| Project/Area Number |
07556064
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (A)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 試験 |
|---|
| Research Field |
Applied animal science
|
|---|
| Research Institution | The University of Tokyo |
|---|
Principal Investigator |
KUROHMARU Masamichi Graduate School of Agricultural and Life Sciences, Associate Professor, 大学院・農学生命科学研究科, 助教授 (00148636)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
HONDO Eiichi Obihiro University of Agriculture and Veterinary Medicine, Department of Veterin, 畜産学部, 助手 (30271745)
OGURA Atsuo National Institute of Health, Department of Veterinary Science, Chief Research S, 実験動物開発室, 主任研究員 (20194524)
OGAWA Kenji The Institute of Physical and Chemical Research, Laboratory, Animal Research Cen, 動物試験室, 研究員 (50251418)
HAYASHI Yoshihiro Graduate School of Agricultural and Life Sciences, Professor, 大学院・農学生命科学研究科, 教授 (90092303)
|
|---|
| Project Period (FY) |
1995 – 1996
|
| Project Status |
Completed (Fiscal Year 1996)
|
| Budget Amount *help |
¥11,900,000 (Direct Cost: ¥11,900,000)
Fiscal Year 1996: ¥5,400,000 (Direct Cost: ¥5,400,000)
Fiscal Year 1995: ¥6,500,000 (Direct Cost: ¥6,500,000)
|
|---|
| Keywords | spermatid / spermatocyte / cultured sertoli cells / NGF / cyclosporin A / follistatin / laminin / elutriation / NGFレセプター / エルトリエーション / DNA合成 |
|---|
| Research Abstract |
1. First, we investigated the effect of extracellular matrix on the adhesion, morphology and proliferation of cultured cells. As a result, Laminin was the most effective among the substrate examined. 2. Mixed cell suspensions, prepared by enzymatic digestion of adult rat testicular tissues, were separated into five fractions according to their differences in sedimentation velocity by centrifugal elutriation, a counterflow centrifugation technique. Through these processes, we obtained viable round spermatids. MTT assay was done at 24 hr intervals after plating to examine the cytotoxic effects of cyclosporin A on round spermatids. Cyclosporin A was shown to have little or no effects on metabolic activity of round spermatids in vitro. 3. We examined whether frozen-thawed mouse round spermatids can fertilize oocytes and contribute to normal embryo development. It was demonstrated that mouse round spermatids can be cryopreserved for production of normal offspring. 4. The expression of NGF receptor was recognized in cultured Sertoli cells by RT-PCR method. In the co-culture of spermatids and NGF-added cultured Sertoli cells, DNA synthesis of spermatids was significantly increased. 5. The appearance of immunoreactive follistatin in rat testis was examined by immunohistochemistry using an antiserum raised against synthetic follistatin peptide. Follistatin immunoreactivity was not found in Sertoli and Leydig cells, while it was clearly detected in the cytoplasm and nucleus of late pachytene spermatocytes. Although the reaction in the cytoplasm desappeared after meiosis, it continued intensely in the nucleus from pachytene spermatocytes to round spermatids. 6. Additionaly, the germ cells of birds and reptiles were studied mainly from a morphological viewpoint.
|
