Immunohistochemistry of secretory particles ('vesiculosomes') from the epithelium of bovine seminal vesicles and ampulla of the vas deferens.
Secretory particles, designated as 'vesiculosomes', were isolated and purified from bovine seminal vesicle fluid. The protein pattern was characterized using one- and two-dimensional gel electrophoresis. Native vesiculosome fractions were used for immunization of female New Zealand rabbits. The resulting vesiculosome antiserum was used for immunohistochemical studies, designed to obtain data on the intracellular formation, the localization and the potential release from secretory cells of the respective proteins present in these particles. The specificity and sensitivity of the antiserum was tested by Western blot analysis and by an ELISA system. Western blot analysis of isolated vesiculosomes separated on SDS-PAGE showed several heterogeneous immunoreactive bands in the range of 16 kD and above 66 kD, indicating a polyvalent mixture of immunoglobulins. Immunohistochemical staining of different bovine tissues, nevertheless, revealed a strong reaction exclusively at the apical portion of secretory cells of the seminal vesicle and the ampulla of the vas deferens and of intraluminal secretion. The antiserum recognizes both secretory granules and plasma membrane proteins of seminal vesicle epithelium, supporting the concept of vesiculosomes to represent cytoplasmic particles formed from plasma membranes surrounding apical 'blebs' of the cells that are released physiologically during secretion.
Renneberg
H
H
Department of Anatomy and Cell Biology, Philipps Universität Marburg, Germany.
Konrad
L
L
Aumüller
G
G
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay; D004848:Epithelium / Q000378:metabolism / Q000648:ultrastructure; D006801:Humans; D007106:Immune Sera; D007150:Immunohistochemistry; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism*; D016253:Microscopy, Immunoelectron; D011817:Rabbits; D012669:Seminal Vesicles / Q000378:metabolism* / Q000648:ultrastructure; D014649:Vas Deferens / Q000378:metabolism* / Q000648:ultrastructure
D007106:Immune Sera; D008565:Membrane Proteins; D000251:Adenosine Triphosphatases