Molecular chaperone calmegin localization to the endoplasmic reticulum of meiotic and post-meiotic germ cells in the mouse testis.
Calmegin is a testis-specific Ca(2+)-binding protein that is homologous to calnexin. Recently, sperm from transgenic mice lacking calmegin have been shown to be infertile. To further characterize calmegin, we analyzed the precise stage of expression and the intracellular localization of this protein in germ cells during mouse spermatogenesis by an immunoperoxidase technique using the anti-calmegin monoclonal antibody TRA369. Light microscopic immunocytochemistry showed that calmegin appeared in early pachytene spermatocytes, with the highest expression in round and elongating spermatids, and disappeared in the maturation phase of spematids at step 15. Immunoelectron microscopy showed that selective localization was found at the endoplasmic reticulum membrane and the nuclear envelope of spermatogenic cells. During the maturation phase, a dramatic reduction in calmegin occurred in the endoplasmic reticulum of the spermatids, suggesting that the major function of calmegin has been completed by the time spermatids reach step 14. In addition, although the immunoreactivity was completely absent in the calmegin-deficient mutant mouse testis, ultrastructural analysis showed that mature sperm from the knockout mice were normal. This suggests that calmegin is not required for the morphogenesis of male germ cells. Thus, our results suggest that calmegin has a major role in mouse spermatogenesis, and also indicate that this protein would be useful as a maker molecule to study the functional role of the endoplasmic reticulum in the process of spermatid differentiation.
Yoshinaga
K
K
Department of Anatomy and Reproductive Cell Biology, Miyazaki Medical College, Japan. kyoshina@post.miyazaki-med.ac.jp
Tanii
I
I
Toshimori
K
K
D000818:Animals; D002135:Calcium-Binding Proteins / Q000032:analysis*; D037281:Calnexin; D004721:Endoplasmic Reticulum / Q000737:chemistry*; D004822:Epididymis; D005854:Germ Cells / Q000737:chemistry* / Q000648:ultrastructure*; D007150:Immunohistochemistry; D008297:Male; D008540:Meiosis; D008565:Membrane Proteins / Q000032:analysis; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D018613:Microscopy, Confocal; D008854:Microscopy, Electron; D018832:Molecular Chaperones / Q000032:analysis*; D010750:Phosphoproteins / Q000032:analysis; D013091:Spermatogenesis; D013094:Spermatozoa / Q000648:ultrastructure; D013737:Testis / Q000166:cytology*
D002135:Calcium-Binding Proteins; D008565:Membrane Proteins; D018832:Molecular Chaperones; D010750:Phosphoproteins; D037281:Calnexin; C087341:calmegin