scholarly journals miR-375 mediates CRH signaling pathway in inhibiting E2 synthesis in porcine ovary

Reproduction ◽  
2017 ◽  
Vol 153 (1) ◽  
pp. 63-73 ◽  
Author(s):  
Chulin Yu ◽  
Meiling Li ◽  
Yue Wang ◽  
Ying Liu ◽  
Chengzhi Yan ◽  
...  
Keyword(s):  
Signaling Pathway ◽  
Granulosa Cells ◽  
Reproductive System ◽  
Follicular Development ◽  
Cell Types ◽  
Signaling System ◽  
Key Factor ◽  
The Relationship

The corticotropin-releasing hormone (CRH) signaling system is involved in numbers of stress-related physiological and pathological responses, including its inhibiting effects on estradiol (E2) synthesis and follicular development in the ovary. In addition, there are reports that microRNAs (miRNAs) can control the function of animal reproductive system. The aim of present study was to investigate the functions of miR-375 and the relationship between miR-375 and CRH signaling molecules in the porcine ovary. First, our common PCR results show that miR-375 and the CRH receptor 1 (CRHR1) are expressed in porcine ovary, whereas CRH receptor 2 (CRHR2) is not detected. We further have located the cell types of miR-375 and CRHR1 by in situ hybridization (ISH), and the results show that miR-375 is located only in the granulosa cells, whereas CRHR1 is positive in all of granulosa cells and oocytes, inferring that miR-375 and CRHR1 are co-localized in granulosa cells. Second, we show that overexpression of miR-375 in cultured granulosa cells suppresses the E2 production, whereas miR-375 knockdown demonstrates the opposite result. Besides, our in vitro results demonstrate that miR-375 mediates the signaling pathway of CRH inhibiting E2 synthesis. Finally, our data show that the action of miR-375 is accomplished by directly binding to the 3′UTR of specificity protein1 (SP1) mRNA to decrease the SP1 protein level. Thus, we conclude that miR-375 is a key factor in regulating E2 synthesis by mediating the CRH signaling pathway.

scholarly journals AKT is involved in granulosa cell autophagy regulation via mTOR signaling during rat follicular development and atresia

Reproduction ◽  
2014 ◽  
Vol 147 (1) ◽  
pp. 73-80 ◽  
Author(s):  
JongYeob Choi ◽  
MinWha Jo ◽  
EunYoung Lee ◽  
DooSeok Choi
Keyword(s):  
Cell Death ◽  
Granulosa Cell ◽  
Granulosa Cells ◽  
Apoptotic Cell ◽  
Follicular Development ◽  
Negative Regulator ◽  
Metabolic Clearance ◽  
Akt Inhibitor ◽  
Western Blots

In this study, we examined whether granulosa cell autophagy during follicular development and atresia was regulated by the class I phosphoinositide-3 kinase/protein kinase B (AKT) pathway, which is known to control the activity of mammalian target of rapamycin (mTOR), a major negative regulator of autophagy. Ovaries and granulosa cells were obtained using an established gonadotropin-primed immature rat model that induces follicular development and atresia. Autophagy was evaluated by measuring the expression level of microtubule-associated protein light chain 3-II (LC3-II) using western blots and immunohistochemistry. The activity of AKT and mTOR was also examined by observing the phosphorylation of AKT and ribosomal protein S6 kinase (S6K) respectively. After gonadotropin injection, LC3-II expression was suppressed and phosphorylation of AKT and S6K increased in rat granulosa cells. By contrast, gonadotropin withdrawal by metabolic clearance promoted LC3-II expression and decreased phosphorylation of AKT and S6K. In addition,in-vitroFSH treatment of rat granulosa cells also indicated inhibition of LC3-II expression accompanied by a marked increase in phosphorylation of AKT and S6K. Inhibition of AKT phosphorylation using AKT inhibitor VIII suppressed FSH-mediated phosphorylation of S6K, followed by an increase in LC3-II expression. Furthermore, co-treatment with FSH and AKT inhibitor increased the levels of apoptosis and cell death of granulosa cells compared with the single treatment with FSH. Taken together, our findings indicated that AKT-mediated activation of mTOR suppresses granulosa cell autophagy during follicular development and is involved in the regulation of apoptotic cell death.

Identification of Gαs messenger ribonucleic acid splice variants in human granulosa cells

1997 ◽  
Vol 18 (1) ◽  
pp. 27-35 ◽  
Author(s):  
G N Europe-Finner ◽  
E Cartwright ◽  
J Bellinger ◽  
H J Mardon ◽  
D H Barlow ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Ovarian Function ◽  
Splice Variants ◽  
Consensus Sequence ◽  
Phosphorylation Site ◽  
Follicular Development ◽  
Protein Isoforms ◽  
Mrna Isoforms ◽  
Messenger Ribonucleic Acid

ABSTRACT Granulosa cells are essential for follicular development and corpus luteum formation and their functions are regulated by gonadotrophins through G protein-coupled receptors. The dominant second messenger pathway involves the stimulation of cyclic AMP formation by Gαs-linked receptors. In this paper we have investigated the expression of Gαs mRNA splice variants in relation to expression of Gαs protein isoforms in granulosa cells obtained from patients undergoing in vitro fertilization. We have carried out ribonuclease protection assays using cRNA riboprobes which are capable of detecting all Gαs mRNA isoforms as well as quantifying total amounts of Gαs mRNA. Granulosa cells express the message for Gαs-Large and Gαs-Small and the presence of two distinct protein products was confirmed by immunoblotting using the antibody RM/1. Moreover, the data show that a significant fraction of Gαs-Large and Gαs-Small mRNAs contain an extra CAG codon. This should generate proteins with an extra serine residue, resulting in Gαs variants with the consensus sequence of a protein kinase C phosphorylation site. These results highlight the possible interaction between different signalling pathways in the control of cAMP production and the need to investigate the relationship between Gαs variants and different adenylyl cyclase isozymes in patients with normal and abnormal ovarian function.

scholarly journals MiRNA-143 mediates the proliferative signaling pathway of FSH and regulates estradiol production

2017 ◽  
Vol 234 (1) ◽  
pp. 1-14 ◽  
Author(s):  
Li Zhang ◽  
XiaoXin Zhang ◽  
Xuejing Zhang ◽  
Yu Lu ◽  
Lei Li ◽  
...  
Keyword(s):  
Signaling Pathway ◽  
Granulosa Cell ◽  
Granulosa Cells ◽  
Loss Of Function ◽  
Cellular Processes ◽  
Cell Functions ◽  
Genes Expression ◽  
Underlying Mechanisms

MicroRNAs (MiRNAs) play important regulatory roles in many cellular processes. MiR-143 is highly enriched in the mouse ovary, but its roles and underlying mechanisms are not well understood. In the current study, we show that miR-143 is located in granulosa cells of primary, secondary and antral follicles. To explore the specific functions of miR-143, we transfected miR-143 inhibitor into primary cultured granulosa cells to study the loss of function of miR-143 and the results showed that miR-143 silencing significantly increased estradiol production and steroidogenesis-related gene expression. Moreover, our in vivo and in vitro studies showed that follicular stimulating hormone (FSH) significantly decreased miR-143 expression. This function of miR-143 is accomplished by its binding to the 3’-UTR of KRAS mRNA. Furthermore, our results demonstrated that miR-143 acts as a negative regulating molecule mediating the signaling pathway of FSH and affecting estradiol production by targeting KRAS. MiR-143 also negatively acts in regulating granulosa cells proliferation and cell cycle-related genes expression. These findings indicate that miR-143 plays vital roles in FSH-induced estradiol production and granulosa cell proliferation, providing a novel mechanism that involves miRNA in regulating granulosa cell functions.

scholarly journals In vitro estimation of rumen fermentable organic matter using enzymes

1996 ◽  
Vol 44 (2) ◽  
pp. 103-110 ◽  
Author(s):  
J.W. Cone ◽  
A.H. Van Gelder ◽  
A.M. Van Vuuren
Keyword(s):  
Organic Matter ◽  
Crude Fibre ◽  
Fermentation Products ◽  
In Situ Method ◽  
Degradation Rates ◽  
Digestible Organic Matter ◽  
Similar Accuracy ◽  
The Relationship

The amount of rumen fermentable organic matter (FOM) can be calculated using tables, taking into account the amount of digestible organic matter, the content of fat and fermentation products, and the amount of starch and protein escaping rumen fermentation, or FOM can be calculated using in situ incubations. An in vitro method is described to predict FOM using amylase and other carbohydrate degrading enzymes. FOM estimated by the enzymic method showed a moderate correlation (Rsuperscript 2 = 0.71) with FOM estimated by the in situ method. The relationship could be improved by separating the high crude fibre samples (Rsuperscript 2 = 0.88) from the other samples (Rsuperscript 2 = 0.77). Because degradation rates with the enzymic method were high compared with the assumed rumen passage rates, it proved that FOM could be predicted with a similar accuracy (Rsuperscript 2 = 0.76 - 0.80) by the undegraded fraction after 24 h.

Temporal changes in the expression of the insulin-like growth factor II gene associated with tissue maturation in the human fetus

Development ◽  
1989 ◽  
Vol 106 (3) ◽  
pp. 543-554 ◽  
Author(s):  
A.L. Brice ◽  
J.E. Cheetham ◽  
V.N. Bolton ◽  
N.C. Hill ◽  
P.N. Schofield
Keyword(s):  
Growth Factor ◽  
Cell Types ◽  
Specific Cell ◽  
Insulin Like Growth Factor ◽  
Vitro Fertilization ◽  
Age Related ◽  
Factor Ii ◽  
Metanephric Blastema

The insulin-like growth factors are broadly distributed in the human conceptus and are thought to play a role in the growth and differentiation of tissues during development. Using in situ hybridization we have shown that a wide variety of specific cell types within tissues express the gene for insulin-like growth factor II at times of development from 18 days to 14 weeks of gestation. Examination of blastocysts produced by in vitro fertilization showed no expression, thus bracketing the time of first accumulation of IGF-II mRNA to between 5 and 18 days postfertilization. The pattern of IGF-II expression shows specific age-related differences in different tissues. In the kidney, for example, expression is found in the cells of the metanephric blastema which is dramatically reduced as the blastema differentiates. The reverse is also seen, and we have noted an increase in expression of IGF-II in the cytotrophoblast layer of the placenta with gestational age. The sites of expression do not correlate with areas of either high mitotic activity or specific types of differentiation, but the observed pattern of expression in the kidney, adrenal glands and liver suggests an explanation for the abnormally high IGF-II mRNA expression in developmental tumours such as Wilms' tumour.

The relationship between erythropoietin-dependent cellular differentiation and colony-forming ability in prenatal haemopoietic tissues

Development ◽  
1975 ◽  
Vol 34 (3) ◽  
pp. 575-588
Author(s):  
R. J. Cole ◽  
T. Regan ◽  
S. L. White ◽  
E. M. Cheek
Keyword(s):  
Cellular Differentiation ◽  
Cell Numbers ◽  
In Vitro Sensitivity ◽  
Foetal Liver ◽  
Rapid Production ◽  
Colony Forming Cell ◽  
Macrophage Colony ◽  
The Relationship

Levels of haem synthesis achieved by foetal liver erythroblasts responding to erythropoietin in vitro are similar in dissociated cell cultures and in cultures of organized tissues. Erythroid colony-forming cells reach maximum numbers on the sixteenth day of gestation. Their presence in foetal liver is associated with the period of most rapid production of erythrocytes, and with in vitro sensitivity to erythropoietin measured as enhanced haem synthesis. It is concluded that at least a proportion of erythroid colony-forming cells in the foetal liver are dependent on erythropoietin in situ and that these cells are separated from the earliest recognizable pro-erythroblast by 1–2 cell divisions. Populations of granulocyte-macrophage colony-forming cells change independently of erythroid colony-forming cell numbers.

RAF1 as Downstream Molecule Mediates the FSH Signaling Pathway to Stimulate E2 Synthesis and Secretion in Mouse Ovarian Granulosa Cells

2020 ◽  
Author(s):  
Xuan Luo ◽  
Hui Liu ◽  
Hongzhou Guo ◽  
Longjie Sun ◽  
Kemian Gou ◽  
...  
Keyword(s):  
Signaling Pathway ◽  
Granulosa Cells ◽  
Regulatory Function ◽  
Hormone Synthesis ◽  
Ovarian Granulosa Cells ◽  
Erk Phosphorylation ◽  
Extracellular Signal ◽  
Key Factor ◽  
Cytochrome P450 Family ◽  
Estradiol Synthesis

Abstract Background: V-raf-leukemia viral oncogene 1 (RAF1) kinase is the key factor in extracellular signal regulated pathway, which transmits signals to the downstream extracellular regulated protein kinases (ERK). Regulatory function of RAF1 has been proved to mediate steroid hormone synthesis, which played an essential physiological function in reproduction and development. Whether RAF1 takes part in the signaling events of gonadotropic hormones follicle-stimulating hormone (FSH) in ovarian is unknown.Results: We found that RAF1 as downstream molecule mediates the FSH signaling pathway to stimulate estradiol (E2) synthesis and secretion in mouse ovarian granulosa cells (GCs). The expression of RAF1 is induced by FSH and the production of E2 is increased in the serum and primary ovarian GCs supernatant, the process of which is blocked by treating with RAF1 inhibitor (N-(2-Methyl-5'-morpholino-6'-((tetrahydro-2H-pyran-4-yl)oxy)-[3,3'-bipyridin]-5-yl)-3(trifluoromethyl) benzamide, RAF709). Inhibition of RAF1 activity by RAF709 decreased ERK phosphorylation, and suppressed the expression of cytochrome P450 family 19 subfamily a member 1 (CYP19A1) which is a major rate-limiting enzyme to participate in the last step of E2 biosynthesis. Conclusion: Our results suggest that RAF1 play a pivotal mediating roles toward E2 production in FSH signaling pathway by inducing the phosphorylation of ERK and promoting the process of estradiol synthesis. RAF1 may be a potential and effective factor to regulate the function of the female mouse reproductive system.

The relationship between in vitro and in situ dry matter disappearance of some Iranian feedstuffs in sheep

2007 ◽  
Vol 2007 ◽  
pp. 210-210
Author(s):  
H. Paya ◽  
A. Taghizadeh ◽  
H. Janmohamadi ◽  
G.A Moghadam
Keyword(s):  
Dry Matter ◽  
Nutrient Requirements ◽  
Feed Ingredients ◽  
The Relationship ◽  
Require Information

Ration formulation systems require information on nutrient requirements of the animal and reliable values for rumen degradable and undegradable fractions of feed ingredients. The in situ nylon-bag technique is widely used to characterize the disappearance of feeds from the rumen (Woods et al., 2002). The objective of this study was determining of relationship between in vitro and in situ dry matter disappearance.

scholarly journals Pentachloronitrobenzene alters progesterone production and primordial follicle recruitment in cultured granulosa cells and rat ovary†

2019 ◽  
Vol 102 (2) ◽  
pp. 511-520
Author(s):  
Yanrong Kuai ◽  
Xiaobo Gao ◽  
Huixia Yang ◽  
Haiyan Luo ◽  
Yang Xu ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Granulosa Cells ◽  
Crop Production ◽  
Follicular Development ◽  
Mitogen Activated Protein Kinase ◽  
Dependent Manner ◽  
Serum Progesterone ◽  
Primordial Follicles ◽  
Progesterone Production

Abstract Pentachloronitrobenzene (PCNB) is an organochlorine fungicide widely used for crop production and has become an environmental concern. Little is known about the effect of PCNB on ovarian steroidogenesis and follicular development. We found that PCNB stimulated Star expression and progesterone production in cultured rat granulosa cells in a dose-dependent manner. PCNB activated mitogen-activated protein kinase (MAPK3/1) extracellulat regulated kinase (ERK1/2), thus inhibition of either protein kinase A (PKA) or MAPK3/1 signaling pathway significantly attenuated progesterone biosynthesis caused by PCNB, suggesting that PCNB induced progesterone production by activating the cyclic adenosine monophosphate (cAMP/PKA) and MAPK3/1 signaling pathways. Further investigation demonstrated that PCNB induced Star expression and altered MAPK3/1 signaling in ovary tissues of immature SD rats treated with PCNB at the dose of 100, 200, or 300 mg/kg by daily gavage for 7 days, while serum progesterone level was dose-dependently decreased. We demonstrated that PCNB exposure accelerated the recruitment of primordial follicles into the growing follicle pool in ovary tissues, accompanied by increased levels of anti-Mullerian hormone (AMH) in both ovary tissues and serum. Taken together, our data demonstrate for the first time that PCNB stimulated Star expression, altered MAPK3/1 signaling and progesterone production in vivo and in vitro, and accelerated follicular development with a concomitant increase in AMH in ovary tissues and serum. Our findings provide novel insight into the toxicity of PCNB to animal ovary function.

scholarly journals hCG Improves Luteal Function and Promotes Progesterone Output through the Activation of JNK Pathway in the Luteal Granulosa Cells of the Stimulated IVF Cycles†

2020 ◽  
Vol 102 (6) ◽  
pp. 1270-1280 ◽  
Author(s):  
Gamze Bildik ◽  
Nazli Akin ◽  
Yashar Esmaeilian ◽  
Francesko Hela ◽  
Kayhan Yakin ◽  
...  
Keyword(s):  
Luteinizing Hormone ◽  
Signaling Pathway ◽  
Granulosa Cells ◽  
Luteal Phase ◽  
Steroidogenic Enzymes ◽  
Jnk Pathway ◽  
Jnk Signaling ◽  
Luteal Function ◽  
Jnk Signaling Pathway

Abstract Human chorionic gonadotropin (hCG) is a luteotropic hormone that promotes the survival and steroidogenic activity of corpus luteum (CL) by acting through luteinizing hormone receptors (LHRs) expressed on luteinized theca and granulosa cells (GCs). Therefore, it is used to support luteal phase in in vitro fertilization (IVF) cycles to improve clinical pregnancy rates and prevent miscarriage. However, the molecular mechanism underlying this action of hCG is not well characterized. To address this question, we designed an in vitro translational research study on the luteal GCs obtained from 58 IVF patients. hCG treatment at different concentrations and time points activated c-Jun N-terminal kinase (JNK) pathway and significantly increased its endogenous kinase activity along with upregulated expression of steroidogenic enzymes (steroidogenic acute regulatory protein (stAR), 3β-Hydroxysteroid dehydrogenase (3β-HSD)) in a dose-dependent manner in the luteal GCs. As a result, in vitro P production of the cells was significantly enhanced after hCG. When JNK pathway was inhibited pharmacologically or knocked-down with small interfering RNA luteal function was compromised, P4 production was declined along with the expression of stAR and 3β-HSD in the cells. Further, hCG treatment after JNK inhibition failed to correct the luteal defect and promote P4 output. Similar to hCG, luteinizing hormone (LH) treatment improved luteal function as well and this action of LH was associated with JNK activation in the luteal GCs. These findings could be important from the perspective of CL biology and luteal phase in human because we for the first time identify a critical role for JNK signaling pathway downstream LHR activation by hCG/LH in luteal GCs. Summary Sentence JNK signaling pathway plays a central role in the upregulated expression of the steroidogenic enzymes StAR and 3b-HSD and augmented progesterone production by hCG/LH in human luteal granulosa cells.

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