INHIBIN GENE EXPRESSION IN THE HUMAN CORPUS LUTEUM

1987 ◽  
Vol 115 (3) ◽  
pp. R21-R23 ◽  
Author(s):  
S.R. Davis ◽  
Z. Krozowski ◽  
R.I. McLachlan ◽  
H.G. Burger

ABSTRACT We report inhibin α- and βA -subunit gene expression in the human corpus luteum and placenta using human α-subunit and bovine βA -subunit nucleic acid probes. In addition, we have demonstrated the presence of immunoreactive and bioactive inhibin in human corpora lutea. Our findings suggest that this tissue is a significant source of inhibin during the luteal phase of the normal human menstrual cycle.

1991 ◽  
Vol 129 (1) ◽  
pp. 155-NP ◽  
Author(s):  
K. B. Smith ◽  
M. R. Millar ◽  
A. S. McNeilly ◽  
P. J. Illingworth ◽  
H. M. Fraser ◽  
...  

ABSTRACT The localization of inhibin α-subunit within the human corpus luteum was investigated. The antiserum used was raised in sheep against the first 1–23 amino acid sequence of the N-terminus of the human inhibin α-subunit. Using the avidin-biotin immunoperoxidase technique, intense immunostaining was localized within the granulosa-lutein cells of the corpus luteum, with absence of staining in the theca-lutein cells and surrounding ovarian tissue. Similar distribution of inhibin α-subunit immunostaining was observed in 12 corpora lutea obtained during the early, mid- and late-luteal phases and no changes in intensity were apparent at these different stages. Negative controls were obtained by applying antiserum which had been preabsorbed overnight with excess inhibin peptide in place of primary antiserum and also normal non-immune sheep serum as a substitute for primary antiserum. These results provide further evidence that the human corpus luteum is a significant source of immunoreactive inhibin during the normal human menstrual cycle. The specific localization within the granulosalutein cells of the corpus luteum suggests that inhibin α-subunit production may originate from a discrete cell population within the human corpus luteum. Journal of Endocrinology (1991) 129, 155–160


1996 ◽  
Vol 148 (1) ◽  
pp. 59-67 ◽  
Author(s):  
W C Duncan ◽  
A S McNeilly ◽  
P J Illingworth

Abstract Tissue inhibitor of metalloproteinases-1 (TIMP-1) is a specific inhibitor of a group of proteolytic enzymes known as matrix metalloproteinases. These enzymes have been widely implicated in the process of tissue remodelling. Extensive remodelling occurs in the corpus luteum during luteolysis unless human chorionic gonadotrophin (hCG) is produced by the early conceptus. This study aimed to investigate the expression and localisation of TIMP-1 in human corpora lutea during the luteal phase of the cycle and after luteal rescue with exogenous hCG to mimic the changes of early pregnancy. Human corpora lutea from the early (n = 4), mid- (n=4) and late (n=4) luteal phases and after luteal rescue by hCG (n=4) were obtained at the time of hysterectomy. Expression of TIMP-1 was investigated in these tissues by Western blotting, immunohistochemistry, Northern blotting and in situ hybridisation. Luteal cells of thecal origin were distinguished from those of granulosa origin by immunostaining for 17α-hydroxylase. A 30 kDa protein consistent with TIMP-1 was detected in human corpora lutea. This protein was localised to the granulosa lutein cells in all tissues examined. TIMP-1 mRNA was found in large quantities in all glands examined and this again localised to the granulosa lutein cells. The expression and localisation of TIMP-1 did not change throughout the luteal phase and was not altered by luteal rescue. The function of this uniform expression of TIMP-1 in the corpus luteum is not clear but these data suggest that the inhibition of structural luteolysis during maternal recognition of pregnancy is not mediated by regulation of TIMP-1 expression. Journal of Endocrinology (1996) 148, 59–67


1977 ◽  
Vol 73 (1) ◽  
pp. 115-122 ◽  
Author(s):  
I. A. SWANSTON ◽  
K. P. McNATTY ◽  
D. T. BAIRD

SUMMARY The concentration of prostaglandin F2α (PGF2α), progesterone, pregnenolone, oestradiol-17β, oestrone, androstenedione and testosterone was measured in corpora lutea obtained from 40 women at various stages of the menstrual cycle. The concentration of PGF2α was significantly higher in corpora lutea immediately after ovulation (26·7 ± 3·9 (s.e.m.) ng/g, P < 0·005) and in corpora albicantia (16·3 ± 3·3 ng/g, P < 0·005) than at any other time during the luteal phase. There was no correlation between the concentration of PGF2α and that of any steroid. The progesterone concentration was highest in corpora lutea just after ovulation (24·9 ± 6·7 μg/g) and in early luteal groups (25·7 ± 6·8 μg/g) but declined significantly (P < 0·05) to its lowest level in corpora albicantia (1·82 ± 0·66 μg/g). The concentration of oestradiol-17β in the corpus luteum and luteal weight were significantly greater during the mid-luteal phase than at any other stage (concentration 282 ± 43 ng/g, P < 0·05; weight 1·86 ± 0·18 g, P < 0·005). The results indicate that regression of the human corpus luteum is not caused by a rise in the ovarian concentration of PGF2α in the late luteal phase of the cycle.


1993 ◽  
Vol 10 (3) ◽  
pp. 245-257 ◽  
Author(s):  
H M Fraser ◽  
S F Lunn ◽  
G M Cowen ◽  
P T K Saunders

ABSTRACT Localization of inhibin/activin subunit mRNAs within the macaque ovary from the immediate pre-ovulatory period of the menstrual cycle, when serum immunoreactive inhibin begins to rise, to day 9 of the luteal phase, when serum inhibin concentrations are maximal, was investigated using in-situ hybridization. Ovaries were studied on the day of the LH surge (day 0) and on days 2, 5, and 9 of the luteal phase by hybridizing frozen tissue sections with radio-labelled riboprobes specific to the inhibin/activin α-, βA-and βB-subunits. After autoradiographic exposure for 10 and 21 days, grain concentrations were quantified by image analysis. Moderate expression of α-, βA- and βB-subunit mRNA was present within the granulosa cells of the pre-ovulatory follicle (day 0). The granulosa-lutein cells of the corpora lutea expressed high levels of α-subunit at days 2, 5 and 9. mRNAs for βA and βB were detected at low but significant levels in all of the corpora lutea. All healthy antral follicles exhibited a high level of expression of βB-subunit mRNA in the granulosa cells. On day 2 after ovulation these follicles also expressed high α- and moderate βA-subunit mRNA. On day 9 the βB-inhibin mRNA in antral follicles was found in association with low expression of the other subunits. Small follicles in ovaries on day 2 expressed moderate α- and low levels of βB-subunit mRNA, while mRNA for βA was absent. α-subunit mRNA expression was present on day 5 while neither βA- nor βB-subunit mRNA was detected. On day 9 a proportion of small follicles expressed α- and βA-subunit mRNA. These results demonstrate that marked differences are present in the levels of expression of the three inhibin/activin subunit genes between follicles and the corpus luteum. The predominance of the βB-subunit mRNA within antral follicles would be consistent with the synthesis of activin. The predominance of the α-subunit combined with the low expression of the β-subunits in the corpus luteum suggests that both biologically active inhibin and free α-subunit are produced by the primate corpus luteum.


1985 ◽  
Vol 104 (1) ◽  
pp. 149-151 ◽  
Author(s):  
M. C. Richardson ◽  
G. M. Masson

ABSTRACT Suspensions of luteal cells were prepared from samples of human corpora lutea obtained during the luteal phase of menstrual cycles. Addition of oxytocin (1 μmol/l) to the various cell preparations had no effect on either basal production of progesterone or on steroidogenic responses to a range of concentrations of gonadotrophin. J. Endocr. (1985) 104, 149–151


Reproduction ◽  
2001 ◽  
pp. 865-873 ◽  
Author(s):  
G Iniguez ◽  
A Villavicencio ◽  
F Gabler ◽  
A Palomino ◽  
M Vega

The presence of insulin-like growth factors (IGF), IGF binding proteins (IGFBP) and IGF receptor type 1 (IGF-IR) in the human corpus luteum was investigated by examining the expression and production of related proteins throughout the lifespan of the corpus luteum and the action of nitric oxide upon their production. The expression of proteins in corpora lutea from the early, mid-and late luteal phases was assessed by immunohisto-chemistry, evaluated by a semi-quantitative analysis and the functional study was performed in corpus luteum explants incubated with nitric oxide donors. IGF-I and -II and IGFBP-1 and -3 were measured in the culture media by specific immunoassays. The results showed that IGF-I and -II, IGFBP-1 to -6 and IGF-IR were detected in the human corpus luteum throughout the luteal phase. Moreover, the expression and production of IGF-I and IGFBP-1 increased progressively from corpora lutea from the early to late luteal phases (P < 0.05), whereas the expression and production of IGFBP-2, -4 and -5 were significantly higher in corpora lutea from the mid-luteal phase (P < 0.05). No differences were observed in the expression of IGF-II, IGFBP-3 and -6 and IGF-IR throughout the lifespan of the corpus luteum. However, functional studies showed that nitric oxide donors elicited a stimulatory action on production of IGF-I in corpora lutea from the early luteal phase (80%) and on production of IGFBP-1 in corpora lutea from the late luteal phase (50%) (P < 0.05), whereas production of IGF-II and IGFBP-3 was not affected by nitric oxide. In conclusion, the components of the IGF-IGFBP system are expressed in the human corpus luteum throughout its lifespan. Nitric oxide regulates IGF-I and IGFBP-1 production, indicating that the growth factors may serve, at least in part, as mediators of the action of nitric oxide in the human corpus luteum.


1984 ◽  
Vol 103 (1) ◽  
pp. 107-110 ◽  
Author(s):  
M. G. Hunter

ABSTRACT Human luteal tissue recovered from varying stages of the luteal phase was minced and incubated for 3 h and the effect of human chorionic gonadotrophin (hCG), prolactin and hCG + prolactin on progesterone and oestradiol production measured. While hCG generally enhanced both progesterone and oestradiol synthesis, prolactin alone at either 20 or 200 μg/l had no significant effect on steroidogenesis. When prolactin was added along with hCG in four of six corpora lutea, however, progesterone production significantly increased and in three of six corpora lutea oestradiol production was increased above that induced by hCG alone. It is concluded that prolactin may play some role in the control of steroidogenesis by the human corpus luteum. J. Endocr. (1984) 103, 107–110


1995 ◽  
Vol 144 (2) ◽  
pp. 201-208 ◽  
Author(s):  
H M Fraser ◽  
S F Lunn ◽  
P F Whitelaw ◽  
S G Hillier

Abstract During the luteal phase of the primate ovulatory cycle the predominant inhibin/activin subunit mRNAs produced by the corpus luteum and antral follicles are those for the α- and βB-subunits respectively. The control of expression of these mRNAs and the resultant nature of the endocrine and paracrine signals which they may potentially generate has yet to be elucidated. Inhibin/activin subunit mRNAs may have a role in both the paracrine regulation of follicular and luteal function and modulation of FSH secretion. The aim of this study was to investigate the expression of inhibin/activin subunit mRNAs following luteal regression induced by either withdrawal of LH support (GnRH antagonist treatment), or by a direct inhibitory action (prostaglandin administration). Marmoset monkeys with regular ovulatory cycles were treated on day 8 and 9 of the luteal phase with either GnRH antagonist, prostaglandin or vehicle (n=3 per group). Ovaries were studied 48 h after onset of treatment (on day 10 of the luteal phase) by hybridizing frozen tissue sections with radiolabelled riboprobes specific to the inhibin/activin α-, βA- and βB-subunit mRNAs. After autoradiographic exposure, grain concentrations were quantified by image analysis. In corpora lutea from control marmosets there was high expression of α-mRNA with only marginal expression of βB-mRNA. Corpora lutea in animals treated with GnRH antagonist or prostaglandin had markedly reduced expression of α-mRNA while βB-mRNA was unchanged. In controls, all healthy antral follicles exhibited a high level of expression of βB-mRNA in the granulosa cells and low expression of α-mRNA in theca cells. This was unaffected by either treatment. βA-mRNA was found at a low level in granulosa cells but was not evident at a significant level in the corpora lutea of any of the groups. These results demonstrate (1) the marmoset corpus luteum is a source of high expression of α-subunit mRNA, (2) this α-mRNA is dependent upon LH support, (3) the process of luteal regression takes place without alteration of βB-mRNA. Antral follicle α- and βB-mRNAs are independent of the process of luteal regression or gonadotrophic withdrawal during the period of the luteal-follicular phase transition. Journal of Endocrinology (1995) 144, 201–208


1997 ◽  
Vol 45 (1) ◽  
pp. 71-77 ◽  
Author(s):  
Firyal S. Khan-Dawood ◽  
Jun Yang ◽  
M. Yusoff Dawood

We have recently shown the presence of E-cadherin and of α- and γ-catenins in human and baboon corpora lutea. These are components of adherens junctions between cells. The cytoplasmic catenins link the cell membrane-associated cadherins to the actin-based cytoskeleton. This interaction is necessary for the functional activity of the E-cad-herins. Our aim therefore was to determine the presence of α-actin in the baboon corpus luteum, to further establish whether the necessary components for E-cadherin activity are present in this tissue. An antibody specific for the smooth muscle isoform of actin, α-actin, was used for these studies. The results using immunohistochemistry show that (a) α-actin is present in steroidogenic cells of the active corpus luteum, theca externa of the corpus luteum, cells of the vasculature, and the tunica albuginea surrounding the ovary. The intensity of immunoreactivity for α-actin varied, with the cells of the vasculature reacting more intensely than the luteal cells. A difference in intensity of immunoreactivity was also observed among the luteal cells, with the inner granulosa cells showing stronger immunoreactivity than the peripheral theca lutein cells. There was no detectable immunoreactivity in the steroidogenic cells of the atretic corpus luteum. However, in both the active and atretic corpora lutea, α-actin-positive vascular cells were dispersed within the tissue. (b) Total α-actin (luteal and non-luteal), as determined by Western blot analyses, does not change during the luteal phase and subsequent corpus luteum demise (atretic corpora lutea). (c) hCG stimulated the expression of α-actin and progesterone secretion by the early luteal phase (LH surge + 1–5 days) and midluteal phase (LH surge + 6–10 days) cells in culture, but only progesterone in the late luteal phase (LH surge + 11–15 days). The data show that α-actin is present in luteal cells and that its expression is regulated by hCG, thus suggesting that E-cadherin may form functional adherens junctions in the corpus luteum.


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