Testis-Like Steroidogenesis: MATERIALS AND METHODS(1)

17 Apr
2013

MATERIALS AND METHODS(1)

Animals

Twenty-three animals were trapped live in the vicinity of Newcastle, England, in late April-early May and mid-November, corresponding to the Northern Hemisphere spring and autumn, respectively. Animals were returned to the laboratory in Aberdeen (approximately 4 h), where they were weighed, killed, and dissected. Ovotestes and testes were frozen on dry ice before storage at -80°C. Plasma was separated from whole blood by centrifugation and stored at -80°C. buy yasmin online

In Vitro Incubations

Ovotestes from spring females were left intact, whereas those from autumn animals were more easily dissected into their respective ovarian and interstitial gland portions; however, it should be noted that while we were successful in removing ovarian tissue from the interstitial gland, the ovarian portion invariably contained some contaminating interstitial gland. Frozen gonads were homogenized individually in 500 ^l of 0.05 M Tris (pH 7.4) with a glass microhomogenizer. The objective of these studies was to determine the ratio of androgens to estrogens in ovotestes of reproductively active and inactive females. Consequently, protein content was not quantified for each sample; rather, gonadal homogenates were diluted such that after the incubation period not all of the precursor steroid was metabolized.

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