scholarly journals Direct Comparison between ICSI-Mediated DNA Transfer and Pronuclear DNA Microinjection for Producing Transgenic Rats

2008 ◽  
Vol 57 (2) ◽  
pp. 145-148 ◽  
Author(s):  
Masumi HIRABAYASHI ◽  
Megumi KATO ◽  
Kazue AMEMIYA ◽  
Shinichi HOCHI
Keyword(s):  
Dna Transfer ◽  
Transgenic Rats ◽  
Dna Microinjection

scholarly journals No Effect of Recombinase-Mediated DNA Transfer on Production Efficiency of Transgenic Rats

2006 ◽  
Vol 55 (2) ◽  
pp. 131-135 ◽  
Author(s):  
Masumi HIRABAYASHI ◽  
Megumi KATO ◽  
Ryosuke KANEKO ◽  
Takahiro HIRABAYASHI ◽  
Mitsuhiro MORITA ◽  
...  
Keyword(s):  
Production Efficiency ◽  
Dna Transfer ◽  
Transgenic Rats

scholarly journals 317 FACTORS AFFECTING PRODUCTION EFFICIENCY OF TRANSGENIC RATS BY ICSI-MEDIATED DNA TRANSFER

2005 ◽  
Vol 17 (2) ◽  
pp. 309
Author(s):  
M. Kato ◽  
S. Hochi ◽  
M. Hirabayashi
Keyword(s):  
Production Efficiency ◽  
Dna Transfer ◽  
Egfp Expression ◽  
Transgenic Rat ◽  
Sprague Dawley ◽  
Transgenic Rats ◽  
Rat Strains ◽  
Exogenous Dna ◽  
Factors Affecting ◽  
Rat Offspring

Successful DNA transfer via intracytoplasmic sperm injection (ICSI) was first reported in mice (Perry et al. 1999, Science 284, 1180), and was recently extended to rats (Kato et al. 2004, Mol. Reprod. Dev. 69, 153). In the present study, factors affecting the production efficiency of transgenic rats by the ICSI-mediated DNA transfer were investigated. Cauda epididymal spermatozoa from Sprague-Dawley rats were sonicated (SO) and/or frozen-thawed (FT) for tail-cutting and membrane-disrupting. The sperm heads were exposed for 1 min to different concentrations (0.02–2.5 ng/μL) of 3.0 kb EGFP DNA solution, and then microinjected into denuded F1 (Donryu × LEW) rat oocytes. The optimal concentration of EGFP DNA was 0.1 ng/μL, as determined by the in vitro developmental competence into morulae/blastocysts and the EGFP expression of the ICSI oocytes. The presumptive 1- or 2-cell stage zygotes were transferred into oviducts of pseudopregnant Wistar females, and the presence of EGFP DNA in the offspring was examined by fluorescence under the 480 nm UV light. The production efficiency of transgenic rat offspring was 2.8% (2/71 zygotes transferred), 1.6% (1/63), and 3.3% (2/61) in the oocytes into which SO-, FT-, and SO+ FT-treated sperm heads were injected, respectively. The founder transgenic rats carrying EGFP DNA transmitted the transgenes to their progeny according to the Mendelian fashion (43.8–54.8%), suggesting the stable incorporation of the transgenes into rat genomes. Four rat strains (F344, LEW, Donryu, and Sprague-Dawley) were compared for their suitability as sperm/oocyte donors in the production of transgenic rats by ICSI with SO + FT-treated and 0.1 ng/μL EGFP DNA-exposed sperm heads. The production efficiency of the transgenic rats in the Sprague-Dawley strain (8.2%, 8/98) was significantly higher than that in LEW strain (0.9%, 1/114), while those in F344 (4.3%, 4/92) and Donryu (4.4%, 5/114) strains were intermediate. Attempts were made to introduce three other DNA constructs (5.0 kb plasmid and 208 kb BAC, both with Fyn gene, and 186 kb BAC with Svet1/IRES-Cre gene) into rat genomes by ICSI with SO + FT-treated and 0.1 ng/μL DNA-exposed sperm heads. PCR analysis showed that the Fyn, Fyn/BAC, and Svet1/IRES-Cre DNA constructs were successfully introduced into Sprague-Dawley rat offspring via ICSI, with production efficiencies of 2.8% (3/109), 0.9% (1/109), and 2.4% (3/125), respectively. These results indicate that transgenic rats can be produced by ICSI-mediated DNA transfer using the various types of exogenous DNA and rat strains with different genetic backgrounds.

Potential of Motor Neurons in the Nucleus Ambiguus ∼ Utilizing Nestin-EGFP Transgenic Rats

2007 ◽  
Vol 58 (2) ◽  
pp. 193-193
Author(s):  
T. Takaoka ◽  
A. Shiotani ◽  
K. Saito ◽  
M. Tomifuji ◽  
Y. Mori ◽  
...  
Keyword(s):  
Motor Neurons ◽  
Nucleus Ambiguus ◽  
Transgenic Rats

Impaired Estrogen Sensitivity in Bone by Inhibiting Both Estrogen Receptor a and b Pathways*

2020 ◽  
Author(s):  
Lungwani Muungo
Keyword(s):  
Bone Mineral Density ◽  
Estrogen Receptor ◽  
Bone Loss ◽  
Bone Turnover ◽  
Bone Mineral ◽  
Female Rats ◽  
Wild Type ◽  
Mineral Density ◽  
Transgenic Rats ◽  
Estrogen Sensitivity

Although it is well established that estrogen deficiencycauses osteoporosis among the postmenopausalwomen, the involvement of estrogen receptor (ER) in itspathogenesis still remains uncertain. In the presentstudy, we have generated rats harboring a dominantnegative ERa, which inhibits the actions of not only ERabut also recently identified ERb. Contrary to our expectation,the bone mineral density (BMD) of the resultingtransgenic female rats was maintained at the same levelwith that of the wild-type littermates when sham-operated.In addition, ovariectomy-induced bone loss wasobserved almost equally in both groups. Strikingly, however,the BMD of the transgenic female rats, after ovariectomized,remained decreased even if 17b-estradiol(E2) was administrated, whereas, in contrast, the decreaseof littermate BMD was completely prevented byE2. Moreover, bone histomorphometrical analysis ofovariectomized transgenic rats revealed that the higherrates of bone turnover still remained after treatmentwith E2. These results demonstrate that the preventionfrom the ovariectomy-induced bone loss by estrogen ismediated by ER pathways and that the maintenanceof BMD before ovariectomy might be compensatedby other mechanisms distinct from ERa and ERbpathways.

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