Apoptosis of Bovine Granulosa Cells After Serum Withdrawal Is Mediated by Fas Antigen (CD95) and Fas Ligand1
Apoptosis of Bovine Granulosa Cells After Serum Withdrawal Is Mediated by Fas Antigen (CD95) and Fas Ligand1
复制标题
Fas 抗原 (CD95) 和 Fas 配体 1 介导血清撤药后牛颗粒细胞凋亡
作者：
Che‐lin Hu;R. Cowan;R. Harman;D. A. Porter;S. Quirk
摘要卵巢卵泡闭锁是由颗粒细胞和卵泡膜细胞凋亡引起的。Fas抗原（Fas），一种细胞表面受体，当被Fas配体（FasL）激活时触发凋亡，可能参与这一过程。Fas通路在介导血清戒断诱导的颗粒细胞凋亡中的可能作用进行了研究。将从5- 10-mm牛卵泡收集的颗粒细胞在含DMEM-F12的血清中培养3天，去除血清，并在去除血清后的不同时间计数活细胞。细胞死亡在血清去除后6小时显著增加（21% ± 7%; P < 0.05 vs. 0 h），并持续增加至24小时（43% ± 6%）。72 h内未观察到细胞死亡进一步增加。磷脂酰丝氨酸的易位的膜联蛋白V结合的细胞膜的外表面的检测表明，细胞凋亡死亡。定量逆转录聚合酶链反应检测显示，在停药后3 h，Fas mRNA水平无明显变化，但FasL mRNA水平增加了4.7倍（P < 0.05）。FasL mRNA在24 h内保持升高，并在48 h恢复至基础水平。免疫组化染色显示Fas和FasL蛋白在3 h内在细胞表面增加，并在12 h（最后检测时间点）内保持升高。FasL与Fas的结合用两种与FasL的细胞外结构域结合的试剂阻断：抗FasL抗体和Fas：Fc，一种由人免疫球蛋白G的Fc部分和人Fas的细胞外结构域组成的嵌合蛋白。在血清去除后24小时，抗FasL抗体和Fas：Fc分别使细胞死亡减少55% ± 10%和34% ± 12%（与无阻断蛋白相比，P < 0.05）。总之，血清撤除诱导的牛颗粒细胞凋亡至少部分由Fas/FasL相互作用介导。这些结果与Fas在自分泌或旁分泌途径中触发卵巢卵泡闭锁的潜在作用一致。
Abstract Ovarian follicular atresia occurs by apoptosis of granulosa and theca cells. The Fas antigen (Fas), a cell surface receptor that triggers apoptosis when activated by Fas ligand (FasL), may be involved in this process. A possible role of the Fas pathway in mediating serum withdrawal-induced apoptosis of granulosa cells was examined. Granulosa cells collected from 5- to 10-mm bovine follicles were cultured in DMEM-F12 containing serum for 3 days, deprived of serum, and live cells were counted at various times after serum withdrawal. Cell death increased significantly 6 h after serum withdrawal (21% ± 7%; P < 0.05 vs. 0 h) and continued to increase until 24 h (43% ± 6%). No further increases in cell death were observed through 72 h. Detection of the translocation of phosphatidylserine to the outer surface of the cell membrane by annexin V binding indicated that cells died by apoptosis. Quantitative reverse transcriptase-polymerase chain reaction assays showed no changes in Fas mRNA levels but a 4.7-fold increase in FasL mRNA 3 h after serum withdrawal (P < 0.05 vs. 0 h). FasL mRNA remained elevated through 24 h and returned to basal levels at 48 h. Immunohistochemical staining showed that both Fas and FasL protein increased on the cell surface within 3 h and remained elevated through 12 h (the last time point tested). Binding of FasL to Fas was blocked with two reagents that bind to the extracellular domain of FasL: an anti-FasL antibody and Fas:Fc, a chimeric protein consisting of the Fc portion of human immunoglobulin G and the extracellular domain of human Fas. Cell death 24 h after serum withdrawal was reduced 55% ± 10% and 34% ± 12% by anti-FasL antibody and Fas:Fc, respectively (P < 0.05 vs. no blocking protein). In conclusion, serum withdrawal-induced apoptosis of bovine granulosa cells is mediated at least partially by Fas/FasL interactions. These results are consistent with a potential role of Fas in an autocrine or paracrine pathway to trigger ovarian follicular atresia.
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影响因子：
4.4
作者：
M. Faris;Niels Kokot;K. Latinis;S. Kasibhatla;Douglas R. Green;G. Koretzky;Andre E. Nel
通讯作者：
M. Faris;Niels Kokot;K. Latinis;S. Kasibhatla;Douglas R. Green;G. Koretzky;Andre E. Nel
影响因子：
3.6
作者：
S. Quirk;R. Cowan;S. Joshi;K. Henrikson;K. Henrikson
通讯作者：
S. Quirk;R. Cowan;S. Joshi;K. Henrikson;K. Henrikson
影响因子：
20.3
作者：
J. Richards
通讯作者：
J. Richards
影响因子：
4.8
作者：
K. I. Tilly;Subhadra Banerjee;Partha P. Banerjee;Jonathan L. Tilly
通讯作者：
K. I. Tilly;Subhadra Banerjee;Partha P. Banerjee;Jonathan L. Tilly
整个月经周期和绝经期人类卵巢内白细胞群的定位和特征。
DOI：
10.1093/oxfordjournals.humrep.a019256
发表时间：
1996
期刊：
Human reproduction (Oxford, England)
影响因子：
--
作者：
Best,CL;Pudney,J;Welch,WR;Burger,N;Hill,JA
通讯作者：
Hill,JA