Gas chromatographic-mass spectrometric analysis of steroids and steroid glucuronides in the seminal vesicle fluid of the African catfish, Clarias gariepinus

Gas chromatographic-mass spectrometric analysis was carried out to identify steroids and steroid glucuronides in the seminal vesicle fluid of African catfish, Clarias gariepinus, collected in the Hula nature reserve (Israel) during the breeding season. Full mass spectra of 5β-pregnane-3α,17α-diol-20-one and cholesterol were obtained. After treatment with β-glucuronidase the following steroid glucuronides were determined by full mass spectra of the corresponding free steroids: etiocholanolone, 5β-androstane-3α,17β-diol-11-one, 5β-pregnane-3α,17α-diol-20-one, and cholesterol. Furthermore, after selected ion monitoring the following steroids and steroid glucuronides could be detected by the presence of at least two characteristic ions at the expected retention time: 5β-androstane-3β,17β-diol, 5β-androstane-3α,17β-diol, etiocholanolone, 5β-androstane-3α,17β-diol-11-one, testosterone, 5β-androstane-3α,17β-diol-glucuronide, and testosterone-glucuronide. These results agree with the hypothesis that steroid glucuronides, synthesized by the seminal vesicles, are excreted with the seminal vesicle fluid into the external environment, where they might function as sex pheromones

Steroidogenesis in ovarian tissue of a viviparous teleost, the guppy Poecilia reticulata

1. 1. Homogenates of ovaries of 3- and 12-month-old guppies were incubated with pregnenolone-7α-3H and progesterone-4-14C, and with androstenedione-1,2-3H, respectively. 2. 2. From the double-labeled experiment, 17α-hydroxypregnenolone, 17α-hydroxyprogesterone, dehydroepiandrosterone, androstenedione and testosterone were isolated and identified by paper- and thin-layer chromatography, derivative formation and by recrystallization to constant specific activity. 3. 3. In the ovary of the guppy the delta-5 pathway in steroidogenesis predominates, as may be concluded from the 3H/14C ratios of the identified metabolites. 4. 4. From the androstenedione-1,2-3H incubation, ovaries of 12-month-old guppies appeared to be able to synthesize steroids with functional groups in the 11-position (11-ketotestosterone and 11β-hydroxytestosterone). 5. 5. The androgen synthesis in ovaries of older guppies may be considered as a possible explanation of the sex conversion occasionally observed

Steroidogenesis in ovarian tissue of a viviparous teleost, the guppy Poecilia reticulata

1. 1. Homogenates of ovaries of 3- and 12-month-old guppies were incubated with pregnenolone-7α-3H and progesterone-4-14C, and with androstenedione-1,2-3H, respectively. 2. 2. From the double-labeled experiment, 17α-hydroxypregnenolone, 17α-hydroxyprogesterone, dehydroepiandrosterone, androstenedione and testosterone were isolated and identified by paper- and thin-layer chromatography, derivative formation and by recrystallization to constant specific activity. 3. 3. In the ovary of the guppy the delta-5 pathway in steroidogenesis predominates, as may be concluded from the 3H/14C ratios of the identified metabolites. 4. 4. From the androstenedione-1,2-3H incubation, ovaries of 12-month-old guppies appeared to be able to synthesize steroids with functional groups in the 11-position (11-ketotestosterone and 11β-hydroxytestosterone). 5. 5. The androgen synthesis in ovaries of older guppies may be considered as a possible explanation of the sex conversion occasionally observed

Aromatase activity in brain and pituitary of immature and mature atlantic salmon (Salmo salar L.) parr

Brain homogenates from Atlantic salmon parr converted tritiated androstenedione to testosterone, estrone, and 17β-estradiol. The formation of estrogens was markedly higher in homogenates of whole brains from mature parr males than from immature parr males. The highest estrogen synthesis was found in the telencephalon and diencephalon. In both of these parts the estrogen production was higher in mature males than in immature males. There was no difference in estrogen formation by pituitaries from immature female and immature male parr, whereas the formation of estrogens and testosterone was higher in the mature male parr

Conservation of apoptosis as an immune regulatory mechanism : effects of cortisol and cortisone on carp lymphocytes

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Hypocorticism and interrenal hyperplasia are not directly related to masculinization in XXmas(-1)/mas(-1) carp, Cyprinus carpio

This study reports on a homozygous XX male strain of common carp (E5), which fail to mount a normal cortisol stress response. Earlier classical genetic analysis had indicated that masculinization of E5 fish was caused by a putative recessive mutation (mas¿1/mas¿1). Hypocorticism in E5 fish was studied to investigate if it was related to masculinization. Head-kidney tissues isolated from E5 fish showed a low cortisol-producing capacity in vitro, and also demonstrated a reduced sensitivity to stimulation with ACTH, when compared with an isogenic XY male carp strain (STD). There was no strain difference in androgen production by head-kidney tissues in vitro. E5 fish exhibited significant hyperplasia of the interrenal tissue (adrenal homologue of teleost fish) located in the head-kidney. Conversion of pregnenolone was significantly lower in E5 head-kidney homogenates, compared to STD homogenates, however, no strain difference was found in the conversion of 17¿-hydroxyprogesterone into cortisol. Gonad homogenates incubated with pregnenolone showed no strain difference in conversion to corticosteroids and androgens. Results indicate that the interrenal hyperplasia and hypocorticism in this strain of carp may be due to a dysfunction of the 17¿-hydroxylase activity of the enzyme P450c17 in the interrenal, but that this defect may not be the primary factor resulting in masculinization of these XX genotype

Steroidogenesis in the testes and seminal vesicles of spawning and non-spawning African catfish, Clarias gariepinus

The in vitro biosynthesis of steroids was studied in testes as well as seminal vesicles of non-spawning pond and spawning feral African catfish, collected during the breeding season. In testes of non-spawners the conversion of [3H]-pregnenolone was directed towards 11-oxygenated androgens and 5β-pregnane-3α,17α,20α-triol, whereas in testes of spawners the production of progesterone and 17α-hydroxyprogesterone prevailed. The seminal vesicles, on the other hand, produced mainly pregnenolone ester, androstenedione and testosterone in non-spawning catfish and 5β-reduced steroids in spawning catfish. Furthermore, in these seminal vesicles, steroid glucuronide synthesis dominated much more than in testes. There was, however, no difference in synthesizing capacity between the two groups. Spawning was accompanied by elevated plasma gonadotropin levels