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.

Abstract 128: Leptin: A Regulator of Aldosterone Synthase Expression & Aldosterone Secretion in Visceral Adipocytes, in Mice

Hypertension ◽  
2016 ◽  
Vol 68 (suppl_1) ◽  
Author(s):  
Eric J Belin de Chantemele ◽  
Anne-Cecile Huby ◽  
P. T Menk ◽  
Weiqin Chen ◽  
Brian Lane ◽  
...  
Keyword(s):  
Adipose Tissue ◽  
Cross Sections ◽  
Leptin Receptor ◽  
Zona Glomerulosa ◽  
Aldosterone Synthase ◽  
Leptin Receptors ◽  
Fat Depots ◽  
Physiological Relevance ◽  
Aldosterone Production

Obesity is associated with inappropriately high aldosterone levels, which contribute to the development of metabolic and cardiovascular disorders. The origin of these high aldosterone levels is incompletely understood. We recently demonstrated that the adipocyte-derived hormone leptin regulates aldosterone synthase (CYP11B2) expression and stimulates aldosterone release from adrenal zona glomerulosa cells. Recent studies demonstrate that adipocytes express CYP11B2 and secrete aldosterone. However, the mechanisms regulating aldosterone release from adipocytes remain unclear. Likewise, whether visceral (Visc) and subcutaneous (SubQ) adipose tissue contribute to a similar extent to aldosterone production is unknown. We tested the hypothesis that leptin increases adipocyte CYP11B2 expression and aldosterone production and investigated whether Visc and SubQ adipose tissues respond similarly to leptin. Immunostaining of mouse adipose tissue cross-sections and isolated mature adipocytes revealed that Visc and SubQ adipose tissue express leptin receptors. Treatment of mouse freshly isolated mature adipocytes, non-differenciated (stromal fraction) and differentiated adipocytes revealed that leptin dose-dependently increased CYP11B2 expression and aldosterone production in Visc adipose tissue only. Although leptin receptor and CYP11B2 levels were similar in SubQ and Visc adipocytes, SubQ adipocytes were unresponsive to leptin. The physiological relevance of these in vitro data was tested by measuring plasma aldosterone levels in mice deprived of adipose tissue (lipodystrophic mice) treated with leptin. Absence of adipose tissue in lipodystrophic mice blunted leptin-induced increases in aldosterone levels (WT-vehicle: 471±82 vs. WT-Leptin: 1699±396, p<0.05; KO-vehicle: 539±71 vs. KO+leptin: 787±156, NS). The human relevance of these data was determined by reporting that CYP11B2 expression gradually increased with body mass index in human mediastinal and omental fat depots. In summary these data strongly suggest that leptin regulates CYP11B2 levels and aldosterone release in visceral adipose tissue and that leptin-induced, adipocyte-derived aldosterone may contribute to obesity-associated hyperaldosteronism.

scholarly journals Leptin Antagonizes the Insulin-Like Growth Factor-I Augmentation of Steroidogenesis in Granulosa and Theca Cells of the Human Ovary1

1999 ◽  
Vol 84 (3) ◽  
pp. 1072-1076 ◽  
Author(s):  
Sanjay K. Agarwal ◽  
Klara Vogel ◽  
Stacy R. Weitsman ◽  
Denis A. Magoffin
Keyword(s):  
Growth Factor ◽  
Follicular Fluid ◽  
Leptin Receptor ◽  
Pcr Analysis ◽  
Insulin Like Growth Factor ◽  
Theca Cells ◽  
Factor I ◽  
Ovarian Cells ◽  
Igf I ◽  
Growth Factor I

There is increasing evidence that leptin is a physiological link between obesity and infertility. Although leptin receptors have been demonstrated in human ovaries, there is no information regarding the effects of leptin on cells from developing ovarian follicles. To test the direct effects of leptin on human ovarian cells, granulosa cells (GC) and theca cells were isolated from the ovaries of regularly cycling women. Serum was obtained at the time of surgery, and follicular fluid was aspirated from the follicles before isolation of the ovarian cells. Leptin concentrations were similar in follicular fluid and serum. RT-PCR analysis demonstrated that the long, signaling form of the leptin receptor was expressed in both theca and GC. In cultured GC, leptin had no effect on estradiol production, alone or in the presence of FSH, but caused a concentration-related inhibition of the insulin-like growth factor I (IGF-I) augmentation of FSH-stimulated estradiol production. The effect of leptin was specific, because there was no effect on progesterone production. In cultured theca cells, leptin did not alter androstenedione production, alone or in the presence of LH. Leptin caused a concentration-related inhibition of the IGF-I augmentation of LH-stimulated androstenedione production. These data demonstrate that leptin can directly inhibit IGF-I action in ovarian theca and GC at concentrations commonly present in obese women.

scholarly journals Role of leptin receptors in granulosa cells during ovulation

Reproduction ◽  
2014 ◽  
Vol 147 (2) ◽  
pp. 221-229 ◽  
Author(s):  
Lisa Dupuis ◽  
Yasmin Schuermann ◽  
Tamara Cohen ◽  
Dayananda Siddappa ◽  
Anitha Kalaiselvanraja ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Leptin Receptor ◽  
Critical Role ◽  
Follicular Development ◽  
Reproductive Function ◽  
Control Treatment ◽  
Maturation Stage ◽  
Leptin Receptors ◽  
Enhancer Binding Protein

Leptin is an important hormone influencing reproductive function. However, the mechanisms underpinning the role of leptin in the regulation of reproduction remain to be completely deciphered. In this study, our objective is to understand the mechanisms regulating the expression of leptin receptor (Lepr) and its role in ovarian granulosa cells during ovulation. First, granulosa cells were collected from superovulated mice to profile mRNA expression of Lepr isoforms (LeprA and LeprB) throughout follicular development. Expression of LeprA and LeprB was dramatically induced in the granulosa cells of ovulating follicles at 4 h after human chorionic gonadotropin (hCG) treatment. Relative abundance of both mRNA and protein of CCAAT/enhancer-binding protein β (Cebpβ) increased in granulosa cells from 1 to 7 h post-hCG. Furthermore, chromatin immunoprecipitation assay confirmed the recruitment of Cebpβ to Lepr promoter. Thus, hCG-induced transcription of Lepr appears to be regulated by Cebpβ, which led us to hypothesise that Lepr may play a role during ovulation. To test this hypothesis, we used a recently developed pegylated superactive mouse leptin antagonist (PEG-SMLA) to inhibit Lepr signalling during ovulation. I.p. administration of PEG-SMLA (10 μg/g) to superovulated mice reduced ovulation rate by 65% compared with control treatment. Although the maturation stage of the ovulated oocytes remained unaltered, ovulation genes Ptgs2 and Has2 were downregulated in PEG-SMLA-treated mice compared with control mice. These results demonstrate that Lepr is dramatically induced in the granulosa cells of ovulating follicles and this induction of Lepr expression requires the transcription factor Cebpβ. Lepr plays a critical role in the process of ovulation by regulating, at least in part, the expression of the important genes involved in the preovulatory maturation of follicles.

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 Leptin receptors in human skeletal muscle

2007 ◽  
Vol 102 (5) ◽  
pp. 1786-1792 ◽  
Author(s):  
Borja Guerra ◽  
Alfredo Santana ◽  
Teresa Fuentes ◽  
Safira Delgado-Guerra ◽  
Alfredo Cabrera-Socorro ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Adipose Tissue ◽  
Molecular Mass ◽  
Protein Level ◽  
Subcutaneous Adipose Tissue ◽  
Human Skeletal Muscle ◽  
Leptin Receptor ◽  
Leptin Receptors ◽  
Subcutaneous Adipose ◽  
Basal Concentration

Human skeletal muscle expresses leptin receptor mRNA; however, it remains unknown whether leptin receptors (OB-R) are also expressed at the protein level. Fourteen healthy men (age = 33.1 ± 2.0 yr, height = 175.9 ± 1.7 cm, body mass = 81.2 ± 3.8 kg, body fat = 22.5 ± 1.9%; means ± SE) participated in this investigation. The expression of OB-R protein was determined in skeletal muscle, subcutaneous adipose tissue, and hypothalamus using a polyclonal rabbit anti-human leptin receptor. Three bands with a molecular mass close to 170, 128, and 98 kDa were identified by Western blot with the anti-OB-R antibody. All three bands were identified in skeletal muscle: the 98-kDa and 170-kDa bands were detected in hypothalamus, and the 98-kDa and 128-kDa bands were detected in thigh subcutaneous adipose tissue. The 128-kDa isoform was not detected in four subjects, whereas in the rest its occurrence was fully explained by the presence of intermuscular adipose tissue, as demonstrated using an anti-perilipin A antibody. No relationship was observed between the basal concentration of leptin in serum and the 170-kDa band density. In conclusion, a long isoform of the leptin receptor with a molecular mass close to 170 kDa is expressed at the protein level in human skeletal muscle. The amount of 170-kDa protein appears to be independent of the basal concentration of leptin in serum.

Inhibin production by porcine granulosa and luteal cells: development and biological validation of a RIA

1989 ◽  
Vol 120 (4) ◽  
pp. 511-518 ◽  
Author(s):  
U. Michel ◽  
H. Jarry ◽  
M. Metten ◽  
W. Wuttke
Keyword(s):  
Detection Limit ◽  
Granulosa Cells ◽  
Follicular Fluid ◽  
Inhibin A ◽  
Serum Free ◽  
Luteal Cells ◽  
Biological Validation ◽  
Ovarian Cells ◽  
Progesterone Secretion ◽  
Porcine Granulosa Cells

Abstract. We describe the development and biological validation of a radioimmunoassay for immuno- and bioactive porcine inhibin. A synthetic 1-32 porcine inhibin peptide was used to raise an antiserum and Tyr-1-32 peptide as tracer. As standard we employed porcine follicular fluid calibrated with the 1-32 α-inhibin. Medium obtained from serum-free cultured porcine granulosa cells was chromatographed on Superose S-12 and Mono-Q. Resulting fractions were analysed for inhibin bio- and immunoreactivity. It is shown that granulosa cells produce at least two types of bioactive inhibins, one being also immunoactive in our RIA. We studied secretion of immunoreactive inhibin from porcine ovarian cells under various conditions: Inhibin secretion from mature and immature granulosa cells can be stimulated by FSH, whereas hCG enhances inhibin secretion only from mature granulosa cells. During extended time of culture, the capability of granulosa cells to secrete inhibin is reduced. In contrast, progesterone secretion from these cells increases; this is due to spontaneous functional luteinization. This assumption is supported by the low inhibin secretion of luteal cells in comparison to granulosa cells. Intracellular inhibin content in luteal cells is below detection limit of the RIA, whereas granulosa cells contain readily detectable amounts of this hormone.

Vaspin, a novel adipokine in woman granulosa cells physiology and PCOS pathogenesis?

2021 ◽  
Author(s):  
Alice Bongrani ◽  
Namya Mellouk ◽  
Christelle Ramé ◽  
Marion Cornuau ◽  
Fabrice Guerif ◽  
...  
Keyword(s):  
Insulin Resistance ◽  
Granulosa Cells ◽  
Follicular Fluid ◽  
Polycystic Ovary ◽  
Normal Weight ◽  
Human Reproduction ◽  
In Vitro Fertilisation ◽  
Obese Women ◽  
Human Ovary

Vaspin is a novel adipokine mainly expressed in visceral adipose tissue and closely related to obesity and insulin-resistance. Currently, data about its ovarian expression are limited to animal models and its role in human reproduction is largely unexplored. Our study’s aims were thento characterise vaspin expression in the human ovary and to study in vitro its effects on granulosa cells physiology. Secondly, we assessed vaspin and its receptor GRP78 variations in granulosa cells and follicular fluid of a cohort of 112 infertile women undergoing an in vitro fertilisation procedure and allocated to 3 groups, each including normal-weight and obese subjects: 34 PCOS patients, 33 women with isolated polycystic ovary morphology (ECHO group) and 45 controls. Vaspin and GRP78 expression in the ovary was assessed by immunohistochemistry, RT-qPCR and Western blot. Granulosa cells and follicular fluid were analysed by RT-qPCR and ELISA, respectively. In vitro, granulosa cells metabolism was studied after stimulation with recombinant human vaspin, with and without a small interfering RNA directed against GRP78. Vaspin was highly expressed in the human ovary and concentration-dependently enhanced granulosa cells steroidogenesis, proliferation and viability through GRP78 (p<0.0001). Vaspin levels in both granulosa cells and follicular fluid were significantly higher in obese women (p<0.0001) and in the normal-weight ECHO group (p<0.001), which also had the highest expression rates of GRP78 (p<0.05).Although further investigation is needed, vaspin appears as a novel modulator of human granulosa cells physiology and possibly plays a role in PCOS pathogenesis, notably protecting from insulin-resistance induced complications.

scholarly journals High-Fat Diet Causes Lipotoxicity Responses in Cumulus–Oocyte Complexes and Decreased Fertilization Rates

Endocrinology ◽  
2010 ◽  
Vol 151 (11) ◽  
pp. 5438-5445 ◽  
Author(s):  
Linda Lin-Yan Wu ◽  
Kylie R. Dunning ◽  
Xing Yang ◽  
Darryl L. Russell ◽  
Michelle Lane ◽  
...  
Keyword(s):  
Er Stress ◽  
Mitochondrial Dysfunction ◽  
Granulosa Cells ◽  
Follicular Fluid ◽  
High Fat Diet ◽  
Marker Genes ◽  
Obese Women ◽  
High Fat ◽  
Fertilization Rates ◽  
Ovarian Cells

In obesity, accumulation of lipid in nonadipose tissues, or lipotoxicity, is associated with endoplasmic reticulum (ER) stress, mitochondrial dysfunction, and ultimately apoptosis. We have previously shown that obese women have increased triglycerides in follicular fluid; thus, the present study examined whether high-fat diet–induced obesity causes lipotoxicity in granulosa cells and the cumulus–oocyte complex (COC). Oocytes of mice fed a high-fat diet had dramatically increased lipid content and reduced mitochondrial membrane potential compared to those of mice fed a control diet. COCs from mice fed a high-fat diet had increased expression of ER stress marker genes ATF4 and GRP78. Apoptosis was increased in granulosa and cumulus cells of mice fed a high-fat diet. Mice fed a high-fat diet also exhibited increased anovulation and decreased in vivo fertilization rates. Thus, lipid accumulation, ER stress, mitochondrial dysfunction, and apoptosis are markedly increased in ovarian cells of mice fed a high-fat diet. ER stress markers were also analyzed in granulosa cells and follicular fluid from women with varying body mass indices (BMI). ATF4 was increased in granulosa cells and [Ca2+] in follicular fluid from obese women compared to nonobese women. These results indicate that lipotoxicity may be occurring in ovarian cells of obese women and may contribute to the reduced pregnancy rates observed in response to obesity.

Butyryl- and acetylcholine-esterases in human IVF-derived follicular fluid and luteinizing granulosa cells

2013 ◽  
Vol 121 (03) ◽  
Author(s):  
J Blohberger ◽  
D Einwang ◽  
D Berg ◽  
U Berg ◽  
S Hecht ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Follicular Fluid ◽  
Human Ivf

A technique for superfusion of isolated granulosa cells. Dynamics of cAMP production and steroidogenesis in response to gonadotropins

1988 ◽  
Vol 117 (4) ◽  
pp. 497-506 ◽  
Author(s):  
Carl Johanson ◽  
Viktor Johanson
Keyword(s):  
Granulosa Cells ◽  
Culture Medium ◽  
Cell Damage ◽  
Adenosine Monophosphate ◽  
Camp Production ◽  
Morphological Examination ◽  
Ovarian Cells ◽  
Temporal Relationships ◽  
Polycarbonate Membranes ◽  
Estradiol 17Β

Abstract. A superfusion model for isolated ovarian cells was developed and characterized in detail. Granulosa cells isolated from pre-ovulatory rat ovarian follicles were placed in superfusion (perifusion) chambers with a volume of 125 μl. Culture medium was pumped through the chambers, collected in 20-min fractions of 600 μl and analysed for cAMP and steroids. Viability was confirmed by morphological examination. The use of polycarbonate membranes to retain the cells in the chambers was abandoned since the membranes caused severe cell damage. The temporal relationships between gonadotropic stimuli and the release of cyclic 3':5'-adenosine monophosphate (cAMP) and steroids was investigated. Within 10 min FSH elicited transient increase in the release of cAMP and progesterone but had no effect on testosterone or estradiol-17β release. Amplitude and duration of the response in cAMP and progesterone release were correlated to concentration and length of the FSH pulse when these parameters were varied within the ranges 1–100 μg/l and 30–270 min, respectively. Compared with the cAMP response, the progesterone response peaked up to 30 min later and lasted 1 to 2 h longer but could not be extended to more than approximately 6 h, not even with longer FSH pulses. These results could indicate a development of desensitization.

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