Immunocytochemical localization of inhibin α-subunit in the human corpus luteum

1991 ◽  
Vol 129 (1) ◽  
pp. 155-NP ◽  
Author(s):  
K. B. Smith ◽  
M. R. Millar ◽  
A. S. McNeilly ◽  
P. J. Illingworth ◽  
H. M. Fraser ◽  
...  
Keyword(s):  
Corpus Luteum ◽  
Ovarian Tissue ◽  
Corpora Lutea ◽  
Immunoperoxidase Technique ◽  
Similar Distribution ◽  
Α Subunit ◽  
Primary Antiserum ◽  
Specific Localization ◽  
Human Menstrual Cycle ◽  
Human Corpus Luteum

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

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
Keyword(s):  
Gene Expression ◽  
Corpus Luteum ◽  
Luteal Phase ◽  
Corpora Lutea ◽  
Subunit Gene ◽  
Nucleic Acid Probes ◽  
Α Subunit ◽  
Normal Human ◽  
Human Menstrual Cycle ◽  
Human Corpus Luteum

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.

STEROID METABOLIC PATHWAYS IN THE OVARY OF THE CHINCHILLA (CHINCHILLA LANIGER)

1972 ◽  
Vol 52 (1) ◽  
pp. 37-50 ◽  
Author(s):  
W. H. TAM
Keyword(s):  
Corpus Luteum ◽  
Metabolic Pathways ◽  
Ovarian Tissue ◽  
Controlling Factor ◽  
Steroid Metabolism ◽  
Interstitial Tissue ◽  
Corpora Lutea ◽  
Luteal Tissue ◽  
Kinetic Investigations ◽  
High Yields

SUMMARY The ovarian tissue components of the pregnant chinchilla were incubated with equimolar amounts of [7α-3H]pregnenolone and [4-14C]progesterone. The greater contribution by [7α-3H]pregnenolone than by [4-14C]progesterone towards the formation of 17α-hydroxyprogesterone and androstenedione, and the relatively high yields of 17α-hydroxypregnenolone and dehydroepiandrosterone showed that both the 4-ene and 5-ene pathways of steroid metabolism were used in the interstitial tissue. No significant amount of 17α-hydroxylation was observed in the primary and accessory corpora lutea. The results of kinetic investigations using [7α-3H]pregnenolone as substrate also demonstrated a precursor—product relationship between dehydroepiandrosterone and androstenedione in the interstitial tissue, but this was not apparent in the luteal tissue. The results indicated that the interstitial tissue was capable of synthesizing progesterone and oestrogens as major products, and that the lack of 17α-hydroxylation in the luteal tissue was a controlling factor ensuring the synthesis of progesterone as its principal hormonal product. A small amount of [4-14C]dehydroepiandrosterone was always isolated with a much larger amount of the tritiated compound. This implied the conversion of 14C-labelled 4-en-3-oxosteroids into 5-ene-3β-hydroxysteroids which has generally been regarded as impossible. The isolation of this product, which may be an artifact, and the possibility that progesterone and oestrogens may be synthesized by different cells (granulosa and theca lutein cells) in the corpus luteum, or that there may be a third pathway for oestrogen synthesis, as suggested by the results of the kinetic experiments, are discussed.

Expression of tissue inhibitor of metalloproteinases-1 in the human corpus luteum after luteal rescue

1996 ◽  
Vol 148 (1) ◽  
pp. 59-67 ◽  
Author(s):  
W C Duncan ◽  
A S McNeilly ◽  
P J Illingworth
Keyword(s):  
Corpus Luteum ◽  
Luteal Phase ◽  
Proteolytic Enzymes ◽  
Specific Inhibitor ◽  
Northern Blotting ◽  
Corpora Lutea ◽  
Tissue Inhibitor Of Metalloproteinases ◽  
Tissue Inhibitor ◽  
Tissue Remodelling ◽  
Human Corpus Luteum

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

Oxytocin may play a role in the control of the human corpus luteum

1982 ◽  
Vol 95 (1) ◽  
pp. 65-70 ◽  
Author(s):  
G. J. S. Tan ◽  
R. Tweedale ◽  
J. S. G. Biggs
Keyword(s):  
Menstrual Cycle ◽  
Corpus Luteum ◽  
Inhibitory Action ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Corpora Lutea ◽  
Luteal Cells ◽  
Progesterone Production ◽  
Human Corpus Luteum

The effects of oxytocin on dispersed luteal cells from human corpora lutea of the menstrual cycle were studied. Oxytocin at a concentration of 4 mi.u./ml produced a slight increase in basal progesterone production. However, higher oxytocin concentrations (400 and 800 mi.u./ml) markedly inhibited both basal and human chorionic gonadotrophin-induced progesterone production. These data provide evidence for an effect of oxytocin on the human corpus luteum. In view of the inhibitory action of oxytocin, increased secretion of this hormone may be important in the demise of the corpus luteum at the end of the menstrual cycle.

PROPERTIES OF HUMAN GONADOTROPHINS ELUTED FROM HUMAN CORPUS LUTEUM AND MOUSE LUTEOMA LH-HCG RECEPTORS

1977 ◽  
Vol 84 (1) ◽  
pp. 142-154 ◽  
Author(s):  
F. E. Cole ◽  
P. C. Arquembourg ◽  
B. F. Rice
Keyword(s):  
Corpus Luteum ◽  
Electrophoretic Mobility ◽  
Present Report ◽  
Corpora Lutea ◽  
Fresh Tissue ◽  
Mouse Tissues ◽  
Tissue Receptors ◽  
Human Corpus Luteum

ABSTRACT Studies were performed to try to determine if gonadotrophins are altered during their interaction with tissue receptors. Immunologic, electrophoretic and binding properties of lactoperoxidase labelled [125I]HLH and [125I]HCG were examined before and after elution from mouse luteoma and human corpora lutea receptor preparations. The anti-HCG used in these studies at a 1:10 000 dilution precipitated 92% of a freshly iodinated [125I]HCG preparation. Receptor eluted [125I]HCG, derived from the same batch of labelled ligand, was virtually quantitatively precipitated by the same dilution of anti-HCG. [125I]HCG eluted from the human corpus luteum was electrophoretically more homogenous when compared to its heterogenous parent labelled preparation and migrated to a position similar to that of native HCG. In Ouchterlony double diffusion experiments against anti-HCG antiserum, corpus luteum eluted [125I]HCG and [125I]HLH showed immunologic identity with each other as well as with native HCG and HLH. Receptor eluted [125I]HCG from the mouse luteoma, following in vivo administration via tail vein injection or after incubation in vitro with labelled hormones, was immunologically indistinguishable from native HCG. The electrophoretic mobility of HCG was retarded when HCG was added to extracts of mouse luteoma, liver and kidney. Eluates of mouse luteoma, applied to Bio-Gel columns previously equilibrated with [125I]HCG showed the ability to concentrate [125I]HCG in the high molecular weight column fractions. Similar results were obtained with columns equilibrated with [125I]TSH and [125I]HGH. [125I]HCG eluted from the mouse luteoma was able to bind to fresh luteoma homogenate but, in contrast to an earlier report with [125I]HCG eluted from rat testis, no enhancement of binding of the eluted [125I]HCG was observed with fresh tissue. These results could be explained by the extraction of non-dialyzable intracellular component during the [125I]HCG elution procedure from the luteoma homogenate which combines with HCG to lower its binding and alter its electrophoretic mobility. This component could be extracted from other mouse tissues and combines with other labelled peptide hormones. Data in the present report support in part the hypothesis that gonadotrophins eluted from mouse luteoma and human corpus luteum are not altered by their interaction with tissue receptors.

CONCENTRATION OF PROSTAGLANDIN F2α AND STEROIDS IN THE HUMAN CORPUS LUTEUM

1977 ◽  
Vol 73 (1) ◽  
pp. 115-122 ◽  
Author(s):  
I. A. SWANSTON ◽  
K. P. McNATTY ◽  
D. T. BAIRD
Keyword(s):  
Menstrual Cycle ◽  
Corpus Luteum ◽  
Luteal Phase ◽  
Prostaglandin F2α ◽  
Corpora Lutea ◽  
Progesterone Concentration ◽  
Late Luteal Phase ◽  
Prostaglandin F ◽  
Human Corpus Luteum

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.

CONVERSION OF ANDROST-4-ENE-3,17-DIONE-4-14C TO OESTROGENS BY SUBCELLULAR FRACTIONS OF THE HUMAN CORPUS LUTEUM OF THE NORMAL CYCLE

1967 ◽  
Vol 56 (2) ◽  
pp. 225-230 ◽  
Author(s):  
Rosalinda B. Arceo ◽  
Kenneth J. Ryan
Keyword(s):  
Subcellular Localization ◽  
Corpus Luteum ◽  
Paper Chromatography ◽  
Specific Activity ◽  
Subcellular Fractions ◽  
Corpora Lutea ◽  
Differential Centrifugation ◽  
Normal Cycle ◽  
Generating System ◽  
Human Corpus Luteum

ABSTRACT The subcellular localization of the aromatizing enzymes from four human corpora lutea was investigated. Subcellular fractions were prepared by differential centrifugation, and incubations of each fraction were carried out with androst-4-ene-3,17-dione-4-14C and a TPNH generating system. Oestrogen metabolites were characterized by phenolic separation, repeated paper chromatography, incubation with freshly prepared placental 17β-ol dehydrogenase, methylation and recrystallization to constant specific activity. The experimental results indicate that the aromatizing enzymes were localized mainly in the microsomal fractions of homogenates of human corpora lutea.

Localization of inhibin/activin subunit mRNAs during the luteal phase in the primate ovary

1993 ◽  
Vol 10 (3) ◽  
pp. 245-257 ◽  
Author(s):  
H M Fraser ◽  
S F Lunn ◽  
G M Cowen ◽  
P T K Saunders
Keyword(s):  
Corpus Luteum ◽  
Granulosa Cells ◽  
Luteal Phase ◽  
Biologically Active ◽  
Corpora Lutea ◽  
Α Subunit ◽  
Antral Follicles ◽  
Subunit Mrna ◽  
High Level ◽  
Low Expression

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.

Lack of direct inhibitory action of oxytocin on progesterone production by dispersed cells from human corpus luteum

1985 ◽  
Vol 104 (1) ◽  
pp. 149-151 ◽  
Author(s):  
M. C. Richardson ◽  
G. M. Masson
Keyword(s):  
Corpus Luteum ◽  
Luteal Phase ◽  
Inhibitory Action ◽  
Corpora Lutea ◽  
Luteal Cells ◽  
Menstrual Cycles ◽  
Progesterone Production ◽  
Human Corpus Luteum ◽  
Dispersed Cells

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

scholarly journals Effect of nitric oxide on the expression of insulin-like growth factors and the insulin-like growth factor binding proteins throughout the lifespan of the human corpus luteum

Reproduction ◽  
2001 ◽  
pp. 865-873 ◽  
Author(s):  
G Iniguez ◽  
A Villavicencio ◽  
F Gabler ◽  
A Palomino ◽  
M Vega
Keyword(s):  
Nitric Oxide ◽  
Growth Factors ◽  
Corpus Luteum ◽  
Luteal Phase ◽  
Nitric Oxide Donors ◽  
Corpora Lutea ◽  
Igf I ◽  
Human Corpus Luteum ◽  
Igfbp 3 ◽  
Insulin Like Growth Factors

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.

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