Induced luteal regression: differential effects on follicular and luteal inhibin/activin subunit mRNAs in the marmoset monkey

1995 ◽  
Vol 144 (2) ◽  
pp. 201-208 ◽  
Author(s):  
H M Fraser ◽  
S F Lunn ◽  
P F Whitelaw ◽  
S G Hillier
Keyword(s):  
Corpus Luteum ◽  
Granulosa Cells ◽  
Luteal Phase ◽  
Gnrh Antagonist ◽  
High Expression ◽  
Corpora Lutea ◽  
Ovulatory Cycle ◽  
Α Subunit ◽  
Luteal Regression ◽  
Antral Follicles

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

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.

Localization of ovine follistatin and α and βA inhibin mRNA in the sheep ovary during the oestrous cycle

1994 ◽  
Vol 12 (2) ◽  
pp. 181-193 ◽  
Author(s):  
D J Tisdall ◽  
N Hudson ◽  
P Smith ◽  
K P McNatty
Keyword(s):  
Gene Expression ◽  
Granulosa Cells ◽  
Ovarian Follicle ◽  
Indirect Evidence ◽  
Oestrous Cycle ◽  
Corpora Lutea ◽  
Α Subunit ◽  
Ovarian Follicle Development ◽  
Antral Follicles

ABSTRACT The sites of follistatin and α and βA inhibin gene expression were examined by in situ hybridization in sheep ovaries during the early and mid-luteal phases (days 3 and 10) of the oestrous cycle and a prostaglandin F2α (PGF2α)-induced follicular phase. Follistatin mRNA was detected in the granulosa cells of preantral, antral and early atretic follicles at all stages of the oestrous cycle, and in the corpora lutea at the early and mid-luteal stages of the cycle. However, only low levels of expression of follistatin were observed in the presumptive preovulatory follicle at 56 h after treatment with PGF2α. Both α and βA inhibin were shown to be expressed in ovaries at all stages of the oestrous cycle. In situ hybridization localized α subunit mRNA to the granulosa cells of most, but not all, healthy antral follicles, and to no other ovarian cell type. In contrast, expression of the βA subunit was confined to a few medium-to-large healthy antral follicles. In antral follicles expressing βA inhibin, mRNAs for α inhibin and follistatin were always detected, but the converse was not true. Unlike follistatin, no α and βA inhibin expression was seen in preantral follicles, developing corpora lutea, or follicles undergoing atresia. These results show that, in the adult sheep ovary, follistatin gene expression is a constitutive event in all growing follicles from the early preantral stage, and also provide indirect evidence of the involvement of follistatin, but not inhibin or activin, in the early stages of ovarian follicle development in sheep.

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.

scholarly journals 11β-Hydroxysteroid dehydrogenase expression and activities in bovine granulosa cells and corpora lutea implicate corticosteroids in bovine ovarian physiology

2007 ◽  
Vol 193 (2) ◽  
pp. 299-310 ◽  
Author(s):  
L M Thurston ◽  
D R E Abayasekara ◽  
A E Michael
Keyword(s):  
Molecular Mass ◽  
Granulosa Cells ◽  
Luteal Phase ◽  
Hydroxysteroid Dehydrogenase ◽  
Follicular Phase ◽  
Corpora Lutea ◽  
Luteal Cells ◽  
Antral Follicles ◽  
Follicle Diameter ◽  
Dependent Type

Cortisol–cortisone metabolism is catalysed by the bi-directional NADP(H)-dependent type 1 11β-hydroxysteroid dehydrogenase (11βHSD1) enzyme and the oxidative NAD+-dependent type 2 11βHSD (11βHSD2). This study related the expression of 11βHSD1 and 11βHSD2 enzymes (mRNA and protein) to net 11-ketosteroid reductase and 11β-dehydrogenase (11β-DH) activities in bovine follicular granulosa and luteal cells. Granulosa cells were isolated from follicles of < 4, 4–8, > 8 and > 12 mm in diameter in either the follicular or luteal phase of the ovarian cycle. Luteal cells were obtained from corpora lutea (CL) in the early non-pregnant luteal phase. Enzyme expression was assessed by reverse transcription-PCR and western blotting, while enzyme activities were measured over 1 h in cell homogenates using radiometric conversion assays with 100 nM [3H]cortisone or [3H]cortisol and pyridine dinucleotide cofactors. Irrespective of follicle diameter, the expression of 11βHSD2 and NAD+-dependent oxidation of cortisol predominated in granulosa cells harvested in the follicular phase. In contrast, in granulosa cells obtained from luteal phase follicles and in bovine luteal cells, expression of 11βHSD1 exceeded that of 11βHSD2 and the major enzyme activity was NADP+-dependent cortisol oxidation. Increasing follicular diameter was associated with progressive increases in expression and activities of 11βHSD2 and 11βHSD1 in follicular and luteal phase granulosa cells respectively. In follicular phase granulosa cells from antral follicles < 12 mm, 11βHSD1 migrated with a molecular mass of 34 kDa, whereas in the dominant follicle, CL and all luteal phase granulosa cells, a second protein band of 68 kDa was consistently detected. In all samples, 11βHSD2 had a molecular mass of 48 kDa, but in large antral follicles (> 8 mm), there was an additional immunoreactive band at 50 kDa. We conclude that 11βHSD2 is the predominant functional 11βHSD enzyme expressed in follicular phase granulosa cells from growing bovine antral follicles. In contrast, in bovine granulosa cells from dominant or luteal phase follicles, and in bovine luteal cells from early non-pregnant CL, 11βHSD1 is the major glucocorticoid-metabolising enzyme. The increasing levels of cortisol inactivation by the combined NADP+- and NAD+-dependent 11β-DH activities suggest a need to restrict cortisol access to corticosteroid receptors in the final stages of follicle development.

Induced luteal regression in the primate: evidence for apoptosis and changes in c-myc protein

1995 ◽  
Vol 147 (1) ◽  
pp. 131-137 ◽  
Author(s):  
H M Fraser ◽  
S F Lunn ◽  
G M Cowen ◽  
P J Illingworth
Keyword(s):  
Corpus Luteum ◽  
Luteal Phase ◽  
Primate Species ◽  
Morphological Evidence ◽  
Molecular Evidence ◽  
Corpora Lutea ◽  
Intense Staining ◽  
Cytoplasmic Staining ◽  
Luteal Regression ◽  
Luteal Cells

Abstract There is increasing molecular evidence that apoptosis is involved in the process of structural luteal regression in non-primate species. Apoptosis is dependent upon the activation of certain proto-oncogenes and c-myc protein has an important regulatory role in this process in some cell types. The aim of the present study was to determine the occurrence and localisation of c-myc protein within the primate corpus luteum, determine changes during induction of luteal regression and examine the corpora lutea for morphological evidence of apoptosis. Ovaries were studied from marmoset monkeys in the late follicular, and in the early, mid and late luteal phases. Luteal regression was induced either by treatment with prostaglandin F2α analogue or GnRH antagonist administered during the mid luteal phase and ovaries obtained 24 and 48 h later. Immunocytochemistry was performed using a monoclonal antibody to the c-myc protein. In pre-ovulatory follicles positive staining was found in the nucleus of a few granulosal cells and in the cytoplasm of thecal cells. c-myc was present in all corpora lutea where it was localised predominantly in the cytoplasm. In early corpora lutea, scattered cells with intense staining were observed in the presence of a majority of moderately or weakly stained cells. In the mid and late luteal phases, corpora lutea were uniformly moderately stained for c-myc. Following induction of luteal regression, nuclear degeneration with condensation and fragmentation indicative of apoptosis was observed. In other luteal cells, increased cytoplasmic volume and dissolution of cellular and nuclear membranes suggested necrosis. After luteal regression the overall intensity of staining for c-myc declined, but was present at high signal concentration in the cytoplasm of those cells whose morphological integrity was best maintained following treatment. In a minority of steroidogenic luteal cells, both nuclear and cytoplasmic staining was observed. These results suggest that after ovulation there appears to be a marked increase in c-myc production in the cytoplasm of the luteal cells of the developing corpus luteum and that c-myc is present throughout the luteal phase. During induced luteal regression c-myc may undergo a transitory rise and transfer to the nucleus and both apoptosis and necrosis occur during the process of luteolysis. Journal of Endocrinology (1995) 147, 131–137

Immunocytochemical demonstration of oxytocin in bovine ovarian tissues

1985 ◽  
Vol 109 (4) ◽  
pp. 537-542 ◽  
Author(s):  
Th. A. M. Kruip ◽  
H. G. B. Vullings ◽  
D. Schams ◽  
J. Jonis ◽  
A. Klarenbeek
Keyword(s):  
Acetic Acid ◽  
Corpus Luteum ◽  
Granulosa Cells ◽  
Ovarian Tissue ◽  
Picric Acid ◽  
Corpora Lutea ◽  
Luteal Cells ◽  
Antral Follicles

Abstract. The presence of oxytocin in ovarian tissue was examined immunocytochemically. Bovine antral follicles and corpora lutea were fixed with glutaraldehyde, picric acid and acetic acid fixative and immuno-stained by the indirect peroxidase-antiperoxydase (PAP) technique. Immunoreactive oxytocin was demonstrated in the granulosa cells of small and large follicles, in the granulosalutein cells of the young corpus luteum and in the large luteal cells of the mature corpus luteum. The regressing corpus luteum was not stainable. It is discussed that these findings additionally support the view that oxytocin is actually synthesized in ovarian tissues.

scholarly journals Immunohistological Localization and Expression of α-Actin in the Baboon (Papio anubis) Corpus Luteum

1997 ◽  
Vol 45 (1) ◽  
pp. 71-77 ◽  
Author(s):  
Firyal S. Khan-Dawood ◽  
Jun Yang ◽  
M. Yusoff Dawood
Keyword(s):  
Corpus Luteum ◽  
Luteal Phase ◽  
Adherens Junctions ◽  
Tunica Albuginea ◽  
Corpora Lutea ◽  
Papio Anubis ◽  
Lh Surge ◽  
Luteal Cells ◽  
Steroidogenic Cells ◽  
E Cadherin

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.

A comparative study of the corpus luteum

1995 ◽  
Vol 7 (3) ◽  
pp. 303 ◽  
Author(s):  
RT Gemmell
Keyword(s):  
Corpus Luteum ◽  
Luteal Phase ◽  
Secretory Granules ◽  
Hormonal Control ◽  
Egg Laying ◽  
Oestrous Cycle ◽  
Corpora Lutea ◽  
Embryonic Diapause ◽  
Alternative Reproductive Strategy ◽  
Uterine Development

The corpus luteum (CL) is a transitory organ which has a regulatory role in reproduction. Sharks, amphibians and reptiles have corpora lutea that produce progesterone which influences the rate of embryonic development. The egg-laying monotremes and the two major mammalian groups, eutherian and marsupial, have a CL that secretes progesterone. Most eutherians have allowed for the uterine development of their young by extending the length of the oestrous cycle and the CL or placenta actively secretes progesterone until birth. Gestation in the marsupial does not extend beyond the length of an oestrous cycle and the major part of fetal development takes place in the pouch. Where the extension of the post-luteal phase in the eutherian has allowed for the uterine development of young, the marsupial has extended the pre-luteal phase of the oestrous cycle and has evolved an alternative reproductive strategy, embryonic diapause. The mechanism for the secretion of hormones from the CL has been controversial for many years. Densely-staining secretory granules have been observed in the CL of sharks, marsupials and eutherians. These granules have been reported to contain relaxin, oxytocin or mesotocin, and progesterone. A hypothesis to suit all available data is that all hormones secreted by the CL are transported within such granules. In conclusion, although there are obvious differences in the mode of reproduction in the two main mammalian groups, it is apparent that there is a great deal of similarity in the hormonal control of regression of the CL and parturition.

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

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