Expression of hck-tr, a truncated form of the src-related tyrosine kinase hck, in bovine spermatozoa and testis.
In bull testicular haploid germ cells, an mRNA encoding for hck was detected in addition to another one encoding for hck-tr, a truncated form of the tyrosine kinase hck. As the transcripts were expressed in spermatids, we tried to determine whether hck-tr is present in mature bovine spermatozoa. Two polyclonal antibodies were produced against peptides specific to the N- and C-terminal portions of the truncated protein. Western blot analyses confirmed the presence of hck-tr in total protein extracts of ejaculated bull spermatozoa, and sub-cellular fractionation experiments suggest its presence in both head and flagellum. The truncated protein appears tightly associated with cytoskeletal elements as it could be extracted only with SDS under reducing conditions. When assessed by indirect immunofluorescence, hck-tr was mostly localized at the acrosomal area of the sperm cell and a similar localization was observed on demembranated spermatozoa. Immunohistochemical studies on testis sections revealed protein expression in spermatocytes as well as in round and elongating spermatids. The results presented in this study clearly show the presence of mRNAs encoding for hck and hck-tr in testicular germ cells; hck-tr being translated during spermatogenesis and expressed on mature ejaculated bull spermatozoa.
Bordeleau
Louis-Jean
LJ
Département d'Obstétrique/Gynécologie, Centre de Recherche en Biologie de la Reproduction, Université Laval and Ontogénie et Reproduction, Centre de recherche du CHUQ-CHUL, Québec, Canada.
Leclerc
Pierre
P
D000177:Acrosome / Q000201:enzymology*; D000818:Animals; D002417:Cattle; D006238:Haploidy; D007527:Isoenzymes / Q000096:biosynthesis; D008297:Male; D009928:Organ Specificity / Q000502:physiology; D051572:Proto-Oncogene Proteins c-hck / Q000096:biosynthesis*; D013082:Sperm Tail / Q000201:enzymology*; D013091:Spermatogenesis / Q000502:physiology*; D013737:Testis / Q000201:enzymology*
D007527:Isoenzymes; D051572:Proto-Oncogene Proteins c-hck