Binding of a major secretory protein from bull seminal vesicles to bovine spermatozoa.
The seminal vesicles synthesize in an androgen-dependent manner a neutral protein of 13.5 kDa molecular weight that makes up about 40% of their secretion ("major protein"). An antiserum against this protein raised in rabbits was used to localize the antigen within the seminal vesicles. In addition to intraluminal secretion of the seminal vesicles and the ampulla of the vas deferens, ejaculated and ampullary spermatozoa revealed an intense immunoreaction, which was restricted to the neck region of the sperm head and the middle piece, while the principal piece of the tail as well as the sperm head were devoid of immunoreactive material. Comparison of spermatozoa taken from the tail of the epididymis with ampullary spermatozoa showed that about 90% of the latter, but only 10-20% of the former presented this distributional pattern of immunoreactive sites. Epididymal epithelium as well as calf seminal vesicle epithelium showed no immunoreactivity with major protein antiserum. Using a pre-embedding staining technique with gold-labeled primary or secondary antibodies, respectively, no immunostaining could be achieved at the ultrastructural level. Incubation experiments of epididymal spermatozoa in EGTA-containing solutions in the absence of calcium resulted in a gradual labilization and eventual loss of the plasma membrane of the sperm middle piece. After removal of (at least part of) the plasma membrane, bound major protein could be visualized immunohistochemically close to the mitochondria of the middle piece using a gold-labeled primary or secondary antibody. The acceptor site for major protein therefore seems to reside inside the plasma membrane of the sperm middle piece.(ABSTRACT TRUNCATED AT 250 WORDS)
Aumüller
G
G
Department of Anatomy and Cell Biology, Philipps University, Marburg, Federal Republic of Germany.
Vesper
M
M
Seitz
J
J
Kemme
M
M
Scheit
K H
KH
D000818:Animals; D000918:Antibody Specificity; D002417:Cattle; D007150:Immunohistochemistry; D008297:Male; D029584:Prostatic Secretory Proteins; D011485:Protein Binding; D014176:Protein Biosynthesis; D011498:Protein Precursors; D011506:Proteins / Q000276:immunology / Q000378:metabolism*; D029589:Seminal Plasma Proteins; D012669:Seminal Vesicles / Q000378:metabolism*; D013094:Spermatozoa / Q000032:analysis / Q000276:immunology / Q000378:metabolism* / Q000648:ultrastructure
D029584:Prostatic Secretory Proteins; D011498:Protein Precursors; D011506:Proteins; D029589:Seminal Plasma Proteins; C043076:beta-microseminoprotein