Spatio-temporal organization of Vam6P and SNAP on mouse spermatozoa and their involvement in sperm-zona pellucida interactions.
ABSTRACT 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.
Article: Evolution of genes involved in gamete interaction: evidence for positive selection, duplications and losses in vertebrates.[show abstract] [hide abstract]
ABSTRACT: Genes encoding proteins involved in sperm-egg interaction and fertilization exhibit a particularly fast evolution and may participate in prezygotic species isolation , . Some of them (ZP3, ADAM1, ADAM2, ACR and CD9) have individually been shown to evolve under positive selection , , suggesting a role of positive Darwinian selection on sperm-egg interaction. However, the genes involved in this biological function have not been systematically and exhaustively studied with an evolutionary perspective, in particular across vertebrates with internal and external fertilization. Here we show that 33 genes among the 69 that have been experimentally shown to be involved in fertilization in at least one taxon in vertebrates are under positive selection. Moreover, we identified 17 pseudogenes and 39 genes that have at least one duplicate in one species. For 15 genes, we found neither positive selection, nor gene copies or pseudogenes. Genes of teleosts, especially genes involved in sperm-oolemma fusion, appear to be more frequently under positive selection than genes of birds and eutherians. In contrast, pseudogenization, gene loss and gene gain are more frequent in eutherians. Thus, each of the 19 studied vertebrate species exhibits a unique signature characterized by gene gain and loss, as well as position of amino acids under positive selection. Reflecting these clade-specific signatures, teleosts and eutherian mammals are recovered as clades in a parsimony analysis. Interestingly the same analysis places Xenopus apart from teleosts, with which it shares the primitive external fertilization, and locates it along with amniotes (which share internal fertilization), suggesting that external or internal environmental conditions of germ cell interaction may not be the unique factors that drive the evolution of fertilization genes. Our work should improve our understanding of the fertilization process and on the establishment of reproductive barriers, for example by offering new leads for experiments on genes identified as positively selected.PLoS ONE 01/2012; 7(9):e44548. · 4.09 Impact Factor