Histochemistry and Cell Biology

, Volume 124, Issue 5, pp 401–407 | Cite as

Immunohistochemical localisation of PDE5 in Leydig and myoid cells of prepuberal and adult rat testis

  • Angela Scipioni
  • Stefania Stefanini
  • Rocco Santone
  • Mauro Giorgi
Original paper
First Online:
Accepted:

Abstract

Expression of phosphodiesterase 5 (PDE5) in the rat testis at several pre and postnatal developmental stages was investigated by immunohistochemistry. The enzyme was localised in vascular smooth muscle cells, as well as in Leydig and peritubular cells. The latter were identified as myoid, based on their immunoreactivity to desmin and α-smooth muscle actin. The presence of PDE5 in myoid cells was confirmed by Western blot analysis and immunohistochemistry performed on highly purified cell fractions, obtained from 16-day-old rats. The expression of PDE5 in these somatic cells of rat testis is discussed in view of the roles played by cGMP signal transduction pathways in the mammalian male reproductive function.

Keywords

Phosphodiesterase 5 Peritubular cells α-SM actin Desmin cGMP 
This is a preview of subscription content, log in to check access.

Notes

Acknowledgements

We thank Prof. Fioretta Palombi and Dr. Francesca Romano (Department of Histology and General Embryology, University of Rome “La Sapienza”) for supplying purified myoid cells. Thanks are also due to Dr. Sandra Moreno (Department of Biology-LIME, University Roma Tre) for helpful suggestions.

References

  1. Adams JC (1992) Biotin amplification of biotin and horseradish peroxidase signals in histochemical stains. J Histochem Cytochem 40:1457–1463PubMedGoogle Scholar
  2. Bauche F, Stephan JP, Touzalin AM, Jegou B (1998) In vitro regulation of an inducible-type NO synthase in the rat seminiferous tubule cells. Biol Reprod 58:431–438PubMedCrossRefGoogle Scholar
  3. Boolell M, Gepi-Attee S, Gingell JC, Allen MJ (1996) Sildenafil, a novel effective oral therapy for male erectile dysfunction. Br J Urol 78:257–261PubMedGoogle Scholar
  4. Conti M, Toscano MV, Petrelli L, Geremia R, Stefanini M (1982) Regulation of follicle-stimulating hormone and dibutyryl adenosine 3′,5′-monophosphate of a phosphodiesterase isoenzyme of the Sertoli cell. Endocrinology 110:1189–1196PubMedGoogle Scholar
  5. Conti M, Iona S, Cuomo M, Swinnen JV, Odeh J, Svoboda ME (1995) Characterization of a hormone-inducible, high affinity adenosine 3′-5′-cyclic monophosphate phosphodiesterase from the rat Sertoli cell. Biochemistry 34:7979–7987PubMedCrossRefGoogle Scholar
  6. Coquil JF, Brunelle G, Guedon J (1985) Occurrence of the methylisobutylxanthine-stimulated cyclic GMP binding protein in various rat tissues. Biochem Biophys Res Commun 127:226–231PubMedCrossRefGoogle Scholar
  7. Davidoff MS, Breucker H, Holstein AF, Seidl K (1990) Cellular architecture of the lamina propria of human seminiferous tubules. Cell Tissue Res 262:253–261PubMedCrossRefGoogle Scholar
  8. Davidoff MS, Middendorff R, Mayer B, deVente J, Koesling D, Holstein AF (1997) Nitric oxide/cGMP pathway components in the Leydig cells of the human testis. Cell Tissue Res 287:161–170PubMedCrossRefGoogle Scholar
  9. Dorrington JH, Roller NF, Fritz IB (1975) Effects of follicle-stimulating hormone on cultures of Sertoli cell preparations. Mol Cell Endocrinol 3:57–70PubMedCrossRefGoogle Scholar
  10. Filippini A, Tripiciano A, Palombi F, Teti A, Paniccia R, Stefanini M, Ziparo E (1993) Rat testicular myoid cells respond to endothelin: characterization of binding and signal transduction pathway. Endocrinology 133:1789–1796PubMedCrossRefGoogle Scholar
  11. Geremia R, d’Agostino A, Monesi V (1978) Biochemical evidence of haploid gene activity in spermatogenesis of the mouse. Exp Cell Res 111:23–30PubMedCrossRefGoogle Scholar
  12. Giordano D, De Stefano ME, Citro G, Modica A, Giorgi M (2001) Expression of cGMP-binding cGMP-specific phosphodiesterase (PDE5) in mouse tissues and cell lines using an antibody against the enzyme amino-terminal domain. Biochim Biophys Acta 1539:16–27PubMedCrossRefGoogle Scholar
  13. Giordano D, Giorgi M, Tata AM, Modica A, Augusti-Tocco G (2004) Expression of PDE5 splice variants during ontogenesis of chick dorsal root ganglia. J Neurosci Res 78:815–823PubMedCrossRefGoogle Scholar
  14. Gravis CJ (1978) Inhibition of spermiation in the syrian hamster using dibutyryl cyclic AMP. Cell Tiss Res 192:241–248CrossRefGoogle Scholar
  15. Hamet P, Coquil JF (1978) Cyclic GMP binding and cyclic GMP phosphodiesterase in rat platelets. J Cyclic Nucleotide Res 4:281–290PubMedGoogle Scholar
  16. Hipkin RW, Moger WH (1991) Interaction between cyclic nucleotide second messenger systems in murine Leydig cells. Mol Cell Endocrinol 82:251–257PubMedCrossRefGoogle Scholar
  17. Hollinger WA, Hwang F (1974) Effect of dibutyryl cyclic AMP on in vitro rat testis DNA, RNA and protein labelling. Endocrinology 94:444–449PubMedCrossRefGoogle Scholar
  18. Jeremy JY, Ballard SA, Naylor AM, Miller MA, Angelini GD (1997) Effects of sildenafil, a type-5 cGMP phosphodiesterase inhibitor, and papaverine on cyclic GMP and cyclic AMP levels in the rabbit corpus cavernosum in vitro. Br J Urol 79:958–963PubMedGoogle Scholar
  19. Kotera J, Yanaka N, Fujishige K, Imai Y, Akatsuka H, Ishizuka T, Kawashima K, Omori K (1997) Expression of rat cGMP-binding cGMP-specific phosphodiesterase mRNA in Purkinje cell layers during postnatal neuronal development. Eur J Biochem 249:434–442PubMedCrossRefGoogle Scholar
  20. Kotera J, Fujishige K, Imai Y, Kawai E, Michibata H, Akatsuka H, Yanaka N, Omori K (1999) Genomic origin and transcriptional regulation of two variants of cGMP-binding cGMP-specific phosphodiesterases. Eur J Biochem 262:866–873PubMedCrossRefGoogle Scholar
  21. Kotera J, Fujishige K, Omori K (2000) Immunohistochemical localization of cGMP-binding cGMP-specific phosphodiesterase (PDE5) in rat tissues. J Histochem Cytochem 48:685–693PubMedGoogle Scholar
  22. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685PubMedCrossRefGoogle Scholar
  23. Lincoln TM, Hall CL, Park CR, Corbin JD (1976) Guanosine 3′:5′-cyclic monophosphate binding proteins in rat tissues. Proc Natl Acad Sci USA 73:2559–2563PubMedCrossRefGoogle Scholar
  24. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275PubMedGoogle Scholar
  25. Maekawa M, Kamimura K, Nagano T (1996) Peritubular myoid cells in the testis: their structure and function. Arch Histol Cytol 59:1–13PubMedCrossRefGoogle Scholar
  26. Means AR, Dedman JR, Tash JS, Tindall JS, Van Sickle M, Welsh MJ (1980) Regulation of testis sertoli cells by follicle-stimulating hormone. Ann Rev Physiol 42:59–70CrossRefGoogle Scholar
  27. Middendorff R, Muller D, Paust HJ, Holstein AF, Davidoff MS (1997a) New aspects of Leydig cell function. Adv Exp Med Biol 424:125–138PubMedGoogle Scholar
  28. Middendorff R, Muller D, Wichers S, Holstein AF, Davidoff MS (1997b) Evidence for production and functional activity of nitric oxide in seminiferous tubules and blood vessels of the human testis. J Clin Endocrinol Metab 82:4154–4161PubMedCrossRefGoogle Scholar
  29. Mukhopadhyay AK, Schumacher M, Leidenberger FA (1986) Steroidogenic effect of atrial natriuretic factor in isolated mouse Leydig cells is mediated by cyclic GMP. Biochem J 239:463–467PubMedGoogle Scholar
  30. O’Bryan MK, Schlatt S, Gerdprasert O, Phillips DJ, de Kretser DM, Hedger MP (2000) Inducible nitric oxide synthase in the rat testis: evidence for potential roles in both normal function and inflammation-mediated infertility. Biol Reprod 63:1285–1293PubMedCrossRefGoogle Scholar
  31. Palombi F, Di Carlo C (1988) Alkaline phosphatase is a marker for myoid cells in cultures of rat peritubular and tubular tissue. Biol Reprod 39:1101–1109PubMedCrossRefGoogle Scholar
  32. Palombi F, Farini D, Salanova M, de Grossi S, Stefanini M (1992) Development and cytodifferentiation of peritubular myoid cells in the rat testis. Anat Rec 233:32–40PubMedCrossRefGoogle Scholar
  33. Purvis K, Hansson V (1980) Hormonal regulation of spermatogenesis. Regulation of target cell response. Int J Androl Suppl 3:82–143CrossRefGoogle Scholar
  34. Ribalkin SD, Rybalkina IG, Shimizu-Albergina M, Tang X, Beavo JA (2003) PDE5 is converted to an activated state upon cGMP binding to the GAF A domain. EMBO J 23:469–478CrossRefGoogle Scholar
  35. Rossi P, Pezzotti R, Conti M, Geremia R (1985) Cyclic nucleotide phosphodiesterases in somatic and germ cells of mouse seminiferous tubules. J Reprod Fertil 74:317–327PubMedCrossRefGoogle Scholar
  36. Sanborn BM, Heindel JJ, Robison GA (1980) The role of cyclic nucleotides in reproductive processes. Ann Rev Physiol 42:37–57CrossRefGoogle Scholar
  37. Sanders DB, Kelley T, Larson D (2000) The role of nitric oxide synthase/nitric oxide in vascular smooth muscle control. Perfusion 15:97–104PubMedGoogle Scholar
  38. Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354PubMedCrossRefGoogle Scholar
  39. Tung PS, Fritz IB (1990) Characterization of rat testicular peritubular myoid cells in culture: alpha-smooth muscle isoactin is a specific differentiation marker. Biol Reprod 42:351–365PubMedCrossRefGoogle Scholar
  40. Virtanen I, Kallajoki M, Narvanen O, Paranko J, Thornell LE, Miettinen M, Lehto VP (1986) Peritubular myoid cells of human and rat testis are smooth muscle cells that contain desmin-type intermediate filaments. Anat Rec 215:10–20PubMedCrossRefGoogle Scholar
  41. Yanaka N, Kotera J, Ohtsuka A, Akatsuka H, Imai Y, Michibata H, Fujishige K, Kawai E, Takebayashi S, Okumura K, Omori K (1998) Expression, structure and chromosomal localization of the human cGMP-binding cGMP-specific phosphodiesterase PDE5A gene. Eur J Biochem 255:391–399PubMedCrossRefGoogle Scholar
  42. Zhao AZ, Yan C, Sonnenburg WK, Beavo JA (1997) Recent advances in the study of Ca2+/CaM-activated phosphodiesterases: expression and physiological functions. Adv Second Messenger Phosphoprotein Res 31:237–251PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Angela Scipioni
    • 1
  • Stefania Stefanini
    • 1
  • Rocco Santone
    • 2
  • Mauro Giorgi
    • 2
  1. 1.Department of Cellular and Developmental BiologyUniversity “La Sapienza”RomeItaly
  2. 2.Department of Basic and Applied BiologyUniversity of L’AquilaL’AquilaItaly

Personalised recommendations