RESUMO
The expression and localization of the human sperm protein hCAP-18/SOB3 were evaluated in human testis and epididymis through in situ hybridization and immunohistochemistry with the use of an anti-recombinant hCAP-18/SOB3 polyclonal antibody. Both hCAP-18/SOB3 messenger RNA and hCAP-18/SOB3 protein were detected in testis germinal cells (from late spermatogonia to spermatozoa) and in the epididymal epithelium. This localization is in agreement with the antimicrobial properties previously described and supports its involvement in zona pellucida binding, as we had previously suggested.
Assuntos
Peptídeos Catiônicos Antimicrobianos/metabolismo , Epididimo/metabolismo , Espermatozoides/metabolismo , Testículo/metabolismo , Catelicidinas , Humanos , Masculino , Distribuição TecidualRESUMO
We screened a mouse germinal cell expression library with a probe derived from Sob1, a human testis-specific cDNA, and identified 2P1, a new mouse cDNA. A database search revealed that 2P1 was 91% identical to ORF1 of E3-3, a rat gene probably involved in the regulation of alternative splicing. Sequencing showed that 2P1 has a destabilization motif in its 3'-untranslated region. Northern blotting showed strong gene expression in the testis and weak expression in the epididymis, with no signal detected in other tissues. RT-PCR analysis confirmed testis and epididymis expression. In situ hybridization revealed that 2P1 mRNA was absent in spermatogonia but expressed in spermatocytes. This last result was confirmed by RT-PCR of FACS isolated primary spermatocytes (pachytene stage). Using RT-PCR, purified spermatids were also shown to express 2P1.
