Abstract
A high performance liquid chromatographic procedure has been used for the purification of rat Sertoli cell secretory protein S70 and S45-535 heterodimeric protein to determine their role during spermatogenesis. These two proteins display binding affinity for each other and appear antigenically related. We have observed that:
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1.
S70 and S45-S35 heterodimeric protein coelute during purification,
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2.
polyclonal antiserum raised against protein S70 recognizes common antigenic determinants in polypeptides S45 and S35, the disulfide-linked components of the heterodimeric protein, and
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3.
a monoclonal antibody that recognizes polypeptide S35 but does not crossreact with either protein S70 or polypeptide S45, immunoprecipitates the S70/S45-S35 heterodimeric protein complex.
In immunofluorescent experiments, antisera raised against protein S70 and polypeptide components of S45-S35 heterodimeric protein immunoreact with two major sperm intracellular structures: the acrosome and periaxonemal outer dense fibers of sperm tail. Immunoreactivity was not detected on the sperm plasma membrane surface of unfixed, living sperm. Outer dense fibers extracted from sperm tails by a combined treatment with cetylthrimethylammonium bromide and 2-mercaptoethanol, yielded a characteristic polypeptide pattern. In immunoblotting experiments, sperm tail polypeptides were recognized by polyclonal antisera raised against Sertoli cell secretory proteins. We conclude that Sertoli cell secretory proteins S70 and S45-S35 heterodimeric protein are antigenically related to each other and to keratin-like polypeptides from sperm tail.
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References
Fawcett DW: The mammalian spermatozoon. Dev Biol 44:394–436, 1975
Calvin HI, Bedford JM: Formation of disulphide bonds in the nucleus and accessory structures of mammalian spermatozoa during epididymal maturation. J Reprod Fert Suppl 13:65–75, 1971
Baccetti B, Pallini V, Burrini G: The accessory fibers of the sperm tail. III. High sulfur and low sulfur components in mammals and cephalopods. J Ultr Res 57:289–308, 1976
Bedford JM, Calvin HI: Changes in -S-S- linked structures of the sperm tail during epididymal maturation, with comparative observations in submammalian species. J Exp Zool 187:181–204, 1974
Olson GE, Sammons DW: Structural chemistry of outer dense fibers of rat sperm. Biol Reprod 22:319–322, 1980
Vera JC, Brito M, Zuvic T, Burzio LO: Polypeptide composition of rat sperm outer dense fibers. A simple procedure to isolate the fibrillar complex. J Biol Chem 259:5970–5977, 1984
O'Brien DA, Bellvé AT: Protein constituents of the mouse spermatozoon. II. Temporal synthesis during spermatogenesis. Dev Biol 75:405–418, 1980
Vera JC, Brito M, Burzio LO: Biosynthesis of rat sperm outer dense fibers during spermiogenesis. In vivo incorporation of [3H]leucine into the fibrillar complex. Biol Reprod 36:193–202, 1987
Irons MJ, Clermont Y: Formation of the outer dense fibers during spermiogenesis in the rat. Anat Rec 202:463–471, 1982
Irons MJ, Clermont Y: Kinetics of fibrous sheath formation in the rat spermatid. Am J Anat 165:121–130, 1982
Irons MJ: Synthesis and assembly of connecting-piece proteins as revealed by autoradiography. J Ultr Res 82:27–34, 1983
Shabanowitz RB, DePhilip RM, Crowell JA, Tres LL, Kierszenbaum AL: Temporal appearance and cyclic behavior of Sertoli cell-specific secretory proteins during the development of the rat seminiferous tubule. Biol Reprod 35:745–760, 1986
Kierszenbaum AL, Abdullah M, Ueda H, Tres L: Spermatogenesis in vitro: Searching for in vivo correlates. In: VB Mahesh, DS Dhindsa, E Anderson, SP Kalra (eds) Regulation of Ovarian and Testicular Function. Plenum Press, New York, 1987, pp 535–560
Kierszenbaum AL, Abdullah M, Ueda H, Tres LL: Antisera raised against Sertoli cell secretory proteins recognize keratin-like polypeptides from sperm tail. J Cell Biol 105:170a, 1987
Ueda H, Abdullah M, Tres LL, Kierszenbaum AL: Immunocytochemical and immunogold electron microscopic localization of two Sertoli cell secretory proteins in developing spermatids. J Cell Biol 105:167a, 1987
Abdullah M, Tres LL, Kierszenbaum AL: Partial purification of three rat Sertoli cell, spermatogenic stage-dependent secretory proteins by HPLC. J Cell Biol 103:483a, 1986
Kierszenbaum AL, Tres LL: The structural and functional cycle of Sertoli cells in culture. In: G Jagiello, HJ Vogel (eds) Bioregulators of Reproduction. Academic Press, New York, 1981, pp 207–228
Tres LL, Smith EP, Van Wyk JJ, Kierszenbaum AL: Immunoreactive sites and accumulation of somatomedin-C in rat Sertoli-spermatogenic cell co-cultures. Exp Cell Res 162:33–50, 1986
Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685, 1970
Shulman M, Wilde CD, Köhler G: A better cell line for making hybridomas secreting specific antibodies. Nature 276:269–270, 1978
Sternberg J, Jeppesen P: Dot-blotting a novel screening assay for antibodies in hybridoma cultures. J Immunol Meth 64:39–43, 1983
Hu P-C, Huang Y-S, Graham JA, Gardner DE: Identification of immunogens of Mycoplasma pneumoniae by protein blotting. Biochem Biophys Res Commun 103:1363–1370, 1981
Towbin H, Staehelin T, Gordon J: Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354, 1979
Burnette WN: ‘Western blotting’: Electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem 112:195–203, 1981
Kierszenbaum AL, DePhilip RM, Spruill WA, Takenaka I: Isolation and culture of rat seminal vesicle epithelial cells. The use of the secretory protein SVS IV as a functional probe. Exp Cell Res 145:293–304, 1983
Wolf DE, Hagopian SS, Lewis RG, Voglmayr JK, Fairbanks G: Lateral regionalization and diffusion of a maturation-dependent antigen in the ram sperm plasma membrane. J Cell Biol 102:1826–1831, 1986
Sylvester SR, Skinner MK, Griswold MD: A sulfated glycoprotein synthesized by Sertoli cells and by epididymal cells is a component of the sperm membrane. Biol Reprod 31:1087–1101, 1984
Griswold MD, Roberts K, Bishop P: Purification and characterization of a sulfated glycoprotein secreted by Sertoli cells. Biochemistry 25:7265–7270, 1986
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Abdullah, M., Tres, L.L., Ueda, H. et al. Antigenic homology between rat sperm tail polypeptides and Sertoli cell secretory proteins. Mol Cell Biochem 81, 165–176 (1988). https://doi.org/10.1007/BF00219319
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DOI: https://doi.org/10.1007/BF00219319