Cell-biological analyses on the mechanism of infertility induced in gene knockout mice and sperm preparation leading to oocyte activation.

主要研究结果如下：1.基因敲除小鼠不育的机制。1)GOPC -/-精子（无顶体头部）不能识别MN 13的贮存程度，但不能识别DNA染色质的破坏程度。这有力地表明MN 13分子在精子头部储存的重要性。2)Gc/ -/-精子的不育（畸形精子）并不反映MN 13的储存程度。3)Caf-1 -/-精子的不育（少弱畸形精子症）本质上是由于支持细胞与生殖细胞之间相互作用的失败。支持细胞中维甲酸X受体β的功能障碍介导了这一失败，导致精子发生在早期精子伸长阶段停止。3)BREK -/-不育主要是由于未成熟的生殖细胞凋亡和早期精子细胞外质特化不良所致。2.精子制备，引发，导致卵母细胞激活。1)免疫球蛋白超家族蛋白质，bigging和MC 31/CE 9，进行了睾丸后修饰，降低了顶体反应后的分子量。这是由于去糖基化。2)通过对顶体内转运途径和MN 9/equatorin行为的分析表明，MN 9/equatorin定位于顶体内膜，由于其赤道段的染色能力而成为评价顶体反应过程中顶体状态的稳定标记。3)核周膜蛋白MN 13由于其顶体后区的染色能力，在精卵融合水平上也是一个稳定的评价精子状态的标志物。3.精子成熟相关的卡米汀。附睾中依赖于肉毒碱的精子成熟被发现依赖于肉毒碱转运蛋白OCTN 2在附睾双头中的表达。

The major results obtained were as follows.1.The mechanisms of the infertility found in gene knockout mice. 1) Inability of GOPC -/- sperm (acrosome-less head) paralleled to the degree of MN13 storage but not to the disruption degree of DNA chromatin. This strongly suggests the importance of MN13 molecule storage in ther sperm head. 2) The infertility of Gc/ -/- sperm (teratozoospermia) did not reflect the degree of MN13 storage. 3) The infertility of Caf-1 -/- sperm (oligo-astheno-teratozoospermia) was essentially due to the failure between Sertoli cell-germ cell interaction. This failure was mediated by the dysfunction of retinoid X receptor beta in Sertoli cells, causing spermiogenesis arrest at early elongating spermatic stage. 3) The infertility of BREK -/- was essentially due to apoptosis in premature germ cells and ill formation of ectoplasmic specialization at early elongating spermatids. 2.Sperm preparation, priming, leading to oocyte activation. 1) Immunoglobulin superfamily proteins, besieging and MC31 /CE9, were posttestically modified, reducing the molecular weight after the acrosome reaction. This was due to deglycosylation. 2) Analyses to detect the intracrosomla transport route to developing acrosomal membranes and the behavior of MN9/equatorin indicated that MN9/equatorin locates to the inner acrosomal membrane and becomes a stable marker to evaluate the acrosome status during acrosome reaction because of the staining ability of the equatorial segment. 3) Perinuclear theca protein MN13 was also a stable marker to evaluate the sperm status at the level of sperm-egg fusion because of the staining ability of the postacrosomal region. 3.Sperm maturation-related camitine. Carnitine-dependent sperm maturation in the epididymis was found to depend on the expression of carnitine transporter OCTN2 in the dital caput epididymis.