Specific localization of RBM1a in the nuclei of all cell types except elongated spermatids within seminiferous tubules of the human

Recent studies have indicated that at least three regions (AZF a-c) on the long arm of the Y-chromosome code for factors are involved in spermatogenes is. One of the candidate genes in the AZFb region is RBM1a, coding for a pr otein with an RNA binding motif In this study, poly clonal antibodies raise d against a 15 amino acid peptide, corresponding to residues 263-304 of the deduced amino acid sequence of RBM1a, has been used to localize the RBM1a protein in the human testis. Immunohistochemistry on normal human testis us ing this RBM1a antibody, localized the antigen to the nuclei of spermatogon ia, primary spermatocytes, and round spermatids but not to the nuclei of el ongated spermatids. The antibody also specifically identified the nuclei of Sertoli cells, although the fluorescence was not as strong as in the germ cell nuclei it identified. No specific fluorescence was seen in the nuclei of either peritubular. endothelial or Leydig cells. Western blot of normal human testicular tissue using the anti-RBM1a antibody gave rise to a single specific band of approximately 55 kDa, corresponding to the expected size of RBM1a. In view of its expression in germ cells, and because RBM1a has an RNA binding domain, RBM1a may be involved in RNA processing, such as RNA s plicing or RNA export which are events necessary for normal spermatogenesis .