Analysis of dymanics and function of Sertoli cell intercellular junction

在2005财政年度，针对支持细胞和精子之间外质特化表达的蛋白质的抗体被微注射到精管腔中。除实验外，对成年雄性小鼠给予雌激素、类雌激素化学物质、抗雌激素（如ICI 182.780）、抗雄激素（如氟他胺），并观察其外质特异性。结果表明，ICI 182.780和氟他胺诱导酪氨酸磷酸化，并在特化位点上表达。另一方面，17 -雌二醇、双酚A和己烯雌酚降低了接触素的表达。这些结果已经发表。2006财年主要进行微注射实验。将抗nectin-2抗体、抗nectin-3抗体、抗afadin抗体微量注入精小管腔内，观察支持细胞与精子间外质特化的超微结构变化。结果表明，抗nectin-2抗体和抗nectin-3抗体的作用相似：特异化处的肌动蛋白层被删除，成熟精子从精细胞上皮脱落（过早精子）。微量注射抗黄花素抗体后，未见超微结构改变。结果表明，这种特化的生物学功能是保存成熟精子直至受精。研究结果正在准备发表。

In fiscal year of 2005, antibodies against proteins, which are expressed at ectoplasmic specialization between Sertoli cell and spermatids, were micro-injected into lumina of seminiferous tubules. In addition to the experiments, estrogens, estrogen like chemicals, anti-estrogen such as ICI 182.780, anti-androgen such as flutamide were administered to adult male mice and the ectoplasmic specializations were examined. As the results, ICI 182.780 and flutamide induced tyrosin phosphorylation of cortactin which is expressed at the specialization. On the other hand, 17beta-estradiol, bisphenol A and diethylstilbestrol decreased expression of cortactin. These results have already been published.In fiscal year of 2006, micro-injection experiments were mainly performed. Anti-nectin-2 antibody, anti-nectin-3 antibody, anti-afadin antibody were micro-injected into lumina of seminiferous tubules, and ultrastructural changes of ectoplasmic specializations between Sertoli cell and spermatids were observed. As the results, anti-nectin-2 antibody and anti-nectin-3 antibody showed similar effects: Actin layers at the specialization were deleted, and mature spermatids were exfoliated from seminiferous epithelia (premature spermiation). No ultrastructural changes were observed after micro-injection of anti-afadin antibody. The results suggest that biological function of the specialization is holding mature spermatids until spermiation. The results are now in preparation for publication.