Studies on Tissue Culture of Equine Ovarian Cell Types: Steroidogenesis

Channing, Cornelia P.; Grieves, Susan A.

doi:10.1677/joe.0.0430391

Cited by 23 publications

(39 citation statements)

References 17 publications

(27 reference statements)

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“…The extraction procedures used for the follicular fluid and ovarian vein plasma were essentially those described by Short (1960), Channing (1969) and Channing & Grieves (1969). The radioactive pregnenolone and androstenedione were rechromatographed on paper immediately before use.…”

Section: Methodsmentioning

confidence: 99%

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Pathways of Steroid Biosynthesis in the Intact Graafian Follicle of Mares in Oestrus

Younglai¹,

Short²

1970

Journal of Endocrinology

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Two mares were operated on during oestrus; in the first animal a mixture of [16-3H]pregnenolone and [4-14C]androstenedione was injected into the largest Graafian follicle; in the second animal, [4-14C]pregnenolone and [7-3H]androstenedione were injected into the follicle. By cannulating the ovarian veins it was possible to measure the rate of transfer of radioactivity from the follicular fluid into the ovarian vein. This ranged from 3 to 9 % of the injected dose per hour. At the end of each experiment, 45-52 min. after the injection of the labelled precursors, the follicular fluid was aspirated. Negligible amounts of radioactivity were recovered from the rest of the ovary and from the 'conjugated' fraction of the ovarian vein plasma and follicular fluid. Within the follicle 6\m=.\2and 12\m=.\2 % of the injected pregnenolone were converted to progesterone. Further metabolism to 17 \g=a\-hydroxyprogesterone, androstenedione and oestradiol-17\g=b\ was very limited.Oestradiol-17\g=b\ isolated from the ovarian vein had a higher specific activity than that from the follicular fluid while the 3H : 14C ratios were similar in both compartments. In addition, the 3H:14C ratios of androstenedione and oestradiol-17\g=b\ in the ovarian vein were similar. Therefore it seems likely that oestradiol-17\g=b\ in the follicular fluid is derived mainly from an extra-follicular source, probably the theca interna cells.

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Section: Methodsmentioning

confidence: 99%

“…Tissue culture studies of equine granulosa cells established that their prin¬ cipal secretory product was indeed progesterone. However, they also secreted small amounts of oestradiol-ß (Channing, 1966;Channing & Grieves, 1969). Thecal cells in culture secreted relatively more oestradiol-17/?…”

Section: Introductionmentioning

confidence: 99%

Pathways of Steroid Biosynthesis in the Intact Graafian Follicle of Mares in Oestrus

Younglai¹,

Short²

1970

Journal of Endocrinology

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“…in the dark followed by extraction, chromatography in system B, scanning and elution of the oestrone region. [16a-3H]20a-Hydroxypregn-4-en-3-one was synthesized by in¬ cubating [16a-3H]pregnenolone with a culture derived from luteal cells and purified as described in a previous communication (Channing & Grieves, 1969). [7a-3H]20/?-Hydroxypregn-4-en-3-one was synthesized from [7a-3H]progesterone by reaction with 20/?-hydroxysteroid dehydrogenase according to Henning & Zander (1962) followed by chromatography in system A, scanning and elution of the 20/?-hydroxypregn-4-en-3-one.…”

Section: Methodsmentioning

confidence: 99%

“…Counting and gas-liquid chromatography were carried out as described previously (Channing & Grieves, 1969). Extraction procedure.…”

Section: Methodsmentioning

confidence: 99%

“…It has been shown previously that cultures of equine granulosa cells harvested from large vascular Graafian follicles undergo hypertrophy and hyperplasia (Chan¬ ning, 1969) in addition to secreting large amounts of hormones characteristic of luteal tissue, namely progesterone and 20a-hydroxypregn-4-en-3-one (Channing, 1966;Channing & Grieves, 1969). The present experiments were designed to deter¬ mine the principal pathways of steroidogenesis in granulosa cell cultures and to see whether they resemble those used by granulosa cells or luteal tissue.…”

Section: Introductionmentioning

confidence: 94%

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Studies on Tissue Culture of Equine Ovarian Cell Types: Pathways of Steroidogenesis

Channing¹

1969

Journal of Endocrinology

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Equine granulosa cell cultures were incubated with various labelled steroid hormone precursors, and the products of these incubations were purified and characterized by a combination of paper chromatography, derivative formation and recrystallization. Sodium acetate, cholesterol and pregnenolone were converted mainly to progesterone in addition to significant amounts of 20\g=a\-hydroxypregn-4-en-3-one and 17-hydroxyprogesterone plus small amounts of dehydroepiandrosterone, androstenedione and oestradiol-17\g=b\.Both androstenedione and testosterone were converted to oestrone and oestradiol-17\g=b\. Testosterone was converted to androstenedione but the converse did not apply. These experiments demonstrated that equine granulosa cells in culture contain all the enzymes for converting acetate to progesterone and oestradiol-17\g=b\ and that the pathways are similar to those of luteal tissue.

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Histochemical study of ovarian hydroxysteroid dehydrogenase activity during normal pseudopregnancy in the rat

Kovarik

Velardo

1979

Anat. Rec.

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ABSTRACT3P-Hydroxysteroid dehydrogenase activity was localized in ovaries removed from rats during normal pseudopregnancy. Ovarian sections were incubated in media containing either pregnenolone (P-one) or dehydroepiandrosterone (DHA). P-one-3P-HSD activity appeared strongest on day 6 whereas DHA-3P-HSD was highest on day 13. NADH, diaphorase activity was localized and observed to be greater than 3P-HSD.Both P-one-and DHA-3P-HSD activities were localized in primary-, secondary-and tertiary follicles; granulosa and theca interna cells have a variable potential for steroidogenesis. Mature tertiary follicles appear most responsive of the follicular population in steroidogenic potential. In atretic follicles, theca interna cells showed a marked ability for potential steroid production and may be one of the more stable sites of steroidogenesis. Interstitial tissue is a major site for potential estrogenic and progestational biosynthesis during the functional life of the ovary. Luteal cells exhibited strong ability for DHA-3P-HSD activity during the ongoing process of pseudopregnancy while P-one-3P-HSD activity declined in these cells as pseudopregnancy progressed. In summary, a t the onset of pseudopregnancy, it appears that ovarian P-one-3P-HSD activity is greater than DHA-3P-HSD activity. Near the termination of this condition, DHA-3P-HSD activity is greater than P-one-3P-HSD activity in the rat.

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Studies on Tissue Culture of Equine Ovarian Cell Types: Steroidogenesis

Cited by 23 publications

References 17 publications

Pathways of Steroid Biosynthesis in the Intact Graafian Follicle of Mares in Oestrus

Pathways of Steroid Biosynthesis in the Intact Graafian Follicle of Mares in Oestrus

Studies on Tissue Culture of Equine Ovarian Cell Types: Pathways of Steroidogenesis

Histochemical study of ovarian hydroxysteroid dehydrogenase activity during normal pseudopregnancy in the rat

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