Characterization of 11β-Hydroxysteroid Dehydrogenase Activity in Testicular Tissue of Control and GnRH-Immunized Boars as a Possible Regulator of Spermatogenesis

Abstract

Glucocorticoids are involved in the regulation of spermatogenesis in the boar testis by initiating apoptosis in early stages of germ cell development. Because cortisol activity is modulated by the 11β-hydroxysteroid dehydrogenase system (11β-HSD), the present study determined both 11β-activating (reductive) and inactivating (oxidative) enzyme activities in testicular tissue preparations of control boars (n = 5), GnRH-immunized boars (n = 5), and immunized, estradiol-infused boars (n = 6) by radioenzyme assay based on the conversion of tritiated cortisol to cortisone in the presence of NAD⁺ (inactivation) or tritiated cortisone to cortisol in the presence of NADPH (activation). The presence of both isoforms, 11β-HSD 1 and 11β-HSD 2, was confirmed by RTPCR in testicular tissue of control boars. Additionally, cortisol, testosterone, estradiol, and LH were determined in blood plasma sampled twice before killing. Immunization led to a drop of LH from 4.3 ± 0.3 pmol/L to 1.0 ± 0.3 pmol/L (testosterone: 11.63 ± 0.83 nmol/L vs. 0.28 ± 0.07 nmol/L; 17β-estradiol: 512.61 ± 47.60 pmol/L vs. 77.05 ± 14.00 pmol/L). Low 11β-HSD reductive activity was found in boars (1 pmol steroid × min^-1 × mg^-1). It decreased to trace amounts in immunized boars (0.07 pmol steroid × min^-1 × mg^-1). Oxidative activity was found in boars with 10.19 ± 2.28 pmol steroid × min^-1 × mg^-1 protein. Immunization led to a sharp decrease (0.08 ± 0.03 pmol × min^-1 × mg^-1). Infusion of 17β-estradiol significantly elevated peripheral estradiol concentrations to 752.07 ± 24.19 pmol/L which still is a physiological concentration in this species. The infusion led to a minimal reductive activity (0.04 pmol steroid × min^-1 × mg^-1), but led to a 6-fold rise of the 11β-HSD oxidative activity to 0.47 ± 0.14 pmol × min^-1 × mg^-1 compared to immunized boars. It is concluded that the 11β-HSD system is involved in the regulation of cortisol activity in the testis and thus in the regulation of spermatogenesis.

Key words

11β-hydroxysteroid dehydrogenase - boar testis - immunization - cortisol - spermatogenesis

References

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