Porcine leptin receptor: Molecular structure and expression in the ovary

Ruiz-Cortés, Zulma Tatiana; Men, Taoyan; Palin, Marie‐France; Downey, B.R.; Lacroix, Dan; Murphy, Bruce D.

doi:10.1002/1098-2795(200008)56:4<465::aid-mrd4>3.3.co;2-h

Cited by 31 publications

(44 citation statements)

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“…Viability of hepatocytes isolated by low speed centrifugation was 89 ± 2.5% by Trypan blue dye exclusion and aliquots were frozen at −80 • C until used. Cytosol and membrane fractions were prepared by centrifugation and differential solubilization in octylglucoside as described by Ruiz-Cortés et al [12]. Purified hepatocytes from the three pigs were thawed on ice, pooled, homogenized by brief exposure to ultrasound in buffer [2 mM octylglucoside, 1 mM DEDTC, 10 mM EDTA in 50 mM Tris, pH 8] and centrifuged at 30,000 g for 60 min at 4 • C. The supernatant (cytosol) was harvested and the particulate fraction was solubilized by ultrasound in 20 mM Tris containing 50 mM EDTA, 0.1 mM DEDTC and 32 mM octylglucoside, and centrifuged at 16,000 g for 15 min at 4 • C. The insoluble pellet was discarded and the supernatant was aspirated and considered to be the membrane fraction.…”

Section: Hepatocyte Preparationcontrasting

confidence: 56%

A gel-based reference map of the porcine hepatocyte proteome

Caperna

Shannon

Garrett

2008

Domestic Animal Endocrinology

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Caperna, Thomas J.; Shannon, Amy E.; and Garrett, Wesley M., "A gel-based reference map of the porcine hepatocyte proteome" (2008 AbstractThe overall goal of our research is to characterize and identify gene expression profiles of porcine hepatic cells. In this study, we have prepared two-dimensional electrophoresis maps of cytosol and membrane fractions from freshly prepared hepatocytes which were pooled from three crossbred pigs (35-69 kg). Following isoelectric focusing with three pH range immobilized pH gradient strips (pH 3-6, 5-8 and 7-10) and staining the second dimension gels with colloidal Coomassie blue, 728 protein spots were picked and digested with trypsin. Extracted tryptic peptides were initially subjected to matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS) analysis for identification of proteins by peptide mass fingerprinting (PMF). Proteins which were not identified by PMF were analyzed by liquid chromatography-tandem MS. Utilizing publicly available databases [NCBInr, Swiss Prot and expressed sequence tags (EST)], 648 proteins were identified. Of those, 282 were unique proteins and greater than 90% of proteins spots contained single proteins. These data represent the first comprehensive proteomic analysis of porcine hepatocytes and will provide a database for future investigations of endocrine regulation of gene expression and metabolic processes in vitro. Published by Elsevier Inc.

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Section: Hepatocyte Preparationcontrasting

confidence: 56%

A gel-based reference map of the porcine hepatocyte proteome

Caperna

Shannon

Garrett

2008

Domestic Animal Endocrinology

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“…Leptin receptor gene expression in the porcine ovary has been characterized and porcine leptin receptor cDNA has been cloned and sequenced (Ruiz-Cortes et al 2000). Leptin receptor levels are detectable in ovarian follicles collected from prepubertal animals, with a sixfold increase in expression in early antral and antral follicles of cycling animals (Gregoraszczuk et al 2007).…”

Section: Introductionsupporting

confidence: 86%

Supraphysiological leptin levels shift the profile of steroidogenesis in porcine ovarian follicles toward progesterone and testosterone secretion through increased expressions of CYP11A1 and 17β-HSD: a tissue culture approach

Gregoraszczuk¹,

Rak²

2013

Reproduction

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Evidence from both clinical and animal studies suggests that exposure to excess androgens results in cyst formation. The present in vitro study assessed the effects of supraphysiological concentrations of leptin (20 and 40 ng/ml) on progesterone (P4), androstenedione androstendione (A4), testosterone and estradiol (E2) secretion by ELISA and the expression of CYP11A1, CYP17, 17β-hydroxysteroid dehydrogenase (17β-HSD) and CYP19 by western blot to answer the question of whether leptin could be independent risk factor for cyst formation in pigs. Small- and medium-sized ovarian follicles were collected from prepubertal and cycling pigs. Increased P4 and testosterone secretions were observed in both small- and medium-sized follicles in prepubertal and cycling animals whereas there was no change in E2 secretion. Leptin treatment resulted in an increase in CYP11A1 and 17β-HSD protein expression but had no effect on CYP17 and CYP19 expression in follicles of either size from prepubertal and cycling pigs. Results of presented data suggest that leptin in elevated doses, by stimulatory effect on CYP11A1 and 17β-HSD protein expression resulting in elevated P4 and testosterone secretions could be an independent risk factor for cyst formation in both prepubertal and cycling pigs.

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“…OB-Rb protein expression in OS is also higher on days 10 to 12 than days 14 to 16 (Smolinska et al, 2007a). Similarly, Ruiz-Cortes et al (2000) demonstrated that both gene and protein expression of the leptin receptor peaked in porcine CL during the mid-luteal phase of the cycle, and was the lowest in regressed CL. Furthermore, leptin receptor mRNA increased as the porcine GC luteinized during 96 h in culture.…”

Section: Discussionmentioning

confidence: 99%

“…Leptin mRNA and protein have been identified in humans in GC and theca cells, CL, oocytes and follicular fluid Cioffi et al, 1997;Loffler et al, 2001;Welt et al, 2003); in mice in GC and theca cells, CL, oocytes and OS Ryan et al, 2002); and in rats in theca cells, CL and oocytes (Archanco et al, 2003;Ryan et al, 2003). OB-Rb gene and protein expression has been found in human GC and theca cells (Cioffi et al, 1997;Karlsson et al, 1997;Agarwal et al, 1999); mouse follicular cells, CL, OS and oocytes (Ryan et al, 2002); rabbit and rat follicular cells, CL and oocytes (Zamorano et al, 1997;Ryan et al, 2003); and porcine GC and theca cells harvested from prepubertal gilts, CL and OS (Ruiz-Cortes et al, 2000 andSmolinska et al, 2007a).…”

Section: Discussionmentioning

confidence: 99%

“…The expression of leptin receptors was observed in human and rat granulosa (GC) and theca cells (Cioffi et al, 1997;Karlsson et al, 1997;Agarwal et al, 1999;Ryan et al, 2003), in luteinized GC and theca cells harvested from prepubertal gilts (in a single in vitro study) and in the porcine corpus luteum (CL) and ovarian stroma (OS; Ruiz-Cortes et al, 2000;Smolinska et al, 2007a). The ovary is also believed to be a site of leptin synthesis.…”

Section: Introductionmentioning

confidence: 99%

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Expression of leptin and its receptor genes in the ovarian follicles of cycling and early pregnant pigs

Smolińska

Kamiński

Siawrys

et al. 2013

Animal

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Leptin is a polypeptide hormone produced primarily by adipocytes. It has been implicated in the regulation of satiety and energy homeostasis. Leptin has been suggested to play a role in reproduction based on its involvement in the regulation of the hypothalamic-pituitary-gonadal axis via endocrine, paracrine and/or autocrine pathways. The aim of the present study was to localize the cellular distribution of leptin and the long isoform of leptin receptor (OB-Rb) genes in porcine ovarian antral follicles and to compare the expression levels of leptin and OB-Rb mRNAs in porcine granulosa cells (GC), theca interna (TIC) and theca externa (TEC) cells during the luteal phase of the estrous cycle and in early pregnancy. The expression of leptin and OB-Rb genes was detected in GC, TIC and TEC. Significantly higher levels of leptin gene expression in GC were observed during the mid-and late-luteal phases of the cycle than on days 30 to 32 of pregnancy. On days 14 to 16 of pregnancy, leptin mRNA expression was higher than that on days 14 to 16 of the cycle. The expression of the OB-Rb gene in GC and TEC increased during pregnancy in comparison with the analyzed luteal phases of the cycle. Our results validate the hypothesis that locally produced leptin plays a role in the regulation of porcine reproduction at the ovarian level and exerts a direct effect on porcine follicles. The differences in OB-Rb gene expression in porcine GC and theca cells also suggest that their sensitivity to leptin varies in the ovaries of pregnant and cyclic pigs.

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Porcine leptin receptor: Molecular structure and expression in the ovary

Cited by 31 publications

References 0 publications

A gel-based reference map of the porcine hepatocyte proteome

A gel-based reference map of the porcine hepatocyte proteome

Supraphysiological leptin levels shift the profile of steroidogenesis in porcine ovarian follicles toward progesterone and testosterone secretion through increased expressions of CYP11A1 and 17β-HSD: a tissue culture approach

Expression of leptin and its receptor genes in the ovarian follicles of cycling and early pregnant pigs

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