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한국재래흑염소에서 발정 및 과배란 유도와 외래유전자 주입에 적합한 1세포기 수정란의 채취
신상태,이두환,김명철,이운규,이철상,한용만,이경광 충남대학교 수의과대학 동물의과학연구소 1998 動物醫科學硏究誌 Vol.6 No.-
Three different treatments for induction of estrus in Korean native black goats were compared: follicle stimulating hormone(FSH, FSH-p^TM), FSH combined with MAP(intravaginal impregnated sponges, Veramix^ⓡ, containing 60 ㎎ medroxy progesterone acetate for 14 days), and FSH combined with progesterone(Ovaron^ⓡ, 10 ㎎ IM for 10 days) and PGF_2α(Lutalyse^ⓡ, 3 ㎎ IM at first FSH injection). FSH for inducing estrus and superovulation was given a total 20 ㎎ intramuscularilly in decreasing dosage injections twice daily over 4 days. The MAPs were withdrawn at the 3rd day of FSH injection. Estrus observations were conducted every 6 hours from last FSH injection for 24 hours by placing the does with fertile male goats. Estrus and superovulation were more successfully induced with treatment of MAP + FSH than other treatments(FSH only, or progesterone + PGF_2α + FSH) (estrus induction; 100 vs 42.8 and 71.4%, ovulation points; 11.4 vs 5.4 and 4.4, respectively). The effect of gonadotropin releasing hormone(GnRH) on the ovulation rate was also examined. However, no difference was observed for inducing ovulation with treatment or dosage(100 ㎍ 200 ㎍) of GnRH. Low midline laparotomies were performed, and then ovarian responses (ovulations and follicular development) were examined by exteriorization of the reproductive tracts. Ova were recovered from oviducts by retrograde flushing 60-146 hours after MAP removal, and were classified the developmental stages. Overall 66.1% (236/357) of recovery rate was obtained from 30 superovulated does. The optimal recovery time of microinjectable 1-cell zygotes was approximately 72-76 hours after MAP removal.
Enhanced SMAD1 Signaling Contributes to Impairments of Early Development in CFC-iPSCs.
Han, Kyu-Min,Kim, Seung-Kyoon,Kim, Dongkyu,Choi, Jung-Yun,Im, Ilkyun,Hwang, Kyu-Seok,Kim, Cheol-Hee,Lee, Beom Hee,Yoo, Han-Wook,Han, Yong-Mahn AlphaMed Press 2015 Stem Cells Vol.33 No.5
<P>Cardio-facio-cutaneous (CFC) syndrome is a developmental disorder caused by constitutively active ERK signaling manifesting mainly from BRAF mutations. Little is known about the role of elevated ERK signaling in CFC syndrome during early development. Here, we show that both SMAD1 and ERK signaling pathways may contribute to the developmental defects in CFC syndrome. Induced pluripotent stem cells (iPSCs) derived from dermal fibroblasts of a CFC syndrome patient (CFC-iPSCs) revealed early developmental defects in embryoid body (EB) development, β-catenin localization, and neuronal differentiation. Both SMAD1 and ERK signalings were significantly activated in CFC-iPSCs during EB formation. Most of the β-catenin was dissociated from the membrane and preferentially localized into the nucleus in CFC-EBs. Furthermore, activation of SMAD1 signaling recapitulated early developmental defects in wild-type iPSCs. Intriguingly, inhibition of SMAD1 signaling in CFC-iPSCs rescued aberrant EB morphology, impaired neuronal differentiation, and altered β-catenin localization. These results suggest that SMAD1 signaling may be a key pathway contributing the pathogenesis of CFC syndrome during early development.</P>
Transmission of Bovine β-Casein/Human Lactoferrin Fusion Gene in Transgenic Cattle
Yong-Mahn Han,Deog-Bon Koo,Jung-Sun Park,Young-Hun Kim,Kea-Joung Lee,Kyung-Kwang Lee 한국동물생명공학회(구 한국동물번식학회) 2005 Reproductive & developmental biology Vol.29 No.4
This study was conducted to test whether the transgenic cattle pass the transgene to their progeny through germ cells, and whether the transgene is expressed in the mammary gland of ransgenic cows. Two male ransgenic calves were born from IVF-derived embryos injected with bovine β-casein/human lactoferrin fusion gene and then grew up to be reproducible. Semen was collected from a transgenic bull after 18 mon of age and then frozen. Bovine oocytes matured in vitro were fertilized with spermatozoa of the transgenic bull and cultured in 50㎕ drops of CRlaa medium supplemented with 3 mg/mL BSA. After 48 h of culture, cleaved embryos were determined for the presence of transgenes by DNA polymerase chain reaction (PCR). Proportion of transgene positives among bovine embryos fertilized with sperm of the transgenic bull was 20.9% (28/134). One of transgenic bulls did not produce transgenic sperm. Out of 34 calves produced from recipient heifers inseminated with semen of the other bull, 3 (8.8%) were transgenic animals (2 females and 1 male). Thus, one transgenic bull showed a low transmission frequency below Mendelian levels in both the IVF-derived embryos and his progeny. It was demonstrated by Southern blot that copy numbers of the transgene in the transgenic progeny enhanced about 1.8 times as compared to those of the founder bull The results demonstrate that the transgenic bull carrying human lactoferrin gene could pass his transgene to the progeny through germ cells, although he is a germ-line mosaic.
Transmission of Bovine $\beta-Casein/Human$ Lactoferrin Fusion Gene in Transgenic Cattle
Han Yong-Mahn,Koo Deog-Bon,Park Jung-Sun,Kim Young-Hun,Lee Kea-Joung,Lee Kyung-Kwang The Korean Society of Animal Reproduction 2005 Reproductive & developmental biology Vol.29 No.4
This study was conducted to test whether the transgenic cattle pass the transgene to their progeny through germ cells, and whether the transgene is expressed in the mammary gland of ransgenic cows. Two male ransgenic calves were born from IVF-derived embryos injected with bovine $\beta-casein/human$ lactoferrin fusion gene and then grew up to be reproducible. Semen was collected from a transgenic bull after 18 mon of age and then frozen. Bovine oocytes matured in vitro were fertilized with spermatozoa of the transgenic bull and cultured in $50\;{\mu}L$ drops of CRlaa medium supplemented with 3 mg/mL BSA. After 48 h of culture, cleaved embryos were determined for the presence of transgenes by DNA polymerase chain reaction (PCR). Proportion of transgene positives among bovine embryos fertilized with sperm of the transgenic bull was $20.9\%$ (28/134). One of transgenic bulls did not produce transgenic sperm. Out of 34 calves produced from recipient heifers inseminated with semen of the other bull, 3 $(8.8\%)$ were transgenic animals (2 females and 1 male). Thus, one transgenic bull showed a low transmission frequency below Mendelian levels in both the IVF-derived embryos and his progeny. It was demonstrated by Southern blot that copy numbers of the transgene in the transgenic progeny enhanced about 1.8 times as compared to those of the founder bull The results demonstrate that the transgenic bull carrying human lactoferrin gene could pass his transgene to the progeny through germ cells, although he is a germ-line mosaic.