Skip to main content
Log in

Changes in ectonucleotidase activities in rat Sertoli cells during sexual maturation

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

Sertoli cell maturation is a complex process involving both morphological and biochemical changes. These cells have previously been shown to be targets for extracellular purine structures such as ATP and adenosine. These compounds evoke responses in rat Sertoli cells through the purinoceptor families, P2X and P2Y and PA1. The signals to purinoceptors are usually terminated by the action of ectonucleotidases. In a previous work, we demonstrated that rat Sertoli cells have ecto-ATPdiphosphohydrolase (EC 3.6.1.5), ecto-5′-nucleotidase (EC 3.1.3.5) and ecto-adenosine deaminase (ecto-ADA) (EC 3.5.4.4) activities. Here we investigated whether some changes occur during rat Sertoli cell maturation in these activities. Rat Sertoli cells obtained from rats of different ages representing the pre pubertal, mid pubertal and ‘young adult’ (10-, 18- and 35-day-old, respectively) were cultured and used for different assays. The nucleotide hydrolysis was estimated by measuring the Pi released using a colorimetric method and by HPLC analysis. ATP and ADP hydrolysis was increased 3-fold during sexual maturation. AMP hydrolysis increased 4-fold in 10- to 35-day-old Sertoli cells. Similar results were obtained when we used other substrates to measure the extracellular hydrolysis of nucleotides (GTP, GDP, GMP and IMP). The ecto-ADA activity showed a 2-fold increase in the specific activity (18- to 35-day-old Sertoli cells). The termination of the purine cascade by adenosine degradation was faster in the 35- than in 18-day-old Sertoli cells. Follicle Stimulating Hormone (FSH) influences on the ectonucleotidase activities were investigated in 10- and 18-day-old Sertoli cells and a significant increase in the ATP and ADP hydrolysis was observed. Our results show an increase in the extracellular purine cascade during the Sertoli cell development, indicating a rise in the purine communication inside the seminiferous tubules with rat sexual maturation.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Magueresse BJ, 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

    Google Scholar 

  2. Orth JM: The role of follicle-stimulating hormone in controlling Sertoli cell proliferation in testis of fetal rats. Endocrinology 115: 1248-1255, 1984

    Google Scholar 

  3. Orth JM: Proliferation of Sertoli cells in fetal and postnatal rats: A quantitative autoradiographic study. Anat Rec 203: 485-492, 1982

    Google Scholar 

  4. Vitale-Calpe R, Fawcett DW, Dym M: The normal development of the blood-testis barrier and the effects of clomiphene and estrogen treatment. Anat Rec 176: 331-344, 1973

    Google Scholar 

  5. Clermont Y, Perey B: Quantitative study of the cell population of the seminiferous tubules in immature rats. Am J Anat 100: 241-267, 1957

    Google Scholar 

  6. Skinner MK: Cell-cell interactions in the testis. Ann NY Acad Sci 513: 158-171, 1987

    Google Scholar 

  7. Murphy KMM, Goodman RR, Snyder SH: Adenosine receptor localization in rat testis: Biochemical and autoradiographic evidence for association with spermatocytes. Endocrinology 113: 1299-1305, 1983

    Google Scholar 

  8. Monaco L, Conti M: Localization of adenosine receptors in rat testicular cells. Biol Reprod 35: 258-266, 1989

    Google Scholar 

  9. Rivkees AS: Localization and characterization of adenosine receptor expression in rat testis. Endocrinology 135: 2307-2313, 1994

    Google Scholar 

  10. 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

    Google Scholar 

  11. Filippini A, Riccioli A, de Cesaris P, Paniccia R, Teti A, Stefanini M, Conti M, Ziparo E: Activation of inositol phospholipids and calcium signaling in rat Sertoli cells by P2 purinergic receptors: Modulation of follicle-stimulating hormone responses. Endocrinology 134: 1537-1545, 1994

    Google Scholar 

  12. Wu WL, So SC, Sun YP, Chung YW, Grima J, Wong PYD, Yan YC, Chan HC: Functional expression of P2U receptors in rat spermatogenic cells: Dual modulation of a Ca2+-activated K+ channel. Biochem Biophys Res Commun 248: 728-732, 1998

    Google Scholar 

  13. Conti M, Boitani C, DeManno DA, Migliaccio S, Monaco L, Szymeczek C: Characterization and function of adenosine receptors in the testis. Ann NY Acad Sci 564: 39-47, 1989

    Google Scholar 

  14. Zimmermann H: Ectonucleotidases: Some recent developments and a note on nomenclature. Drug Dev Res 52: 44-56, 2001

    Google Scholar 

  15. Cunha RA, Almeida T, Ribeiro JA: Modification by arachidonic acid of extracellular adenosine metabolism and neuromodulatory action in the rat hippocampus. J Biol Chem 275: 37572-37581, 2000

    Google Scholar 

  16. Müller J, Rocha JBT, Battastini AMO, Sarkis JJF, Dias RD: Ontogeny of ATP and ADP hydrolysis by cerebral cortex synaptosomes from rats. Braz J Med Biol Res 23: 935-939, 1990

    Google Scholar 

  17. Müller J, Rocha JBT, Battastini AMO, Sarkis JJF, Dias RD: Postnatal development of ATPase-ADPase activities in synaptosomal fraction from cerebral cortex of rats. Neurochem Int 23: 471-477, 1993

    Google Scholar 

  18. Zimmermann H: 5′-nucleotidase: Molecular structure and functional aspects. Biochem J 285: 345-365, 1992

    Google Scholar 

  19. Franco R, Casadó V, Ciruela F, Saura C, Mollol J, Canela EI, Lluis C: Cell surface adenosine deaminase: Much more than an ectoenzyme. Progr Neurobiol 52: 283-294, 1997

    Google Scholar 

  20. Casali EA, da Silva T, Gelain DP, Battastini AMO, Sarkis JJF, Bernard EA: Ectonucleotidases activities in Sertoli cells from immature rats. Braz J Med Biol Res 34: 1247-1256, 2001

    Google Scholar 

  21. Wagle JR, Heindel JJ, Steiberger A, Sanborn BM: Effect of hypotonic treatment on Sertoli cell purity and function in culture. In Vitro Cell Dev Biol 22: 325-331, 1986

    Google Scholar 

  22. Palombi F, Di Carlo C: Alkaline phosphatase is a marker for myoid cells in cultures of peritubular and tubular tissue. Biol Reprod 39: 1101-1109, 1988

    Google Scholar 

  23. Griswold MD, Solari A, Tung PS, Fritz IB: Stimulation by follicle-stimulating hormone of DNA synthesis and of mitosis in cultured Sertoli cells prepared from testes of immature rats. Mol Cell Endocrinol 7: 151-165, 1977

    Google Scholar 

  24. Chan K, Delfert D, Junger KD: A direct colorimetric assay for the Ca+2-ATPase activity. Anal Biochem 157: 375-380, 1986

    Google Scholar 

  25. Voelter W, Zech K, Arnold P, Ludwig G: Determination of selected pyrimidines, purines and their metabolites in serum and urine by reversed-phase ion-pair. J Chromatogr 199: 345-354, 1980

    Google Scholar 

  26. Trams GE, Lauter CJ: On the sidedness of plasma membrane enzymes. Biochem Biophys Acta 345: 180-197, 1974

    Google Scholar 

  27. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ: Protein measurement with the Folin phenol reagent. J Biol Chem 193: 265-275, 1951

    Google Scholar 

  28. Casali EA, Gelain DP, Souza LF, Kaiser GRRF, Wink MR, Battastini AMO, Sarkis JJF: Study of ecto-adenosine deaminase and apyrase immunodetection in cultures of Sertoli cells from immature rats. Drug Special Issue from Purine 2000: Biochemical, Pharmacological and Clinical Perspectives. Abstracts from the International Symposium, Madrid, Spain. Dev Res 50 (abstr 258): 2000, 113 pp

  29. Hodgen GD, Sherins RJ: Enzymes as markers of testicular growth and development in the rat. Endocrinology 93: 985-989, 1973

    Google Scholar 

  30. Sickle MV, Oberwetter JM, Birnbaumer L, Means AR: Developmental changes in the hormonal regulation of rat testis Sertoli cell adenylyl cyclase. Endocrinology 109: 1270-1280, 1981

    Google Scholar 

  31. Rich KA, Bardin CW, Gunsalus GL, Mather JP: Age-dependent pattern of androgen-biding protein secretion from rat Sertoli cells in primary culture. Endocrinology 133: 2284-2293, 1983

    Google Scholar 

  32. Sanborn BM, Wagle JR, Steinberger A, Greer-emmert D: Maturational and hormonal influences on Sertoli cell function. Endocrinology 118: 1700-1709, 1986

    Google Scholar 

  33. Skinner MK, Schlitz SM, Anthony CT: Regulation of Sertoli cell differentiated function: Testicular transferrin and androgen-binding protein expression. Endocrinology 124: 3015-3024, 1989

    Google Scholar 

  34. Kohring K, Zimmermann H: Upregulation of ecto-5′-nucleotidase in human neuriblastoma SH-SY5Y cells on differentiation by retinoic acid or phorbolester. Neurosci Lett 258: 127-130, 1998

    Google Scholar 

  35. Heilbronn A, Maienschein V, Carstensen K, Gann W, Zimmermann H: Crucial role of ecto-5′-nucleotidase in differentiation and survival of developing neural cells. NeuroReport 7: 257-261, 1995

    Google Scholar 

  36. Franco R, Maloot J, Casadó V, Lluis C, Canela EI, Saura CA, Blanco J, Ciruela F: Ecto-adenosine deaminase: An ecto-enzyme and costimulatory protein acting on a variety of cell surface receptors. Drug Devel Res 45: 261-268, 1998

    Google Scholar 

  37. Lu Q, Porter D, Cui X, Sanborn BM: Ecto-ATPase mRNA is regulated by FSH in Sertoli cells. J Androl 22: 289-301, 2001

    Google Scholar 

  38. Meng J, Zhang F, Huhtaniemi I, Pakarinen P: Characterization and developmental expression of a testis-specific adenosine deaminase mRNA in the mouse. J Androl 18: 88-95, 1997

    Google Scholar 

  39. Bonan CD, Schetinger MRC, Battanstini AMO, Sarkis JJF: Ectonu-cleotidases and synaptic plasticity: Implications in physiological and pathological condition. Drug Dev Res 52: 57-65, 2001

    Google Scholar 

  40. Kegel B, Braun N, Heine P, Maliszewski CR, Zimmermann H: An ecto-ATPase and an ecto-ATPdiphosphohydrolase are expressed in rat brain. Neuropharmacology 36: 1189-1200, 1997

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Casali, E.A., de Souza, L.F., Gelain, D.P. et al. Changes in ectonucleotidase activities in rat Sertoli cells during sexual maturation. Mol Cell Biochem 247, 111–119 (2003). https://doi.org/10.1023/A:1024150619490

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1024150619490

Navigation