AIRE1A might be involved in cyclin B2 degradation in testicular lysates.

The autoimmune regulator gene Aire shows predominant expression in thymus and other immunologically relevant tissues, and is assigned the major function of programming autoreactive T-cell deletion. However, the expression of this gene in tissues outside the immune system raises a question about its possible function beyond the T-cell deletion dogma. We detected Aire in mouse testis, and the expression of AIRE protein was remarkably high in postmeiotic germ cells. Sequencing results indicate that testis expressed Aire variant 1a. AIRE could be detected in spermatozoa, with heavy localization on the principal acrosomal domains. Mouse oocytes stained negatively for AIRE before fertilization, but stained positively for AIRE 30 min after fertilization. In the zygote, the levels of AIRE correlated negatively with cyclin B2 levels. Goat testicular lysates spiked with recombinant human AIRE exhibited augmented cyclin B2 degradation in the presence of protease inhibitors, which was inhibited by MG-132, indicating the operation of proteasomal pathways. Thus, this study identifies a correlation between the presence of AIRE and proteasomal breakdown of cyclin B2, which leads us to speculate that cyclin B2 could be a target of AIRE's E3-ubiquitin ligase activity.

Spatio-temporal organization of Vam6P and SNAP on mouse spermatozoa and their involvement in sperm-zona pellucida interactions.

Acrosomal assembly during spermatogenesis and acrosome reaction during sperm-oocyte interaction are unique events of vesicle synthesis, transport, and fusion leading to fertilization. SNARE complex formation is essential for membrane fusion, and vesicle-associated (v-) SNARE intertwines with target membrane (t-) SNARE to form a coiled coil that bridges two membranes and facilitates fusion. We detected messages of Vam6P and SNAP in mammalian testis and epididymis. Vam6P and SNAP were detected in a temporally organized fashion on the spermatozoa from testis and epididymis, which showed accumulation on the principal acrosomal domains during capacitation. Vam6P and SNAP were shed off from the principal acrosomal domain after acrosome reaction, but the equatorial and the post-acrosomal domains retained these proteins. Antibodies to VAMP and SNAP inhibited sperm-zona pellucida interaction, suggesting their possible involvement in sperm membrane vesiculation.

Impaired E-cadherin expression in human spermatozoa in a male factor infertility subset signifies E-cadherin-mediated adhesion mechanisms operative in sperm-oolemma interactions.

Cadherins comprise a family of calcium-dependent glycoproteins that function in mediating cell-cell adhesion in virtually all solid tissues of multicellular organisms. We have examined the presence of a cadherin on spermatozoon and its possible involvement in sperm-oocyte interaction. Spermatozoa from fertile human subjects showed the presence of E-cadherin on its head domains, detectable only after permeabilization of the surface membranes. On the contrary, spermatozoa from oligozoospermic subjects did not possess E-cadherin on their principal acrosomal and equatorial domains. Immunoprecipitation and Western blot studies also showed the presence of E-cadherin in spermatozoa from fertile males and its absence in oligozoospermic males. Using RT-PCR, we detected E-cadherin message in the round cells of fertile males, which was absent in the cells from oligozoospermic males. The presence of anti-E-cadherin antibody brought about quantitative reduction in the sperm-oocyte binding in vitro. These observations indicate the possibility of the interplay of a cadherin-dependent homophilic and/or heterophilic adhesion interaction between spermatozoa and oocyte during fertilization. The absence of a key adhesion molecule in a human male infertility disorder points towards genetic defects causing failure in gamete interactions.