis elaborates three distinctions between the One World, One Health agenda and its transgenic counterpoint. The first concerns the conceptualization of outbreaks and the forms of surveillance that support disease control efforts. The second addresses the nature of the interspecies interface, and the relative role of humans and animals in preventing pathogen transmission. The third axis of comparison considers the proprietary dimensions of transgenic animals and their implications for the assumed public health ethos of One Health programs. We argue that the fundamental difference between these two approaches to infectious disease control can be summarized as one between strategies of containment and strategies of competition. While One World, One Health programs seek to establish an equilibrium in the human-animal interface in order to contain the circulation of pathogens across species, transgenic strategies deliberately trigger a new ecological dynamic by introducing novel animal varieties designed to out-compete pathogen-carrying hosts and vectors. In other words, while One World, One Health policies focus on introducing measures of inter-species containment, transgenic approaches derive their prophylactic benefit from provoking new cycles of intra-species competition between GM animals and their wild-type counterparts. The coexistence of these divergent health protection strategies, we suggest, helps to elucidate enduring tensions and concerns about how humans should relate to, appraise, and intervene on animals and their habitats. PMID:24961736

Lezaun, Javier; Porter, Natalie

2015-03-01

341

Transgenic Vegetable Breeding for Nutritional Quality and Health Benefits  

Directory of Open Access Journals (Sweden)

Full Text Available Vegetables are essential for well-balanced diets. About 3 billion people in the world are malnourished due to imbalanced diets. Vegetables can contribute to the prevention of malnutrition disorders. Genetic engineering enables vegetable breeders to incorporate desired transgenes into elite cultivars, thereby improving their value considerably. It further offers unique opportunities for improving nutritional quality and bringing other health benefits. Many vegetable crops have been genetically modified to improve traits such as higher nutritional status or better flavour, and to reduce bitterness or anti-nutritional factors. Transgenic vegetables can be also used for vaccine delivery. Consumers could benefit further from eating more nutritious transgenic vegetables, e.g. an increase of crop carotenoids by metabolic sink manipulation through genetic engineering appears feasible in some vegetables. Genetically engineering carrots containing increase Ca levels may boost Ca uptake, thereby reducing the incidence of Ca deficiencies such as osteoporosis. Fortified transgenic lettuce with zinc will overcome the deficiency of this micronutrient that severely impairs organ function. Folates deficiency, which is regarded as a global health problem, can also be overcomed with transgenic tomatoes with folate levels that provide a complete adult daily requirement. Transgenic lettuce with improved tocopherol and resveratrol composition may prevent coronary disease and arteriosclerosis and can contribute to cancer chemopreventative activity. Food safety and health benefits can also be enhanced through transgenic approaches, e.g. rural African resource-poor consumers will benefit eating cyanide-free cultivars of cassava. Biotechnology-derived vegetable crops will succed if clear advantages and safety are demonstrated to both growers and consumers.

João Silva Dias

2012-09-01

342

Loss of Responses to Visual But Not Electrical Stimulation in Ganglion Cells of Rats With Severe Photoreceptor Degeneration  

OpenAIRE

Retinal implants are intended to help patients with degenerative conditions by electrically stimulating surviving cells to produce artificial vision. However, little is known about how individual retinal ganglion cells respond to direct electrical stimulation in degenerating retina. Here we used a transgenic rat model to characterize ganglion cell responses to light and electrical stimulation during photoreceptor degeneration. Retinas from pigmented P23H-1 rats were compared with wild-type re...

Sekirnjak, Chris; Hulse, Clare; Jepson, Lauren H.; Hottowy, Pawel; Sher, Alexander; Dabrowski, Wladyslaw; Litke, A. M.; Chichilnisky, E. J.

2009-01-01

343

Glyoxalase-1 overexpression reduces endothelial dysfunction and attenuates early renal impairment in a rat model of diabetes  

DEFF Research Database (Denmark)

In diabetes, advanced glycation end-products (AGEs) and the AGE precursor methylglyoxal (MGO) are associated with endothelial dysfunction and the development of microvascular complications. In this study we used a rat model of diabetes, in which rats transgenically overexpressed the MGO-detoxifying enzyme glyoxalase-I (GLO-I), to determine the impact of intracellular glycation on vascular function and the development of early renal changes in diabetes.

Brouwers, Olaf; Niessen, Petra M G

2014-01-01

344

CENTRAL ANGIOTENSIN-(1-7) IMPROVES VAGAL FUNCTION INDEPENDENT OF BLOOD PRESSURE IN HYPERTENSIVE (MREN2)27 RATS  

OpenAIRE

Hypertensive transgenic (mRen2)27 rats with overexpression of the mRen2 gene have impaired baroreflex sensitivity (BRS) for heart rate control and high NADPH oxidase and kinase-to-phosphatase signaling activity in medullary tissue compared to normotensive Hannover Sprague-Dawley control rats. They also exhibit insulin resistance at a young age. To determine whether blocking angiotensin (Ang) II actions, supplementing Ang-(1-7) or scavenging reactive oxygen species (ROS) in brain differentiall...

Nautiyal, Manisha; Shaltout, Hossam A.; Lima, Daniel C.; Donascimento, Kenia; Chappell, Mark C.; Diz, Debra I.

2012-01-01

345

Transgênicos sem maniqueísmo Transgenics without manichaeism  

Directory of Open Access Journals (Sweden)

Full Text Available Vivemos em uma época marcada pela hegemonia da ciência e da tecnologia, carregada de questões à espera de respostas, para que o futuro da humanidade seja alcançado de forma segura e sustentável. O desenvolvimento de processos agroindustriais - especificamente, a produção de alimentos - com tecnologia de DNA recombinante tem trazido perspectivas de bons lucros apenas para os grandes conglomerados da biotecnologia e para produtores rurais com alto grau de desenvolvimento tecnológico. Discordamos de uma moratória para a tecnologia do DNA recombinante. Além disso, afirmações de ausência de risco podem levar a conclusões precipitadas, pois pouquíssimos testes foram realizados até hoje. É premente que se estabeleça uma política nacional de biossegurança, que envolva a sociedade civil organizada e todos os órgãos de governo (federal e estaduais responsáveis pela fiscalização, e que se implantem um programa de biovigilância e um código de ética de manipulações genéticas.We live in an era characterized by the hegemony of science and technology, an era fraught with questions awaiting answers which would enable a safe and sustainable future for humankind. The development of agro-industrial processes - food products in particular - through recombinant DNA technology has enhanced the profit prospects of the few big biotechnology companies and of large-scale farmers who have access to the latest technological developments. We thus oppose a moratorium on recombinant DNA technology. Moreover, hasty statements about risk-free transgenics may be misleading in the absence of extensive safety tests. There is a pressing need for the establishment of a biosafety policy in this country involving the organized civil society and every government agency responsible for monitoring such matters. There is also the need to put in place a bio-surveillance and a code of ethics regarding genetic manipulation.

Silvio Valle

2000-10-01

346

Transgênicos sem maniqueísmo / Transgenics without manichaeism  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: Portuguese Abstract in portuguese Vivemos em uma época marcada pela hegemonia da ciência e da tecnologia, carregada de questões à espera de respostas, para que o futuro da humanidade seja alcançado de forma segura e sustentável. O desenvolvimento de processos agroindustriais - especificamente, a produção de alimentos - com tecnologi [...] a de DNA recombinante tem trazido perspectivas de bons lucros apenas para os grandes conglomerados da biotecnologia e para produtores rurais com alto grau de desenvolvimento tecnológico. Discordamos de uma moratória para a tecnologia do DNA recombinante. Além disso, afirmações de ausência de risco podem levar a conclusões precipitadas, pois pouquíssimos testes foram realizados até hoje. É premente que se estabeleça uma política nacional de biossegurança, que envolva a sociedade civil organizada e todos os órgãos de governo (federal e estaduais) responsáveis pela fiscalização, e que se implantem um programa de biovigilância e um código de ética de manipulações genéticas. Abstract in english We live in an era characterized by the hegemony of science and technology, an era fraught with questions awaiting answers which would enable a safe and sustainable future for humankind. The development of agro-industrial processes - food products in particular - through recombinant DNA technology ha [...] s enhanced the profit prospects of the few big biotechnology companies and of large-scale farmers who have access to the latest technological developments. We thus oppose a moratorium on recombinant DNA technology. Moreover, hasty statements about risk-free transgenics may be misleading in the absence of extensive safety tests. There is a pressing need for the establishment of a biosafety policy in this country involving the organized civil society and every government agency responsible for monitoring such matters. There is also the need to put in place a bio-surveillance and a code of ethics regarding genetic manipulation.

Silvio, Valle.

2000-10-01

347

Development of transgenic watermelon resistant to Cucumber mosaic virus and Watermelon mosaic virus by using a single chimeric transgene construct.  

Science.gov (United States)

Watermelon, an important fruit crop worldwide, is prone to attack by several viruses that often results in destructive yield loss. To develop a transgenic watermelon resistant to multiple virus infection, a single chimeric transgene comprising a silencer DNA from the partial N gene of Watermelon silver mottle virus (WSMoV) fused to the partial coat protein (CP) gene sequences of Cucumber mosaic virus (CMV), Cucumber green mottle mosaic virus (CGMMV) and Watermelon mosaic virus (WMV) was constructed and transformed into watermelon (cv. Feeling) via Agrobacterium-mediated transformation. Single or multiple transgene copies randomly inserted into various locations in the genome were confirmed by Southern blot analysis. Transgenic watermelon R(0) plants were individually challenged with CMV, CGMMV or WMV, or with a mixture of these three viruses for resistance evaluation. Two lines were identified to exhibit resistance to CMV, CGMMV, WMV individually, and a mixed inoculation of the three viruses. The R(1) progeny of the two resistant R(0) lines showed resistance to CMV and WMV, but not to CGMMV. Low level accumulation of transgene transcripts in resistant plants and small interfering (si) RNAs specific to CMV and WMV were readily detected in the resistant R(1) plants by northern blot analysis, indicating that the resistance was established via RNA-mediated post-transcriptional gene silencing (PTGS). Loss of the CGMMV CP-transgene fragment in R1 progeny might be the reason for the failure to resistant CGMMV infection, as shown by the absence of a hybridization signal and no detectable siRNA specific to CGMMV in Southern and northern blot analyses. In summary, this study demonstrated that fusion of different viral CP gene fragments in transgenic watermelon contributed to multiple virus resistance via PTGS. The construct and resistant watermelon lines developed in this study could be used in a watermelon breeding program for resistance to multiple viruses. PMID:22203520

Lin, Ching-Yi; Ku, Hsin-Mei; Chiang, Yi-Hua; Ho, Hsiu-Yin; Yu, Tsong-Ann; Jan, Fuh-Jyh

2012-10-01

348

Transgenic resistance to Citrus tristeza virus in grapefruit.  

Science.gov (United States)

Grapefruit (Citrus paradisi) transgenic plants transformed with a variety of constructs derived from the Citrus tristeza virus (CTV) genome were tested for their resistance to the virus. Most transgenic lines were susceptible (27 lines), a few were partially resistant (6 lines) and only one line, transformed with the 3' end of CTV was resistant. Transgene expression levels and siRNA accumulation were determined to identify whether the resistance observed was RNA-mediated. The responses were varied. At least one resistant plant from a partially resistant line showed no steady-state transgene mRNA, siRNA accumulation and no viral RNA, implicating posttranscriptional gene silencing (PTGS) as the mechanism of resistance. The most resistant line showed no transgene mRNA accumulation and promoter methylation of cytosines in all contexts, the hallmark of RNA-directed DNA methylation and transcriptional gene silencing (TGS). The variety of responses, even among clonally propagated plants, is unexplained but is not unique to citrus. The genetics of CTV, host response or other factors may be responsible for this variability. PMID:17882423

Febres, Vicente J; Lee, Richard F; Moore, Gloria A

2008-01-01

349

A selectively terminable transgenic rice line expressing human lactoferrin.  

Science.gov (United States)

Human lactoferrin (hLF) is a multifunctional milk protein which could be utilized for promoting human health. Transgenic rice has been used as a bioreactor for mass production of recombinant hLF. However, one major concern over such transgenic rice is the risk of its unintended spreading into environment and into our food supplies. Here we report the development of selectively terminable transgenic rice expressing human lactoferrin in seeds. These transgenic rice plants could be selectively terminated by bentazon, a common herbicide used for rice weed control. The hLF expression cassette was constructed into a T-DNA containing the RNA interference cassette suppressing the expression of the rice gene CYP81A6 which detoxifies herbicide bentazon, and the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) cassette which confers to glyphosate tolerance. A transgenic line, named as G281, was identified for its high sensitivity to bentazon, high tolerance to glyphosate, and high expression of hLF. Southern analysis suggested G281 is a single copy insertion event. Field tests demonstrated that G281 plants can be completely killed by a single spray of bentazon at 1000 mg/L, which is safe to regular rice and represents only half of the dose recommended by manufacturer for rice field weed control. Therefore, any G281 contaminations in regular rice could be selectively terminated to make sure it will not enter food or feed supplies. PMID:20433928

Lin, Chaoyang; Nie, Peng; Lu, Wei; Zhang, Qing; Li, Jing; Shen, Zhicheng

2010-11-01

350

Efficient soluble protein production on transgenic silkworms expressing cytoplasmic chaperones.  

Science.gov (United States)

Baculovirus expression systems (BES) are widely used for recombinant protein production in lepidopteran cells or larvae. However, even in BES, the insolubility of recombinant proteins sometimes makes their expression difficult. In this study, to improve the solubility and yield of foreign proteins, we constructed transgenic silkworms using silkworm heat-shock proteins, Hsp70 and Hsp40, or Hsc70 and Hsp90 co-chaperone Hop. In these transgenic silkworms, the expression levels of the transgenes were under the control of a UAS.hsp mini-promoter driven by a Gal4NFkBp65 activator. When the transgenic silkworm with HSP70 and 40 (TGS-HSP70/40) was infected with BmNPV carrying mC3d and Gal4NFkBp65 under the control of baculovirus polyhedrin or p10 promoters, respectively, the soluble fraction of the His- or His.GST-tagged mC3d increased significantly. Similarly, the transgenic silkworm with HSC70 and HOP (TGS-HOP7) was effective for the expression of a steroid hormone receptor, USP2. In conclusion, the His-tagged baculovirus expression system featuring the chaperone effect TGS-HSP70/40 and TGS-HOP7 silkworms is effective for increasing the yields of soluble and functional foreign gene products. PMID:20496148

Hong, Sun Mee; Yamashita, Jun; Mitsunobu, Hitoshi; Uchino, Keiro; Kobayashi, Isao; Sezutsu, Hideki; Tamura, Toshiki; Nakajima, Hideki; Miyagawa, Yoshitaka; Lee, Jae Man; Mon, Hiroaki; Miyata, Yoshihiko; Kawaguchi, Yutaka; Kusakabe, Takahiro

2010-08-01

351

Neutralizing antibodies against rotavirus produced in transgenically labelled purple tomatoes.  

Science.gov (United States)

Edible fruits are inexpensive biofactories for human health-promoting molecules that can be ingested as crude extracts or partially purified formulations. We show here the production of a model human antibody for passive protection against the enteric pathogen rotavirus in transgenically labelled tomato fruits. Transgenic tomato plants expressing a recombinant human immunoglobulin A (hIgA_2A1) selected against the VP8* peptide of rotavirus SA11 strain were obtained. The amount of hIgA_2A1 protein reached 3.6 ± 0.8% of the total soluble protein in the fruit of the transformed plants. Minimally processed fruit-derived products suitable for oral intake showed anti-VP8* binding activity and strongly inhibited virus infection in an in vitro virus neutralization assay. In order to make tomatoes expressing hIgA_2A1 easily distinguishable from wild-type tomatoes, lines expressing hIgA_2A1 transgenes were sexually crossed with a transgenic tomato line expressing the genes encoding Antirrhinum majus Rosea1 and Delila transcription factors, which confer purple colour to the fruit. Consequently, transgenically labelled purple tomato fruits expressing hIgA_2A1 have been developed. The resulting purple-coloured extracts from these fruits contain high levels of recombinant anti-rotavirus neutralizing human IgA in combination with increased amounts of health-promoting anthocyanins. PMID:22070155

Juárez, Paloma; Presa, Silvia; Espí, Joaquín; Pineda, Benito; Antón, María T; Moreno, Vicente; Buesa, Javier; Granell, Antonio; Orzaez, Diego

2012-04-01

352

Design rules for efficient transgene expression in plants.  

Science.gov (United States)

Sustained expression of transgenes in specified developmental patterns is commonly needed in plant biotechnology, but obstructed by transgene silencing. Here, we present a set of gene design rules, tested on the silencing-susceptible beetle luc and bacterial ims genes, expressed in sugarcane. Designs tested independently or in combination included removal of rare codons, removal of RNA instability sequences, blocking of likely endogenous sRNA binding sites and randomization of non-rare codons. Stable transgene expression analyses, on multiple independent lines per construct, showed greatest improvement from the removal of RNA instability sequences, accompanied by greatly reduced transcript degradation evident in northern blot analysis. We provide a set of motifs that readily can be eliminated concurrently with rare codons and undesired structural features such as repeat sequences, using Gene Designer 2.0 software. These design rules yielded 935- and 5-fold increased expression in transgenic callus, relative to the native luc and ims sequences; and gave sustained expression under the control of sugarcane and heterologous promoters over several years in greenhouse and field trials. The rules can be applied easily with codon usage tables from any plant species, providing a simple and effective means to achieve sustained expression of otherwise silencing-prone transgenes in plants. PMID:24854834

Jackson, Mark A; Sternes, Peter R; Mudge, Stephen R; Graham, Michael W; Birch, Robert G

2014-09-01

353

Handmade cloned transgenic sheep rich in omega-3 Fatty acids.  

Science.gov (United States)

Technology of somatic cell nuclear transfer (SCNT) has been adapted worldwide to generate transgenic animals, although the traditional procedure relies largely on instrumental micromanipulation. In this study, we used the modified handmade cloning (HMC) established in cattle and pig to produce transgenic sheep with elevated levels of omega-3 (n-3) fatty acids. Codon-optimized nematode mfat-1 was inserted into a eukaryotic expression vector and was transferred into the genome of primary ovine fibroblast cells from a male Chinese merino sheep. Reverse transcriptase PCR, gas chromatography, and chromosome analyses were performed to select nuclear donor cells capable of converting omega-6 (n-6) into n-3 fatty acids. Blastocysts developed after 7 days of in vitro culture were surgically transplanted into the uterus of female ovine recipients of a local sheep breed in Xinjiang. For the HMC, approximately 8.9% (n ?=925) of reconstructed embryos developed to the blastocyst stage. Four recipients became pregnant after 53 blastocysts were transplanted into 29 naturally cycling females, and a total of 3 live transgenic lambs were produced. Detailed analyses on one of the transgenic lambs revealed a single integration of the modified nematode mfat-1 gene at sheep chromosome 5. The transgenic sheep expressed functional n-3 fatty acid desaturase, accompanied by more than 2-folds reduction of n-6/n-3 ratio in the muscle (pmethod, HMC showed an equivalent efficiency but proved cheaper and easier in operation. PMID:23437077

Zhang, Peng; Liu, Peng; Dou, Hongwei; Chen, Lei; Chen, Longxin; Lin, Lin; Tan, Pingping; Vajta, Gabor; Gao, Jianfeng; Du, Yutao; Ma, Runlin Z

2013-01-01

354

Reversible methylation and silencing of homologous transgenes in tobacco plants  

International Nuclear Information System (INIS)

Homology dependent gene silencing in transgenic plants occurs frequently when multiple copies of a transgene or a transgene with homology to an endogenous plant gene are present in a plant nucleus. The multiple copies can be arranged in cis on the same DNA molecule, or they can be present on different DNA molecules, either in allelic or non-allelic locations (trans inactivation). Although the phenomenon of silencing is well established, the mechanism by which it occurs are not completely understood. At present, different silencing systems can be distinguished by three major features: (1) the region of homology involved in the interaction (promoter or protein coding region); (2) the level at which silencing occurs (transcriptional or post-transcriptional); and (3) the degree of meiotic heritability of the silenced/methylated states after segregation of two interacting homologous loci in progeny. Interactions that lead to the silencing and methylation of partially homologous transgenes in tobacco have been studied. The transgenes share homology only in promoter regions; both the nopaline synthase promoter and the 35S promoter of the cauliflower mosaic virus have been used to drive the expression of various selectable and biochemical marker genes. The properties of these silencing systems are discussed, with particular reference to the features mentioned above. (author). 13 refs

355

Expression and inheritance of multiple transgenes in rice plants.  

Science.gov (United States)

The ability to control integration, inheritance, and expression of multiple transgenes is a prerequisite for manipulating biosynthetic pathways and complex agronomic characteristics in plants. One hundred and twenty-five independent transgenic rice plants were regenerated after cobombarding embryogenic tissues with a mixture of 14 different pUC-based plasmids. Eighty-five percent of the R0 plants contained more than two, and 17% more than nine, of the target genes. Plants containing multiple transgenes displayed normal morphologies and 63% set viable seed. Multigene cotransformation efficiency was correlated with the ratio in which the plasmids were mixed with respect to the selectable marker. All target genes had an equal chance of integration, indicating that the nature of the coding region had no effect on the efficiency of integration. Three plant lines containing 11, 10, and 9 transgenes, respectively, were analyzed for patterns of integration and inheritance until the R3 generation. Integration of multiple transgenes occurred at either one or two genetic loci, with inheritance conforming to a 3:1 Mendelian ratio. Coexpression of four marker genes was investigated until the R2 generation. PMID:9831036

Chen, L; Marmey, P; Taylor, N J; Brizard, J P; Espinoza, C; D'Cruz, P; Huet, H; Zhang, S; de Kochko, A; Beachy, R N; Fauquet, C M

1998-11-01

356

Handmade cloned transgenic piglets expressing the nematode fat-1 gene  

DEFF Research Database (Denmark)

Abstract Production of transgenic animals via somatic cell nuclear transfer (SCNT) has been adapted worldwide, but this application is somewhat limited by its relatively low efficiency. In this study, we used handmade cloning (HMC) established previously to produce transgenic pigs that express the functional nematode fat-1 gene. Codon-optimized mfat-1 was inserted into eukaryotic expression vectors, which were transferred into primary swine donor cells. Reverse transcriptase PCR (RT-PCR), gas chromatography, and chromosome analyses were performed to select donor clones capable of converting n-6 into n-3 fatty acids. Blastocysts derived from the clones that lowered the n-6/n-3 ratio to approximately 1:1 were transferred surgically into the uteri of recipients for transgenic piglets. By HMC, 37% (n=558) of reconstructed embryos developed to the blastocyst stage after 7 days of culture in vitro, with an average cell number of 81±36 (n=14). Three recipients became pregnant after 408 day-6 blastocysts were transferred into four naturally cycling females, and a total of 14 live offspring were produced. The nematode mfat-1 effectively lowered the n-6/n-3 ratio in muscle and major organs of the transgenic pig. Our results will help to establish a reliable procedure and an efficient option in the production of transgenic animals.

Zhang, Peng; Zhang, Yidi

2012-01-01

357

Glycinebetaine synthesizing transgenic potato plants exhibit enhanced tolerance to salt and cold stresses  

International Nuclear Information System (INIS)

Abiotic stresses are the most important contributors towards low productivity of major food crops. Various attempts have been made to enhance abiotic stress tolerance of crop plants by classical breeding and genetic transformation. Genetic transformation with glycinebetaine (GB) synthesizing enzymes' gene(s) in naturally non accumulating plants has resulted in enhanced tolerance against variety of abiotic stresses. Present study was aimed to evaluate the performance of GB synthesizing transgenic potato plants against salt and cold stresses. Transgenic potato plants were challenged against salt and cold stresses at whole plant level. Transgenic lines were characterized to determine the transgene copy number. Different parameters like integrity, chlorophyll contents, tuber yield and vegetative biomass were studied to monitor the stress tolerance of transgenic potato plants. The results were compared with Non-transgenic (NT) plants and statistically analyzed to evaluate significant differences. Multi-copy insertion of expression cassette was found in both transgenic lines. Upon salt stress, transgenic plants maintained better growth as compared to NT plants. The tuber yield of transgenic plants was significantly greater than NT plants in salt stress. Transgenic plants showed improved membrane integrity against cold stress by depicting appreciably reduced ion leakage as compared to NT plants. Moreover, transgenic plants showed significantly less chlorophyll bleaching than NT plants upon cold stress. In addition, NT plants accumulated significantly less biomass, and yielded fewer tubers as compared to transgenic plants after cold stress treatment. The study will be a committed step for field evaluation of transgenic plants with the aim of commercialization. (author)

358

Nucleus-targeted Dmp1 transgene fails to rescue dental defects in Dmp1 null mice.  

Science.gov (United States)

Dentin matrix protein 1 (DMP1) is essential to odontogenesis. Its mutations in human subjects lead to dental problems such as dental deformities, hypomineralization and periodontal impairment. Primarily, DMP1 is considered as an extracellular matrix protein that promotes hydroxyapatite formation and activates intracellular signaling pathway via interacting with ?v?3 integrin. Recent in vitro studies suggested that DMP1 might also act as a transcription factor. In this study, we examined whether full-length DMP1 could function as a transcription factor in the nucleus and regulate odontogenesis in vivo. We first demonstrated that a patient with the DMP1 M1V mutation, which presumably causes a loss of the secretory DMP1 but does not affect the nuclear translocation of DMP1, shows a typical rachitic tooth defect. Furthermore, we generated transgenic mice expressing (NLS)DMP1, in which the endoplasmic reticulum (ER) entry signal sequence of DMP1 was replaced by a nuclear localization signal (NLS) sequence, under the control of a 3.6?kb rat type I collagen promoter plus a 1.6?kb intron 1. We then crossbred the (NLS)DMP1 transgenic mice with Dmp1 null mice to express the (NLS)DMP1 in Dmp1-deficient genetic background. Although immunohistochemistry demonstrated that (NLS)DMP1 was localized in the nuclei of the preodontoblasts and odontoblasts, the histological, morphological and biochemical analyses showed that it failed to rescue the dental and periodontal defects as well as the delayed tooth eruption in Dmp1 null mice. These data suggest that the full-length DMP1 plays no apparent role in the nucleus during odontogenesis. PMID:25105818

Lin, Shu-Xian; Zhang, Qi; Zhang, Hua; Yan, Kevin; Ward, Leanne; Lu, Yong-Bo; Feng, Jian-Quan

2014-09-01

359

Establishment of a novel, eco-friendly transgenic pig model using porcine pancreatic amylase promoter-driven fungal cellulase transgenes.  

Science.gov (United States)

Competition between humans and livestock for cereal and legume grains makes it challenging to provide economical feeds to livestock animals. Recent increases in corn and soybean prices have had a significant impact on the cost of feed for pig producers. The utilization of byproducts and alternative ingredients in pig diets has the potential to reduce feed costs. Moreover, unlike ruminants, pigs have limited ability to utilize diets with high fiber content because they lack endogenous enzymes capable of breaking down nonstarch polysaccharides into simple sugars. Here, we investigated the feasibility of a transgenic strategy in which expression of the fungal cellulase transgene was driven by the porcine pancreatic amylase promoter in pigs. A 2,488 bp 5'-flanking region of the porcine pancreatic amylase gene was cloned by the genomic walking technique, and its structural features were characterized. Using GFP as a reporter, we found that this region contained promoter activity and had the potential to control heterologous gene expression. Transgenic pigs were generated by pronuclear microinjection. Founders and offspring were identified by PCR and Southern blot analyses. Cellulase mRNA and protein showed tissue-specific expression in the pancreas of F1 generation pigs. Cellulolytic enzyme activity was also identified in the pancreas of transgenic pigs. These results demonstrated the establishment of a tissue-specific promoter of the porcine pancreatic amylase gene. Transgenic pigs expressing exogenous cellulase may represent a way to increase the intake of low-cost, fiber-rich feeds. PMID:25063310

Lin, Y S; Yang, C C; Hsu, C C; Hsu, J T; Wu, S C; Lin, C J; Cheng, W T K

2015-02-01

360

Proteomic analysis of plasma from a Tau transgenic mouse.  

Science.gov (United States)

The neurofibrillary tangles (NFTs) formed by the accumulation of abnormal tau filaments have been shown to be involved in Alzheimer's disease (AD) brain degeneration. In this study, a tau transgenic mouse (pNSE/htau23) model was used to monitor changes in protein levels and to search for novel biomarker candidates suitable for the early diagnosis of AD before onset of clinical symptoms. Plasma samples from 2-month (n=13, asymptomatic) and 4-month (n=7, symptomatic) tau transgenic mice were compared to the control group (n=8) by 2-dimensional gel electrophoresis (2-DE) coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS). Three proteins, ATP synthase, Adenosine kinase and Regucalcin showed significantly decreased levels in the plasma of tau transgenic mouse, which was further confirmed by Western blotting. This study suggests that these proteins could be used as candidate biomarkers for early diagnosis of AD in combination with previously discovered protein biomarkers. PMID:22406198

Kim, Yoon-Ha; Lee, Eun-Kyung; Park, Seung-Ah; Kim, Nam-Hee; Kim, Chan-Wha

2012-06-01

361

Establishment and detection of HBV transgenic mice with YMDD mutation  

Directory of Open Access Journals (Sweden)

Full Text Available Objective To establish the hepatitis B virus(HBV transgenic mice with YMDD mutation,and provide an animal model for research of HBV prevention and therapeutic approach.Methods 1.3 copies HBV genome containing YMDD mutation associated with lamivudine resistance was injected into the zygote of FVB/N female mice by microinjection.Integration and passage of exogenous gene in transgenic mice was confirmed by PCR.The expression of HBsAg in liver and kidney tissues in transgenic mice was identified by ELISA and immunohistochemistry.Results A total of 3401 zygotes were injected and 269 F0 pups were born.PCR analysis indicated that 33 out of 269 pups were positive,and the integrating rate of exogenous gene was 12.3% in F0.Fluorescent quantitative PCR showed that HBV DNA was weakly positive in serum samples in 9 transgenic mice,less than 103copies/ml.The expression of HBsAg in transgenic mice was observed in liver and kidney tissues by immunohistochemistry,and it was higher in kidney than in liver.The target gene was detected by PCR in 27.6% of 47 F1 offsprings.The expression of HBsAg could be observed in liver and kidney tissues in F1,which was similar to that in F0.Conclusion 1.3 copies HBV transgenic mice model containing YMDD mutation associated with lamivudine resistance is successfully produced by microinjection,and HBsAg expression can be transmitted through germline cells.

Yu-qin YOU

2011-09-01

362

A Wt1-Dmrt1 Transgene Restores DMRT1 to Sertoli Cells of Dmrt1?/? Testes: A Novel Model of DMRT1-Deficient Germ Cells1  

Science.gov (United States)

ABSTRACT DMRT1 is an evolutionarily conserved transcriptional factor expressed only in the postnatal testis, where it is produced in Sertoli cells and germ cells. While deletion of Dmrt1 in mice demonstrated it is required for postnatal testis development and fertility, much is still unknown about its temporal- and cell-specific functions. This study characterized a novel mouse model of DMRT1-deficient germ cells that was generated by breeding Dmrt1-null (Dmrt1?/?) mice with Wt1-Dmrt1 transgenic (Dmrt1+/?;tg) mice, which express a rat Dmrt1 cDNA in gonadal supporting cells by directing it from the Wilms tumor 1 locus in a yeast artificial chromosome transgene. Like Dmrt1?/? mice, male Dmrt1?/? transgenic mice (Dmrt1?/?;tg) were infertile, while female mice were fertile. Immunohistochemistry and Western blot analysis showed transgenic DMRT1 expressed in supporting cells of the newborn gonads of both sex and in Sertoli cells of the testis afterbirth. Sertoli cells were evaluated by electron microscopy, revealing that maturation of Dmrt1?/?;tg Sertoli cells was incomplete. Morphological analysis of testes from 42-day-old mice showed that, compared to Dmrt1?/? mice, Dmrt1?/?;tg mice have improved seminiferous tubule structure, with lumens present in many. Immunohistochemistry of the polarity markers ESPIN and NECTIN-2 showed that DMRT1 in Sertoli cells is required for NECTIN-2 expression and influences organization of ectoplasmic specializations. Further functional analyses of the transgene on a Dmrt1?/? background showed that it did not rescue the decrease in Dmrt1?/? testis size, but when expressed on a wild-type background, exogenous DMRT1 prevented the normal age-related decline in testis size and enhanced sperm progressive motility. The studies suggest that DMRT1 in Sertoli cells regulates tubule morphology, spermatogenesis, and sperm function via its effects on Sertoli cell maturation and polarity. Furthermore, expression and function of transgenic DMRT1 in Sertoli cells establishes a novel mouse model of DMRT1-deficient germ cells generated by breeding Dmrt1-null mice with Wt1-Dmrt1 transgenic mice (rescue; Dmrt1?/?;tg). PMID:23255335

Agbor, Valentine A.; Tao, Shixin; Lei, Ning; Heckert, Leslie L.

2012-01-01

363

A Site-Specific Recombinase-Based Method to Produce Antibiotic Selectable Marker Free Transgenic Cattle  

OpenAIRE

Antibiotic selectable marker genes have been widely used to generate transgenic animals. Once transgenic animals have been obtained, the selectable marker is no longer necessary but raises public concerns regarding biological safety. The aim of this study was to prepare competent antibiotic selectable marker free transgenic cells for somatic cell nuclear transfer (SCNT). PhiC31 intergrase was used to insert a transgene cassette into a “safe harbor” in the bovine genome. Then, Cre recombin...

Yu, Yuan; Wang, Yongsheng; Tong, Qi; Liu, Xu; Su, Feng; Quan, Fusheng; Guo, Zekun; Zhang, Yong

2013-01-01

364

ELITE TRANSGENIC LINES OF BASMATI-370 REVEALED HIGH LEVEL OF LODGING RESISTANCE UNDER FIELD CONDITIONS  

OpenAIRE

Lodging decreases crop yield, photosynthesis and grain quality. Three transgenic lines of Basmati-370 containing two Bt genes (cry1Ac & cry2A) were evaluated in the field along with non-transgenic parent as control. The experiment was repeated for consecutive two years. Transgenic rice revealed tremendous morphological variations associated with lodging such as short stature, more number of nodes, less internodal length, etc. The transgenic plants were 33% short in stature while average numbe...

Mahmood-ur-Rahman; Noor Muhammad; Ahmad Ali Shahid, Tayyab Husnain

2012-01-01

365

Male mating strategy and the introgression of a growth hormone transgene  

OpenAIRE

Escaped transgenic organisms (GMO's) may threaten the populations of their wild relatives if able to hybridize with each other. The introgression of a growth enhancement transgene into a wild Atlantic salmon population may be affected by the transgene's effects not only on fitness parameters, but also on mating behaviour. Large anadromous GMO males are most preferred in mating, but a transgene can also give the large sneakers a reproductive advantage over the smaller wild individuals. With a ...

Valosaari, Kata-riina; Aikio, Sami; Kaitala, Veijo

2008-01-01

366

Tolerance induced by physiological levels of secreted proteins in transgenic mice expressing human insulin.  

OpenAIRE

We have used transgenic mice to study immune tolerance to autologous, non-MHC encoded proteins that are expressed at physiological levels in the circulation. The transgenic mice used in these studies express the human preproinsulin gene and synthesize human proinsulin. Human and mouse insulin are secreted from the pancreatic islets of transgenic mice in response to normal physiological stimuli, such as glucose. Our data demonstrate that the transgenic mice have acquired tolerance to human ins...

Whiteley, P. J.; Lake, J. P.; Selden, R. F.; Kapp, J. A.

1989-01-01

367

Detection of transgenic cp4 epsps genes in the soil food web  

OpenAIRE

The persistence and movement of transgenic DNA in agricultural and natural systems is largely unknown. This movement poses a threat of horizontal gene transfer and possible proliferation of genetically modified DNA into the general environment. To assess the persistence of transgenic DNA in a field of Roundup Ready® corn, we quantified the presence of the transgene for glyphosate tolerance within a soil food web. Using quantitative real-time PCR, we identified the cp4 epsps transgene in bulk...

Hart, Miranda M.; Powell, Jeff R.; Gulden, Robert H.; Levy-booth, David J.; Dunfield, Kari E.; Peter Pauls, K.; Swanton, Clarence J.; Klironomos, John N.; Trevors, Jack T.

2009-01-01

368

Transgene Repeat Arrays Interact with Distant Heterochromatin and Cause Silencing in Cis and Trans  

OpenAIRE

Tandem repeats of Drosophila transgenes can cause heterochromatic variegation for transgene expression in a copy-number and orientation-dependent manner. Here, we demonstrate different ways in which these transgene repeat arrays interact with other sequences at a distance, displaying properties identical to those of a naturally occurring block of interstitial heterochromatin. Arrays consisting of tandemly repeated white transgenes are strongly affected by proximity to constitutive heterochrom...

Dorer, D. R.; Henikoff, S.

1997-01-01

369

Biodiversity versus transgenic sugar beet: the one euro question  

OpenAIRE

The decision on whether to release transgenic crops in the EU is subject to irreversibility, uncertainty and flexibility. We analyse the case of herbicide-tolerant sugar beet and assess whether the EU’s 1998 de facto moratorium on transgenic crops for sugar beet was correct from a cost–benefit perspective, using a real option approach. We show that the decision was correct, providing households on average value the possible annual irreversible costs of herbicide-tolerant sugar beet at 1 o...

Demont, M.; Wesseler, J. H. H.; Tollens, E.

2004-01-01

370

A strategy to study tyrosinase transgenes in mouse melanocytes  

OpenAIRE

[Results] Here, we describe the efforts towards obtaining melanocyte cultures from our Tyr transgenic mice. We have bred our Tyr transgenic mice into Tyr c-32DSD mutant background, lacking the endogenous Tyr locus. In these conditions, we failed to obtain immortalised melanocytes. We decided to include the inactivation of the Ink4a-Arf locus to promote melanocyte immortalisation. For this purpose, we report the segregation of the Ink4a-Arf null allele from the brown (Tyrp1b) mutation in mice....

Lavado, Alfonso J.; Matheu, Ander; Serrano, Manuel; Montoliu, Llui?s

2005-01-01

371

Neutralizing antibodies against rotavirus produced in transgenically labelled purple tomatoes  

OpenAIRE

Edible fruits are inexpensive biofactories for human health-promoting molecules that can be ingested as crude extracts or partially purified formulations. We show here the production of a model human antibody for passive protection against the enteric pathogen rotavirus in transgenically labelled tomato fruits. Transgenic tomato plants expressing a recombinant human immunoglobulin A (hIgA_2A1) selected against the VP8* peptide of rotavirus SA11 strain were obtained. The amount of hIgA_2A1 pro...

Jua?rez, Paloma; Presa, Silvia; Espi?, Joaqui?n; Pineda, Benito; Anto?n, Mari?a T.; Moreno, Vicente; Buesa, Javier; Granell, Antonio; Orzaez, Diego

2012-01-01

372

Detecting adventitious transgenic events in a maize center of diversity  

Scientific Electronic Library Online (English)

Full Text Available SciELO Chile | Language: English Abstract in english Background: The genetic diversity of maize in Peru includes several landraces (within race clusters) and modern open pollinated and hybrid cultivars that are grown by farmers across various regions, thereby making this country a secondary center of diversity for this crop. A main topic of controvers [...] y in recent years refers to the unintended presence of transgenic events in locally grown cultivars at main centers of crop diversity. Peru does not yet have biosafety regulations to control or permit the growing of genetically modified crops. Hence, the aim of this research was to undertake a survey in the valley of Barranca, where there were recent claims of authorized transgenic maize grown in farmers fields as well as in samples taken from feed storage and grain or seed trade centers. Results: A total of 162 maize samples (134 from fields, 15 from local markets, eight from the collecting centers of poultry companies, from the local trading center and four samples from seed markets) were included for a qualitative detection by the polymerase chain reaction (PCR) of Cauliflower Mosaic Virus (CaMV) 35S promoter (P35S) and nopaline synthase terminator (Tnos) sequences, as well as for six transgenic events, namely BT11, NK603, T25, 176, TC1507 and MON810. The 134 maize samples from farmers fields were negative for Cry1Ab delta-endotoxin insecticidal protein and enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) using lateral flow strips. The PCR analysis did not detect any of the six transgenic events in samples from farmers fields, local markets, seed trading shops and the local collecting center. There were four transgenic events (T25, NK603, MON810 and TC1507) in grain samples from the barns of poultry companies. Conclusions: This research could not detect, at the 95% probability level, transgenes in farmers' fields in the valley of Barranca. The four transgenic events in grain samples from barns of poultry companies were not surprising because Peru imports maize, mainly for animal feed, from Argentina and the United States that are known for growing transgenic maize.

Luis Fernando Rimachi, Gamarra; Jorge Alcántara, Delgado; Yeny Aquino, Villasante; Rodomiro, Ortiz.

2011-07-15

373

[Generation of sugar beet transgenic plants expressing bar gene].  

Science.gov (United States)

The parameters of transformation using Agrobacterium tumefaciens EHA 105 for 5 domestic sorts and lines of sugar beet (Beta vulgaris L. var. saccharifera (Alef) Krass) were optimized. The system of transgenic tissue selection based on resistance to phosphinothricin, allowing to avoid the appearing of chimeric shoots among initial transformants was developed. The transgenic plants of sugar beet sorts Ramonskaya single seed 47, L'govskaya single seed 52 and RMS 73, and LBO 17 and LBO 19 lines expressing the gene of phosphinothricin acetyl transferase bar have been obtained. The resistance of these sorts and lines to the effect of phosphinothricin in vitro has been shown. PMID:20198924

Mishutkina, Ia V; Kamionskaia, A M; Skriabin, K G

2010-01-01

374

Neuron Loss in Transgenic Mouse Models of Alzheimer's Disease  

Directory of Open Access Journals (Sweden)

Full Text Available Since their initial generation in the mid 1990s, transgenic mouse models of Alzheimers's disease (AD have been proven to be valuable model systems which are indispensable for modern AD research. Whereas most of these models are characterized by extensive amyloid plaque pathology, inflammatory changes and often behavioral deficits, modeling of neuron loss was much less successful. The present paper discusses the current achievements of modeling neuron loss in transgenic mouse models based on APP/A? and Tau overexpression and provides an overview of currently available AD mouse models showing these pathological alterations.

Oliver Wirths

2010-01-01

375

[Cotton laccase gene overexpression in transgenic Populus alba var. pyramidalis and its effects on the lignin biosynthesis in transgenic plants].  

Science.gov (United States)

Using petioles as explants, a cotton laccase cDNA (GaLA C1) was introduced into Populus alba var. pyramidalis by A. tumefaciens-mediated transformation. PCR and Southern blot analysis indicated that transgene was stably integrated into the genome of transformants. Enzyme assay showed that laccase activity was obviously increased in transformants. As compared with untransformed control, total lignin content in all tested transgenic lines was elevated in varying degrees (as highest as 21.5%). Histochemical staining of lignin further confirmed that overexpressing GaLA C1 could result in increased lignin content in transformants. Together, our data strongly suggested that GaLA C1 may participate in lignin synthesis and this is the first direct transgenic evidence for the involvement of plant laccases in lignification. PMID:18464585

Wang, Ji; Zhu, Mu Lan; Wei, Zhi Ming

2008-02-01

376

Genetic and Molecular Analysis of Transgenic Rice cv. Rojolele Expressing Lactoferrin  

Directory of Open Access Journals (Sweden)

Full Text Available In a previous study, human lactoferrin gene have introduced into Javanica rice cv. Rojolele by Agrobacterium-mediated transformation. Lactoferrin (LF is an 80 kDa iron-binding glycoprotein that has been proposed to have many biological roles such as protection against microbial and virus infection. This study aims to analyze the integration and level of lactoferrin gene expression of transgenic rice cv. Rojolele. The study also aims to examine the genetic character of transgenic rice expressing recombinant lactoferrin. Stability expression of recombinant lactoferrin transgenic rice seeds over generations were analyzed by ELISA, while the integration stability of recombinant hLF gene in transgenic plants performed by PCR. The mitotic time, cell cycle and chromosome characterization of transgenic and non-transgenic rice cv. Rojolele were determined. Chromosome characterization of the trangenic and non transgenic rice cv. Rojolele was investigated to determine the genetic variation. All of the above efforts were aimed to evaluate the genetically engineered rice containing recombinant lactoferrin as a nutraceutical food. The results showed that the expression was stable through three consecutive generations. The expression of the hLF gene increased during grain-filling period. The active time of mitotic cells of transgenic rice rojolele was longer than the cells of non-transgenic rice. In addition, the cycle cell of transgenic and non-transgenic rojolele contained prophase, prometaphase, metaphase, anaphase, telophase and interphase. The result showed that all of the transgenic lines had diploid (2n chromosome number = 24.

Diah Rachmawati

2014-02-01

377

Maintenance and distribution of transgenic mice susceptible to human viruses: memorandum from a WHO meeting.  

OpenAIRE

This Memorandum discusses the use of transgenic mice in poliovirus research and the potential risks to public health. General and specific recommendations are given concerning the maintenance, containment and transport of transgenic animals which are susceptible to pathogenic human viruses, with special attention to transgenic mice susceptible to polioviruses.

1993-01-01

378

[Transgenic tobacco (Nicotiana tabacum SR1) plants expressing the gene coding for Serratia marcescens nuclease].  

Science.gov (United States)

The gene coding for the secreted Serratia marcescens endonuclease was fused with the mannopine synthase promoter of Agrobacterium tumefaciens Ti plasmid and transferred to Nicotiana tabacum SR1 plants. The promoter is leaf- and root-specific. The resulting transgenic plants demonstrated elevated nuclease activity. The level of the transgene product was determined in the transgenic lines. PMID:11898619

Trifonova, E A; Komarova, M L; Syrnik, O A; Kochetov, A V; Shumny?, V K

2002-02-01

379

Production of transgenic pigs over-expressing the antiviral gene Mx1.  

Science.gov (United States)

The myxovirus resistance gene (Mx1) has a broad spectrum of antiviral activities. It is therefore an interesting candidate gene to improve disease resistance in farm animals. In this study, we report the use of somatic cell nuclear transfer (SCNT) to produce transgenic pigs over-expressing the Mx1 gene. These transgenic pigs express approximately 15-25 times more Mx1 mRNA than non-transgenic pigs, and the protein level of Mx1 was also markedly enhanced. We challenged fibroblast cells isolated from the ear skin of transgenic and control pigs with influenza A virus and classical swine fever virus (CFSV). Indirect immunofluorescence assay (IFA) revealed a profound decrease of influenza A proliferation in Mx1 transgenic cells. Growth kinetics showed an approximately 10-fold reduction of viral copies in the transgenic cells compared to non-transgenic controls. Additionally, we found that the Mx1 transgenic cells were more resistant to CSFV infection in comparison to non-transgenic cells. These results demonstrate that the Mx1 transgene can protect against viral infection in cells of transgenic pigs and indicate that the Mx1 transgene can be harnessed to develop disease-resistant pigs. PMID:25408889

Yan, Quanmei; Yang, Huaqiang; Yang, Dongshan; Zhao, Bentian; Ouyang, Zhen; Liu, Zhaoming; Fan, Nana; Ouyang, Hongsheng; Gu, Weiwang; Lai, Liangxue

2014-01-01

380

COORDINATED PATTERNS OF GENE EXPRESSION FOR SKELETAL MUSCLE HYPERTROPHY IN TRANSGENIC MICE EXPRESSING MYOSTATIN PROPEPTIDE  

Science.gov (United States)

Previously, we have generated myostatin propeptide transgenic mice. Expression of the propeptide transgene at 5% of beta-actin mRNA level results in 17-30% increase in growth rate, and 22-44% increase in carcass weight at 9 weeks of age. The total mass of the main muscles of transgenic mice increase...

381

BIODIVERSITY (COMMUNICATIONS ARISING): MAIZE TRANSGENE RESULTS IN MEXICO ARE ARTEFACTS  

Science.gov (United States)

Quist and Chapela's conclusion that the transgenes they claim to have detected in native maize in Oaxaca, Mexico, are predominantly reassorted and inserted into a "diversity of genomic contexts" seems to be based on an artefact arising from the inverse polymerase chain reaction (i-PCR) they used to ...

382

Body composition of transgenic pigs expressing the myostatin pro domain  

Science.gov (United States)

Previous results have shown that male mice expressing a myostatin pro domain construct (MLC-Pro) have increased body weight, more total body lean mass, and lower percentage of total body fat. Founder transgenic (TG) pigs were generated by standard pronuclear microinjection techniques using the sam...

383

Transgenic cotton age effects on lepidopteran larvae mortality  

Science.gov (United States)

Leaves from plants containing various transgenic traits (Non-Bt, Bollgard, Bollgard II, and WideStrike)were assayed for bioactivity in the laboratory against bollworm, Helicoverpa zea (Boddie), beet armyworm, Spodoptera frugiperda (J. E. Smith), and cabbage looper, Trichoplusia ni (Hubner). About 5...

384

Modulation of Alzheimer?s Pathology in Transgenic Mouse Models  

OpenAIRE

Die Alzheimer-Krankheit (AK) ist eine fortschreitende neurodegenerative Erkrankung, welche die höchste Prävalenz unter den Demenzen aufweist. Die neuropathologischen Erkennungszeichen der AK bestehen aus extrazellulären Ablagerungen des Amyloid-ß-Peptids (Aß) und neurofibrillären Bündeln aus filamentösen Aggregaten des hyperphosphorylierten Proteins Tau. Zur näheren Untersuchung der Alzheimer Pathogenese wurden transgene Mäuse generiert, welche das mutierte humane Amyloid Precourser...

Duma, Carmen Cecilia

2009-01-01

385

Developing Transgenic Citrus for Resistance to Huanglongbing and Citrus Canker  

Science.gov (United States)

Huanglongbing (HLB) and Citrus Bacterial Canker (CBC) are serious threats to citrus production, and resistant transgenic citrus is desirable. Genes for antimicrobial peptides (AMPs) with diverse promoters have been used to generate thousands of rootstock and scion transformants. D35S::D4E1 transfor...

386

Transgenic RNAi in mouse oocytes: The first decade.  

Czech Academy of Sciences Publication Activity Database

Ro?. 134, 1-2 (2012), s. 64-68. ISSN 0378-4320 Institutional research plan: CEZ:AV0Z50520514 Institutional support: RVO:68378050 Keywords : RNAi * oocyte * transgene * silencing Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 1.897, year: 2012

Malík, Radek; Svoboda, Petr

2012-01-01

387

Early Parkinson's disease symptoms in ?-synuclein transgenic monkeys.  

Science.gov (United States)

Parkinson's disease (PD) is an age-dependent neurodegenerative disease that can be caused by genetic mutations in ?-synuclein (?-syn) or duplication of wild-type ?-syn; PD is characterized by the deposition of ?-syn aggregates, indicating a gain of toxicity from accumulation of ?-syn. Although the major neuropathologic feature of PD is the degeneration of dopaminergic (DA) neurons in the substantia nigra, non-motor symptoms including anxiety, cognitive defect and sleep disorder precede the onset of motor impairment, and many clinical symptoms of PD are not caused by degeneration of DA neurons. Non-human primate models of PD are important for revealing the early pathology in PD and identifying effective treatments. We established transgenic PD rhesus monkeys that express mutant ?-syn (A53T). Six transgenic A53T monkeys were produced via lentiviral vector expressing A53T in fertilized monkey eggs and subsequent embryo transfer to surrogates. Transgenic A53T is expressed in the monkey brain and causes age-dependent non-motor symptoms, including cognitive defects and anxiety phenotype, without detectable sleeping disorders. The transgenic ?-syn monkeys demonstrate the specific early symptoms caused by mutant ?-syn and provide insight into treatment of early PD. PMID:25552648

Niu, Yuyu; Guo, Xiangyu; Chen, Yongchang; Wang, Chuan-En; Gao, Jinquan; Yang, Weili; Kang, Yu; Si, Wei; Wang, Hong; Yang, Shang-Hsun; Li, Shihua; Ji, Weizhi; Li, Xiao-Jiang

2015-04-15

388

Development of transgenic chickens expressing enhanced green fluorescent protein  

International Nuclear Information System (INIS)

In this work we demonstrated the successful production of transgenic chickens expressing the enhanced green fluorescence protein (EGFP) gene. Replication-defective recombinant retroviruses produced from vesicular stomatitis virus G glycoprotein pseudotyped retrovirus vector system were injected beneath the blastoderm of non-incubated chicken embryos (stage X). From 129 injected eggs, 13 chicks hatched after 21 days of incubation. All hatched chicks were found to express vector-encoded EGFP gene, which was under the control of the Rous sarcoma virus promoter and boosted post-transcriptionally by woodchuck hepatitis virus post-transcriptional regulatory element sequence. Green fluorescent signals, indicative of the EGFP gene expression, were detected in various body parts, including head, limb, eye, toe, and several internal organs. Genomic incorporation of the transgene was also proven by Southern blot assay. Our results show the exceptional versatile effectiveness of the EGFP gene as a marker in the gene expression-related studies which therefore would be very helpful in establishing a useful transgenic chicken model system for studies on embryo development and for efficient production of transgenic chickens as bioreactors

389

Transgenes and national boundaries - The need for international regulation.  

Science.gov (United States)

What happens when one nation cultivates a transgenic crop variety but neighboring nations do not? Using alfalfa as a case study, we argue that the potential for international transgene flow is substantial, and therefore, the need for international cooperation in regulatory decisions concerning transgenic crops is imperative. Alfalfa (Medicago sativa, L.) is the major forage crop in North America. Recently, genetically modified (GM) alfalfa received a moratorium on further cultivation in the US on the grounds that the approvals were based on inadequate environmental impact assessments. With their deep root system, symbiotic nitrogen fixation, prolific seed production and prolonged dormancy, alfalfa plants are capable of establishing self-perpetuating (feral) populations in unmanaged environments. Given what is known about alfalfa pollen dispersal, such feral populations could facilitate gene flow between GM and non-GM fields. The border between the US and Canada, particularly in farming areas, is very narrow (border and found both alfalfa fields and potentially feral alfalfa plants in the ditches along the border. Our survey results provide evidence of the possibility of cross-border transgene flow, suggesting a need for international co-operative risk assessment initiatives between the US and Canada. Such situations could occur for other crops, in other international border regions as well. PMID:20028616

Bagavathiannan, Muthukumar; Van Acker, Rene

2009-01-01

390

Efficient expression of transgenes in adult zebrafish by electroporation  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background Expression of transgenes in muscle by injection of naked DNA is widely practiced. Application of electrical pulses at the site of injection was demonstrated to improve transgene expression in muscle tissue. Zebrafish is a precious model to investigate developmental biology in vertebrates. In this study we investigated the effect of electroporation on expression of transgenes in 3–6 month old adult zebrafish. Results Electroporation parameters such as number of pulses, voltage and amount of plasmid DNA were optimized and it was found that 6 pulses of 40 V·cm-1 at 15 ?g of plasmid DNA per fish increased the luciferase expression 10-fold compared to controls. Similar enhancement in transgene expression was also observed in Indian carp (Labeo rohita. To establish the utility of adult zebrafish as a system for transient transfections, the strength of the promoters was compared in A2 cells and adult zebrafish after electroporation. The relative strengths of the promoters were found to be similar in cell lines and in adult zebrafish. GFP fluorescence in tissues after electroporation was also studied by fluorescence microscopy. Conclusion Electroporation after DNA injection enhances gene expression 10-fold in adult zebrafish. Electroporation parameters for optimum transfection of adult zebrafish with tweezer type electrode were presented. Enhanced reporter gene expression upon electroporation allowed comparison of strengths of the promoters in vivo in zebrafish.

Rao S Hari

2005-10-01

391

Transgenic plants: from first successes to future applications.  

Science.gov (United States)

This dialogue was held between the Guest Editors of the Special Issue on "Plant Transgenesis" of the Int. J. Dev. Biol. and Marc De Block. He was one of the first scientists worldwide to obtain transgenic plants transformed with the chimeric selectable marker genes encoding neomycin phosphotransferase and bialaphos that confer resistance against the antibiotic kanamycin and the herbicide Basta®/glufosinate, respectively at the Department of Genetics of Ghent University and, later on, at the spin-off company, Plant Genetic Systems. Today, these two genes are still the most frequently utilized markers in transgene technology. Marc De Block chose to work on the improvement of crops in an industrial environment to help realize the production of superior seeds or products. He was part of the team that developed the male sterility/restorer system in canola (Brassica napus var. napus) that led to the first hybrid lines to be commercialized as successful products of transgene technology. In more than 30 years of research, he developed transformation procedures for numerous crops, designed histochemical, biochemical and physiological assays to monitor plant performance, and made original and innovative contributions to plant biology. Presently, he considers transgenic research part of the toolbox for plant improvement and essential for basic plant research. PMID:24166429

Van Lijsebettens, Mieke; Angenon, Geert; De Block, Marc

2013-01-01

392

Tilapia chromosomal growth hormone gene expression accelerates growth in transgenic zebrafish (Danio rerio)  

Scientific Electronic Library Online (English)

Full Text Available Gene transfer is economically important and model fish species has produced a great impact in modern biology and biotechnology. Transgenic zebrafish (Danio rerio) were generated through the co-injection of a GFP-expressing plasmid and an "all fish" transgene composed by the carp beta-actin promoter [...] and the chromosomal tilapia (Oreochromis hornorum) growth hormone gene. The GFP expression was a good indicator of stable transformation and allowed for high efficiency selection of transgenic fish. Transgenic F1 zebrafish grew 20% faster than full sibling non-transgenic controls.

Reynold, Morales; María Teresa, Herrera; Amílcar, Arenal; Asterio, Cruz; Oscar, Hernández; Rafael, Pimentel; Isabel, Guillén; Rebeca, Martínez; Mario P, Estrada.

2001-08-15

393

Transgene integration - an analysis in autotransgenic Labeo rohita Hamilton (Pisces: Cyprinidae)  

OpenAIRE

Transgenic Labeo rohita founder population was analyzed for the presence of autotransgene having histone 3 promoter and growth hormone (GH) cDNA (LRH3-GHcDNA) or total GH gene (LRH3-GH2.8) by PCR with transgene specific primers. Transgene specific amplification was seen with LRH3-GHcDNA in five out of seven individuals and all three fishes with LRH3-GH2.8, indicating their transgenic nature. Transgene integration was also studied by Southern hybridization of DNA isolated from blood of the tra...

Rajesh, R.; Majumdar, K. C.

2005-01-01

394

Options for development of transgenic pigs with enhanced performance traits  

International Nuclear Information System (INIS)

Full text: Traditional breeding practices have yielded a slow but steady genetic improvement of domestic animals. Unfortunately, these practices often do not enable the separation of desirable production traits from undesirable traits, and furthermore, do not enable the transfer of advantageous genetic traits from one species to another. Transgenic methodologies surmount these barriers, and transgenic pigs have been developed that have a variety of novel enhanced performance traits, the capability to serve as factories for the production of pharmaceuticals, and soon may provide a reliable supply of organs for xenotransplantation. This presentation will focus primarily on the expression of novel performance traits, since they have the potential to provide the greatest benefit to farmers in countries with a less well developed agricultural infrastructure. The first hurdle in the development of animals with novel production characteristics is the availability of reliable methods for the production of transgenic animals. This requires the combination of a suitable transgenic technique and an appropriate genetic construct. Classic pronuclear microinjection, the original method for producing transgenic animals may soon be surpassed by the more convenient sperm-mediated transgenesis, use of a retroviral vector system, or by techniques involving nuclear transfer. Despite the complexity involved in generating transgenic animals, a variety of interesting performance enhancing gety of interesting performance enhancing genes have been introduced into pigs. These include porcine growth hormone and IGF1 to enhance growth characteristics and carcass quality, ?-lactalbumin to enhance the growth of nursing piglets, plant oleate desaturase (http://www.newscientist.com/hottopics/gm/gm.jsp?id=ns99991841) to increase the proportion of unsaturated fatty acids in tissues, and phytase to enhance plant phosphorus utilization. No information is available on the performance or meat quality characteristics of pigs expressing the desaturase gene, however, expression of growth hormone, ?- lactalbumin and phytase genes in pigs has been shown to be efficacious. As an example, phytase in transgenic pigs enable practically complete utilization of the phosphorus in cereal grain as compared to less than half by non-transgenic pigs. This new trait would be particularly beneficial in a production environment without extensive infrastructure characteristic of modern intensive Western-style agriculture. In addition to bypassing the need for expensive supplemental phosphorus, it also could help avoid environmental problems associated with intensive agriculture. In addition to genes already tested that enhance performance characteristics of pigs, other genes of interest include those coding for plant cell wall hydrolases, genes coding for disease resistance, and yet others that could improve protein utilization. Expression of a novel gene in a pig is only the first step in bringing a new line of transgenic pigs into the pork production system. Once a transgenic pig has been documented to perform according to expectation, and several generations have passed to ensure that the novel gene is stably inherited and expression maintained, governmental regulatory requirements must be satisfied to ensure that the pig has no deleterious effect on the environment, and that the meat is safe for human consumption. The food safety standards may vary among countries, but those established by the FAO (http://www.fao.org/es/ESN/food/riskbiotechpapersen.stm) are usually taken as the primary requirement and country specific requirement overlaid. Meeting these standards dictates that, at least initially, only traits with great utility will be funded at a sufficiently high level to afford the essential testing to meet national and international safety requirements. (author)

395

Galanin impairs performance on learning and memory tasks: findings from galanin transgenic and GAL-R1 knockout mice.  

Science.gov (United States)

Galanin (GAL) impairs performance on cognitive tasks when administered centrally to rats. GAL transgenic (GAL-tg) mice overexpressing endogenous GAL show deficits on the probe trial of the Morris water maze spatial learning task, on the social transmission of food preference olfactory memory task, and on the trace cued fear conditioning emotional learning and memory task. Knockout mice deficient in the GAL-R1 receptor subtype were normal on most memory tasks, while showing a small deficit in trace cued fear conditioning, suggesting a selective role for the GAL-R1 in aversive memories, and implicating other GAL receptor subtypes in spatial learning and olfactory social memory. The growing body of rodent literature implicating excess GAL in cognitive impairment is relevant to the overexpression of GAL in the basal forebrain during the progression of Alzheimer's disease. PMID:15944016

Rustay, Nathan R; Wrenn, Craige C; Kinney, Jefferson W; Holmes, Andrew; Bailey, Kathleen R; Sullivan, Timothy L; Harris, Ashley P; Long, Katharine C; Saavedra, Maria C; Starosta, Grzegorz; Innerfield, Caitlin E; Yang, Rebecca J; Dreiling, Jennifer L; Crawley, Jacqueline N

2005-06-01

396

Accumulation of transgene-derived siRNAs is not sufficient for RNAi-mediated protection against Citrus tristeza virus in transgenic Mexican lime.  

Science.gov (United States)

Mexican lime plants transformed with the 3'-terminal 549 nucleotides of the Citrus tristeza virus (CTV) genome in sense, antisense and intron-hairpin formats were analysed for transgene-derived transcript and short interfering RNA (siRNA) accumulation, and for CTV resistance. Propagations from all sense, antisense and empty-vector transgenic lines were susceptible to CTV, except for a single sense-line plant with a complex transgene integration pattern that showed transgene-derived siRNAs in association with low levels of the transgene-derived transcript. In contrast, nine of 30 intron-hairpin lines showed CTV resistance, with 9%-56% of bud-propagated plants, depending on the line, remaining uninfected on graft inoculation, and the others being susceptible. Although resistance was always associated with the presence of transgene-derived siRNAs, their level in different sense and intron-hairpin transformants was variable irrespective of the response to CTV infection. In intron-hairpin lines with single transgene integration, CTV resistance was correlated with low accumulation of the transgene-derived transcript rather than with high accumulation of transgene-derived siRNAs. PMID:20078774

López, Carmelo; Cervera, Magdalena; Fagoaga, Carmen; Moreno, Pedro; Navarro, Luis; Flores, Ricardo; Peña, Leandro

2010-01-01

397

A strategy to study tyrosinase transgenes in mouse melanocytes  

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Full Text Available Abstract Background A number of transgenic mice carrying different deletions in the Locus Control Region (LCR of the mouse tyrosinase (Tyr gene have been developed and analysed in our laboratory. We require melanocytes from these mice, to further study, at the cellular level, the effect of these deletions on the expression of the Tyr transgene, without potential interference with or from the endogenous Tyr alleles. It has been previously reported that it is possible to obtain and immortalise melanocyte cell cultures from postnatal mouse skin. Results Here, we describe the efforts towards obtaining melanocyte cultures from our Tyr transgenic mice. We have bred our Tyr transgenic mice into Tyr c-32DSD mutant background, lacking the endogenous Tyr locus. In these conditions, we failed to obtain immortalised melanocytes. We decided to include the inactivation of the Ink4a-Arf locus to promote melanocyte immortalisation. For this purpose, we report the segregation of the Ink4a-Arf null allele from the brown (Tyrp1b mutation in mice. Finally, we found that Ink4a-Arf +/- and Ink4a-Arf -/- melanocytes had undistinguishable tyrosine hydroxylase activities, although the latter showed reduced cellular pigmentation content. Conclusion The simultaneous presence of precise genomic deletions that include the tyrosinase locus, such as the Tyr c-32DSD allele, the Tyr transgene itself and the inactivated Ink4a-Arf locus in Tyrp1B genetic background appear as the crucial combination to perform forthcoming experiments. We cannot exclude that Ink4a-Arf mutations could affect the melanin biosynthetic pathway. Therefore, subsequent experiments with melanocytes will have to be performed in a normalized genetic background regarding the Ink4a-Arf locus.

Serrano Manuel

2005-04-01

398

Functional screening of an asthma QTL in YAC transgenic mice  

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While large numbers of quantitative trait loci (QTLs) contributing to genetically complex conditions have been discovered, few causative genes have been identified. This is mainly due to the large size of QTLs and the subtle connection between genotype and quantitative phenotype associated with these conditions. While large numbers of quantitative trait loci (QTLs) contributing to genetically complex conditions have been discovered, few causative genes have been identified. This is mainly due to the large size of QTLs and the subtle connection between genotype and quantitative phenotype associated with these conditions. To screen for genes contributing to an asthma QTL mapped to human chromosome 5q33, the authors characterized a panel of large-insert 5q31 transgenics based on studies demonstrating that altering gene dosage frequently affects quantitative phenotypes normally influenced by that gene. This panel of human YAC transgenics, propagating a one megabase interva2048 chromosome 5q31 containing 23 genes, was screened for quantitative changes in several asthma-associated phenotypes. Multiple independent transgenic lines with altered IgE response to antigen treatment shared a 180 kb region containing 5 genes, including human interleukin 4 (IL4) and interleukin 13 (IL13), which induce IgE class switching in B cells5. Further analysis of these mice and mice transgenic for only murine Il4 and Il13 demonstrated that moderate changes in murine Il4 and Il13 expression affect asthma-associated phenotypes in vivo. This functional screen of large-insert transgenics enabled them to sift through multiple genes in the 5q3 asthma QTL without prior consideration of assumed individual gene function and identify genes that influence the QTL phenotype in vivo.

Symula, Derek J.; Frazer, Kelly A.; Ueda, Yukihiko; Denefle, Patrice; Stevens, Mary E.; Wang, Zhi-En; Locksley, Richard; Rubin, Edward M.

1999-07-02

399

Inheritance and segregation of virus and herbicide resistance transgenes in sugarcane.  

Science.gov (United States)

Transgenic sugarcane parents containing multiple copies of herbicide resistance ( bar) and Sorghum mosaic virus (SrMV) resistance ( hut) genes were crossed with non-transgenic sugarcane varieties. Segregation of the transgenes in the progeny was determined using Southern blot analysis; herbicide resistance and SrMV resistance were assessed using bioassays. The segregation data were used to infer linkage relationships between transgenes in the parent plants. In two of the parents, all transgene insertions were linked in one position in the genome, although some recombination between insertion events did occur. In the third parent, insertion had occurred in two independent, unlinked loci. Analysis of progeny of this parent indicated that rearrangement or mutation occurred in both loci, resulting in non-parental transgene DNA fragments in some progeny. Most transgenic progeny containing the bar gene showed resistance to herbicide. SrMV inoculation indicated that a fairly high proportion of the transgenic progeny showed susceptibility. As the post-transcriptional gene silencing mechanism responsible for the virus resistance phenotype may be reset during meiosis, phenotypic screening of older plants may be a more reliable indication of virus resistance than screening young seedlings. This is the first report of transgene segregation in sugarcane, and we have demonstrated that transgenic sugarcane parents showing stable inheritance of transgenes can be effectively used in breeding programs. PMID:12582639

Butterfield, K.; Irvine, E.; Valdez Garza, M.; Mirkov, E.

2002-04-01

400

Full-term development of rat after transfer of nuclei from two-cell stage embryos.  

Science.gov (United States)

Cloning technology would allow targeted genetic alterations in the rat, a species which is yet unaccessible for such studies due to the lack of germline-competent embryonic stem cells. The present study was performed to examine the developmental ability of reconstructed rat embryos after transfer of nuclei from early preimplantation stages. We observed that single blastomeres from two-cell embryos and zygotes reconstructed by pronuclei exchange can develop in vitro until morula/blastocyst stage. When karyoplasts from blastomeres were used for the reconstruction of embryos, highest in vitro cleavage rates were obtained with nuclei in an early phase of the cell cycle transferred into enucleated preactivated oocytes or zygotes. However, further in vitro development of reconstructed embryos produced from blastomere nuclei was arrested at early cleavage stages under all conditions tested in this study. In contrast, immediate transfer to foster mothers of reconstructed embryos with nuclei from two-cell embryos at an early stage of the cell cycle in preactivated enucleated oocytes resulted in live newborn rats, with a general efficiency of 0.4%-2.2%. The genetic origin of the cloned offspring was verified by using donor nuclei from embryos of Black Hooded Wistar rats and transgenic rats carrying an ubiquitously expressed green fluorescent protein transgene. Thus, we report for the first time the production of live cloned rats using nuclei from two-cell embryos. PMID:16807380

Popova, Elena; Bader, Michael; Krivokharchenko, Alexander

2006-10-01

401

Non-Homologous End Joining Plays a Key Role in Transgene Concatemer Formation in Transgenic Zebrafish Embryos  

Directory of Open Access Journals (Sweden)

Full Text Available This study focused on concatemer formation and integration pattern of transgenes in zebrafish embryos. A reporter plasmid based on enhanced green fluorescent protein (eGFP driven by Cytomegalovirus (CMV promoter, pCMV-pax6in-eGFP, was constructed to reflect transgene behavior in the host environment. After removal of the insertion fragment by double digestion with various combinations of restriction enzymes, linearized pCMV-pax6in-eGFP vectors were generated with different combinations of 5?-protruding, 3?-protruding, and blunt ends that were microinjected into zebrafish embryos. Repair of double-strand breaks (DSBs was monitored by GFP expression following religation of the reporter gene. One-hundred-and-ninety-seven DNA fragments were amplified from GFP-positive embryos and sequenced to analyze the repair characteristics of different DSB end combinations. DSBs involving blunt and asymmetric protruding ends were repaired efficiently by direct ligation of blunt ends, ligation after blunting and fill-in, or removed by cutting. Repair of DSBs with symmetric 3?-3? protrusions was less efficient and utilized template-directed repair. The results suggest that non-homologous end joining (NHEJ was the principal mechanism of exogenous gene concatemer formation and integration of transgenes into the genome of transgenic zebrafish.

Jun Dai, Xiaojuan Cui, Zuoyan Zhu, Wei Hu

2010-01-01

402

Establishment of a DsRed-Monomer-Harboring ICR Transgenic Mouse Model and Effects of the Transgene on Tissue Development.  

Science.gov (United States)

DsRed-monomer is an enhanced red fluorescent protein that may serve as a marker for studies in biotechnology and cell biology. Since the ICR mouse strain is a widely utilized outbred strain for oncology, toxicology, vaccine development and for aging studies, the objective of this study was to produce a DsRed-monomer transgenic mouse by means of pronuclear micro-injection of a vector driven by the cytomegalovirus (CMV) enhancer/chicken beta-actin promoter. Four transgenic mice were successfully produced, one of which expressed the DsRed-monomer protein in every tissue, although at varying levels. High expression levels were observed in the heart, pancreas and muscle. Moreover, amniotic fluid-derived progenitor cells, which also expressed the DsRed-monomer protein, could be collected from the DsRed-monomer- harboring ICR mice. As compared to wild-type mice, a few biochemical and histological dissimilarities were found in the DsRed-monomer transgenic mice, including the presence of intra-cytoplasmic eosinophilic threadlike materials in the acinar cells. Taken together, transgenic mice stably expressing DsRed-monomer can be produced using pronuclear micro-injection; however, expression of the DsRed-monomer gene or its insertion position may lead to minor influences. PMID:25687489

Chou, Chih-Jen; Peng, Shao-Yu; Wan, Cho-Hua; Chen, Sou-Fu; Cheng, Winston Teng-Kui; Lin, Kun-Yi; Wu, Shinn-Chih

2015-02-28

403

Bt-transgenic cotton is more sensitive to CeO? nanoparticles than its parental non-transgenic cotton.  

Science.gov (United States)

Because genetically modified crops are developing widely in the world while nanoparticles (NPs) are being synthesized and applied in various fields, they will have many opportunities for interactions in the future. The effects of NPs on genetically modified crops therefore require investigation. In the present study, CeO2 NPs were revealed to have toxic effects on root biomass of Bt 29317 at 100 and 500 mg L(-1), but had no toxic effects on Jihe 321. Besides, we also studied the effects of CeO2 NPs on nutrient element uptake in transgenic cotton, and found that contents of most nutrient elements (Fe, Ca, Mg, Zn and Na) in roots of Bt 29317 were affected at lower NP concentrations (100 mg L(-1)) compared with Jihe 321. In addition, ICP-MS analysis revealed that CeO2 NPs were more heavily adsorbed by roots of Bt 29317 than Jihe 321, whereas fewer CeO2 NPs were transported from roots to shoots of Bt 29317 than its non-transgenic counterpart. These data confirm that Bt 29317 is more sensitive to CeO2 NPs than its parental non-transgenic cotton, indicating that nanomaterials are potentially more detrimental to transgenic plants than conventional ones. PMID:24793293

Li, Xuguang; Gui, Xin; Rui, Yukui; Ji, Weikang; Van Nhan, Le; Yu, Zihan; Peng, Shengnan

2014-06-15

404

Acute and chronic systemic CB1 cannabinoid receptor blockade improves blood pressure regulation and metabolic profile in hypertensive (mRen2)27 rats  

OpenAIRE

We investigated acute and chronic effects of CB1 cannabinoid receptor blockade in renin?angiotensin system?dependent hypertension using rimonabant (SR141716A), an orally active antagonist with central and peripheral actions. In transgenic (mRen2)27 rats, a model of angiotensin II?dependent hypertension with increased body mass and insulin resistance, acute systemic blockade of CB1 receptors significantly reduced blood pressure within 90 min but had no effect in Sprague?Dawley rats. No...

Schaich, Chris L.; Shaltout, Hossam A.; Brosnihan, K. Bridget; Howlett, Allyn C.; Diz, Debra I.

2014-01-01

405

FAS-Dependent Cell Death in ?-Synuclein Transgenic Oligodendrocyte Models of Multiple System Atrophy  

DEFF Research Database (Denmark)

Multiple system atrophy is a parkinsonian neurodegenerative disorder. It is cytopathologically characterized by accumulation of the protein p25? in cell bodies of oligodendrocytes followed by accumulation of aggregated ?-synuclein in so-called glial cytoplasmic inclusions. p25? is a stimulator of ?-synuclein aggregation, and coexpression of ?-synuclein and p25? in the oligodendroglial OLN-t40-AS cell line causes ?-synuclein aggregate-dependent toxicity. In this study, we investigated whether the FAS system is involved in ?-synuclein aggregate dependent degeneration in oligodendrocytes and may play a role in multiple system atrophy. Using rat oligodendroglial OLN-t40-AS cells we demonstrate that the cytotoxicity caused by coexpressing ?-synuclein and p25? relies on stimulation of the death domain receptor FAS and caspase-8 activation. Using primary oligodendrocytes derived from PLP-?-synuclein transgenic mice we demonstrate that they exist in a sensitized state expressing pro-apoptotic FAS receptor, which makes them sensitive to FAS ligand-mediated apoptosis. Immunoblot analysis shows an increase in FAS in brain extracts from multiple system atrophy cases. Immunohistochemical analysis demonstrated enhanced FAS expression in multiple system atrophy brains notably in oligodendrocytes harboring the earliest stages of glial cytoplasmic inclusion formation. Oligodendroglial FAS expression is an early hallmark of oligodendroglial pathology in multiple system atrophy that mechanistically may be coupled to ?-synuclein dependent degeneration and thus represent a potential target for protective intervention.

Kragh, Christine L; Fillon, Gwenaëlle

2013-01-01

406

Immune selection of tumor cells in TCR ?-chain transgenic mice.  

Science.gov (United States)

The concept of immunological surveillance implies that immunogenic variants of tumor cells arising in the organism can be recognized by the immune system. Tumor progression is provided by somatic evolution of tumor cells under the pressure of the immune system. The loss of MHC Class I molecules on the surface of tumor cells is one of the most known outcomes of immune selection. This study developed a model of immune selection based on the immune response of TCR 1d1 single ?-chain transgenic B10.D2(R101) (K(d)I(d)D(b)) mice to allogeneic EL4 (H-2(b)) thymoma cells. In wild-type B10.D2(R101) mice, immunization with EL4 cells induced a vigorous CTL response targeted to the H-2K(b) molecule and results in full rejection of the tumor cells. In contrast, transgenic mice developed a compromised proliferative response in mixed-lymphocyte response assays and were unable to reject transplanted allogeneic EL4 cells. During the immune response to EL4 cells, CD8(+) T-lymphocytes with endogenous ?-chains accumulated predominantly in the spleen of transgenic mice and only a small part of the T-lymphocytes expressing transgenic ?-chains became CD8(+)CD44(+)CD62L(-) effectors. Then, instead of a full elimination of tumor cells as in wild-type mice, a reproducible prolonged equilibrium phase and subsequent escape was observed in transgenic mice that resulted in death of 90% of the mice in 40-60 days after grafting. Prolonged exposure of tumor cells to the pressure of the immune system in transgenic mice in vivo resulted in a stable loss of H-2K(b) molecules on the EL4 cell surface. Genetic manipulation of the T-lymphocyte repertoire was sufficient to reproduce the classic pattern of interactions between tumor cells and the immune system, usually observed in reliable syngeneic models of anti-tumor immunity. This newly-developed model could be used in further studies of immunoregulatory circuits common for transplantational and anti-tumor immune responses. PMID:24308870

Silaeva, Yulia Yu; Grinenko, Tatyana S; Vagida, Murad S; Kalinina, Anastasia A; Khromykh, Ludmila M; Kazansky, Dmitry B

2014-10-01

407

Similar incidence of K-ras mutations in lung carcinomas of FVB/N mice and FVB/N mice carrying a mutant p53 transgene.  

Science.gov (United States)

Mutated p53 genes are capable of complementing activated ras genes in the transformation of primary rat embryo fibroblasts in vitro. Mutations in both genes have also been found in several human cancers, including lung carcinomas. We generated transgenic mice containing a p53 construct with a missense mutation in exon 5 (ala135val) to study the role of p53 mutations in lung tumorigenesis, and to facilitate identification of other genetic events that might complement p53 mutations in mouse lung carcinogenesis. The p53 transgenic lines exhibited a higher frequency of lethal lung tumors than the parental FVB/N strain. We examined the spontaneously-arising lung carcinomas from mice expressing the mutated p53 transgene for K-ras mutations using single-stranded conformation polymorphism (SSCP) and/or direct sequencing approaches. Fifteen of 29 (52%) carcinomas contained mutations in the K-ras oncogene. Six of 15 of the K-ras mutations were in codon 61 and 9/15 were in codon 12. Subsequent analysis of spontaneous lung carcinomas from mice of the FVB/N parental strain showed that 9/12 (75%) carcinomas examined contained K-ras mutations. Two of these were in codon 12, one in codon 13, and 6 were in codon 61. These results demonstrate that the frequency of ras mutations does not differ between the p53 FVB/N transgenic mice and their parental FVB/N strain but suggest that a high frequency of mutations K-ras can be correlated with lung tumorigenesis in both groups of mice. PMID:9230291

Shafarenko, M; Mahler, J; Cochran, C; Kisielewski, A; Golding, E; Wiseman, R; Goodrow, T

1997-07-01

408

Instability of transgenes in plants and its implications for plant breeding  

International Nuclear Information System (INIS)

A major commitment is being made to the exploitation of transgenes for crop improvement. Numerous studies involving many transgenes and plant species indicate that transgenes are often silent, or display variable expression during development or between sexual generations. Transgene expression is affected by the number of transgenes, the genetic background and the environment. Instability in expression of chalcone synthase genes in petunia in the presence of a transgene(s) encoding chalcone synthase under the control of the cauliflower mosaic virus 35S promoter is described. Chalcone synthase is a key enzyme in floral anthocyanin production. Such transgenic plants vary in floral phenotype. Some have purple flowers, others white flowers and many have flowers with non-random white and purple sectors. Flower type is characteristic of a particular transformant. Somatic variants are observed frequently. Loss of purple pigment production is associated with post-transcriptional loss of mRNA from trans and endogenous chalcone synthase genes (co-suppression). Studies on the structure of the chalcone synthase RNAs produced in transgenic plants, amplified using polymerase chain reaction, have revealed several kinds of aberration. It is therefore postulated that aberrant RNAs provoke chalcone synthase RNA turnover. A speculative scheme to explain how RNA degradation occurs, involving antisense RNA, is presented. The implications of transgene behaviour and instability in plant breene behaviour and instability in plant breeding are discussed. (author). 27 refs, 3 figs

409

Effects of Transgenic Rice on Life History Traits of Daphnia magna in Life Table Experiments  

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Full Text Available To investigate the impacts of transgenic rice on freshwater organisms, we conducted two life tableexperiments using Daphnia magna for fifteen and twenty days, respectively. We examined life history traits suchas population growth rates (r, reproductive rates (R0, generation times, and survivorship. In the first experiment,we used non-drought-stressed transgenic and non-transgenic rice harvested in 2005. In the second study, weused non-transgenic and transgenic rice harvested in 2006 following drought stress. Each experiment involvedthree treatments in which D. magna neonates were fed with Selenastrum capricornutum (control treatment andS. capricornutum with 5% aqueous extracts of non-transgenic rice (N-T and transgenic rice (T. In the firstexperiment, D. magna showed reduced population growth rates and lowered fecundity in the N-T and Ttreatments. In the second experiment, D. magna receiving both transgenic and non-transgenic rice extractsshowed very high mortality, low population growth rates and reproduction rates. We could not detect anysignificant negative effects of extracts from transgenic rice on D. magna life history traits at 95%.

Nam, Sungjin

2007-11-01

410

[Enhancement of artemisinin biosynthesis in transgenic Artemisia annua L. by overexpressed HDR and ADS genes].  

Science.gov (United States)

Artemisnin is a novel sesquiterpene lactone with an internal peroxide bridge structure, which is extracted from traditional Chinese herb Artemisia annua L. (Qinghao). Recommended by World Health Organization, artemisinin is the first-line drug in the treatment of encephalic and chloroquine-resistant malaria. In the present study, transgenic A. annua plants were developed by overexpressing the key enzymes involved in the biosynthetic pathway of artemisinin. Based on Agrobacterium-mediated transformation methods, transgenic plants of A. annua with overexpression of both HDR and ADS were obtained through hygromycin screening. The genomic PCR analysis confirmed six transgenic lines in which both HDR and ADS were integrated into genome. The gene expression analysis given by real-time quantitative PCR showed that all the transgenic lines had higher expression levels of HDR and ADS than the non-transgenic control (except ah3 in which the expression level of ADS showed no significant difference compared with control); and the HPLC analysis of artemisinin demonstrated that transgenic A. annua plants produced artemisinin at significantly higher level than non-transgenic plants. Especially, the highest content of artemisinin was found in transgenic line ah70, in which the artemisinin content was 3.48 times compared with that in non-transgenic lines. In summary, overexpression of HDR and ADS facilitated artemisinin biosynthesis and this method could be applied to develop transgenic plants of A. annua with higher yield of artemisinin. PMID:25518337

Wang, Ya-Xiong; Long, Shi-Ping; Zeng, Li-Xia; Xiang, Li-En; Lin, Zhi; Chen, Min; Liao, Zhi-Hua

2014-09-01

411

Quantitative analysis of tetracycline-inducible expression of the green fluorescent protein gene in transgenic chickens.  

Science.gov (United States)

The use of transgenic farm animals as "bioreactors" to address the growing demand for biopharmaceuticals, both in terms of increased quantity and greater number, represents a key development in the advancement of medical science. However, the potential for detrimental side-effects as a result of uncontrolled constitutive expression of foreign genes in transgenic animals is a well-recognized limitation of such systems. Previously, using a tetracycline-inducible expression system, we demonstrated the induction of expression of a transgene encoding green fluorescent protein (GFP) in transgenic chickens by feeding with doxycycline, a tetracycline derivative; expression of GFP reverted to pre-induction levels when the inducer was removed from the diet. As a proof of principle study, however, quantitative assessment of expression was not possible, as only one G0 and one G1 transgenic chicken was obtained. In the current study, a sufficient number of G2 and G3 transgenic chickens were obtained, and quantification analysis demonstrated up to a 20-fold induction of expression by doxycycline. In addition, stable transmission of the transgene without any apparent genetic modifications was observed through several generations. The use of an inducible expression system that can be regulated by dietary supplementation could help mitigate the physiological disruption that can occur in transgenic animals as a result of uncontrolled constitutive expression of a transgene. Importantly, these results also support the use of the retroviral system for generating transgenic animals with minimal risk in terms of biosafety. PMID:22850941

Koo, Bon Chul; Kwon, Mo Sun; Roh, Ji Yeol; Kim, Minjee; Kim, Jin-Hoi; Kim, Teoan

2012-01-01

412

Conditional expression of a myocardium-specific transgene in zebrafish transgenic lines.  

Science.gov (United States)

To develop the first heart-specific tetracycline (Tet)-On system in zebrafish, we constructed plasmids in which the cardiac myosin light chain 2 promoter of zebrafish was used to drive the reverse Tet-controlled transactivator (rtTA) and the green fluorescent protein (GFP) reporter gene was preceded by an rtTA-responsive element. In the zebrafish fibroblast cell-line, rtTA-M2, one of rtTA's derivatives, demonstrated the highest increase in luciferase activity upon doxycycline (Dox) induction. We then generated two germ lines of transgenic zebrafish: line T03 was derived from microinjection of a plasmid containing rtTA-M2 and a plasmid containing a responsive reporter gene, whereas line T21 was derived from microinjection of a single dual plasmid. Results showed that line T21 was superior to line T03 in terms of greater GFP intensity after induction and with of minimal leakiness before induction. The photographic images of induced GFP in the heart of F2 larvae showed that the fluorescent level of GFP was dose-responsive. The level of GFP expressed in the F3 3 days postfertilization larvae that were treated with Dox for 1 hr decreased gradually after the withdrawal of the inducer; and the fluorescent signal disappeared after 5 days. The GFP