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Restart of steroidogenesis in dogs during recrudescence of testicular function following downregulation with a GnRH-agonist implant

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Abstract

To date, no details are available concerning the restart of steroidogenesis following the downregulation of testicular endocrine and germinative function by gonadotrophin-releasing hormone (GnRH)-agonist implants. This restart was assessed by determining the expression of steroidogenic acute regulatory (StAR) protein, cytochrome P450 side-chain cleavage enzyme (P450scc) and cytochrome P450 17α-hydroxylase,17,20-lyase (P450c17). The re-establishment of steroidogenesis was initiated by the removal of the GnRH-agonist implant (18.5 mg azagly nafarelin, Gonazon) at 5 months after treatment. Testes were removed at 3-week intervals (weeks 0–24) and four groups were formed according to the stage of spermatogenesis as revealed by the most developed germ cells observed (developmental group [DG] spermatocytes to DG elongated spermatids). Five dogs served as untreated controls. Positive immunostaining for StAR, P450scc and P450c17 was restricted to Leydig cells. Western blot indicated the specifity of the respective antibodies with hints of a expression of canine-specific P450scc and P450c17 proteins. A significant effect of group was observed for a percentage of the immunopositive area (PIA) as an indicator of active Leydig cells for StAR (P<0.05), P450scc (P<0.001) and P450c17 (P<0.001), with PIA being lowest for the DG spermatocytes. With regard to the strength of the immunopositive signal, a significant effect of group was found for P450scc (P<0.01) and P450c17 (P<0.05), with the lowest intensity being observed in DG spermatocytes. At the mRNA level, the upregulation from DG spermatocytes to DG round spermatids was clearly evident but was only significant for P450scc (P<0.05). Thus, downregulation affects the whole cascade of steroidogenesis, whereas withdrawal of inhibition results in a rapid restart, in part indicating a rebound phenomenon.

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References

  • Amory JK, Bremner W (2001) Endocrine regulation of testicular function in men: implications for contraceptive development. Mol Cell Endocrinol 182:175–179

    Article  PubMed  CAS  Google Scholar 

  • Bustamante E, Soper JW, Pedersen PL (1977) A high-yield preparative method for isolation of rat liver mitochondria. Anal Biochem 80:401–408

    Article  PubMed  CAS  Google Scholar 

  • Clark BJ, Wells J, King SR, Stocco DM (1994) The purification, cloning, and expression of a novel luteinizing hormone-induced mitochondrial protein in MA-10 mouse Leydig tumor cells. Characterization of the steroidogenic acute regulatory protein (StAR). J Biol Chem 269:28314–28322

    PubMed  CAS  Google Scholar 

  • Goericke-Pesch S, Spang A, Schulz M, Özalp G, Bergmann M, Ludwig C, Hoffmann B (2009) Recrudescence of spermatogenesis in the dog following downregulation using a slow release GnRH agonist implant. Reprod Domest Anim 44 (Suppl 2):302–308

    Article  PubMed  Google Scholar 

  • Goericke-Pesch S, Wilhelm E, Hoffmann B (2010a) Hormonelle downregulation der Hodenfunktion bei Rüde und Kater; eine retrospektive studie. Prakt Tierarztl 91:563–570

    Google Scholar 

  • Goericke-Pesch S, Wilhelm E, Ludwig C, Desmoulins PO, Driancourt MA, Hoffmann B (2010b) Evaluation of the clinical efficacy of Gonazon® implants in the treatment of reproductive pathologies, behavioural problems and suppression of reproductive function in the male dog. Theriogenology 73:920–926

    Article  PubMed  CAS  Google Scholar 

  • Hall PF (1988) Testicular steroid synthesis: organisation and regulation. In: Knobil E (ed) The physiology of reproduction. Raven, New York, pp 975–994

    Google Scholar 

  • Hoffmann B, Büsges F, Baumgärtner W (2004) Immunohistochemical detection of CD4-, CD8- and MHC II-expressing immune cells and endoglin in the canine corpus luteum at different stages of dioestrus. Reprod Domest Anim 39:391–395

    Article  PubMed  CAS  Google Scholar 

  • Junaidi A, Williamson PE, Cummins JM, Martin GB, Blackberry MA, Trigg TE (2003) Use of a new drug delivery formulation of the gonadotrophin-releasing hormone analogue deslorelin for reversible long-term contraception in male dogs. Reprod Fertil Dev 15:317–322

    Article  PubMed  CAS  Google Scholar 

  • Junaidi A, Williamson PE, Martin GB, Stanton PG, Blackberry MA, Cummins JM, Trigg TE (2007) Pituitary and testicular endocrine responses to exogenous gonadotrophin-releasing hormone (GnRH) and luteinising hormone in male dogs treated with GnRH agonist implants. Reprod Fertil Dev 19:891–898

    Article  PubMed  CAS  Google Scholar 

  • Junaidi A, Williamson PE, Trigg TE, Cummins JM, Martin GB (2009a) Morphological study of the effects of the GnRH superagonist deslorelin on the canine testis and prostate gland. Reprod Domest Anim 44:757–763

    Article  PubMed  CAS  Google Scholar 

  • Junaidi A, Williamson PE, Martin GB, Blackberry MA, Cummins JM, Trigg TE (2009b) Dose–response studies for pituitary and testicular function in male dogs treated with the GnRH superagonist, deslorelin. Reprod Domest Anim 44:725–734

    Article  PubMed  CAS  Google Scholar 

  • Kowalewski MP, Hoffmann B (2008) Molecular cloning and expression of StAR protein in the canine corpus luteum during dioestrus. Exp Clin Endocrinol Diabetes 116:158–161

    Article  PubMed  CAS  Google Scholar 

  • Kowalewski MP, Schuler G, Taubert A, Engel E, Hoffmann B (2006) Expression of cyclooxygenase 1 and 2 in the canine corpus luteum during diestrus. Theriogenology 66:1423–1430

    Article  PubMed  CAS  Google Scholar 

  • Ludwig C, Desmoulins PO, Driancourt MA, Goericke-Pesch S, Hoffmann B (2009) Reversible downregulation of endocrine and germinative testicular function (hormonal castration) in the dog with the GnRH-agonist azagly-nafarelin as a removable implant "Gonazon"; a preclinical trial. Theriogenology 71:1037–1045

    Article  PubMed  CAS  Google Scholar 

  • Miller WL (1987) Structures of genes encoding steroidogenic enzyme. J Steroid Biochem 27:759–766

    Article  PubMed  CAS  Google Scholar 

  • Miller WL (2002) Androgen biosynthesis from cholesterol to DHEA. Mol Cell Endocrinol 198:7–14

    Article  PubMed  CAS  Google Scholar 

  • Payne AH, Hales DB (2004) Overview of steroidogenic enzymes in the pathway from cholesterol to active steroid hormones. Endocr Rev 25:947–970

    Article  PubMed  CAS  Google Scholar 

  • Payne AH, O’Shaughnessy PJ (1996) Structure, function and regulation of steroidogenic enzymes in the Leydig cell. In: Payne AH, Hardy MP, Russsell LD (eds) The Leydig cell. Cache River Science, St. Louis, pp 260–285

    Google Scholar 

  • Peterson JK, Moran F, Conley AJ, Bird IM (2001) Zonal expression of endothelial nitric oxide synthase in sheep and rhesus adrenal cortex. Endocrinology 142:5351–5363

    Article  PubMed  CAS  Google Scholar 

  • Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 29:e45

    Article  PubMed  CAS  Google Scholar 

  • Riesenbeck A, Klein R, Hoffmann B (2002) Downregulation, a new and reversible approach to eliminate testicular function in the dog. Prakt Tierarztl 83:512–520

    Google Scholar 

  • Roby KF, Larsen D, Deb S, Soares MJ (1994) Generation and characterization of antipeptide antibodies to rat cytochrome P-450 side-chain cleavage enzyme. Mol Cell Endocrinol 79:13–20

    Article  Google Scholar 

  • Rone MB, Fan J, Papadopoulos V (2009) Cholesterol transport in steroid biosynthesis: role of protein-protein interactions and implications in disease states. Biochim Biophys Acta 1791:646–658

    Article  PubMed  CAS  Google Scholar 

  • Sachs L (1982) Applied statistics—a handbook of techniques. Springer, Heidelberg Berlin New York

    Google Scholar 

  • Saez JM (1994) Leydig cells: endocrine, paracrine, and autocrine regulation. Endocr Rev 15:574–626

    PubMed  CAS  Google Scholar 

  • Schuler G, Özalp GR, Hoffmann B, Harada N, Browne P, Conley AJ (2006) Reciprocal expression of 17alpha-hydroxylase-C17,20-lyase and aromatase cytochrome P450 during bovine trophoblast differentiation: a two-cell system drives placental oestrogen synthesis. Reproduction 131:669–679

    Article  PubMed  CAS  Google Scholar 

  • Sheskin D (2007) Handbook of parametric and nonparametric statistical procedures, 4th edn. Chapman & Hall/CRC, Boca Raton

    Google Scholar 

  • Stocco DM, Clark BJ (1996a) Regulation of the acute production of steroids in steroidogenic cells. Endocr Rev 17:221–244

    PubMed  CAS  Google Scholar 

  • Stocco DM, Clark BJ (1996b) Role of the steroidogenic acute regulatory protein (StAR) in steroidogenesis. Biochem Pharmacol 51:197–205

    Article  PubMed  CAS  Google Scholar 

  • Stocco DM, McPhaul MJ (2006) Physiology of testicular steroidogenesis. In: Neil JD (ed) Knobil and Neills physiology of reproduction. Elsevier/Academic Press, Amsterdam/New York, pp 977–1016

    Chapter  Google Scholar 

  • Trigg TE, Wright PJ, Armour AF, Williamson PE, Junaidi A, Martin GB, Doyle AG, Walsh J (2001) Use of a GnRH analogue implant to produce reversible long-term suppression of reproductive function in male and female domestic dogs. J Reprod Fertil Suppl 57:255–261

    PubMed  CAS  Google Scholar 

  • van der Molen HJ, Eik-Nes NB (1971) Biosynthesis and secretion of steroids by the canine testis. Biochem Biophys Acta 248:343–362

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

The authors gratefully thank Dr. Gabi Itter (Sanofi Aventis, Frankfurt) for providing the Beagle dogs. They also greatly appreciate the provision of the Gonazon implants by Intervet Pharma R&D, Angers Technopole, France and the good cooperation of those people who took the dogs into their custody. We are also grateful to Prof. D.M. Stocco, Texas Technical University Health Sciences Center, Lubbock, Texas, USA for kindly providing the StAR antibody and to Prof. A. Conley, UC Davis, California, USA, for kindly providing the P450c17 antibody.

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Authors and Affiliations

  1. Clinic for Obstetrics, Gynecology and Andrology of Large and Small Animals, Justus-Liebig-University Giessen, Frankfurter Strasse 106, 35392, Giessen, Germany

    M. Gentil, B. Hoffmann, A. Spang & S. Goericke-Pesch

  2. Unit for Biomathematics and Data Processing, Justus-Liebig-University Giessen, Frankfurter Strasse 95, 35392, Giessen, Germany

    K. Failing

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  1. M. Gentil
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  2. B. Hoffmann
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  3. A. Spang
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  4. K. Failing
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  5. S. Goericke-Pesch
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Correspondence to S. Goericke-Pesch.

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Gentil, M., Hoffmann, B., Spang, A. et al. Restart of steroidogenesis in dogs during recrudescence of testicular function following downregulation with a GnRH-agonist implant. Cell Tissue Res 350, 513–523 (2012). https://doi.org/10.1007/s00441-012-1506-5

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