Abstract
The way in which the human epididymis modifies spermatozoa during their sojourn in this structure might be clarified by knowledge of the nature of its secretions. We have examined the presence of several lysosomal hydrolases in human epididymal tissue and fluids, and their synthesis and secretion by monolayer cultures. Tissues were obtained from men undergoing orchidectomy for prostatic carcinoma. The enzymes cathepsin D and acid α-glucosidase were localised in the lysosomes of epithelial cells from the corpus epididymidis, by an immunocytochemical technique. Cathepsin D was also found in epithelial cells of the efferent ducts within lysosomes, apical vesicles and multivesicular bodies. No immunolocalisation of acid glucosidase in the efferent ducts or on the microvilli of the corpus was demonstrable. Cathepsin D, β-hexosaminidase (N-acetylglucosaminidase) and α-glucosidase were measurable in the luminal fluid from the human corpus epididymidis; β-hexosaminidase was secreted into the culture medium by confluent monolayers of epididymal and efferent duct cells. Immunoprecipitation of cell extracts and culture medium of these cultures incubated with 35S-methionine revealed that the precursors of cathepsin D and β-hexosaminidase were synthesized and secreted by such monolayers. Thus, active lytic enzymes are secreted by the human epididymis and could modify sperm membranes.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Barrett AJ (1977) Cathepsin D and other carboxy proteinases. In: Barrett AJ (ed) Proteinases in mammalian cells and tissues. Elsevier/North Holland, Amsterdam, pp 209–248
Brown WJ, Goodhouse J, Farquhar MG (1986) Mannose 6-phosphate receptors for lysosomal enzymes cycle between the Golgi complex and endosome. J Cell Biol 103:1235–1247
Cooper TG (1986) The epididymis, sperm maturation and fertilisation. Springer, Berlin Heidelberg New York
Cooper TG (1992) Epididymal proteins and sperm maturation. In: Nieschlag E, Habenicht U-F (eds) Spermatogenesis, fertilization, contraception. Molecular, cellular and endocrine events in male reproduction, Schering Foundation Workshop 4. Springer, Berlin Heidelberg New York, pp 285–318
Cooper TG, Yeung CH, Meyer R (1989) Immature rat epididymal epithelial cells grown in static primary monolayer culture on permeable supports. I. Vectorial secretion. Cell Tissue Res 256:567–572
Cooper TG, Yeung CH, Meyer R, Schulze H (1990a) Maintenance of human epididymal epithelial cell function in monolayer culture. J Reprod Fert 90:81–91
Cooper TG, Yeung CH, Nashan D, Jockenhövel F, Nieschlag E (1990b) Improvement in the assessment of human epididymal function by the use of inhibitors on the assay of α-glucosidase in seminal plasma. Int J Androl 13:297–305
Figura KA von, Hasilik A (1986) Lysosomal enzymes and their receptors. Ann Rev Biochem 55:167–193
Fransen JAM, Ginsel LA, Cambier PH, Klumperman J, Oude Elferink RPJ (1988) Immunocytochemical demonstration of the lysosomal enzyme α-glucosidase in the brush border of the intestinal epithelial cells. Eur J Cell Biol 47:72–80
Geuze H, Slot JW, Strous GJAM, Hasilik A, Figura K von (1985) Possible pathways for lysosomal enzyme delivery. J Cell Biol 101:2253–2262
Gieselmann V, Hoflack B, Simons K, Mellman I, Kornfeld S (1983) The mannose 6-phosphate receptor and the biogenesis of lysosomes. Cell 52:329–341
Grässel S, Roeling A, Hasilik A (1989) Immunoprecipitation of labeled antigens with Eupergit C1Z. Anal Biochem 180:72–78
Griffiths G, Hoflack B, Simons K, Mellman I, Kornfeld S (1988) The mannose 6-phosphate receptor and the biogenesis of lysosomes. Cell 52:329–341
Guérin JF, Czyba JC (1987) Hydrolytic enzymes detected in fluid and in spermatozoa from different portions of human epididymis. In: Spera G, De Kretser DM (eds) Morphological basis of human reproductive function. Plenum Press London, pp 197–200
Hasilik A, Neufeld EF (1980) Biosynthesis of lysosomal enzymes in fibroblasts. Phosphorylation of mannose residues. J Biol Chem 255:4937–4945
Hasilik A, Figura K von, Conzelmann E, Nehrkorn H, Sandhoff K (1982) Lysosomal enzyme precursors in human fibroblasts. Activation of cathepsin D precursor in vitro and activity of β-hexosaminidase A precursor towards ganglioside Gm2. Eur J Biochem 125:317–321
Holstein AF (1969) Morphologische Studien am Nebenhoden des Menschen. In: Bargmann W, Doerr W (eds) Zwanglose Abhandlungen aus dem Gebiet der normalen und pathologischen Anatomie. Thieme, Stuttgart, pp 1–91
Jones T (1974) Absorption and secretion in the cauda epididymidis of the rabbit and the effects of degenerating spermatozoa on epididymal plasma after castration. J Endocrinol 63:157–165
Klumperman J, Fransen JAM, Boekestijn TC, Oude Elferink RPJ, Matter K, Hauri H-P, Tager JM, Ginsel LA (1991) Biosynthesis and transport of lysosomal α-glucosidase in the human colon carcinoma cell line Caco-2: secretion from the apical surface. J Cell Sci 100:339–347
Klumperman J, Fransen JAM, Tager JM, Ginsel LA (1992) The cation-independent mannose 6-phosphate receptor is not involved in the polarized secretion of lysosomal α-glucosidase from Caco-2 cells. Eur J Cell Biol 57:147–154
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage. Nature 227:680–685
Lane RD, Federman D, Flora JL, Beck BL (1986) Computer-assisted determination of protein concentrations from dye-binding and bicinchoninic acid protein assays performed in microtiter plates. J Immunol Methods 92:261–270
Laskey RA, Mills AD (1975) Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem 56:335–341
Mayorga LS, Bertini F (1982) Effect of androgens on the activity of acid hydrolases in rat epididymis. Int J Androl 5:345–352
Mayorga LS, Bertini F (1985) The origin of some acid hydrolases of the fluid of the rat cauda epididymidis. J Androl 6:243–245
Moore HDM, Pryor JP (1981) The comparative ultrastructure of the epididymis in monkeys and man: a search for a suitable animal model for studying epididymal physiology in Primates. Am J Primatol 1:241–250
Paquin R, Chapdelaine P, Dube JY, Tremblay RR (1984) Similar biochemical properties of human seminal plasma and epididymal α-1,4-glucosidase. J Androl 5:277–282
Raczek S, Yeung CH, Hertle L, Schulze H, Cooper TG (1992) Appearance and endocytic activity of epithelial cells from human efferent ducts in primary monolayer culture. Cell Tissue Res 270:513–519
Roth J, Bendayan M, Orci L (1982) The protein A-gold (pAg) technique. A qualitative and quantitative approach for antigen localization on thin sections. In: Butlock GR, Petrisz P (eds) Techniques in immunocytochemistry. Academic Press, New York, pp 107–133
Sahagian GG (1984) The mannose 6-phosphate receptor function. Biosynthesis and translocation. Biol Cell 51:207–214
Schirren C (1982) Gewebsentnahme aus dem Nebenhoden zu diagnostischen Zwecken? Andrologia 14:461–462
Vendrely E, Dadoune P (1988) Quantitative ultrastructural analysis of the principal cells in the human epididymis. Reprod Nutr Dev 28:1225–1235
Völker W, Frick B, Robeneck H (1985) A simple device for low temperature polymerization of Lowicryl K4M. J Microsc 138:91–93
Yeung CH, Cooper TG, Meyer R (1989) Immature rat epididymal epithelial cells grown in static primary monolayer culture on permeable supports. II. Ultrastructure and histochemistry. Cell Tissue Res 256:573–580
Yeung CH, Cooper TG, Senge T (1990) Histochemical localization and quantification of α-glucosidase in the epididymis of men and laboratory animals. Biol Reprod 42:669–676
Yeung CH, Cooper TG, Bergmann M, Schulze H (1991) Organization of tubules in the human caput epididymidis and the ultrastructure of their epithelia. Am J Anat 191:261–279
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Raczek, S., Yeung, C.H., Hasilik, A. et al. Immunocytochemical localisation of some lysosomal hydrolases, their presence in luminal fluid and their directional secretion by human epididymal cells in culture. Cell Tissue Res 280, 415–425 (1995). https://doi.org/10.1007/BF00307815
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00307815