Arnault Emilie, Tosca Lucie, Courtot Anne-Marie, Doussau Mireille, Pesty Arlette, Finaz Catherine, Allemand Isabelle, Lefèvre Brigitte
UMR-S 566, CEA, DSV/iRCM/SCSR/LGAG, INSERM, Université Denis Diderot-Paris 7, Université Paris-Sud, Fontenay aux Roses, France.
Dev Dyn. 2008 Dec;237(12):3892-903. doi: 10.1002/dvdy.21793.
Several studies have shown that apoptotic pathways control fragmentation of unfertilized ovulated oocyte, induced by doxorubicin. But very few have investigated the basis of this process, from prophase I to later stages. Our results revealed the presence of caspase-2(L), caspase-9, and caspase-3 in their zymogen and cleaved forms in the oocyte before meiosis resumption. Caspase-2(L) and caspase-9 were detected in the nucleus of GV-oocytes in a distribution related to chromatin configuration. The inhibition of caspase activity by Z-VAD-fmk accelerated the transition from metaphase I to metaphase II, and caspase-9 and caspase-3 were detected along the meiotic spindle. Surprisingly, Western blot analysis revealed that the three cleaved caspases were present in similar amounts in healthy and fragmented oocytes and caspase inhibition did not prevent doxorubicin-induced apoptosis. Our results suggest that, if cleaved, caspases may be dispensable for final oocyte death and they could be involved in regulating the maturation process.
多项研究表明,凋亡途径控制着阿霉素诱导的未受精排卵卵母细胞的碎片化。但很少有研究从减数分裂前期I到后期探究这一过程的基础。我们的结果显示,在减数分裂恢复前,卵母细胞中存在半胱天冬酶-2(L)、半胱天冬酶-9和半胱天冬酶-3的酶原形式和裂解形式。在GV期卵母细胞的细胞核中检测到半胱天冬酶-2(L)和半胱天冬酶-9,其分布与染色质构型有关。Z-VAD-fmk对半胱天冬酶活性的抑制加速了从减数分裂中期I到减数分裂中期II的转变,并且在减数分裂纺锤体上检测到了半胱天冬酶-9和半胱天冬酶-3。令人惊讶的是,蛋白质印迹分析显示,在健康和碎片化的卵母细胞中,三种裂解的半胱天冬酶含量相似,并且半胱天冬酶抑制并不能阻止阿霉素诱导的细胞凋亡。我们的结果表明,如果发生裂解,半胱天冬酶对于卵母细胞最终死亡可能是可有可无的,并且它们可能参与调节成熟过程。