scholarly journals Cell death during natural and induced luteal regression in mares

Reproduction ◽  
2002 ◽  
pp. 67-77 ◽  
Author(s):  
MO Al-Zi'abi ◽  
HM Fraser ◽  
ED Watson
Keyword(s):  
Cell Death ◽  
Luteal Phase ◽  
Apoptotic Cell ◽  
Dense Body ◽  
Smooth Endoplasmic Reticulum ◽  
Corpora Lutea ◽  
Luteal Regression ◽  
Luteal Cells ◽  
Dense Bodies ◽  
Marked Accumulation

In mares, little information is available on the type of cell death that occurs during natural and induced luteal regression. Corpora lutea were collected from mares in the early luteal phase, days 3-4 (n = 4); mid-luteal phase, day 10 (n = 5); early regression, day 14 (n = 4); late regression, day 17 (n = 4); and 12 and 36 h (n = 3 per group) after PGF2alpha administration on day 10. Histological and ultrastructural sections were examined and TUNEL was used to detect DNA fragmentation. In early luteal regression, there were more pyknotic luteal cells and extracellular round dense bodies compared with the mid-luteal phase. By late regression, there was a significant decline (P < 0.01) in the number of round dense body clusters and a marked accumulation of lipid. Twelve and 36 h after PGF2alpha administration, changes were similar to those seen in natural regression, but there was also a marked infiltration of neutrophils. Accumulation of lipid was not apparent until 36 h after PGF2alpha administration. Ultrastructural examination revealed rarefaction and distortion of the mitochondrial cristae in most of the luteal cells by the mid-luteal phase. Luteal cells showed shrinkage, accumulation of lipid with foamy appearance, and disruption in both smooth endoplasmic reticulum and mitochondria during natural and induced regression. Some luteal cells showed fragmented or pyknotic chromatin characteristic of apoptosis. Other luteal cells showed crenation of the nuclear membrane and shrinkage of the nucleus, features not characteristic of apoptotic cell death. In late regression, capillaries were obstructed by swollen endothelial cells and round dense bodies. These results show that structural regression may be initiated as early as the mid-luteal phase, and is clearly visible by day 14 in natural regression and 12 h after induced regression. Apoptosis did appear to be involved in luteolysis in the equine corpus luteum, but non-apoptotic changes were also observed in some luteal cells during regression. Accumulation of lipid was a late feature of luteal regression.

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

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.

Growth hormone and insulin-like growth factor-I action on progesterone secretion by porcine corpora lutea isolated at various periods of the luteal phase

2003 ◽  
Vol 51 (2) ◽  
pp. 197-208 ◽  
Author(s):  
Anna Ptak ◽  
Ewa L. Gregoraszczuk ◽  
J. Rząsa
Keyword(s):  
Growth Hormone ◽  
Growth Factor ◽  
Luteal Phase ◽  
Actinomycin D ◽  
Corpora Lutea ◽  
Luteal Cells ◽  
Factor I ◽  
Progesterone Secretion ◽  
Igf I ◽  
Porcine Luteal Cells

This study was conducted to investigate the interactions between growth hormone (GH) and insulin-like growth factor-I (IGF-I) on progesterone (P4) secretion by porcine luteal cells cultured in vitro. Cells isolated from corpora lutea (CL) collected at three different periods of the luteal phase (CL1 - early luteal phase; CL2 - middle luteal phase and CL3 - late luteal phase) were incubated with different doses of GH (10, 100 or 200 ng/ml). After 48 h cultures were terminated and the media were frozen until further P4 concentration analysis. GH (100 ng/ml) increased P4 secretion by CL1 and CL2 and had no effect on CL3. In separate studies these cells were treated for 48 h with IGF-I alone or with GH combined with IGF-I. IGF-I alone increased basal P4 secretion only by cells collected from CL1 while concurrent treatment with GH had no effect on P4 secretion by any type of CL. To investigate the possible mechanism of GH and IGF-I mediated induction of P4 secretion, an inhibitory study was conducted. In this experiment, luteal cells collected from CL1 were cultured in the absence or presence of cycloheximide (an inhibitor of protein synthesis) or actinomycin D (an inhibitor of DNA transcription). Cycloheximide or actinomycin D completely blocked the stimulatory effect of both GH and IGF-I on P4 production but did not reduce basal progesterone secretion suggesting involvement of gene transcription and translation in the GH and IGF-I action on luteal cells. Additionally, the activity of 3ß-hydroxysteroid dehydrogenase (3ß-HSD) under the influence of GH added alone or together with IGF was measured by the conversion of pregnenolone to progesterone. Stimulation of P4 secretion in P5-treated cells in GH-stimulated cultures was not observed, however, high stimulatory effect was noted in IGF-I treated cultures. In conclusion, the present studies indicate that there is direct and cycle stage dependent influence of GH and IGF-I on steroidogenesis in porcine luteal cells. It is suggested that both IGF and GH may exert some regulatory action during CL development in the pig.

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 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.

scholarly journals Expression of matrix metalloproteinases to induce the expression of genes associated with apoptosis during corpus luteum development in bovine

2018 ◽  
Author(s):  
Sang Hwan Kim ◽  
Ji Hye Lee ◽  
Jong Taek Yoon
Keyword(s):  
Cell Death ◽  
Matrix Metalloproteinases ◽  
Programmed Cell Death ◽  
Luteal Phase ◽  
Expression Patterns ◽  
Basal Membrane ◽  
Luteal Cell ◽  
Luteal Cells ◽  
Expression Of Genes ◽  
Lutein Cell

Here we investigated the expressions of apoptosis-associated genes known to induce programmed cell death through mRNA expressions of two matrix metalloproteinases (MMPs) that are involved in the degradation of collagen and basal membrane in luteal cells cultured in the treatment media. Our results show that the activity of MMP-2 gelatinase was higher in the CL2 and CL1 of luteal phase, was gradually decreased in the CH2 and CH3 of luteal phase. In particular, the expressions of P4-r and survival-associated genes (IGFr, PI3K, AKT, and mTOR) were strongly induced during CL3 stage, whereas the levels of these genes in CL were lower during CL2 and CL1 stages. And in the cultured lutein cell analyzed result, we found that as MMPs increase, genes related to apoptosis ( 20α-HSD and Casp-3) also increase. In other words, the results for P4-r and survival-related gene expression patterns in the luteal cells were contrary to the MMPs activation results. These results indicate that active MMPs are differentially expressed to induce the expression of genes associated with programmed cell death from the degrading luteal cells. Therefore, our results suggest that the MMPs activation may lead to luteal cell development or death.

scholarly journals Prostaglandin E2 functions as a luteotrophic factor in the dog

Reproduction ◽  
2013 ◽  
Vol 145 (3) ◽  
pp. 213-226 ◽  
Author(s):  
Mariusz P Kowalewski ◽  
Barbara Fox ◽  
Aykut Gram ◽  
Alois Boos ◽  
Iris Reichler
Keyword(s):  
Luteal Phase ◽  
Regulatory Protein ◽  
Regulatory Mechanisms ◽  
Prostaglandin E ◽  
Luteal Regression ◽  
Luteal Cells ◽  
Progesterone Secretion ◽  
Steroidogenic Acute Regulatory ◽  
Post Implantation

The luteal phase in dogs is governed by many poorly understood regulatory mechanisms. Functioning of the corpus luteum (CL) is unaffected by hysterectomy. Recently, the role of prostaglandins in regulating canine CL function was addressed suggesting a luteotrophic effect of prostaglandin E2 (PGE2) during the early luteal phase. However, compelling functional evidence was lacking. The potential of PGE2 to stimulate steroidogenesis was tested in canine primary luteal cells isolated from developing CL of non-pregnant dogs. In addition, the luteal expression of prostaglandin transporter (PGT) and steroidogenic acute regulatory protein (STAR) was demonstrated and characterized in CL from non-pregnant bitches during the course of dioestrus as well as from pregnant animals during the pre-implantation, post-implantation and mid-gestation periods of pregnancy and during luteolysis; the luteal expression of PGE2 receptors (EP2 and EP4) has been investigated at the protein level throughout pregnancy. Our findings show that PGE2 is an activator of STAR expression in canine luteal cells from early luteal phase, significantly up-regulating STAR promoter activity and protein expression resulting in increased steroidogenesis. The 3βHSD (HSD3B2) and P450scc (CYP11A1) expression remained unaffected by PGE2 treatment. The expression of PGT was confirmed in CL during both pregnancy and dioestrus and generally localized to the luteal cells. After initial up-regulation during the earlier stages of the CL phase, its expression declined towards the luteal regression. Together with the demonstration of EP2 and EP4 throughout pregnancy, and the decline in EP2 at prepartum, our findings further support our hypothesis that intra-luteal PGE2 may play an important role in regulating progesterone secretion in the canine CL.

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

Prostaglandin F2α- and phorbol 12-myristate-13-acetate-stimulated progesterone production by cultured human luteal cells in the mid-luteal phase: prostaglandin F2α increases cytosolic Ca2+ and inositol phosphates

1992 ◽  
Vol 133 (3) ◽  
pp. 451-458 ◽  
Author(s):  
T. Endo ◽  
H. Watanabe ◽  
H. Yamamoto ◽  
S. Tanaka ◽  
M. Hashimoto
Keyword(s):  
Luteal Phase ◽  
Inositol Phosphates ◽  
Prostaglandin F2α ◽  
Free Calcium ◽  
Corpora Lutea ◽  
Luteal Cells ◽  
Progesterone Production ◽  
Kinase C ◽  
Prostaglandin F ◽  
Human Corpus Luteum

ABSTRACT While prostaglandin F2α (PGF2α) has been thought to be a natural luteolysin in non-primates, a luteolytic effect in the human corpus luteum is less evident. We therefore investigated the action of PGF2α on monolayer cultures of human luteal cells obtained from mid-luteal phase corpora lutea. PGF2α increased basal and human chorionic gonadotrophin (hCG)-stimulated progesterone production by human cultured luteal cells. A potent tumour-promoting phorbol ester, phorbol 12-myristate-13-acetate (PMA), also stimulated progesterone production by cultured human luteal cells. Although human luteal cells were incubated for 24 h with PMA, hCG was still able to stimulate the production of progesterone by PMA-pretreated cells. However, PMA pretreatment blocked the ability of PGF2α to stimulate progesterone production. It is possible that the luteotrophic effect of PGF2α may be mediated, in part, by the activation of protein kinase C. Addition of PGF2α to suspensions of human luteal cells preincubated with myo-[2-3H]inositol promoted an increase in labelled inositol phosphates. PGF2α also rapidly increased intracellular free Ca2+ in human luteal cells loaded with the fluorescent Ca2+ probe, fura-2. We conclude that PGF2α and PMA stimulate progesterone production and that PGF2α increases the intracellular free calcium and inositol phosphates of human cultured luteal cells in the mid-luteal phase. Journal of Endocrinology (1992) 133, 451–458

Ultrastructure of Luteal Cells in Fully Formed and Regressing Corpora Lutea During Pregnancy and Lactation in the Marsupials Isoodon Macrourus and Perameles Nasuta.

1980 ◽  
Vol 28 (2) ◽  
pp. 195 ◽  
Author(s):  
DE Hollis ◽  
AG Lyne
Keyword(s):  
Endoplasmic Reticulum ◽  
Lipid Droplets ◽  
Smooth Endoplasmic Reticulum ◽  
Cell Junction ◽  
Corpora Lutea ◽  
Cholesterol Crystals ◽  
Luteal Cells ◽  
Pregnancy And Lactation ◽  
Advanced Stages ◽  
Spherical Nuclei

Corpora lutea (CL) collected from 23 bandicoots (I. macrourus and P. nasuta), from day 5 of pregnancy (gestation 12.5 days) to day 53 of lactation (lactation c. 60 days), were examined with the electron microscope. The luteal cells of fully formed CL (from day 5 of pregnancy to day 44 of lactation) were large and contained spherical nuclei with distinct nucleoli. The amount of heterochromatin increased during the latter part of this period. The cytoplasm contained numerous lipid droplets and abundant smooth endoplasmic reticulum (SER). Granular endoplasmic reticulum (GER) was less common. Mitochondria were most numerous from day 16 to day 44 of lactation and some of them contained large osmiophilic inclusions. Several types of granules and inclusions were present in the cytoplasm. During pregnancy, small dense-cored granules were common in P. nasuta and sparse in I. macrourus. They were still present in small numbers during early lactation in P. nasuta but were absent throughout lactation in I. macrourus. A special type of cell junction associated with endoplasmic reticulum was present between the luteal cells in P. nasuta but not in I. macrourus. The luteal cells of regressing CL on days 48, 50 and 53 of lactation were markedly reduced in size, with small irregularly shaped nuclei containing clumps of heterochromatin and indistinct nucleoli. The cells still contained numerous lipid droplets, and osmiophilic inclusions were still present in some of the mitochondria, which were reduced in number (and absent in the animal at day 50). Lancet-shaped spaces, which probably initially contained cholesterol crystals extracted by solvents during processing, were present in some of the regressing luteal cells. Organelles, including SER and GER, were either sparse or unrecognizable in luteal cells at advanced stages of regression. In general, the ultrastructural features of the luteal cells in fully formed and regressing CL of bandicoots were similar to those described in active and regressing CL of eutherian mammals.

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