1988 Fiscal Year Final Research Report Summary
In vitro analysis of the function in various premature follicles isolated enzymatically from rat ovaries.
| Project/Area Number |
62560294
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
基礎獣医学
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| Research Institution | University of Miyazaki |
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Principal Investigator |
ETOH Teiichi Miyazaki Univ., Facult. Agriculture, 農学部獣医学科, 教授 (70081510)
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| Co-Investigator(Kenkyū-buntansha) |
MURAKAMI Noboru Miyazaki Univ., Facult. Agriculture, 農学部獣医学科, 助手 (80150192)
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| Project Period (FY) |
1987 – 1988
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| Keywords | follicle culture / Method of follicle separation / follicular development / follicnlar function / follicular steroid / PMSG / 新生児卵巣 |
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| Research Abstract |
A single follicle was isolated from rat ovary by digestion with enzymes and filtration with stainless steel wire gauze. The isolation was made use of the method applied in hamster by Roy c Greenwald. The isolated follicles were observed with phase-contrast or scanning-elect-ron microscopes and cultured individually for 4 days in two differentmethod in vitro, such as liquid medium or the same medium on agarose gel seat. The results are as follows. 1.The co-relation between diameter of follicles and the including ova was estimated. With increasing of the follicular diameter to 200 m, the ovular diameter increased to 70 m,but thereafter, the ovular diameter persisted in constant in spite of the follicular growth. The follicles were coated by basement membrane having many small projections, but shortly after the bigin-ing of incuvation, these were decoated the membrane and became a cluster of global cells. 2.Both preantral and small antral follicles in the liquid medium grew to monolayer c
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ellular seat holding cumulus cells and ovum in centre, and sometimes those cumulus cells changed the shape as similar as large antral follicle. And also, PMSG stimulated the changes and progest-erone production as well. 3. Follicles cultured in the medium on agarose gel seat did not grow clearly. This result indicated that the liquid was suitable for follicle incuvation than the agarose one. 4. The small follicles were relatively less sensitive to PMSG than the larger follicles. Ovaries from neonate rats, which were containing exclusively primary and preant-ral follicles, were placed in organ culture medium for 4 to 8 days in order to confirm vi-ability in in vitro circumstance. The follicles in these ovaries were maintained for as long time as 8 days histologically and produced a large amount of 4-androstene-3,17-dione, a precureser of estrogen, in limited period of 5 to 9 days in age. It is suggested that, from these experiments, enzymatically dissociated follicles even in neonate ovaries may be able to stimulate to growth in vitro and this incuvation system may be a useful tool for invest-igating very complex intraovarian endocrine mechanism. Less
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