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首页> 外文期刊>Molecular reproduction and development >Multiparametric study of atresia in ewe antral follicles: histology, flow cytometry, internucleosomal DNA fragmentation, and lysosomal enzyme activities in granulosa cells and follicular fluid.
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Multiparametric study of atresia in ewe antral follicles: histology, flow cytometry, internucleosomal DNA fragmentation, and lysosomal enzyme activities in granulosa cells and follicular fluid.

机译：母羊窦卵泡闭锁的多参数研究：组织学，流式细胞术，核小体间DNA片段化以及颗粒细胞和卵泡液中的溶酶体酶活性。

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The differential quantitative participation of apoptosis and necrosis in ewe antral follicles of 2 sizes, classified into 4 stages of atresia using macroscopic, histologic and steroid quantification methods was assessed. Annexin V binding and propidium iodide uptake was used to detect healthy live cells (Annexin V negative/propidium iodide negative), early apoptotic cells (Annexin V+/propidium iodide-) and necrotic or late apoptotic cells (propidium iodide+). We used internucleosomal DNA fragmentation as a quantitative estimate of apoptosis. Presence and distribution of lysosomal enzymes in follicular fluid and granulosa cells was used as a measure of necrotic cell death. DNA flow cytometry and gel electrophoresis were positively correlated with the progression of atresia, small atretic follicles tend to have higher percentages of internucleosomal cleaved DNA than follicles >6 mm. Annexin/propidium iodide binding also indicated that apoptosis and necrosis increase with atresia progression, generally apoptosis outweighs necrosis in small follicles. Acid phosphatase and glucosaminidase in follicular fluid of 3-6 mm follicles showed no significant modifications between healthy and initially atretic follicles and only a small but significant increase in activity in advanced atretic follicles. Lysosomal enzyme activity in follicles >6 mm in diameter showed positive correlation between atresia stages and the activities of acid phosphatase and glucosaminidase in follicular fluid. A similar size-differential behaviour was found in free or membrane-bound lysosomal enzyme activity of granulosa cells. Necrosis, but principally apoptosis, were present during all stages of follicular maturation, indicating that growth and maturation of ovarian follicles involves a continuous renewal of granulosa cells, regulated by apoptosis. It is suggested that mechanisms regulating this equilibrium participate in the final destiny, whether ovulation or atresia of ovarian follicles.
机译：使用宏观，组织学和类固醇定量方法评估了2种大小的母羊窦卵泡中凋亡和坏死的差异定量参与，分为2个闭锁阶段。 Annexin V结合和碘化丙锭摄取用于检测健康的活细胞（Annexin V阴性/碘化丙啶阴性），早期凋亡细胞（Annexin V + /碘化丙啶-）和坏死或晚期凋亡细胞（碘化丙啶+）。我们使用核小体间DNA片段化作为凋亡的定量评估。卵泡液和颗粒细胞中溶酶体酶的存在和分布被用作坏死细胞死亡的量度。 DNA流式细胞术和凝胶电泳与闭锁的进展呈正相关，小的闭锁卵泡往往比> 6 mm的卵泡具有更高的核糖体间切割DNA百分比。膜联蛋白/碘化丙啶的结合还表明，随着闭锁的进行，细胞凋亡和坏死增加，通常在小卵泡中细胞凋亡超过坏死。 3-6 mm卵泡的卵泡液中的酸性磷酸酶和氨基葡萄糖苷酶在健康的和最初的无定形卵泡之间没有显示出明显的变化，而在晚期的无定形卵泡中，活性只有很小的但显着的增加。直径> 6 mm的卵泡中的溶酶体酶活性显示，闭锁阶段与卵泡液中酸性磷酸酶和氨基葡萄糖苷酶的活性呈正相关。在颗粒细胞的游离或膜结合的溶酶体酶活性中发现了相似的大小差异行为。在卵泡成熟的所有阶段都存在坏死，但主要是细胞凋亡，表明卵巢卵泡的生长和成熟涉及由细胞凋亡调节的颗粒细胞的连续更新。建议调节这种平衡的机制参与最终的命运，无论是卵泡的排卵还是闭锁。

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