Seasonal variation in tissue estrogen-2/4-hydroxylases (EH) and in vitro effects of steroids on ovarian EH activity in the catfish Heteropneustes fossilis

• Published in: Steroids Vol. 75; no. 13; pp. 1097 - 1105
• Main Authors: Chourasia, T.K., Joy, K.P.
• Format: Journal Article
• Language: English
• Published: Kidlington Elsevier Inc 12.12.2010; Elsevier
• Subjects: Animals; Aryl Hydrocarbon Hydroxylases - metabolism; Biological and medical sciences; Brain - cytology; Brain - drug effects; Brain - enzymology; Catecholestrogens; Catfish; Catfishes; Cytochrome P-450 CYP1A1 - metabolism; Cytochrome P-450 CYP1B1; Female; Fundamental and applied biological sciences. Psychology; In Vitro Techniques; Intracellular Space - drug effects; Intracellular Space - enzymology; Liver - cytology; Liver - drug effects; Liver - enzymology; Organ Specificity; Ovary - cytology; Ovary - drug effects; Ovary - enzymology; Reproducibility of Results; Seasonal variation; Seasons; Steroid effects; Steroid Hydroxylases - metabolism; Steroids - pharmacology; Tissue estrogen hydroxylases; Vertebrates: endocrinology; Tissue estrogen hydroxylases; Catecholestrogens; Catfish; Steroid effects; Seasonal variation; Steroid; Ovary; Female genital system; Estrogen; Sex steroid hormone; In vitro; Ovarian hormone
• ISSN: 0039-128X; 1878-5867
• DOI: 10.1016/j.steroids.2010.07.004

▶ EH activity was the highest in microsomes of liver >brain >ovary. ▶ The enzyme showed seasonal changes with the highest activity in spawning phase. ▶ In oocyte follicles, strong activity was elicited by the follicular layer. ▶ Steroids showed varying degree of suppressive effects, P4 and 2-hydroxy-E2 were highly effective. A radiometric assay was used to measure microsomal EH activity from tritiated H 2O formed during the conversion of [2,4 3H] estradiol-17β into catecholestrogens in the microsomal fractions of liver, brain and ovary of the catfish Heteropneustes fossilis. The validation data show that enzyme activity increased with incubation time, and substrate and cofactor (NADPH) concentrations, elicited temperature optima of 30–37 °C and pH optima of 6.8–7.8. EH activity was strongly NADPH-dependent and in its absence only 13.48% activity was recorded. Liver recorded the highest enzyme activity, followed by brain and ovary. EH activity showed a significant seasonal variation with the peak activity in spawning phase and the lowest activity in resting phase. In the ovary, the follicular layer (theca and granulosa) elicited the highest activity over that of the denuded oocytes. Modulatory effects of steroids on ovarian enzyme activity were further demonstrated. The incubation of postvitellogenic follicles with 1, 10 or 100 nM concentrations of various steroids for 24 h produced varied effects on EH activity. Progesterone and 2-hydroxyestradiol-17β elicited strong suppressive effects on enzyme activity. Estrogens (E 1, E 2 and E 3) suppressed the activity in a concentration-dependent manner. Among the progestins tested, 17,20α-dihydroxy-4-pregnen-3-one, the isomer of 17,20β-dihydroxy-4-pregnen-3-one (a teleost maturation-inducing steroid) showed the lowest depressing effect. Among androgens, the testosterone metabolite 11-ketotestosterone (functional teleost androgen) showed a high suppressing effect. Corticosteroids elicited low activity with cortisol suppressed the activity at higher concentrations. The study will form a basis to understand the physiological role of catecholestrogens in ovarian functions.