Follicular fluid stimulation of steroidogenesis in immature granulosa cells in vitro

Ledwitz-Rigby, Florence; Rigby, B W

doi:10.1016/0303-7207(79)90059-5

Cited by 23 publications

(8 citation statements)

References 14 publications

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“…Since all the cultures were grown in the presence of this dose of FSH (close to a saturating dose of FSH) the effect of FSH occurring in follicular fluid or in partially purified fractions, such as lOOng/ml in the second Sephadex peak should be essentially nondetectable. The findings of LedwitzRigby and Rigby [7] also support our conclu sion that the stimulatory activity in LFF is not due to FSH and LH activity purified from follicular fluid. They observed clear-cut stimulation of progesterone secretion with 50% LFF in the presence of high concentra tions of exogenous FSH and LH (2 pg/ml and 0.1 pg/ml, respectively) in immature porcine granulosa cell cultures.…”

Section: Discussionsupporting

confidence: 78%

“…The changes in gonadotropin receptors are accompanied by an increased ability of LH and a decreased ability of FSH to stimulate progesterone se cretion and synthesis of cyclic 3'-5'-AMP [3,4], The control of these maturation processes appear to involve not only the pituitary hor mones (LH and FSH) along with local intraovarian steroids (androgens, estrogens, and progcstins) but also nonsteroidal factors. Several recently described nonsteroidal ovar ian components include oocyte maturation inhibitor, LH receptor binding inhibitor, FSH receptor binding inhibitor, luleinization inhibitor and inhibin-F [5][6][7][8]. Ledwilz-Rigby et al [9] and Rigby et al [10] have demon strated that addition of follicular fluid from large porcine follicles (LFF) to cultures of granulosa cells from small follicles stimu lated progesterone secretion as well as aromatization of androgen [11].…”

Section: Introductionmentioning

confidence: 99%

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Stimulatory Action of Follicular Fluid Components on Maturation of Granulosa Cells from Small Porcine Follicles

Kolena¹,

Channing²

1985

Horm Res

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Follicular fluid obtained from large (6-12 mm) porcine follicles (LFF) was investigated to determine its stimulatory activity on progesterone secretion and on follicle stimulating hormone (FSH) induction of ¹²⁵I-human chorionic gonadotropin (hCG)-luteinizing hormone (LH) binding sites in porcine granulosa cells in a 4-day culture. Incubation of granulosa cells harvested from small porcine follicles (1-2 mm) with 50% LFF led to stimulation of LH/hCG binding sites and progesterone secretion. After partial purification of pooled LFF or proteins precipitated with 90% ethanol on Sephadex G-100 the greatest stimulatory activity was found in the second protein peak eluted from the column. Chromatography of part of the active fraction on DEAE Sephacel using a continuous gradient of NH4HCO₃ yielded seven protein fractions. The second fraction, which eluted early, contained the majority of the stimulatory activity which was purified about 32-fold compared to native LFF. In contrast, addition of follicular fluid recovered from small porcine follicles inhibited FSH induction of LH/hCG binding sites and progesterone secretion. It can be concluded, that maturation of granulosa cells from small follicles may be enhanced by protein(s) present in LFF, but not in fluid recovered from less mature follicles.

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Section: Discussionsupporting

confidence: 78%

Section: Introductionmentioning

confidence: 99%

Stimulatory Action of Follicular Fluid Components on Maturation of Granulosa Cells from Small Porcine Follicles

Kolena¹,

Channing²

1985

Horm Res

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“…Follicular fluid from immature pig follicles has been shown to inhibit progesterone secretion in cultures of pig granulosa cells from large preovulatory follicles (Ledwitz-Rigby et al, 1977,1981Ledwitz-Rigby & Rigby, 1979). "Luteinization inhibitors" have also been reported from bovine (Amsterdam, Riesel, Mintz, Shemesh & Salomon, 1979;Shemesh, 1979;Bernard, 1973Bernard, ,1975 and human (Hillensjöef a/., 1978;Kraiem, Druker & Lunenfield, 1978;Kumari, Vohra & Rayhavan, 1983) follicular fluid.…”

Section: Discussionmentioning

confidence: 91%

“…However, components contained within pig follicular fluid also seem to influence granulosa cell maturation. Ledwitz-Rigby & Rigby (1979) observed that follicular fluid obtained from small follicles inhibited basal and LH-stimulated progesterone secretion, whereas follicular fluid from large follicles enhanced granulosa cell progesterone secretion. Ledwitz-Rigby, Rigby, Long, Stewart & McLean (1981) also reported that follicular fluid obtained from large follicles enhanced granulosa cell conversion of androgen to oestrogen.…”

Section: Introductionmentioning

confidence: 92%

Follicular fluid modulation of functional LH receptor induction in pig granulosa cells

Osteen¹,

Anderson²,

Reichert³

et al. 1985

Reproduction

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Follicular fluid was collected from small (1\p=n-\2mm), medium (3\p=n-\5mm) and large (6\p=n-\12mm) follicles of pigs, treated with charcoal to remove steroids, and tested for effects on the induction of functional LH/hCG receptors in cultures of granulosa cells from small antral pig follicles. Granulosa cells were cultured for 2, 4 or 6 days in Medium 199 + 10% pig serum. Granulosa cells cultured in the presence of purified human FSH ( 0 \ p = n -\ 1 \g=m\g/ml, LER 8/117), insulin (1 mU/ml), cortisol (0\m=.\01\g=m\g/ml) and thyroxine (10\m=-\7 m) accumulated a 4-to 8-fold increase in LH/hCG receptors compared to control cultures. The amounts of cyclic AMP and progesterone secreted after exposure to ovine LH (1 \g=m\g/ml:NIH-S19) were also increased 2\p=n-\3-foldand 80\p=n-\100\x=req-\ fold, respectively. Exposure to FSH alone resulted in lower amounts of LH/hCG receptors with a concomitant decrease in optimum LH responses.Addition of 12\m=.\5\p=n-\50% follicular fluid obtained from small (1\p=n-\2mm) follicles led to a dose-dependent inhibition of the FSH plus insulin, cortisol and thyroxine induction of LH/hCG receptors after 4 days of culture. Fluid from medium follicles showed reduced ability to inhibit LH/hCG receptor induction, and fluid from large follicles exerted only a slight inhibition or no inhibition of receptor induction. Fluid from medium-sized and large follicles exerted a progressive dose-dependent stimulation of progesterone secretion by the granulosa cell cultures. The inhibitory activity was precipitated primarily with 70% ethanol and to a lesser degree by 36 and 90% ethanol. These studies demonstrate that induction of functional LH/hCG receptors in cultures of pig granulosa cells from immature follicles is enhanced by including insulin, cortisol and thyroxine, in addition to FSH, in the culture medium, and that follicular fluid modulates both receptor induction and progesterone secretion as a function of follicular maturation.

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“…The plausible local regulating mechanism may lie in a local follicular luteinization inhibitor, which can inhibit FSH induction of LH receptors in cultured porcine granulosa cells. This inhibitor is present in immature but not large mature porcine follicles (20,26,27). In other studies we have examined luteinization inhibitor in some of these same human follicles and find it to be essentially absent in the preovulatory follicles containing mature oocytes (12 follicles).…”

Section: Discussionmentioning

confidence: 95%

Decline of follicular oocyte maturation inhibitor coincident with maturation and achievement of fertilizability of oocytes recovered at midcycle of gonadotropin-treated women.

Channing

Cheng-quan

Jones

et al. 1983

Proc. Natl. Acad. Sci. U.S.A.

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To examine whether a decline in follicular oocyte maturation inhibitor (OMI) is associated with attainment of oocyte maturation and fertilizability, OMI was measured in follicular fluid (FF) of 39 follicles of 20 normal women given human menopausal gonadotrophin and human chorionic gonadotropin to induce follicular growth and maturation. Oocytes were aspirated per laparoscope, the fluid was saved, and the egg was observed, incubated, and inseminated with the husband's sperm. Concepti that developed to the 4-to 8-cell stage were transferred to the uterus and the women were followed for pregnancy. OMI activity in each FF was measured by using cultured cumulus-enclosed porcine oocytes (30-40 oocytes per FF sample). Estrogen, progesterone, and A4-androstenedione were measured in FF by radioimmunoassay. The FF of 13 preovulatory follicles yielding oocytes that were mature and fertilizable had significantly less OMI activity (mean ± SEM) (0.58 ± 0.10 unit/ml) compared to follicles yielding immature oocytes (2.8 ± 0.56 units/ml; n = 9), atretic oocytes (5.5 ± 2.5 units/ml; n = 7), or preovulatory oocytes with fractured zonae (1.9 ± 0.63 units/ml; n = 7). The estrogen concentration (mean ± SEM) of prevulatory follicles yielding mature fertilizable eggs or mature eggs with fractured zonae was greater (396 ± 34 ng/ml; n = 20) compared to follicles yielding immature or atretic eggs (203 ± 59 ng/ml; n = 9 and 97 ± 47 ng/ml; n = 7, respectively; P < 0.05). Progesterone concentration (mean + SEM; ng/ml) of FF was generally elevated in all preovulatory follicles (635 ± 53) compared to immature or atretic follicles (230 -64 and-76 ± 17, respectively; P < 0.05). It may be concluded that in normal follicle maturation there is a decline in OMI in the follicle containing an oocyte that becomes mature and fertilizable. There is also an increase in estrogen, progesterone, and follicle size. It is also possible to have an abnormal follicle maturation when there is an increase in size as well as FF, estrogen, and progesterone, but without a decline in OMI-a situation which can lead to production of a nonfertilizable oocyte.It is well documented that the human oocyte completes its meiotic maturation within the dominant preovulatory follicle within 30-48 hr prior to ovulation (1, 2). It is believed that the preovulatory gonadotropin surge brings this maturation about; yet the mechanism is poorly understood. Oocyte maturation only occurs within the dominant mature follicle and not in the adjacent less mature follicles. Several investigators in this laboratory have partially purified an oocyte maturation inhibitor (OMI) from immature porcine follicles, which may play a role in keeping the oocyte in the immature dictyate state (3-6).Whether a decline in OMI at the time of ovulation is the cause of oocyte maturation is not known. Furthermore, the mechanisms responsible for choosing the dominant follicle to be ovulated are not well understood. The purpose of this study was to measure changes in OMI in preovulatory human follicles and...

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Follicular fluid stimulation of steroidogenesis in immature granulosa cells in vitro

Cited by 23 publications

References 14 publications

Stimulatory Action of Follicular Fluid Components on Maturation of Granulosa Cells from Small Porcine Follicles

Stimulatory Action of Follicular Fluid Components on Maturation of Granulosa Cells from Small Porcine Follicles

Follicular fluid modulation of functional LH receptor induction in pig granulosa cells

Decline of follicular oocyte maturation inhibitor coincident with maturation and achievement of fertilizability of oocytes recovered at midcycle of gonadotropin-treated women.

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