Summary
Immunoelectron microscopy of the rat seminal vesicle was performed using specific antibodies to secretory proteins. Proteins were precipitated from rat seminal vesicle secretion and were separated by SDS—polyacrylamide gel electrophoresis. Among the great number of bands the two most prominent bands were selected and designated SVS II and IV. Their apparent molecular weights were 48 kDa and 16.5 kDa respectively. The bands were excised from the gels and used for antibody production in rabbits. The respective antisera were used for immunohistochemical studies both at the light and electron microscopic levels in the rat seminal vesicle and the different prostatic lobes in infantile, adult and castrated animals. A positive immunoreaction was observed in seminal vesicle and lateral prostatic epithelium of the intact adult rat, while it was lacking in prepubertal and castrated animals. The subcellular distribution of both proteins was clearly different: SVS II was exclusively confined to the electron dense core of the secretory vacuoles, while SVS IV was detected only in the clear halo surrounding the central granule. It is suggested that the spatial arrangement of both proteins in the seminal vesicle secretion vacuole reflects a particular functional significance of each of these proteins. These proteins may serve as a tool in the study of regulation of androgendependent protein synthesis.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
ANDERSON, D. & PETERSON, C. (1981) High resolution electophoresis of proteins in SDS polyacrylamide gels. InElecrophoresis '81, pp. 41–8. Berlin, New York: De Gruyter.
AUMÜLLER, G. (1979) Prostate gland and seminal vesicles. InHandbuch der Mikroskopischen Anatomie des Menschen (edited by OKSCHE, A. and VOLLRATH, L.), Vol VII/7. Berlin, Heidelberg, New York: Springer.
AUMÜLLER, G., SEITZ, J., HEYNS, W. & FLICKINGER, C. J. (1982) Intracellular localization of Prostatic Binding Protein (PBP) in rat prostate by light and electron microscopic immunocytochemistry.Histochemistry 76, 497–516.
BASKIN, D. G., ERLANDSEN, S. L. & PARSONS, J. A. (1979) Influence of hydrogen peroxide or alcoholic sodium hydroxide on the immunocytochemical detection of growth hormone and prolactin after osmium fixation.J. Histochem. Cytochem. 27, 1290–2.
DRAVLAND, E. & JOSHI, M. S. (1981) Sperm-coating antigens secreted by the epididymis and seminal vesicle of the rat.Biol. Reprod. 25, 649–58.
FLICKINGER, C. J. (1974) Synthesis, intracellular transport and release of secretory protein in the seminal vesicle of the rat, as studied by electron microscope radioautography.Anat. Rec. 180, 407–26.
HARRIS, S. E., MAUSSON P.-E., TULLY, D. B. & BURKHART, B. (1983) Seminal vesicle secretion IV gene: Allelic difference due to a series of 20-base-pair direct tandem repeats within an intron.Proc. natn. Acad. Sci. USA 80, 6460–4.
HIGGINS, S. J., BURCHELL, J. M. & MAINWARING, W. I. P. (1976) Androgen-dependent synthesis of basic secretory proteins by the rat seminal vesicle.Biochem. J. 158, 271–82.
HIGGINS, S. J. & FULLER, F. M. (1981) Effects of testosterone on protein sythesis in rat seminal vesicles analysed by two-dimensional gel electrophoresis.Mol. Cell Endocrinol. 24, 85–101.
HIGGINS, S. J., SMITH, S. E. & WILSON, J. (1982) Development of secretory protein synthesis in the seminal vesicles and ventral prostate of the male rat.Mol. Cell. Endocrinol. 27, 55–65.
KIERSZENBAUM, A. L., DE PHILIP, R. M., SPRUILL, W. A. & TAKENAKA, J. (1983) Isolation and culture of rat seminal vesicle epithelial cells. The use of the secretory protein SVS IV as a functional probe.Expl. Cell. Res. 145, 293–304.
KOENIG, H., LU, C. Y. & BAKAY, R. (1976) Testosterone and 6-N, 2′-O-dibutyryladenosine 3′:5′ cyclic monophosphate stimulate protein and lysosomal enzyme secretion in rat seminal vesicle.Biochem, J. 158, 543–7.
MANN, T. & LUTWAK-MANN, C. A. (1981)Male Reproductive Function and Semen. Themes and Trends in Physiology, Biochemistry and Investigative Andrology. Berlin: Springer.
MATA, L. R. & MAUNSBACH, A. B. (1982) Absorption of secretory protein by the epithelium of hamster seminal vesicle as studied by electron microscope autoradiography.Biol. Cell. 46, 65–75.
MURAKAMI, M., SUGITA, A., SHIMADA, T. & YOSHIMURA, T. (1978) Scanning electron microscope observation of the seminal vesicle in the Japanese monkey, with special reference to intraluminar spermiophagy by macrophages.Arch. Histol. Jap. 41, 275–83.
OSTROWSKI, M. C., KISTLER, M. K. & KISTLER, W. S. (1979) Purification and cell-free synthesis of a major protein from rat seminal vesicle secretion. A potential marker for androgen action.J. biol. Chem. 254, 383–90.
OSTROWSKI, M. C., KISTLER, M. K. & KISTLER, W. S. (1982) Effect of castration on the synthesis of seminal vesicle secretory protein IV in the rat.Biochemistry 21, 3525–9.
RIVA, A., COSSU, M., USAI, E. & TESTA-RIVA, F. (1981) Spermatophagy by epithelial cells of the seminal vesicle and of the ampulla ductus deferentis in man: a scanning and transmission EM study. InOligozoospermia: Recent Progress in Andrology (edited by FRAJESE, F.) pp. 45–53. New York: Raven Press.
SLOT, J. W. & GEUZE, H. J. (1981) Sizing of protein A—colloidal gold probes for immunoelectron microscopy.J. Cell. Biol. 90, 533–6.
STERNBERGER, L. A., HARDY, P. H., CUCULIS, J. J. & MEYER, H.-G. (1970) The unlabelled antibody enzyme method of immunohistochemistry. Properties and preparation of soluble antigen—antibody complex (horseradish peroxidase—anti horseradish peroxidase) and its use in identification of spirochetes.J. Histochem. Cytochem. 18, 315–33.
STERNBERGER, L. A. (1979)Immunocytochemistry.2nd edn. New York: John Wiley.
TENG, C. T. & HARRIS, S. E. (1983) The seminal vesicle secretion gene; detection of S1 nuclease-sensitive sites in supercoiled plasmid p SVS 3.3.DNA 2, 105–11.
THOMPSON, S. A., ROWLEY, D. R. & HEIDGER, P. M. (1979) Fine structural studies of rat seminal vesicle in castrated and intact animals following estrogen treatment.Am. J. Anat. 154, 525–44.
TOTH, M. & ZAKÁR, T. (1977) Studies on the binding of androgens to the cytosol proteins and nuclei of the rat seminal vesicle.Acta biochim. biophys. Acad. Sci. Hung. 12, 1–14.
TOTH, M. & HERTELENDY, F. (1979) Androgen binding and androgen effect in the rat seminal vesicles: studies with testosterone and 19-nortestosterone.J. Steroid Biochem. 11, 1091–7.
TOWBIN, H., STAEHELIN, T. & GORDON, J. (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications.Proc. natn. Acad. Sci. USA 76, 4350–4.
VENABLE, J. H. & COGGESHALL, R. (1965) A simplified lead citrate stain for use in electron microscopy.J. Cell. Biol. 25, 407.
VENEZIALE, C. M., BURNS, J. M., LEWIS, J. C. & BUCHI, K. A. (1977) Specific protein synthesis in isolated epithelium of guinea pig seminal vesicle. Effects of castration and androgen replacement.Biochem. J. 166, 167–73.
WILLIAMS-ASHMAN, H. G. (1984) Transglutaminases and the clotting of mammalian seminal fluids.Mol. Cell. Biochem. 58, 51–61.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Aumüller, G., Seitz, J. Immunoelectron microscopic evidence for different compartments in the secretory vacuoles of the rat seminal vesicles. Histochem J 18, 15–23 (1986). https://doi.org/10.1007/BF01676193
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF01676193