Summary
The expression of cytokeratin- and vimentin-type intermediate filaments was studied by means of immunohistochemistry in Sertoli cells cultured on two types of reconstituted basement membrane in two-compartment culture chambers. In situ, the Sertoli cells of 17-day-old rats contained only vimentin intermediate filaments. During culture, a gradual reorganization of intermediate filaments accompanied by an increased cytokeratin immunoreactivity was observed. After 6 days, Sertoli cells contained both cytokeratin and vimentin, and the same cytokeratin type as in fetal and newborn testis was revealed by electrophoresis and immunoblotting. The present study shows that the isolation and culture of Sertoli cells causes, even in an improved culture system qualitative changes in the expression of intermediate filament proteins.
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Ailenberg M, Tung PS, Pelletier M, Fritz IB (1988) Modulation of Sertoli cell functions in two-chamber assembly by peritubular cells and extracellular matrix. Endocrinology 122:2604–2618
Bergmann M, Dierichs R (1983) Postnatal formation of the blood-testis barrier in the rat with special reference to the initiation of meiosis. Anat Embryol 168:269–275
Bradford M (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
Byers SW, Hadley MA, Djakiew D, Dym M (1986) Growth and characterization of polarized monolayers of epididymal epithelial cells and Sertoli cells in dual environment culture chamber. J Androl 7:59–68
Chapin R, Phelps J, Miller B, Gray T (1987) Alkaline phosphatase histochemistry discriminates peritubular cells in primary rat testicular cell culture. J Androl 8:155–161
Clermont Y, Perey B (1957) Quantitative study of the cell population of the seminiferous tubules in immature rats. Am J Anat 100:241–268
Dym M (1977) The role of the Sertoli cell in spermatogenesis. In: Yates RD, Gordon M (eds) Male reproductive system. Masson, St Paul, Minn, pp 155–169
Gilula NB, Fawcett DW, Aoki A (1976) The Sertoli cell occluding junctions and gap junctions in mature and developing mammalian testis. Dev Biol 50:142–168
Guillou F (1990) Characterization of a new superfusion, two-compartement culture system for Sertoli cells. Influence of extracellular matrix on cell permeability. Dynamics of transferrin secretion. J Androl (in press)
Guillou F, Martinat N, Combarnous Y (1986) Study of the superactivity of equine follicle-stimulating hormone in in vitro stimulation of rat Sertoli cells. Biochim Biophys Acta 887:196–203
Hadley MA, Byers SW, Suarez-Quian CA, Kleinman HK, Dym M (1985) Extracellular matrix regulates Sertoli cell differentiation, testicular cord formation, and germ cell development in vitro. J Cell Biol 101:1511–1522
Holthöfer H, Miettinen A, Paasivuo R, Lehto VP, Linder E, Alfthan O, Virtanen I (1983) Cellular origin and differentiation of renal carcinomas. Lab Invest 49:317–326
Holthöfer H, Miettinen A, Lehto VP, Lehtonen E, Virtanen I (1984) Expression of vimentin and cytokeratin types of intermediate filament proteins in developing and adult human kidneys. Lab Invest 50:552–559
Janecki A, Steinberger A (1987) Vectorial secretion of transferrin and androgen binding protein in Sertoli cell cultures: effect of extracellular matrix, peritubular myoid cells and medium composition. Mol Cell Endocrinol 52:125–135
Keski-Oja J, Lehto VP, Virtanen I (1981) Keratin filaments of mouse epidermal cells are rapidly affected by epidermal growth factor. J Cell Biol 90:537–541
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–683
Magro G, Charpentier G, Magre S (1988) Cytokeratin expression and testicular morphogenesis. Biol Cell 64:Suppl 4a
Miettinen M, Franssila K, Lehto VP, Paasivuo R, Virtanen I (1984) Expression of intermediate filament proteins in thyroid gland and thyroid tumors. Lab Invest 50B 262–270
Miettinen M, Virtanen I, Talerman A (1985) Intermediate filament proteins in human testis and testicular germ cell tumors. Am J Pathol 120:402–410
Moll R, Franke WW, Schiller DL, Geiger B, Krepler R (1982) The catalog of human cytokeratins: patterns of expression in normal epithelia, tumors and cultured cells. Cell 31:11–24
Orth JM (1982) Proliferation of Sertoli cells in fetal and postnatal rats: a quantitative autoradiographic study. Anat Rec 203:485–492
Paranko J, Virtanen I (1986) Epithelial and mesenchymal cell differentiation in the fetal rat genital ducts: changes in the expression of cytokeratin and vimentin type of intermediate filaments and desmosomal plaque proteins. Develop Biol 117:135–145
Paranko J, Kallajoki M, Pelliniemi LT, Lehto VP, Virtanen I (1986) Transcent coexpression of cytokeratin and vimentin in differentiating rat Sertoli cells. Develop Biol 117:35–44
Paulin D, Lilienbaum A, Duprey P, Zhenlin L, Vicart P (1990) Regulatory elements of the human vimentin gene: activation during proliferation. Reprod Nutr Dev (in press)
Ramaekers FCS, Haag D, Kant A, Moesker O, Jap PHK, Vooijs GP (1983a) Coexpression of keratin- and vimentin-type intermediate filaments in human metastatic carcinoma cells. Proc Natl Acad Sci USA 80:2618–2622
Ramaeker FCS, Huysmans A, Moesker O, Kant A, Jap P, Herman C, Vooijs P (1983b) Monoclonal antibody to keratin filaments, specific for glandular epithelia and their tumor. Lab Invest 49:353–361
Setchell BP (1978) Fluid secretion and the entry of substance into the tubules. In: Setchell BP (eds) The mammalian testis. Cornell University Press, Ithaca, pp 233
Spruill WA, Steiner AL, Tres LL, Kierszenbaum AL (1983a) Follicle-stimulating hormone-dependent phosphorylation of vimentin in cultures of rat Sertoli cells. Proc Natl Acad Sci USA 80:993–997
Spruill WA, Zysk JR, Tres LL, Kierszenbaum AL (1983b) Calcium/calmodulin-dependent phosphorylation of vimentin in rat Sertoli cells. Proc Natl Acad Sci USA 80:760–764
Tung PS, Fritz IB (1984) Extracellular matrix promotes rat Sertoli cell histotypic expression in vitro. Biol Reprod 30:213–229
Van Vorstenbosch CJAHV, Spek E, Colenbrander B, Wensing CJG (1984b) Sertoli cell development of pig testis in the fetal and neonatal period. Biol Reprod 31:565–577
Virtanen L, Lehto VP, Lehtonen E, Vartio T, Stenman S, Kurki P, Wager O, Small JV, Dahl D, Badley RA (1981) Expression of intermediate filaments in cultured cells. J Cell Sci 50:45–63
Virtanen J, Miettinen M, Lehto VP, Kariniemi AL, Paasivuo R (1985) Diagnostic application of monoclonal antibodies to intermediate filaments. Ann N Y Acad Sci 455:635–648
Virtanen J, Heikinheimo K, Hormia M, Kivelä T, Laitinen L, Thornell LE (1989) Expression of intermediate filaments (IF) in tissues and cultured cells. Int J Dev Biol 33:55–61
Waites GMH, Gladwell RT (1982) Physiological significance of fluid secretion in the testis and blood-testis barrier. Physiol Rev 62:624–671
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Guillou, F., Monet-Kuntz, C., Fontaine, I. et al. Expression of fetal-type intermediate filaments by 17-day-old rat Sertoli cells cultured on reconstituted basement membrane. Cell Tissue Res 260, 395–401 (1990). https://doi.org/10.1007/BF00318642
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DOI: https://doi.org/10.1007/BF00318642