Project/Area Number |
10460128
|
Research Category |
Grant-in-Aid for Scientific Research (B).
|
Allocation Type | Single-year Grants |
Section | 一般 |
Research Field |
Basic veterinary science/Basic zootechnical science
|
Research Institution | Shinshu University |
Principal Investigator |
ONO Tamao Fac.of Agriculture Shinshu University Associate Professor, 農学部, 助教授 (10177264)
|
Co-Investigator(Kenkyū-buntansha) |
TABUCHI Akira Fac.of Agr., Shinshu Univ. Associate Professor, 農学部, 助教授 (50236725)
HIRAMATSU Kohzy Fac.of Agr., Shinshu Univ. Associate Professor, 農学部, 助教授 (80238386)
TAKAGI Yuji Fac.of Agr., Shinshu Univ. Associate Professor, 農学部, 助教授 (20226757)
SHIMADA Kiyoshi Nagoya Univ. Professor, 院生命農学研究科, 教授 (40065579)
TSUDZUKI Masaoki Hiroshima Univ. Associate Professor, 生物生産学部, 助教授 (70212058)
|
Project Period (FY) |
1998 – 2000
|
Project Status |
Completed (Fiscal Year 2000)
|
Budget Amount *help |
¥10,600,000 (Direct Cost: ¥10,600,000)
Fiscal Year 2000: ¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1999: ¥2,800,000 (Direct Cost: ¥2,800,000)
Fiscal Year 1998: ¥5,600,000 (Direct Cost: ¥5,600,000)
|
Keywords | Japanese quail / chicken / embryo development / gonad / primordial germ cells / chimera / antibodies / PCR |
Research Abstract |
Interspecific transfers of PGCs have been performed and the transferred PGCs migrated into gonadal tissues of the host animal, but there was no evidence that those PGCs differentiated into functional gametes thereafter. If their PGCs became functional, endangered species could be rescued by transferring their PGCs into the embryos of a more prolific species. In order to directly investigate the transferred PGCs in the recipient animal, germ cell markers are required that can distinguish between cells of exogenous and endogenous origins. In this study, isolated quail and PGCs were transferred into chick embryos, and vise vasa. A progeny test and an analysis of the sperm DNA were performed to examine the donor-derived-gamete. To distinguish between quail and chick PGCs, we evaluated three monoclonal antibodies (mAbs), QCR1, QB2 and 2C9. QCR1 and QB2 were quail-specific mAbs and 2C9 was a chick-specific mAb, showing development-specific expression. Transferred quail PGCs settled in the ch
… More
ick embryonic gonads and constituted 14.2% of total PGCs in the gonad. A transfused male hatchling contained two populations of spermatogonia, derived from exogenous quail and endogenous chick PGCs. Among 111 chick embryos transfused with quail PGCs, 22 (19.8%) hatched, and 6 males and 2 females reached maturity. In PCR analysis, quail-PGC-derived W-chromosomal sperm was observed in transfused roosters. Transferred chick PGCs settled in the quail embryonic gonads and constituted 5.6% of total PGCs in the gonad. Offspring from crosses between PGC-transferred female quail and roosters exhibited PGCs positively labeled with mAb QCR1, indicating that these were hybrids resulting from the fertilization of female quail ova by sperm of roosters. In chick PGC-transfused quail, the chicken Z-chromosome-specific band was detected by PCR in 7-day embryos, the gonadal tissues of 7-day hatchlings, and the spermatozoa of mature males. We have also designed PCR primers distinguishable between quail and chicken genomic DNA.Chicken genome specific band was observed in DNA of semen taken from chimeric quail. Less
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