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Role of the platelet‐activating factor and its receptor in the proliferative regulation of bovine ovarian granulosa cells

机译:血小板活化因子及其受体在牛卵巢颗粒细胞增殖调控中的作用

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摘要

Proliferation of granulosa cells and their withdrawal from the cell cycle may regulate follicular ovulation. Antagonists of platelet‐activating factor (PAF) and its receptor (PAFr) inhibit follicle rupture. : Thus, PAF and PAFr may be involved in proliferative regulation of granulosa cells; however, expression of PAFr in these cells is unknown. : The aim of this study was to investigate the presence of PAFr and the effect of PAF on proliferation of cultured bovine granulosa cells using real‐time polymerase chain reaction to assay steady‐state level of mRNA, immunocytochemistry to quantify PAFr protein and proliferating cell nuclear antigen protein by flow cytometry. : We found that granulosa cells express PAFr transcripts and protein. PAF presence did not change the concentration of PAFr mRNA or PAFr protein. Granulosa cells responded to PAF doses of 10 and 50 n with increasing proportions of cells entering G /G phase, as well as a significant expansion of total cell numbers. Rise in G /G ‐phase cells was accompanied by a decline in proliferating cell nuclear antigen protein expression, and these effects could be suspended by simultaneous PAFr blockage. The results provide clear evidence for expression of PAFr in bovine granulosa cells and its functional involvement in PAF/PAFr‐mediated stimulation of cell recruitment. : PAF antagonists are suggested to disturb this regulative activity of PAF and to contribute in this way to blockage of ovulation.
机译:颗粒细胞的增殖及其从细胞周期中退出可能调节卵泡排卵。血小板活化因子(PAF)及其受体(PAFr)的拮抗剂可抑制卵泡破裂。 :因此,PAF和PAFr可能参与颗粒细胞的增殖调控;然而,PAFr在这些细胞中的表达是未知的。 :这项研究的目的是使用实时聚合酶链反应测定mRNA的稳态水平,免疫细胞化学法定量PAFr蛋白和增殖细胞核,研究PAFr的存在以及PAF对培养的牛颗粒细胞增殖的影响。流式细胞仪检测抗原蛋白。 :我们发现颗粒细胞表达PAFr转录本和蛋白质。 PAF的存在不会改变PAFr mRNA或PAFr蛋白的浓度。颗粒细胞对10和50 n PAF剂量有反应,进入G / G期的细胞比例增加,并且总细胞数显着增加。 G / G期细胞的上升伴随着增殖性细胞核抗原蛋白表达的下降,并且这些作用可以通过同时的PAFr阻断而中止。结果为牛颗粒细胞中PAFr的表达及其在PAF / PAFr介导的细胞募集刺激中的功能涉及提供了明确的证据。 :建议PAF拮抗剂干扰PAF的这种调节活性,并以这种方式有助于排卵。

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