scholarly journals KIRREL is differentially expressed in adipose tissue from ‘fertil+’ and ‘fertil−’ cows: in vitro role in ovary?

Reproduction ◽  
2018 ◽  
Vol 155 (2) ◽  
pp. 181-196
Author(s):  
S Coyral-Castel ◽  
C Ramé ◽  
J Cognié ◽  
J Lecardonnel ◽  
S Marthey ◽  
...  
Keyword(s):  
Adipose Tissue ◽  
Dairy Cows ◽  
Granulosa Cells ◽  
Tiling Array ◽  
Bovine Chromosome ◽  
Bovine Kidney ◽  
Ovarian Cells ◽  
Expression Of Genes ◽  
Trait Locus

We have previously shown that dairy cows carrying the ‘fertil−’ haplotype for one quantitative trait locus affecting female fertility located on the bovine chromosome three (QTL-F-Fert-BTA3) have a significantly lower conception rate and body weight after calving than cows carrying the ‘fertil+’ haplotype. Here, we compared by Tiling Array the expression of genes included in the QTL-F-Fert-BTA3 in ‘fertil+’ and ‘fertil−’ adipose tissue one week after calving when plasma non-esterified fatty acid concentrations were greater in ‘fertil−’ animals. We observed that thirty-one genes were overexpressed whereas twelve were under-expressed in ‘fertil+’ as compared to ‘fertil−’ cows (P < 0.05). By quantitative PCR and immunoblot we confirmed that adipose tissue KIRREL mRNA and protein were significantly greater expressed in ‘fertil+’ than in ‘fertil−’. KIRREL mRNA is abundant in bovine kidney, adipose tissue, pituitary, and ovary and detectable in hypothalamus and mammary gland. Its expression (mRNA and protein) is greater in kidney of ‘fertil+’ than ‘fertil−’ cows (P < 0.05). KIRREL (mRNA and protein) is also present in the different ovarian cells with a greater expression in granulosa cells of ‘fertil+’ than ‘fertil−’ cows. In cultured granulosa cells, recombinant KIRREL halved steroid secretion in basal state (P < 0.05). It also decreased cell proliferation (P < 0.05) and in vitro oocyte maturation (P < 0.05). These results were associated to a rapid increase in MAPK1/3 and MAPK14 phosphorylation in granulosa cells and to a decrease in MAPK1/3 phosphorylation in oocyte. Thus, KIRREL could be a potential metabolic messenger linking body composition and fertility.

scholarly journals The Differential Metabolomes in Cumulus and Mural Granulosa Cells from Human Preovulatory Follicles

2021 ◽  
Author(s):  
Er-Meng Gao ◽  
Bongkoch Turathum ◽  
Ling Wang ◽  
Di Zhang ◽  
Yu-Bing Liu ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
Granulosa Cells ◽  
Quantitative Polymerase Chain Reaction ◽  
Cumulus Cells ◽  
Cholesterol Transport ◽  
Vitro Fertilization ◽  
Expression Of Genes ◽  
Preovulatory Follicles ◽  
Morphological Differences

AbstractThis study evaluated the differences in metabolites between cumulus cells (CCs) and mural granulosa cells (MGCs) from human preovulatory follicles to understand the mechanism of oocyte maturation involving CCs and MGCs. CCs and MGCs were collected from women who were undergoing in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) treatment. The differences in morphology were determined by immunofluorescence. The metabolomics of CCs and MGCs was measured by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) followed by quantitative polymerase chain reaction (qPCR) and western blot analysis to further confirm the genes and proteins involved in oocyte maturation. CCs and MGCs were cultured for 48 h in vitro, and the medium was collected for detection of hormone levels. There were minor morphological differences between CCs and MGCs. LC-MS/MS analysis showed that there were differences in 101 metabolites between CCs and MGCs: 7 metabolites were upregulated in CCs, and 94 metabolites were upregulated in MGCs. The metabolites related to cholesterol transport and estradiol production were enriched in CCs, while metabolites related to antiapoptosis were enriched in MGCs. The expression of genes and proteins involved in cholesterol transport (ABCA1, LDLR, and SCARB1) and estradiol production (SULT2B1 and CYP19A1) was significantly higher in CCs, and the expression of genes and proteins involved in antiapoptosis (CRLS1, LPCAT3, and PLA2G4A) was significantly higher in MGCs. The level of estrogen in CCs was significantly higher than that in MGCs, while the progesterone level showed no significant differences. There are differences between the metabolomes of CCs and MGCs. These differences may be involved in the regulation of oocyte maturation.

scholarly journals A rapid and robust method for the cryopreservation of human granulosa cells

2021 ◽  
Author(s):  
Sarah Beschta ◽  
Katja Eubler ◽  
Nancy Bohne ◽  
Ignasi Forne ◽  
Dieter Berg ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Large Scale ◽  
Ovarian Function ◽  
Oocyte Retrieval ◽  
Cell Contact ◽  
Cellular Model ◽  
Preovulatory Follicle ◽  
Human Granulosa Cells ◽  
Ovarian Cells

AbstractHuman primary granulosa cells (GCs) derived from women undergoing oocyte retrieval can be cultured and used as a cellular model for the study of human ovarian function. In vitro, they change rapidly, initially resembling cells of the preovulatory follicle and then cells of the corpus luteum. They are derived from individual patients, whose different medical history, lifestyle and age lead to heterogeneity. Thus, cells can rarely be ideally matched for cellular experiments or, if available, only in small quantities. We reasoned that cryopreservation of human GCs may be helpful to improve this situation. Previous studies indicated the feasibility of such an approach, but low survival of human GCs was reported, and effects on human GC functionality were only partially evaluated. We tested a slow freezing protocol (employing FCS and DMSO) for human GCs upon isolation from follicular fluid. We compared cryopreserved and subsequently thawed cells with fresh, non-cryopreserved cells from the same patients. About 80% of human GCs survived freezing/thawing. No differences were found in cell morphology, survival rate in culture, or transcript levels of mitochondrial (COX4, OPA1, TOMM20), steroidogenic (CYP11A1, CYP19A1) or cell–cell contact genes (GJA1) between the two groups in cells cultured for 1–5 days. A proteomic analysis revealed no statistically significant change in the abundance of a total of 5962 proteins. The two groups produced comparable basal levels of progesterone and responded similarly to hCG with elevation of progesterone. Taken together, our results show this to be a rapid and readily available method for the cryopreservation of human GCs. We anticipate that it will allow future large-scale experiments and may thereby improve cellular studies with human ovarian cells.

scholarly journals Serum deprivation affects secretory activity of cultured porcine ovarian follicles and granulosa cells and their response to hormones

2009 ◽  
Vol 54 (No. 10) ◽  
pp. 455-460
Author(s):  
A.V. Sirotkin
Keyword(s):  
Granulosa Cells ◽  
Ovarian Follicle ◽  
Secretory Activity ◽  
Ovarian Follicles ◽  
Serum Deprivation ◽  
Ovarian Cells ◽  
Igf I ◽  
Granulose Cells ◽  
Vitro Model

The aim of the present study is to understand the hormonal mechanisms of the effect of malnutrition on ovarian follicle functions. For this purpose, we examined the effect of malnutrition/serum deprivation, addition of metabolic hormones and gonadotropin (IGF-I, leptin and FSH) and their combination on the release of progesterone (P<sub>4</sub>), testosterone (T), estradiol (E<sub>2</sub>) and insulin-like growth factor I (IGF-I) by cultured whole ovarian follicles and on P<sub>4</sub> and IGF-I output by cultured granulosa cells isolated from porcine ovaries. It was observed that in ovarian follicles cultured with nutrients/serum addition of IGF-I reduced release of P<sub>4</sub>, but not of T or E<sub>2</sub>. Exogenous leptin reduced output of E<sub>2</sub>, but not of P<sub>4</sub> or T, and increased IGF-I output. No significant effect of FSH on release of steroid hormones by isolated follicles was found. Serum deprivation did not affect release of P<sub>4</sub>, but reduced output of T and E<sub>2</sub>, and promoted IGF-I release by cultured ovarian follicles. Addition of hormones failed to prevent the effect of malnutrition on the secretory activity of cultured ovarian follicles. In cultured granulose cells, all the tested hormones promoted release of both P<sub>4</sub> and IGF-I. Food restriction/serum deprivation reduced both P<sub>4</sub> and IGF-I output. Additions of either IGF-I, leptin and FSH prevented the inhibitory action of malnutrition on both P<sub>4</sub> and IGF-I release. The present observations (1) confirm the involvement of the hormones IGF-I, leptin and FSH in the control of secretory activity of ovarian cells, (2) demonstrate, that both isolated ovarian granulosa cells and whole follicles cultured in the absence of serum nutrients could be an adequate in-vitro model for studying the effect of malnutrition on ovarian secretory functions, and (3) suggest, that malnutrition could affect ovarian functions through changes in the release of ovarian hormones.

Lpin1 in human visceral and subcutaneous adipose tissue: similar levels but different associations with lipogenic and lipolytic genes

2010 ◽  
Vol 299 (2) ◽  
pp. E308-E317 ◽  
Author(s):  
Merce Miranda ◽  
Xavier Escoté ◽  
María J. Alcaide ◽  
Esther Solano ◽  
Victòria Ceperuelo-Mallafré ◽  
...  
Keyword(s):  
Adipose Tissue ◽  
Metabolic Profile ◽  
Low Grade ◽  
Mrna Levels ◽  
Cross Sectional ◽  
Expression Of Genes ◽  
Close Relationship ◽  
Few Data ◽  
Vitro Analysis

LPIN1 is a gene with important effects on lipidic and metabolic homeostasis. Human subcutaneous LPIN1 expression levels in adipose tissue are related with a better metabolic profile, including insulin sensitivity markers. However, there are few data on the regulation of LPIN1 in visceral adipose tissue (VAT). Our aim was to perform a cross-sectional analysis of VAT compared with subcutaneous (SAT) LPIN1 expression in a well-characterized obese cohort, its relation with the expression of genes involved in lipid metabolism, and the in vitro response to lipogenic and lipolytic stimuli. A downregulation of total LPIN1 mRNA expression in subjects with obesity was found in VAT similarly to that in SAT. Despite similar total LPIN1 mRNA levels in SAT and VAT, a close relationship with clinical parameters and with many lipogenic and lipolytic genes was observed primarily in SAT depot. As shown in the in vitro analysis, the low-grade proinflammatory environment and the insulin resistance associated with obesity may contribute to downregulate LPIN1 in adipose tissue, leading to a worse metabolic profile.

scholarly journals Mechanisms subserving the trophic actions of insulin on ovarian cells. In vitro studies using swine granulosa cells

1983 ◽  
Vol 72 (3) ◽  
pp. 1046-1057 ◽  
Author(s):  
Johannes D. Veldhuis ◽  
Lisa A. Kolp ◽  
Michael E. Toaff ◽  
Jerome F. Strauss ◽  
Lawrence M. Demers
Keyword(s):  
Granulosa Cells ◽  
In Vitro Studies ◽  
Ovarian Cells

scholarly journals FSH, Oxytocin and IGF-I Regulate the Expression of Sirtuin 1 in Porcine Ovarian Granulosa Cells

2020 ◽  
pp. 461-466
Author(s):  
A SIROTKIN ◽  
P DEKANOVÁ ◽  
A HARRATH
Keyword(s):  
Granulosa Cells ◽  
Intracellular Signaling ◽  
Sirtuin 1 ◽  
Signaling System ◽  
Factor I ◽  
Ovarian Granulosa Cells ◽  
Ovarian Cells ◽  
Igf I ◽  
Hormonal Action

The involvement of the mTOR system/enzyme sirtuin 1 (SIRT1) intracellular signaling system in the control of ovarian functions and its role in mediating hormonal action on the ovary has been proposed, but this hypothesis should be supported by a demonstrated influence of hormones on mTOR/SIRT1. Therefore, the aim of our in vitro experiments was to examine the effect of the known hormonal regulators of ovarian functions, such as follicle-stimulating hormone (FSH), oxytocin (OT) and insulin-like growth factor I (IGF-I), on mTOR/SIRT1. The accumulation of SIRT1 in porcine ovarian granulosa cells cultured with and without these hormones (at doses of 1, 10 or 100 ng.ml-1) was evaluated using immunocytochemistry. It was observed that the addition of FSH (at 10 ng.ml-1 but not at 1 or 100 ng/ml) and OT (at all tested doses) increased the expression of SIRT1 in ovarian cells. In addition, 100 ng.ml-1, but not at 1 or 10 ng.ml-1, of IGF-I decreased SIRT1 accumulation. Our observations are the first demonstration that hormones can directly regulate the ovarian mTOR/SIRT1 system and that this system could mediate the action of hormonal regulators on the ovary.

The Expression of Genes Involved in Ovulation Is Regulated by Angiotensin II in Granulosa Cells In Vitro.

2008 ◽  
Vol 78 (Suppl_1) ◽  
pp. 80-81
Author(s):  
Valerio M. Portela ◽  
Paulo B.D. Goncalves ◽  
Joao F.C. de Oliveira ◽  
Christopher A. Price
Keyword(s):  
Angiotensin Ii ◽  
Granulosa Cells ◽  
Expression Of Genes

scholarly journals Expression of Functional Leptin Receptors in the Human Ovary1

1997 ◽  
Vol 82 (12) ◽  
pp. 4144-4148 ◽  
Author(s):  
Cecilia Karlsson ◽  
Kajsa Lindell ◽  
Eva Svensson ◽  
Christina Bergh ◽  
Peter Lind ◽  
...  
Keyword(s):  
Adipose Tissue ◽  
Granulosa Cells ◽  
Follicular Fluid ◽  
Leptin Receptor ◽  
Biological Response ◽  
Human Ovary ◽  
Leptin Receptors ◽  
Short Isoforms ◽  
Reverse Transcription Pcr ◽  
Ovarian Cells

The size of body fat stores is known to influence fertility, indicating a link between adipose tissue and the reproductive system. Studies in mice have identified the adipocyte-derived hormone, leptin (Ob protein), as a possible mediator of this effect. The aim of this study was to investigate the possibility that leptin may have direct effects on the human ovary. To probe this hypothesis we first analyzed the expression of leptin receptors in the human ovary. Transcripts encoding both the long and short isoforms of the leptin receptor were present in human granulosa cells and thecal cells; however, the short isoforms were expressed at much higher levels. Immunoreactive leptin was present in follicular fluid at levels similar to those found in serum. ob gene expression, however, was undetectable in the ovary, as determined by reverse transcription-PCR, whereas it was easily detected in adipose tissue. To determine whether leptin could induce a biological response in ovarian cells, we examined the effect of leptin on estradiol production in cultured granulosa cells. Leptin (100 ng/mL) inhibited LH (0.1 ng/mL)-stimulated estradiol production. In contrast, leptin had no effect on estradiol production in the absence of LH. In conclusion, this study has demonstrated that the leptin receptor is expressed in the human ovary, that leptin is present in follicular fluid, and that leptin can induce a biological response in ovarian cells. These results suggest that leptin may have a direct effect on the human ovary.

Differential expression of signal transduction factors in ovarian follicle development: a functional role for betaglycan and FIBP in granulosa cells in cattle

2008 ◽  
Vol 33 (2) ◽  
pp. 193-204 ◽  
Author(s):  
N. Forde ◽  
M. Mihm ◽  
M. J. Canty ◽  
A. E. Zielak ◽  
P. J. Baker ◽  
...  
Keyword(s):  
Signal Transduction ◽  
Granulosa Cells ◽  
Ovarian Follicle ◽  
Growth Trajectories ◽  
Dominant Follicle ◽  
Quantitative Real Time Pcr ◽  
Differential Growth ◽  
Ovarian Follicle Development ◽  
Expression Of Genes

Ovarian follicles develop in groups yet individual follicles follow different growth trajectories. This growth and development are regulated by endocrine and locally produced growth factors that use a myriad of receptors and signal transduction pathways to exert their effects on theca and granulosa cells. We hypothesize that differential growth may be due to differences in hormonal responsiveness that is partially mediated by differences in expression of genes involved in signal transduction. We used the bovine dominant follicle model, microarrays, quantitative real-time PCR and RNA interference to examine this. We identified 83 genes coding for signal transduction molecules and validated a subset of them associated with different stages of the follicle wave. We suggest important roles for CAM kinase-1 and EphA4 in theca cells and BCAR1 in granulosa cells for the development of dominant follicles and for betaglycan and FIBP in granulosa cells of regressing subordinate follicles. Inhibition of genes for betaglycan and FIBP in granulosa cells in vitro suggests that they inhibit estradiol production in regressing subordinate follicles.

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