scholarly journals Secreted Protein, Acidic and Rich in Cysteine (SPARC) and Thrombospondin in the Developing Follicle and Corpus Luteum of the Rat

1998 ◽  
Vol 46 (9) ◽  
pp. 1043-1049 ◽  
Author(s):  
P. Bagavandoss ◽  
E. Helene Sage ◽  
Robert B. Vernon
Keyword(s):  
Corpus Luteum ◽  
Granulosa Cells ◽  
Molecular Mechanisms ◽  
Basement Membranes ◽  
Oocyte Nucleus ◽  
Variable Degree ◽  
Matricellular Proteins ◽  
Luteal Cells ◽  
Postovulatory Follicle ◽  
Thecal Cells

In adult mammals, growth of new vasculature from extant blood vessels (angiogenesis) is rare in the absence of pathology. However, nonpathogenic angiogenesis occurs in the cycling ovary when the avascular postovulatory follicle transforms into a highly vascularized corpus luteum (CL). To improve our understanding of molecular mechanisms that regulate nonpathogenic vascular growth, we characterized the expression of two secreted matricellular proteins associated with angiogenesis, SPARC and thrombospondin (TSP), in postovulatory preluteal follicles and CL of hormone-primed immature rats. By indirect immunofluorescence with specific antibodies, we found SPARC in the cytoplasm of granulosa cells and thecal cells of preluteal follicles, in connective tissue cells of the ovarian interstitium, and in the oocyte nucleus. Administration of a luteinizing stimulus (chorionic gonadotropin) increased the expression of SPARC in granulosa cells. TSP was prominent in the basement membranes of growing follicles. Many cells in the early vascularizing CL expressed both SPARC and TSP. Neovascularization of CL was accompanied by expression of SPARC in nascent vessels and concentration of TSP in central avascular areas. In mature CL, steroidogenic luteal cells expressed both SPARC and TSP. Luteal cells of regressing CL retained SPARC to a variable degree but did not express TSP. The observed changes in expression of SPARC and TSP during development of the CL support distinct roles for these matricellular proteins in nonpathological morphogenesis and angiogenesis.

Effect of cortisol on neurophysin I/oxytocin and peptidyl glycine-α-amidating mono-oxygenase mRNA expression in bovine luteal and granulosa cells

2013 ◽  
Vol 16 (2) ◽  
pp. 231-239
Author(s):  
A. Ziolkowska ◽  
J. Mlynarczuk ◽  
J. Kotwica
Keyword(s):  
Mrna Expression ◽  
Granulosa Cells ◽  
Molecular Mechanisms ◽  
Hormone Secretion ◽  
Oestrous Cycle ◽  
Luteal Cells ◽  
Ru 486 ◽  
Ovarian Cells ◽  
Key Genes ◽  
Ovary Function

Abstract Cortisol stimulates the synthesis and secretion of oxytocin (OT) from bovine granulosa and luteal cells, but the molecular mechanisms of cortisol action remain unknown. In this study, granulosa cells or luteal cells from days 1-5 and 11-15 of the oestrous cycle were incubated for 4 or 8 h with cortisol (1x10-5, 1x10-7 M). After testing cell viability and hormone secretion (OT, progesterone, estradiol), we studied the effect of cortisol on mRNA expression for precursor of OT (NP-I/OT) and peptidyl glycine-α-amidating mono-oxygenase (PGA). The influence of RU 486 (1x10-5 M), a progesterone receptor blocker and inhibitor of the glucocorticosteroid receptor (GR), on the expression for both genes was tested. Cortisol increased the mRNA expression for NP-I/OT and PGA in granulosa cells and stimulated the expression for NP-I/OT mRNA in luteal cells obtained from days 1-5 and days 11-15 of the oestrous cycle. Expression for PGA mRNA was increased only in luteal cells from days 11-15 of the oestrous cycle. In addition, RU 486 blocked the cortisol-stimulated mRNA expression for NP-I/OT and PGA in both types of cells. These data suggest that cortisol affects OT synthesis and secretion in bovine ovarian cells, by acting on the expression of key genes, that may impair ovary function.

scholarly journals Is the canine corpus luteum an insulin-sensitive tissue?

2016 ◽  
Vol 231 (3) ◽  
pp. 223-233 ◽  
Author(s):  
Liza Margareth Medeiros de Carvalho Sousa ◽  
Renata dos Santos Silva ◽  
Vanessa Uemura da Fonseca ◽  
Rafael Magdanelo Leandro ◽  
Thiago Senna Di Vincenzo ◽  
...  
Keyword(s):  
Glucose Uptake ◽  
Corpus Luteum ◽  
Molecular Mechanisms ◽  
Glucose Transporter ◽  
Hypoxia Inducible Factor ◽  
Glucose Disposal ◽  
Luteal Cell ◽  
Luteal Function ◽  
Luteal Cells ◽  
Glut4 Expression

This study aimed to determine in the canine corpus luteum throughout the dioestrus (1) the influence of insulin on glucose uptake; (2) the regulation of genes potentially involved; and (3) the influence of hypoxia on glucose transporter expression and steroidogenesis, after treatment with cobalt chloride (CoCl2). Glucose uptake by luteal cells increased 2.7 folds (P < 0.05) in response to insulin; a phenomenon related to increased expression of glucose transporter (GLUT) 4 and phosphorylation of protein kinase B (AKT). The gene expression of insulin receptor and SLC2A4 (codifier of GLUT4) genes after insulin stimulation increased on day 20 post ovulation (p.o.) and declined on day 40 p.o. (P < 0.05). Regarding potentially involved molecular mechanisms, the nuclear factor kappa B gene RELA was upregulated on days 30/40 p.o., when SLC2A4 mRNA was low, and the interleukin 6 (IL6) gene was upregulated in the first half of dioestrus, when SLC2A4 mRNA was high. CoCl2 in luteal cell cultures increased the hypoxia-inducible factor HIF1A/HIF1A and the SLC2A4/GLUT4 expression, and decreased progesterone (P4) production and hydroxyl-delta-5-steroid dehydrogenase 3 beta (HSD3B) mRNA expression (P < 0.05). This study shows that the canine luteal cells are responsive to insulin, which stimulates glucose uptake in AKT/GLUT4-mediated pathway; that may be related to local activity of RELA and IL6. Besides, the study reveals that luteal cells under hypoxia activate HIF1A-modulating luteal function and insulin-stimulated glucose uptake. These data indicate that insulin regulates luteal cells’ glucose disposal, participating in the maintenance and functionality of the corpus luteum.

scholarly journals Terminal differentiation of human granulosa cells as luteinization is reversed by activin-A through silencing of Jnk pathway

2020 ◽  
Vol 6 (1) ◽  
Author(s):  
Gamze Bildik ◽  
Nazli Akin ◽  
Yashar Esmaeilian ◽  
Francesko Hela ◽  
Ceren Sultan Yildiz ◽  
...  
Keyword(s):  
Corpus Luteum ◽  
Signaling Pathway ◽  
Granulosa Cells ◽  
Molecular Mechanisms ◽  
Terminal Differentiation ◽  
Reproductive Technologies ◽  
Activin A ◽  
Jnk Pathway ◽  
Jnk Signaling ◽  
Jnk Signaling Pathway

Abstract Molecular mechanisms underlying luteinization (terminal differentiation of granulosa and theca cells after ovulation) and luteolysis (demise of corpus luteum) are poorly understood in human ovary. Here we report that activin-A, after binding to its cognate receptors induces a functional luteolytic state and reverses luteinization phenotype by downregulating the expression of the steroidogenic enzymes, LH receptor and VEGF and reducing estradiol (E2) progesterone (P4) production and upregulating FSH receptor and cyclin D1 expression in human primary luteinized granulosa cells. Further, this action of activin-A involves downregulation of JNK signaling pathway and is opposite to that of human chorionic gonadotropin (hCG), which acts as a luteotropic hormone and improves luteal function through the activation of JNK pathway in the same cell type. Reversal of luteinization phenotype in luteal granulosa cells by activin-A potentially makes this hormone an attractive candidate for use under certain clinical situations, where induction of luteolysis and rapid reduction of endogenous sex steroid levels are beneficial such as ovarian hyperstimulation syndrome (OHSS), in which the ovaries hyper-respond to gonadotropin stimulation by producing too many growing follicles along with development of ascites, pleural effusion, and hemo-concentrations as a result of increased vascular permeability and leakage of intravascular volume into third spaces. Our work unveils a previously undefined role for activin-A and JNK signaling pathway in human corpus luteum biology, that might have a direct clinical impact in assisted reproductive technologies.

107 Localization of Kisspeptin Immunoreactivity in the Cat Ovary on Different Reproductive Stages

2018 ◽  
Vol 30 (1) ◽  
pp. 193
Author(s):  
P. Tanyapanyachon ◽  
O. Amelkina ◽  
K. Chatdarong
Keyword(s):  
Corpus Luteum ◽  
Granulosa Cells ◽  
Ovarian Surface Epithelium ◽  
Domestic Cat ◽  
Primary Antibody ◽  
Surface Epithelium ◽  
Domestic Cats ◽  
Luteal Cells ◽  
Theca Cells ◽  
Steroidogenic Cells

Kisspeptin (Kp) is considered one of the main regulators of the reproductive axis, exerting its effects via stimulating GnRH expression in the hypothalamus. Apart from its central localization in the hypothalamus, the presence of Kp has been reported in the ovary, with possible local function. To date, very little is known about the ovarian Kp in the domestic cat. Therefore, our aim was to investigate the presence and localization of Kp at different reproductive stages in domestic cat ovaries. Twenty ovaries were collected from free-ranging domestic cats (body weight 2.7–4.5 kg) after routine ovariohysterectomy. Reproductive stages were classified by ovarian gross morphology, vaginal cytology, and blood progesterone level. Ovarian samples were grouped into inactive (n = 6), follicular (n = 8), and luteal stages (n = 6). Tissues were fixed in 4% paraformaldehyde and processed routinely. Immunohistochemistry was performed using polyclonal rabbit Kp-10 primary antibody (AB9754; Millipore, Billerica, MA, USA) at 1:500 at 4°C overnight. Immunoreactive cells were identified by avidin-biotin-peroxidase system. Rat hypothalamic tissue was used as a positive control. Primary antibody was substituted with PBS and normal rabbit IgG as the negative and isotypic negative controls, respectively. In addition, primary antibody was incubated with metastin overnight and applied for preabsorption test. Negative, isotypic negative, and preabsorption tests showed no staining. Immunoreactive Kp was detected in the ovaries of all reproductive stages with no obvious changes in localization or intensity of staining between stages. Kisspeptin was present in the cytoplasm of oocytes, granulosa cells, and theca cells of preantral (primordial, primary, and secondary) follicles and antral follicles. Interestingly, in most follicles, Kp staining was more prominent in theca cells and oocytes compared with granulosa cells. In corpus luteum, Kp was localised in the cytoplasm of luteal cells, with more intense staining on the periphery of corpus luteum compared with the middle in 3 luteal samples, whereas the rest of the samples demonstrated homogeneous staining distribution. Apart from oocytes and steroidogenic cells, Kp was also present in the cytoplasm of cells of the ovarian surface epithelium. Our study for the first time demonstrated the presence and localization of Kp in the ovary of the domestic cats. The localization of Kp in the cat oocyte is similar to previous reports on hamsters and dogs, indicating a possible function in oocyte development. The staining in steroidogenic cells, mainly theca cells and luteal cells, is in good agreement with studies on hamsters, rats, humans, and marmosets, suggesting the possible local involvement of Kp in steroidogenesis. In addition, Kp staining in the ovarian surface epithelium suggests a possible role in the ovarian remodeling after ovulatory defects, as reported in humans and marmosets. This research was funded by the RGJ PhD program PHD/01882556; RG 7/2559.

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 Analysis of TGFB1, CD105 and FSP1 expression in human granulosa cells during a 7-day primary in vitro culture

2020 ◽  
Vol 8 (4) ◽  
pp. 152-157
Author(s):  
Artur Bryja ◽  
Wojciech Pieńkowski ◽  
Katarzyna Stefańska ◽  
Błażej Chermuła ◽  
Rut Bryl ◽  
...  
Keyword(s):  
In Vitro Culture ◽  
Corpus Luteum ◽  
Expression Profile ◽  
Granulosa Cells ◽  
Luteal Cells ◽  
Human Granulosa Cells

AbstractThe human granulosa cells (GCs) surround the oocyte and form the ovarian follicle’s proper architecture.These sub-populations include mural granulosa cells, antral granulosa cells, and cumulus granulosa cells.Their main functions are to support the oocyte’s growth (cumulus granulosa cells) and estradiol production (mural granulosa cells). After ovulation, the granulosa cells transform into the luteal cells of the corpus luteum and produce progesterone.Our study investigated the expression profile of three genes: TGFB1, CD105, and FSP1 during a 7-day in vitro culture. The analysis was conducted using the RT-qPCR technique.Changes in the expression of CD105 and FSP1 could be observed during the 7-day in vitro culture. In the case of TGFB, the expression remained at a similar level, with no statistically significant differences observed.Running title: Expression of TGFB1, CD105 and FSP1 in granulosa cells

scholarly journals Morphometric Estimation of the Numbers of Granulosa Cells in Preovulatory Follicles of the Ewe

1987 ◽  
Vol 40 (4) ◽  
pp. 451
Author(s):  
JD O'Shea ◽  
PJ Wright ◽  
KE Davis
Keyword(s):  
Light Microscopy ◽  
Corpus Luteum ◽  
Mitotic Index ◽  
Granulosa Cells ◽  
Prostaglandin Analogue ◽  
Previous Estimate ◽  
Luteal Cells ◽  
Preovulatory Follicles ◽  
Morphometric Methods ◽  
Lines Of Evidence

Several lines of evidence suggest that follicular granulosa cells give rise to the large luteal cells of the corpus luteum in the sheep. To further investigate this suggestion, numbers of granulosa cells in preovulatory follicles were estimated by morphometric methods for comparison with a previous estimate of numbers of large luteal cells (9.6 � 0.9 x 106). Preovulatory follicles from five Corriedale ewes were obtained after synchronization of the oestrous cycle with the prostaglandin analogue cloprostenol. Morphometry was undertaken using light microscopy of plastic-embedded tissue sectioned at Il-'m. Mitotic index in the membrana granulosa was 0.05 � s.e.m. 0.05"70. Mean follicular diameter was 6.25 � 0.25 mm and there were 7.68 � 0.53 x 106 granulosa cells per follicle. These results demonstrate a similarity between the number of granulosa cells per follicle and the number of large luteal cells per corpus luteum and thus support the hypothesis that large luteal cells are derived from granulosa cells.

Regulation of Bovine Corpus Luteum Function

1995 ◽  
Author(s):  
Rina Meidan ◽  
Robert Milvae
Keyword(s):  
Arachidonic Acid ◽  
Corpus Luteum ◽  
Granulosa Cells ◽  
Luteal Cells ◽  
Bovine Corpus Luteum ◽  
Pure Cell ◽  
Igf Binding ◽  
Gonadotropin Surge ◽  
Follicular Fluids

The main goal of this research plan was to elucidate regulatory mechanisms controlling the development, function of the bovine corpus luteum (CL). The CL contains two different sterodigenic cell types and therefore it was necessary to obtain pure cell population. A system was developed in which granulosa and theca interna cells, isolated from a preovulatory follicle, acquired characteristics typical of large (LL) and small (SL) luteal cells, respectively, as judged by several biochemical and morphological criteria. Experiments were conducted to determine the effects of granulosa cells removal on subsequent CL function, the results obtained support the concept that granulosa cells make a substaintial contribution to the output of progesterone by the cyclic CL but may have a limited role in determining the functional lifespan of the CL. This experimental model was also used to better understand the contribution of follicular granulosa cells to subsequent luteal SCC mRNA expression. The mitochondrial cytochrome side-chain cleavage enzyme (SCC), which converts cholesterol to pregnenolone, is the first and rate-limiting enzyme of the steroidogenic pathway. Experiments were conducted to characterize the gene expression of P450scc in bovine CL. Levels of P450scc mRNA were higher during mid-luteal phase than in either the early or late luteal phases. PGF 2a injection decreased luteal P450scc mRNA in a time-dependent manner; levels were significantly reduced by 2h after treatment. CLs obtained from heifers on day 8 of the estrous cycle which had granulosa cells removed had a 45% reduction in the levels of mRNA for SCC enzymes as well as a 78% reduction in the numbers of LL cells. To characterize SCC expression in each steroidogenic cell type we utilized pure cell populations. Upon luteinization, LL expressed 2-3 fold higher amounts of both SCC enzymes mRNAs than SL. Moreover, eight days after stimulant removal, LL retained their P4 production capacity, expressed P450scc mRNA and contained this protein. In our attempts to establish the in vitro luteinization model, we had to select the prevulatory and pre-gonadotropin surge follicles. The ratio of estradiol:P4 which is often used was unreliable since P4 levels are high in atretic follicles and also in preovulatory post-gonadotropin follicles. We have therefore examined whether oxytocin (OT) levels in follicular fluids could enhance our ability to correctly and easily define follicular status. Based on E2 and OT concentrations in follicular fluids we could more accurately identify follicles that are preovulatory and post gonadotropin surge. Next we studied OT biosynthesis in granulosa cells, cells which were incubated with forskolin contained stores of the precursor indicating that forskolin (which mimics gonadotropin action) is an effective stimulator of OT biosynthesis and release. While studying in vitro luteinization, we noticed that IGF-I induced effects were not identical to those induced by insulin despite the fact that megadoses of insulin were used. This was the first indication that the cells may secrete IGF binding protein(s) which regonize IGFs and not insulin. In a detailed study involving several techniques, we characterized the species of IGF binding proteins secreted by luteal cells. The effects of exogenous polyunsaturated fatty acids and arachidonic acid on the production of P4 and prostanoids by dispersed bovine luteal cells was examined. The addition of eicosapentaenoic acid and arachidonic acid resulted in a dose-dependent reduction in basal and LH-stimulated biosynthesis of P4 and PGI2 and an increase in production of PGF 2a and 5-HETE production. Indomethacin, an inhibitor of arachidonic acid metabolism via the production of 5-HETE was unaffected. Results of these experiments suggest that the inhibitory effect of arachidonic acid on the biosynthesis of luteal P4 is due to either a direct action of arachidonic acid, or its conversion to 5-HETE via the lipoxgenase pathway of metabolism. The detailed and important information gained by the two labs elucidated the mode of action of factors crucially important to the function of the bovine CL. The data indicate that follicular granulosa cells make a major contribution to numbers of large luteal cells, OT and basal P4 production, as well as the content of cytochrome P450 scc. Granulosa-derived large luteal cells have distinct features: when luteinized, the cell no longer possesses LH receptors, its cAMP response is diminished yet P4 synthesis is sustained. This may imply that maintenance of P4 (even in the absence of a Luteotropic signal) during critical periods such as pregnancy recognition, is dependent on the proper luteinization and function of the large luteal cell.

HISTOCHEMICAL OBSERVATIONS ON THE GRANULOSA AND THECA INTERNA DURING FOLLICULAR DEVELOPMENT AND CORPUS LUTEUM FORMATION AND REGRESSION IN THE AMERICAN OPOSSUM

1968 ◽  
Vol 40 (2) ◽  
pp. 237-241 ◽  
Author(s):  
S. S. GURAYA
Keyword(s):  
Corpus Luteum ◽  
Granulosa Cells ◽  
Follicular Development ◽  
Follicle Cell ◽  
Luteal Cell ◽  
Histochemical Change ◽  
Luteal Cells ◽  
Lipid Granules ◽  
American Opossum ◽  
Theca Interna

SUMMARY A study has been made of the histochemical changes which occur during follicular growth and formation and regression of the corpus luteum in the ovary of the American opossum. The granulosa cells show abundant cytoplasmic RNA. Some lipid bodies consisting of phospholipids are sparsely distributed among the granulosa cells. After ovulation, the granulosa cells undergo 'luteinization' to form the large luteal cells. The most striking histochemical change involved in the differentiation (or luteinization) of the granulosa follicle cell into a luteal cell is the development of abundant diffuse lipoproteins throughout the cytoplasm. Fine lipid granules consisting of phospholipids are also formed in the cytoplasm of luteal cells. The stromal elements of the theca interna, which contain some sparsely scattered phospholipid granules, do not show any histochemical change during corpus luteum formation. With the regression of luteal cells, coarse lipid granules consisting of cholesterol and cholesterol esters, triglycerides and some phospholipids accumulate abundantly in the cytoplasm. Some of these regressing luteal cells continue to persist in the ovarian stroma for some time.

Enzymic Correlates of Development, Secretory Function and Regression of Follicles and Corpora Lutea in the Bovine Ovary. PART II: Formation, Development and Involution of Corpora Lutea

1968 ◽  
Vol 59 (2_Suppl) ◽  
pp. S35-S51 ◽  
Author(s):  
B. L. Lobel ◽  
E. Levy
Keyword(s):  
Corpus Luteum ◽  
Vascular System ◽  
Hydrolytic Enzymes ◽  
Sharp Decrease ◽  
Cellular Growth ◽  
Corpora Lutea ◽  
Cell Nuclei ◽  
Steroid Synthesis ◽  
Luteal Cells ◽  
Vascular Walls

ABSTRACT Activities of various hydrolases and dehydrogenases were studied during the formation, development and involution of cyclic corpora lutea and in the corpora lutea of early pregnancy. At 24 hours postovulation the luteal cells, whether of granulosal or thecal origin, contained demonstrable levels of Δ5-3β-hydroxysteroid dehydrogenase and the NADP and NADPH2 diaphorases. During the period of proliferation and cellular growth, enzymic activities in the luteal cells were moderate at first, and then increased. In the mature corpus luteum, activities of the dehydrogenases occurred in all luteal cells but were most intense in the large polymorphic luteal cells. Activities of hydrolytic enzymes, low in the immediate postovulatory period, increased with the development of the vascular system. Enzymic characteristics of corpora lutea of gestation were similar to those of cyclic corpora, except for phosphorylase activity which was observed in luteal cells in gestational corpora, but confined to the vascular walls in cyclic corpora. No increase in activities of 17β- and 20β-hydroxysteroid dehydrogenases (above those seen in pre-ovulatory follicles) were observed after incubation of sections of either mature cyclic or gestational corpora. Involution of cyclic corpora lutea began with degenerative changes in the blood vessels: pyknosis of the endothelial cell nuclei and a sudden decline in activities of hydrolytic enzymes in the vascular walls. Subsequently, the luteal cells showed a sharp decrease in activities of the dehydrogenases as well as other signs of regressive change. The cytochemical findings are discussed in relation to biochemical observations on steroid synthesis by the bovine corpus luteum.

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