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

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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.