REGULATION OF CUMULUS CELL STEROIDOGENESIS BY THE PORCINE OOCYTE AND PRELIMINARY CHARACTERIZATION OF OOCYTE-PRODUCED FACTOR(S)
REGULATION OF CUMULUS CELL STEROIDOGENESIS BY THE PORCINE OOCYTE AND PRELIMINARY CHARACTERIZATION OF OOCYTE-PRODUCED FACTOR(S)
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DOI：
10.1095/biolreprod53.3.670
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发表时间：
1995-09-01
影响因子：
3.6
通讯作者：
ALAK, BM
中科院分区：
文献类型：
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作者：
COSKUN, S;UZUMCU, M;ALAK, BM
本研究旨在调查猪卵母细胞是否产生影响卵丘和壁颗粒细胞类固醇产生的因子，并表征此类因子的生化性质和作用模式。从 2-5 毫米的卵泡中收集猪卵丘-卵母细胞复合物 (COG)，并完整培养或在卵母细胞切除后培养 48 小时。然后测量培养基中的类固醇水平。通过培养裸卵母细胞 48 小时获得的条件培养基经过热处理或木炭提取，并用于培养完整和卵母细胞切除的 COG。与完整 COC 培养物相比，卵母细胞切除培养物中 FSH 刺激的孕酮、20α-OH-孕酮和雌二醇显着较高。与对照相比，与颗粒细胞一起培养的裸露卵母细胞显着抑制黄体酮的产生。此外，用不同数量的裸露卵母细胞（0至300）调节的培养基显着抑制卵母细胞切除的COC产生黄体酮，其方式取决于卵母细胞数量。木炭提取而非热处理显着消除了条件培养基对切除卵母细胞的COG产生孕酮的抑制作用。切除卵母细胞的COC孕酮产生的增加并不伴随着cAMP形成的类似增加。庚醇是一种间隙连接阻断剂，不会改变完整 COG 产生的黄体酮。 总之，猪卵母细胞分泌一种抑制卵丘和壁颗粒细胞类固醇生成的因子。该因子对热稳定，但可通过木炭提取。该因子似乎不会通过间隙连接转移至体细胞，其作用是在 cAMP 形成的下游。
This study was designed to investigate whether porcine oocytes produce a factor(s) that influences cumulus and mural granulosa cell steroid production and to characterize the biochemical nature and mode of action of a such factor(s). Porcine cumulus-oocyte complexes (COG) were collected from 2-5-mm follicles and cultured either intact or after oocytectomy for 48 h. Steroid levels were then measured in the culture media. Conditioned media, obtained by culturing denuded oocytes for 48 h, were subjected to heat treatment or charcoal extraction and utilized to culture intact and oocytectomized COG.FSH-stimulated progesterone, 20 alpha-OH-progesterone, and estradiol were significantly higher in oocytectomized vs. intact COC cultures. Denuded oocytes cultured with granulosa cells significantly inhibited progesterone production compared to control. Also, media conditioned with different numbers of denuded oocytes (0 to 300) significantly inhibited progesterone production by oocytectomized COC in a manner dependent on oocyte number. Charcoal extraction, but not heat treatment, significantly removed the inhibitory effect of the conditioned media on progesterone production by oocytectomized COG, Increased progesterone production by oocytectomized COC was not accompanied by a similar increase in cAMP formation. Heptanol, a gap junction blocker, did not alter progesterone production by intact COG.In conclusion, porcine oocytes secrete a factor(s) that inhibits cumulus and mural granulosa cell steroidogenesis. This factor(s) is heat stable but extractable by charcoal. The factor(s) appears not to be transferred to somatic cells via gap junctions, and its effect is downstream of cAMP formation.