Characterization of GALNTL5 gene sequence and expression in ovine testes and sperm

The polypeptide N-acetylgalactosaminyltransferase-like protein 5 (GALNTL5), which belongs to the polypeptide N-acetylgalactosaminyltransferase (pp-GalNAc-T) gene family, is a newly identified gene that is specifically expressed in testis and involved in spermatogenesis. However, there is no data showing the existence of GALNTL5 in ram testis at various developmental stages and its influence on sperm motility. Therefore, the objectives of the present study were to evaluate the presence of GALNTL5 in the testis of 3-24 months (M) ram and to investigate the expression of GALNTL5 in spermatozoa with different motilities. We detected a 1602 bp cDNA fragment of GALNTL5 that included a 1326 bp coding sequence, encoding 441 amino acids and 90 and 185 bp of the 5' and 3' untranslated regions, respectively. The GALNTL5 amino acid sequence showed 51.87-83.48% identity with the sequences of proteins from other species. It was detected exclusively in the testis and the levels of both the mRNA and protein were progressively increased with age. Immunohistochemistry further revealed that GALNTL5 specifically localized in the elongating spermatids and spermatozoa, and it was demonstrated to be strongly concentrated in the head, neck, and mid-piece region of spermatozoa by immunocytochemistry. The sperm density and the percentage of live sperm in the high motility group (≥80%) were significantly higher than in the low motility group (≤50%), and the reverse trend was observed with the abnormal sperm. Western blot analysis showed that the protein expression of PGK2, ALDOA, and GALNTL5 were significantly higher in the high motility group than in the low motility group. Overall, the data suggest that GALNTL5 is an important functional molecule during spermatogenesis. Moreover, it is the first to suggest that the expression level of GALNTL5 is positively correlated with the sperm motility.