Production of Transgenic Rats Using Young Sprague-Dawley Females Treated with PMSG and hCG.

Hirabayashi, Masumi; Ito, Kazumi; Sekimoto, Akiyo; Hochi, Shinichi; Ueda, Masatsugu

doi:10.1538/expanim.50.365

Cited by 29 publications

(28 citation statements)

References 20 publications

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“…This result is probably caused by age difference of sensitivities to exogenous hormones. In this study, the doses of injected PMSG and hCG were uniformly 150 and 75 IU/ kg body weight, respectively, while it was previously reported that, using SD strain rats in the pre-pubertal stage, the effect of superovulation was the highest when both PMSG and hCG were injected at the dose of 300 IU/kg body weight [6]. According to that report, the superovulatory response should be improved by adjustment of the dose of injected gonadotrophins.…”

Section: Discussionmentioning

confidence: 77%

Estrous Stage- and Animal Age-Independent Superovulation in the BrlHan:WIST@Jcl(GALAS) Rat

Sotomaru

Kamisako²,

Hioki

2005

Exp. Anim.

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In most strains of rats, the effects of treatment for the induction of superovulationapplication of reproductive biotechnology techniques in rats. Since rats are the most widely used experimental animals next to mice and numerous experimental data using rats have been accumulated in research fields such as medicine, pharmacology, physiology, nutrition, psychology and genetics, there is an urgent demand for the application of reproductive technology in rats.In most strains of rats, the effect of treatment for superovulation is strongly influenced by the stage of the estrous cycle [5]. There are two main protocols to induce superovulation. One is to emphasize efficacy of endogenous gonadotrophins, in which rats at the diestrus stage are selected to initiate exogenous hormonal

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Section: Discussionmentioning

confidence: 77%

Estrous Stage- and Animal Age-Independent Superovulation in the BrlHan:WIST@Jcl(GALAS) Rat

Sotomaru

Kamisako²,

Hioki

2005

Exp. Anim.

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“…Rats were superovulated by intraperitoneal i n j e c t i o n s o f 1 5 0 I U / k g e q u i n e c h o r i o n i c gonadotrophin (eCG; Nippon Zenyaku, Co., Fukushima, Japan) at 09:00 on the first diestrous day and 75 IU/kg human chorionic gonadotrophin (hCG; Sankyo, Co., Tokyo, Japan) after an interval of 48 h, as described previously [10]. The rats were then copulated at 18:00 with fertile Wistar male rats.…”

Section: Collection Of Zygotes / Embryosmentioning

confidence: 99%

Donor and Recipient Rat Strains Affect Full-Term Development of One-Cell Zygotes Cultured to Morulae/Blastocysts

Kato

Ishikawa

Hochi

et al. 2004

Journal of Reproduction and Development

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Abstract. The present study was conducted to examine the developmental potential to offspring of rat embryos cultured from 1-cell to morula/blastocyst stage. Pronuclear zygotes from Wistar × Wistar or (SD × DA) × Wistar strains were cultured in modified rat 1-cell embryo culture medium (mR1ECM) for 96 h in 5% CO2 in air at 37 C. The proportion of the 3-way cross hybrid zygotes developing into morula/blastocyst stage (74%) was higher than that of the Wistar zygotes (66%). Day-5 morulae/ blastocysts developed in vitro were transferred into Day-3 or -4 pseudopregnant recipients of Wistar or SD × DA strain. The transfer of cultured embryos resulted in the birth of offspring at 13-59%, while that of non-cultured control blastocysts showed birth rates of 35-65%. The best offspring rate of cultured embryos (59%) was obtained when the hybrid 1-cell zygotes were cultured in mR1ECM medium and transferred into the 2-days earlier uteri of SD × DA recipients. These results suggest that genetic background of recipients as well as donors is a possible factor affecting full-term development of rat morulae/blastocysts derived from 1-cell stage zygotes cultured in vitro. Key words: Embryo transfer, Hybrid, In vitro culture, mR1ECM, Rat zygotes (J. Reprod. Dev. 50: [191][192][193][194][195] 2004) chini and Bavister [1] reported that glucose and phosphate are responsible for the 2-cell block of hamster embryos cultured in 5% O 2 , 5% CO 2 , and 90% N2. Based on that observation, 1-cell stage zygotes of mice [2], rats [3][4][5], and cattle [6,7] were f o u n d t o b e c a p a b l e o f o v e r c o m i n g t h e i r developmental blocks and of developing into the blastocyst stage when cultured in phosphate-and glucose-free hamster embryo culture medium-1, HECM-1 [1]. As for rats, Miyoshi et al. [5] reported that 80% of 1-cell stage zygotes cultured in modified HECM-1 medium of lowered osmolarity, named as rat 1-cell embryo culture medium (R1ECM), developed into morulae and blastocysts after 96 h. The same group [8] also demonstrated that morulae and blastocysts produced by in vitro f e r t i l i z a t i o n i n 0 . 4 % B S A / 1 1 0 m M N a C ls u p p l e m e n t e d m o d i f i e d R 1 E C M m e d i u m (mR1ECM) and in vitro culture in 0.01% PVA/80 mM NaCl-supplemented modified R1ECM medium had developmental potential to full-term. But, the rate of offspring production from cultured 1-cell rat embryos was not high (13-28%) and more than half of the recipient rats failed to maintain pregnancy [8,9]. While cultured embryos from Wistar strain rats have been transferred into the 1-day younger uteri of recipient Wistar rats (as common protocol), the suitability of rat strains for embryo recipients as well as zygote donors, and of Accepted for publication: December 12, 2003 Correspondence: M. Hirabayashi (e-mail: mhirarin@nips.ac.jp) or S. Hochi KATO et al. 192 embryo-recipient synchronization is poorly understood in rat offspring production from cultured embryos.The present study was conducted to examine the full-term developmenta...

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“…Our first priority was to design a working protocol to determine the ovulation kinetics of three rat strains, Wistar, ACI/N and Fisher, by using a single dose of PMSG and hCG. In spite of the time for oocyte collection reported in the literature being variable (Mukumoto et al 1995, Hirabayashi et al 2001, Krivokharchenko et al 2003, our results demonstrated that 24 h after hCG injection was the most adequate time to harvest the oocytes in the three rat strains. These data agree with other reports (Tain et al 2000, Popova et al 2002.…”

Section: Discussionmentioning

confidence: 57%

“…Production of ES cells in rats is somehow limited by the fact that superovulation protocols have not been optimized for the production of high-quality embryos. Although superovulation protocols for rat have already been published, they mainly focused on the production of either oocytes or zygotes (Mukumoto et al 1995, Tain et al 2000, Hirabayashi et al 2001, Corbin & McCabe 2002, Popova et al 2002, Krivokharchenko et al 2003, and the techniques for the production of morulae or blastocysts are still not well defined (Tain et al 2001, Ishigame et al 2004. Since both morulae and blastocysts are the source of ES cells (Evans & Kaufman 1981, Capecchi 1989, Iannaccone et al 1994, Ouhibi et al 1995, Thomson et al 1998, Vassilieva et al 2000, it would be useful to improve the superovulation protocols that allow obtaining embryos suitable for ES cell production.…”

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confidence: 99%

Rat embryo quality and production efficiency are dependent on gonadotrophin dose in superovulatory treatments

et al. 2006

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SummaryThe present study was performed to optimize a superovulation protocol in rats in order to produce a large number of good-quality embryos suitable to develop rat embryonic stem (rES) cells. We first evaluated the ovulation kinetics of three rat strains: Wistar, Fisher and ACI/N. Animals (n ¼ 30 per strain) were treated with 50 IU of pregnant mare serum gonadotrophin (PMSG), and ovulation was induced with 50 IU of human chorionic gonadotrophin (hCG) 50 h apart. Next, we evaluated the dose-response curves of PMSG and hCG in Wistar rats in order to obtain the highest number of embryos. The parameters evaluated for superovulation efficiency were: percentage of mated females, percentage of pregnant females and the average number of embryos collected per female. The results of these experiments suggested that the best dose combination was 50 IU for each hormone. Subsequent experiments, again with Wistar rats, were designed to test which of four hormonal combination treatments (30/30, 30/50, 50/30, and 50/50 IU of PMSG/hCG) will produce the largest numbers of good-quality embryos. Embryo quality was evaluated by embryo development uniformity, embryo morphology, embryo survival in an in vitro culture and embryo ability to generate rES-like cells. Results from these experiments showed that 30/50 IU of PMSG/hCG was the treatment that induced the best embryo quality. In conclusion, our results indicated that, in Wistar rats, the most appropriate hormonal combination dose for superovulation protocols with high number of good-quality embryos was 30 IU of PMSG and 50 IU of hCG given 50 h apart. We are performing further studies with rES-like cells produced with the present methodology to evaluate if they are able to participate in the production of germ-line chimeras.

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Production of Transgenic Rats Using Young Sprague-Dawley Females Treated with PMSG and hCG.

Cited by 29 publications

References 20 publications

Estrous Stage- and Animal Age-Independent Superovulation in the BrlHan:WIST@Jcl(GALAS) Rat

Estrous Stage- and Animal Age-Independent Superovulation in the BrlHan:WIST@Jcl(GALAS) Rat

Donor and Recipient Rat Strains Affect Full-Term Development of One-Cell Zygotes Cultured to Morulae/Blastocysts

Rat embryo quality and production efficiency are dependent on gonadotrophin dose in superovulatory treatments

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