Abstract
It has been long postulated that extracellular purines can modulate the function of the male reproductive system by interacting with different purinergic receptors of Sertoli and germinative cells. Many authors have described the biological changes induced by extracellular ATP and/or adenosine in these cells, and some hypothetical models for paracrine communication mediated by purines were proposed; however, the cellular source(s) of these molecules in seminiferous tubules remains unknown. In this study, we demonstrated for the first time that Sertoli cells are able to release ATP (0.3 nmol/mg protein) and adenosine (0.1 nmol/mg protein) in the extracellular medium, while germinative and myoid peritubular cells are able to secrete adenosine (0.02 and 0.37 nmol/mg protein, respectively). Indeed, all the three types of cells were able to release inosine at significant concentrations (about 0.4 nmol/mg protein). This differential secretion depending on the cellular type suggests that these molecules may be involved in the paracrine regulation and/or control of the maturation processes of these cells.
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Dubyak GR, El-Moatassim C: Signal transduction via P2-purinergic receptors for extracellular ATP and other nucleotides. Am J Physiol 265: C577-C606, 1993
Di Virgilio F, Chiozzi P, Ferrari D, Falzoni S, Sanz JM, Morelli A, Torboli M, Bolognesi G, Baricordi R: Nucleotide receptors: An emerging family of regulatory molecules in blood cells. Blood 97: 587-600, 2000
Katzur AC, Koshimizu TA, Tomic M, Schultze-Mosgau A, Ortmann O, Stojilkovic SS: Expression and responsiveness of P2Y2 receptors in human endometrial cancer cell lines. J Clin Endocrinol Metab 84: 4085-4091, 1999
Rivkees SA: Localization and characterization of adenosine receptor expression in rat testis. Endocrinology 135: 2307-2313, 1994
Conti M, Boitani C, Demanno D, Migliaccio S, Monaco L, Szymeczek C: Characterization and function of adenosine receptors in the testis. Ann NY Acad Sci 564: 39-47, 1989
Monaco L, Conti M: Localization of adenosine receptors in rat testicular cells. Biol Reprod 35: 258-266, 1986
Stiles GL, Pierson G, Sunay S, Parsons WJ: The rat testicular adenosine A1 receptor-adenylate cyclase system. Endocrinology 119: 1845-1851, 1986
Monaco L, DeManno DA, Martin MW, Conti M: Adenosine inhibition of the hormonal response in the Sertoli cell is reversed by Pertussis toxin. Endocrinology 122: 2692-2698, 1988
Filippini A, Riccioli A, De Cesaris P, Paniccia R, Teti A, Stefanini M, Conti M, Ziparo E: Activation of inositol phospholipid turnover and calcium signaling in rat Sertoli cells by P2-purinergic receptors: Modulation of follicle-stimulating hormone responses. Endocrinology 134: 1537-1545, 1994
Meroni SB, Cánepa DF, Pellizzari EH, Schteingart HF, Cigorraga SB: Effects of purinergic agonists on aromatase and gamma-glutamyl transpeptidase activities and on transferrin secretion in cultured Sertoli cells. J Endocrinol 157: 275-283, 1998
Foresta C, Rossato M, Bordon P, Di Virgilio F: Extracellular ATP activates different signaling pathways in rat Sertoli cells. Biochem J 311: 269-274, 1995
Wong PY: Control of anion and fluid secretion by apical P2-purinoceptors in the rat epididymis. Br J Pharmacol 95: 1315-1321, 1988
Wu WL, So SC, Sun YP, Chung YW, Grima J, Wong PY, Yan YC, Chan HC: Functional expression of P2U receptors in rat spermatogenic cells: Dual modulation of a Ca(2+)-activated K+ channel. Biochem Biophys Res Commun 248: 728-732, 1998
Minelli A, Allegrucci C, Piomboni P, Manucci R, Lluis C, Franco R: Immunolocalization of A1 adenosine receptors in mammalian spermatozoa. J Histochem Cytochem 48: 1163-1171, 2000
Fenichel P, Gharib A, Emiliozzi C, Donzeau M, Menezo Y: Stimulation of human sperm during capacitation in vitro by an adenosine agonist with specificity for A2 receptors. Biol Reprod 54: 1405-1411, 1996
Fraser LR: Adenosine and its analogues, possibly acting at A2 receptors, stimulate mouse sperm fertilizing ability during early stages of capacitation. J Reprod Fertil 89: 467-476, 1990
Fraser LR, Duncan AE: Adenosine analogues with specificity for A2 receptors bind to mouse spermatozoa and stimulate adenylate cyclase activity in uncapacitated suspensions. J Reprod Fertil 98: 187-194, 1993
Glass R, Bardinini M, Robson T, Burnstock G: Expression of nucleotide P2X receptor subtypes during spermatogenesis in the adult rat testis. Cells Tissues Organs 169: 377-387, 2001
Jégou B, Sharpe RM: Paracrine mechanisms in testicular control. In: D. de Krester (ed). Molecular Biology of the Male Reproductive System. Academic Press, San Diego, 1993, pp 271-310
Dym M, Fawcett DW: The blood-testis barrier in the rat and the physiological compartmentation of the seminiferous epithelium. Biol Reprod 3: 308-326, 1970
Plöen L, Setchell BP: Blood-testis barriers revisited. A homage to Lennart Nicander. Int J Androl 15: 1-4, 1992
Jutte NHPM, Grootegoed JA, Rommerts FFG, van der Molen HJ: Exogenous lactate is essential for metabolic activities in isolated rat spermatocytes and spermatids. J Reprod Fert 62: 399-405, 1981
Mita M, Price JM, Hall PF: Stimulation by follicle-stimulating hormone of synthesis of lactate by Sertoli cells from rat testis. Endocrinology 110: 1535-1541, 1982
Jutte NHPM, Jansen R, Grootegoed JA, Rommerts FFG, van der Molen HJ: FSH stimulation of the production of pyruvate and lactate by rat Sertoli cells may be involved in hormonal regulation of spermatogenesis. J Reprod Fert 68: 219-226, 1983
Jutte NHPM, Eikvar L, Levy FO, Hansson VM: Metabolism of palmitate in cultured rat Sertoli cells. J Reprod Fert 73: 497-503, 1985
Le Magueresse B, Jégou B: In vitro effects of germ cells on the secretory activity of Sertoli cells recovered from rats of different ages. Endocrinology 122: 1672-1680, 1988
Verhoeven G, Cailleau J: Testicular peritubular cells secrete a protein under androgen control that inhibits induction of aromatase activity in Sertoli cells. Endocrinology 123: 2100-2110, 1988
Welsh MJ, Ireland ME: The second messenger pathway for germ cell-mediated stimulation of Sertoli cells. Biochem Biophys Res Commun 184: 217-224, 1992
Onoda M, Djakiew D: A24,500 Da protein derived from rat germ cells is associated with Sertoli cell secretory function. Biochem Biophys Res Commun 197: 688-695, 1993
Grima J, Pineau C, Bardin CW, Cheng CY: Rat Sertoli cell clusterin, α2-macroglobulin, and testins: Biosynthesis and differential regulation by germ cells. Mol Cell Endocrinol 89: 127-140, 1992
Shubhada S, Glinz M, Lamb DJ: Sertoli cell secreted growth factor — cellular origin, paracrine and endocrine regulation of secretion. J Androl 14: 99-109, 1993
Pineau C, Syed V, Bardin CW, Jégou B, Cheng CY: Identification and partial purification of a germ cell factor that stimulates transferrin secretion by Sertoli cells. Recent Prog Horm Res 48: 539-542, 1993
Norton JH, Vigne JL, Skinner MK: Regulation of Sertoli cell differentiation by the testicular paracrine factor PmodS: Analysis of common signal transduction pathways. Endocrinology 134: 149-157, 1994
Casali EA, da Silva TR, Gelain DP, Kaiser GRRF, Battastini AMO, Sarkis JJF, Bernard EA: Ectonucleotidase activities in Sertoli cells from immature rats. Braz J Med Biol Res 34: 1247-1256, 2001
Laleveé N, Rogier C, Becq F, Joffre M: Acute effects of adenosine triphosphates, cyclic 3′,5′-adenosine monophosphates, and follicle-stimulating hormone on cytosolic calcium level in cultured immature rat Sertoli cells. Biol Reprod 61: 343-352, 1999
Shabanowitz RB, Kierszenbaum AL: Newly synthesized proteins in seminiferous intertubular and intratubular compartments of the rat testis. Biol Reprod 35: 179-190, 1986
Rocha AB, Guma FCR, Casali EA, Scherer GS, Elena MA, Bernard EA: Influence of the biomatrix on the response of Sertoli cells to FSH. Arch Physiol Biochem 105: 473-477, 1997
Tung PS, Fritz IB: Extracellular matrix promotes rat Sertoli cell histotypic expression in vitro. Biol Reprod 30: 213-229, 1984
Syed V, Hecht NB: Up-regulation and down-regulation of genes expressed in cocultures of rat Sertoli cells and germ cells. Mol Reprod Dev 47: 380-389, 1997
Meistrich ML, Longtin J, Brock WA, Grimes SR Jr, Mace ML: Purification of rat spermatogenic cells and preliminary biochemical analysis of these cells. Biol Reprod 25: 1065-1077, 1981
Ciccarelli R, Di Iorio P, Giuliani P, D'alimonte I, Ballerini P, Caciagli F, Rathbone M: Rat cultured astrocytes release guanine-based purines in basal conditions and after hypoxia/hypoglycemia. Glia 25: 93-98, 1999
Cunha RA, Sebastião AM, Ribeiro JA: Separation of adenosine-triphosphate and its degradation products in innervated muscle of the frog by reverse phase high-performance liquid-chromatography. Chromatographia 28: 610-612, 1989
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ: Protein measurement with folinphenol reagent. J Biol Chem 193: 265-275, 1951
Jin X, Shepperd RK, Duling BR, Linden J: Inosine binds to A3 receptors and stimulates mast cell degranulation. J Clin Invest 100: 2849-2857, 1997
Tilley SL, Wagoner VA, Salvatore CA, Jacobson MA, Koller BH: Adenosine and inosine increase cutaneous vasopermeability by activating A3 receptors on mast cells. J Clin Invest 105: 361-367, 2000
Gao Z, Li BS, Day YJ, Linden J: A3 adenosine receptor activation triggers phosphorilation of protein kinase B and protects rat basophilic leukemia 2H3 mast cells from apoptosis. Mol Pharmacol 59: 76-82, 2001
Lu Q, Porter LD, Cui X, Sanborn BM: Ecto-ATPase mRNA is regulated by FSH in Sertoli cells. J Androl 22: 289-301, 2001
Monaco L, Toscano MV, Conti M: Purine modulation of the hormonal response of the rat Sertoli cell in culture. Endocrinology 115: 1616-1624, 1984
Burnstock G, Willians M: P2 purinergic receptors: Modulation of cell function and therapeutic potential. J Pharmacol Exp Ther 295: 862-869, 2000
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Gelain, D.P., de Souza, L.F. & Bernard, E.A. Extracellular purines from cells of seminiferous tubules. Mol Cell Biochem 245, 1–9 (2003). https://doi.org/10.1023/A:1022857608849
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DOI: https://doi.org/10.1023/A:1022857608849