Isolation of inhibin from ovine follicular fluid

Leversha, Lorraine; Robertson, David; Vos, F.L. de; Morgan, Francis J.; Hearn, Milton T. W.; Wettenhall, Richard E.H.; Findlay, Jock K.; Burger, H. G.; Kretser, David Morritz de

doi:10.1677/joe.0.1130213

Cited by 59 publications

(23 citation statements)

References 11 publications

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“…For members of the TGFsuperfamily, the ability to produce sufficient quantities that are biologically active has been difficult. Earlier work relied upon the purification from follicular and rete testes fluid (Ling et al 1985, Robertson et al 1986, Leversha et al 1987). For recombinant production of inhibin and activin, the production of one (inhibin) leads to the production of the antagonist protein (activin) (Mason et al 1996).…”

Section: Discussionmentioning

confidence: 99%

Production and purification of recombinant human inhibin and activin

Pangas¹,

Woodruff²

2002

Journal of Endocrinology

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Inhibin and activin are protein hormones with diverse physiological roles including the regulation of pituitary FSH secretion. Like other members of the transforming growth factor-gene family, they undergo processing from larger precursor molecules as well as assembly into functional dimers. Isolation of inhibin and activin from natural sources can only produce limited quantities of bioactive protein.To purify large-scale quantities of recombinant human inhibin and activin, we have utilized stably transfected cell lines in self-contained bioreactors to produce protein. These cells produce approximately 200 µg/ml per day total recombinant human inhibin.Conditioned cell media can be purified through column chromatography resulting in dimeric mature 32-34 kDa inhibin A and 28 kDa activin A. The purified recombinant proteins maintain their biological activity as measured by traditional in vitro assays including the regulation of FSH in rat anterior pituitary cultures and the regulation of promoter activity of the activin-responsive promoter p3TP-luc in tissue culture cells. These proteins will be valuable for future analysis of inhibin and activin function and have been distributed to the US National Hormone and Peptide Program.

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Section: Discussionmentioning

confidence: 99%

Production and purification of recombinant human inhibin and activin

Pangas¹,

Woodruff²

2002

Journal of Endocrinology

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“…Inhibin is mainly produced by granulosa cells and is detectable in follicular fluids with multiple molecular forms in female mammals [3,[22][23][24][25]. After injections of inhibin antiserum or NGS, Ig G enters the follicular fluid of antral follicles in the ovary, and this makes it possible for inhibin antibodies, one kind of Ig G, to combine with inhibins in follicular fluid and have local functions in follicular development.…”

Section: Discussionmentioning

confidence: 99%

Ovarian Localization of Immunoglobulin G and Inhibin .ALPHA.-Subunit in Guinea Pigs after Passive Immunization against the Inhibin .ALPHA.-Subunit.

Shi

Mochida

Сузукі

et al. 2000

Journal of Reproduction and Development

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Abstract. The experiment was conducted to elucidate ovarian localization of immunoglobulin G (Ig G) and inhibin α-subunit and follicular development in guinea pigs after passive immunization against the inhibin α-subunit. Estrus was synchronized in laboratory guinea pigs by implanting progesterone containing-tubes. After removal of the progesterone implants, the guinea pigs were injected i.v. with 1.0 ml inhibin antiserum or normal goat serum (NGS). The results showed that Ig G mainly entered the follicular fluid of antral follicles, indicating that possibly inhibin antibodies could be a local regulator affecting follicular development in guinea pigs. The results also showed that passive immunization against inhibin increased the number of follicles and plasma concentrations of estradiol before ovulation, but did not change the plasma concentrations of follicle-stimulating hormone (FSH) or the numbers of follicles stained with inhibin antiserum. The present study first reports that the distribution of Ig G after injections of inhibin antiserum and confirms our previous hypothesis that only dominant follicles could be stained for inhibin α-subunit. The results also imply that only one injection of inhibin antiserum could not increase the ovulation rate in guinea pigs.

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“…The inhibin assay method used was that described for sheep plasma by Findlay et al (1990). The standard, a pool of charcoal-stripped ovine follicular fluid diluted in ovariectomized ewe plasma, contained 1.04 nmol equivalent inhibin ml~, by calibration with a reference standard preparation of pure 31 kDa ovine inhibin (Leversha et al, 1987). This antibody also recognizes the inhibin pro-aC subunit, but not aN.…”

Section: Hormone Assaysmentioning

confidence: 99%

Pituitary and ovarian function in ewes immunized against the amino-terminal peptide ( N) of the inhibin 43-subunit

Russell¹,

Doughton²,

Tsonis³

et al. 1994

Reproduction

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Immunization of ewes against the amino-terminal peptide (alpha N) of the pro-alpha-subunit of inhibin has been shown to reduce fertility, thought to be due to disruption of ovulation. The aims of this study were to examine the effects of active immunization of ewes against alpha N on circulating concentrations of FSH, LH and on ovarian inhibin and progesterone, and to relate these observations to number of corpora lutea and oocyte recovery rates. Ewes were immunized against one or both of two recombinant full length bovine-alpha N immunogens (FP1 and FP2). Three experiments were performed in which jugular venous plasma was sampled from control and immunized ewes: (1) hourly across the oestrous surge of gonadotrophins (Expt 1); (2) daily for one entire oestrous cycle, and in the subsequent cycle, oviducts were flushed to recover ovulated eggs (Expt 2); and (3) samples were taken at 10 min intervals during the follicular and luteal phases (Expt 3). Binding of 125I-labelled alpha N1-26 to serum was greater (P < 0.05) in immunized groups than in controls for all experiments. The number of eggs per corpus luteum recovered from the oviducts was lower (P < 0.05) in the alpha N-immunized groups (39%) than in controls (88%). There were more (P < 0.05) corpora lutea per ewe in FP2 immunized groups 4 (1.8 +/- 0.45) and 5 (1.75 +/- 0.5) than in the control group (1.13 +/- 0.13), but no increase in group 3 (FP1; 1.4 +/- 0.24).(ABSTRACT TRUNCATED AT 250 WORDS)

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Isolation of inhibin from ovine follicular fluid

Cited by 59 publications

References 11 publications

Production and purification of recombinant human inhibin and activin

Production and purification of recombinant human inhibin and activin

Ovarian Localization of Immunoglobulin G and Inhibin .ALPHA.-Subunit in Guinea Pigs after Passive Immunization against the Inhibin .ALPHA.-Subunit.

Pituitary and ovarian function in ewes immunized against the amino-terminal peptide ( N) of the inhibin 43-subunit

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