Molecular mechanism of apoptosis of reproductive cells and fertilization

1)为了了解导致MRL/LPR小鼠卵泡闭锁的分子机制，我们研究了细胞凋亡信号转导途径。结果表明，MRL/+小鼠卵母细胞和MRL/LPR是通过Fas受体激活caspase-3。相反，我们发现突变的Fas受体在MRL/LPR小鼠卵母细胞中的异常表达和功能障碍与无法激活caspase级联(尤其是caspase-3)和诱导核DNA片段化有关。这些结果表明，在缺乏启动caspase级联反应的Fas受体缺陷的情况下，mrl/lpr小鼠卵母细胞没有发生凋亡诱导，提示未能诱导卵泡闭锁。2)6种凋亡诱导核苷(ANs)释放到57细胞培养上清液中。更多来自人蜕膜组织的)通过物理化学方法相结合的方法分离。随后，我们证明了AINS可以诱导人白血病细胞Molt4和癌细胞BeWo/GCIY的凋亡，但不能诱导人成纤维细胞WI-38的凋亡。该药物还可诱导人前列腺癌PC3细胞、雌激素不敏感的人乳腺癌MDA-MB-435细胞和人胃癌细胞发生凋亡。细胞凋亡的特征是DNA链断裂和caspase级联激活，尤其是caspase-3。在荷瘤的SCID小鼠体内给药，由于肿瘤细胞的凋亡，最终抑制了肿瘤的生长。3)利用糖探针，我们在猪精子裂解液中发现了几种与寡糖探针结合的蛋白质。根据部分氨基酸序列分析，其中一个是ADAM4的同源物。我们克隆了它的cDNA，并在酵母中以分泌型形式表达了它的重组蛋白。功能分析表明，该蛋白具有与整合素的黏附活性，提示其在受精过程中起作用。较少

1) To understand the molecular mechanisms leading to ovarian follicular atresia in MRL/lpr mouse, we examined the apoptotic signalling pathway. The results indicated that MRL/+ murine oocytes and MRL/lpr is through the Fas receptor followed by the activation of caspase-3. In contrast, we found that the aberrant expression and dysfunction of the mutant Fas receptor in MRL/lpr murine oocytes caused by insertion of the early transposable element into the Fas gene were associated with an inability to activate the caspase cascade (especially caspase-3) and to induce nuclear DNA fragmentation. These findings indicate that the induction of apoptosis in MRL/lpr murine oocytes did not occur in the presence of a defective Fas receptor lacking the death domain to trigger the caspase cascade, suggesting a failure to induce ovarian follicular atresia.2) Six apoptosis-inducing nucleosides (AINs) released into cell culture media of 57.DR-NS (the CD57^+HLA-DR-<bright> natural suppressor cell line deri … More ved from human decidual tissue) were isolated by the combination of physicochemical procedures. Subsequently, we demonstrated that AINs could induce apoptosis 'in human leukemia Molt4 and carcinoma BeWo/GCIY cells but not human fibroblast WI-38 cells. The AINs also induced apoptosis in human prostate cancer PC3 cells, estrogen-non-responsive human breast carcinoma MDA-MB-435 cells, and human gastric carcinoma cells. Apoptosis was characterized by DNA strand breaks and activation of the caspase cascade, especially caspase-3. The administration of AINs into tumor bearing SCID mice culminated in suppression of tumor growth due to apoptosis of tumor cells.3) By using sugar probes, we found several proteins in boar sperm lysate which bind to oligosaccharide probes. One of them was a homologue of ADAM4 based on the analysis of partial amino acid sequence. We cloned its cDNA, and produced its recombinant protein in a secreted form in yeast. Functional analysis indicated that the protein has an adhesion activity to integrin, suggesting its role in fertilization. Less