scholarly journals Association of β-arrestin1 and p53-MDM2 signaling in the development of missed abortion

2020 ◽  
Author(s):  
Ting Liu ◽  
Yuyan Ma ◽  
Qihui Yin ◽  
Huanyu Zhou ◽  
Yan Fang
Keyword(s):  
G Protein ◽  
Cell Invasion ◽  
Positive Staining ◽  
Mrna Levels ◽  
Healthy Controls ◽  
G Protein Coupled Receptor ◽  
Missed Abortion ◽  
Real Time Quantitative Pcr ◽  
Elevated Expression ◽  
G Protein Coupled

Abstract BackgroundMissed abortion is a nonviable pregnancy before the 20th week of gestation with retained products of conception. The definite etiology and pathogenesis are not fully understood. β-arrestin1, the important dynamic multitask scaffold protein, it play an important regulatory role in interacting with G protein-coupled receptor (GPCR) to mediate its homologous desensitization and internalization. Recent studies have demonstrated that p53/Mdm2-mediated ubiquitination of the IGF-1R maybe closely related to G protein-coupled receptor kinases (GRK)/β-arrestin1 system. Our previous studies have confirmed that the elevated expression of p53 and Mdm2 may be responsible for apoptosis during missed abortion. However, there was no information surrounding β-arrestin1 in missed abortion.MethodsThe mRNA levels of β-arrestin1 in villous samples of 31 missed abortion patients and 31 healthy controls were determined by real-time quantitative PCR. Immunohistochemistry was used to explore the expression and location of β-arrestin1, p53, Mdm2, VEGF, HIF-lα in trophoblasts. We up-regulated and silenced the expression of β-arrestin1 by lentiviral transfection, transwell assays were performed to examine the influences of β-arrestin1 expression on cell invasion. Furthermore, we explored the expression of several important proteins which may be related to β-arrestin1.Resultsβ-arrestin1 mRNA levels in the villous samples from women with missed abortion were found to be dramatically lower than in women who had normal pregnancies. The immunohistochemistry results showed that β-arrestin1 positive staining was significantly lower in missed abortion group than that in normal pregnancies group. Furthermore, the patients with missed abortion showed significantly higher levels of p53, Mdm2 and HIF-lα, lower level of VEGF than healthy controls by immunohistochemistry. The protein expressions of p-ERK、p-AKT、p-p53 in HRT-8 cells were significantly downregulated by reducing β-arrestin1 expression, while the expression of p-NF-ΚB、p-Mdm2 were enhanced. Overexpression of β-arrestin1 exhibited the adverse effect. The loss expression of β-arrestin1 expression was significantly related with decreased cell invasion ability.ConclusionOur data indicated that β-arrestin1 could play an important role in maintaining the maternal-fetal tolerance, the decreased β-arrestin1 expression in the villous samples may be related to the development of missed abortion.

scholarly journals G Protein-Coupled Receptor 30 Expression Is Required for Estrogen Stimulation of Primordial Follicle Formation in the Hamster Ovary

Endocrinology ◽  
2008 ◽  
Vol 149 (9) ◽  
pp. 4452-4461 ◽  
Author(s):  
Cheng Wang ◽  
Eric R. Prossnitz ◽  
Shyamal K. Roy
Keyword(s):  
G Protein ◽  
Fluorescein Isothiocyanate ◽  
Primordial Follicle ◽  
Mrna Levels ◽  
G Protein Coupled Receptor ◽  
Ovary Cells ◽  
Primordial Follicles ◽  
Protein Levels ◽  
G Protein Coupled

Estradiol-17β (E2) plays an important role in the formation and development of primordial follicles, but the mechanisms remain unclear. G protein-coupled receptor 30 (GPR30) can mediate a rapid and transcription-independent E2 signaling in various cells. The objectives of this study were to examine whether GPR30 was expressed in the neonatal hamster ovary and whether it could mediate estrogen action during the formation of primordial follicles. GPR30 mRNA levels decreased from the 13th day of gestation (E13) through the second day of postnatal (P2) life, followed by steady increases from P3 through P6. Consistent with the changes in mRNA levels, GPR30 protein expression decreased from E13 to P2 followed by a significant increase by P7, the day before the first appearance of primordial follicles in the hamster ovary. GPR30 was expressed both in the oocytes and somatic cells, although the expression in the oocytes was low. GPR30 protein was located primarily in the perinuclear endoplasmic reticulum, which was also the site of E2-BSA-FITC (E2-BSA-fluorescein isothiocyanate) binding. E2 or E2-BSA increased intracellular calcium in neonatal hamster ovary cells in vitro. Exposure to GPR30 small interfering RNA in vitro significantly reduced GPR30 mRNA and protein levels in cultured hamster ovaries, attenuated E-BSA binding to cultured P6 ovarian cells, and markedly suppressed estrogen-stimulated primordial follicle formation. These results suggest that a membrane estrogen receptor, GPR30, is expressed in the ovary during perinatal development and mediates E2 action on primordial follicle formation.

scholarly journals Developmental expression of the G protein-coupled receptor 54 and three GnRH mRNAs in the teleost fish cobia

2007 ◽  
Vol 38 (2) ◽  
pp. 235-244 ◽  
Author(s):  
J Shaik Mohamed ◽  
Abby D Benninghoff ◽  
G Joan Holt ◽  
Izhar A Khan
Keyword(s):  
G Protein ◽  
Teleost Fish ◽  
Expression Patterns ◽  
Developmental Expression ◽  
Mrna Levels ◽  
G Protein Coupled Receptor ◽  
Early Puberty ◽  
Cdna Sequences ◽  
Gnrh Release ◽  
G Protein Coupled

The cDNAs of the G protein-coupled receptor 54 (GPR54) and three prepro-gonadotropin-releasing hormones, GnRH-I (seabream GnRH), GnRH-II (chicken GnRH-II), and GnRH-III (salmon GnRH) were isolated and cloned from the brain of the teleost fish cobia, Rachycentron canadum. The cobia GPR54 cDNA was 95 and 51–56% identical to those of tilapia and mammalian models respectively. The GnRH cDNA sequences of cobia showed strong identities to those of tilapia, Atlantic croaker, red drum, and the seabass and seabream species. The real-time quantitative RT-PCR methods allowed detection of all three GnRH mRNAs on the first day after hatching (DAH). The GnRH-I mRNA levels, which were the lowest among the three GnRHs, increased gradually with two distinct peaks in larvae at 3 and 4 DAH. On the other hand, GnRH-II and GnRH-III mRNAs were significantly higher in larvae at 2 and 6 DAH compared with those on the preceding days. In addition, significant peaks of all the three GnRH mRNAs were observed in the brains of 26-day-old fish. The finding of higher GnRH-I and GnRH-II mRNAs in males than females at 153 DAH may be related to early puberty observed during the first year in laboratory-reared male cobia. Moreover, this study demonstrates for the first time the expression of GPR54 mRNA during larval development in a vertebrate species. The concomitant expression patterns of GPR54 and GnRH mRNAs during different stages of larval and juvenile developments, and during early puberty in male cobia suggest a potential relationship between GPR54 and multiple GnRHs during these stages of development consistent with the role of GPR54 in controlling GnRH release in mammals. The increase in GPR54 and GnRH mRNAs observed during early puberty in cobia is consistent with a similar change reported in pubertal rats. This finding together with the localization of GPR54 mRNAs on GnRH neurons in fish and mammals suggests that the GPR54–GnRH interactions may be conserved in different vertebrate groups.

Targeted overexpression of G protein-coupled receptor kinase-2 in osteoblasts promotes bone loss

2005 ◽  
Vol 288 (4) ◽  
pp. E826-E834 ◽  
Author(s):  
Liming Wang ◽  
Shiguang Liu ◽  
L. Darryl Quarles ◽  
Robert F. Spurney
Keyword(s):  
Bone Formation ◽  
Trabecular Bone ◽  
Bone Remodeling ◽  
G Protein ◽  
Mrna Levels ◽  
G Protein Coupled Receptor ◽  
Mineral Density ◽  
Trabecular Thickness ◽  
Osteoclastic Activity ◽  
G Protein Coupled

To investigate the role of G protein-coupled receptor kinases (GRKs) in regulating bone formation in vivo, we overexpressed the potent G protein-coupled receptor (GPCR) regulator GRK2 in osteoblasts, using the osteocalcin gene-2 promoter to target expression to osteoblastic cells. Using the parathyroid hormone (PTH) receptor as a model system, we found that overexpression of GRK2 in osteoblasts attenuated PTH-induced cAMP generation by mouse calvaria ex vivo. This decrease in GPCR responsiveness was associated with a reduction in bone mineral density (BMD) in transgenic (TG) mice compared with non-TG littermate controls. The decrease in BMD was most prominent in trabecular-rich lumbar spine and was not observed in cortical bone of the femoral shaft. Quantitative computed tomography indicated that the loss of trabecular bone was due to a decrease in trabecular thickness, with little change in trabecular number. Histomorphometric analyses confirmed the decrease in trabecular bone volume and demonstrated reduced bone remodeling, as evidenced by a decrease in osteoblast numbers and osteoblast-mediated bone formation. Osteoclastic activity also appeared to be reduced because urinary excretion of the osteoclastic activity marker deoxypyridinoline was decreased in TG mice compared with control animals. Consistent with reduced coupling of osteoblast-mediated bone formation to osteoclastic bone resorption, mRNA levels of both osteoprotegrin and receptor activator of NF-κB ligand were altered in calvaria of TG mice in a pattern that would promote a low rate of bone remodeling. Taken together, these data suggest that enhancing GRK2 activity and consequently reducing GPCR activity in osteoblasts produces a low bone-turnover state that reduces bone mass.

scholarly journals Adropin acts in brain to inhibit water drinking: potential interaction with the orphan G protein-coupled receptor, GPR19

2016 ◽  
Vol 310 (6) ◽  
pp. R476-R480 ◽  
Author(s):  
Lauren M. Stein ◽  
Gina L. C. Yosten ◽  
Willis K. Samson
Keyword(s):  
G Protein ◽  
Peptide Hormone ◽  
Inhibitory Effect ◽  
Potential Interaction ◽  
Male Rats ◽  
Mrna Levels ◽  
G Protein Coupled Receptor ◽  
Medial Basal Hypothalamus ◽  
Water Drinking ◽  
G Protein Coupled

Adropin, a recently described peptide hormone produced in the brain and liver, has been reported to have physiologically relevant actions on glucose homeostasis and lipogenesis, and to exert significant effect on endothelial function. We describe a central nervous system action of adropin to inhibit water drinking and identify a potential adropin receptor, the orphan G protein-coupled receptor, GPR19. Reduction in GPR19 mRNA levels in medial basal hypothalamus of male rats resulted in the loss of the inhibitory effect of adropin on water deprivation-induced thirst. The identification of a novel brain action of adropin and a candidate receptor for the peptide should extend and accelerate the study of the potential therapeutic value of adropin or its mimetics for the treatment of metabolic disorders.

scholarly journals Inhibition of vascular smooth muscle G protein-coupled receptor kinase 2 enhances α1D-adrenergic receptor constriction

2008 ◽  
Vol 295 (4) ◽  
pp. H1695-H1704 ◽  
Author(s):  
Heather Irina Cohn ◽  
David M. Harris ◽  
Stephanie Pesant ◽  
Michael Pfeiffer ◽  
Rui-Hai Zhou ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
G Protein ◽  
Adrenergic Receptor ◽  
Plasma Norepinephrine ◽  
Mrna Levels ◽  
Receptor Kinase ◽  
G Protein Coupled Receptor ◽  
G Protein Coupled

G protein-coupled receptor kinase 2 (GRK2) is a serine/theorinine kinase that phosphorylates and desensitizes agonist-bound G protein-coupled receptors. GRK2 is increased in expression and activity in lymphocytes and vascular smooth muscle (VSM) in human hypertension and animal models of the disease. Inhibition of GRK2 using the carboxyl-terminal portion of the protein (GRK2ct) has been an effective tool to restore compromised β-adrenergic receptor (AR) function in heart failure and improve outcome. A well-characterized dysfunction in hypertension is attenuation of βAR-mediated vasodilation. Therefore, we tested the role of inhibition of GRK2 using GRK2ct or VSM-selective GRK2 gene ablation in a renal artery stenosis model of elevated blood pressure (BP) [the two-kidney, one-clip (2K1C) model]. Use of the 2K1C model resulted in a 30% increase in conscious BP, a threefold increase in plasma norepinephrine levels, and a 50% increase in VSM GRK2 mRNA levels. BP remained increased despite VSM-specific GRK2 inhibition by either GRK2 knockout (GRK2KO) or peptide inhibition (GRK2ct). Although βAR-mediated dilation in vivo and in situ was enhanced, α1AR-mediated vasoconstriction was also increased. Further pharmacological experiments using α1AR antagonists revealed that GRK2 inhibition of expression (GRK2KO) or activity (GRK2ct) enhanced α1DAR vasoconstriction. This is the first study to suggest that VSM α1DARs are a GRK2 substrate in vivo.

High tumoral levels of Kiss1 and G-protein-coupled receptor 54 expression are correlated with poor prognosis of estrogen receptor-positive breast tumors

2007 ◽  
Vol 14 (3) ◽  
pp. 691-702 ◽  
Author(s):  
Didier Marot ◽  
Ivan Bieche ◽  
Chantal Aumas ◽  
Stéphanie Esselin ◽  
Céline Bouquet ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Estrogen Receptor ◽  
G Protein ◽  
Breast Tumors ◽  
Metastasis Suppressor ◽  
Mrna Levels ◽  
Breast Cancers ◽  
G Protein Coupled Receptor ◽  
Primary Tumors ◽  
G Protein Coupled

KiSS1 is a putative metastasis suppressor gene in melanoma and breast cancer-encoding kisspeptins, which are also described as neuroendocrine regulators of the gonadotropic axis. Negative as well as positive regulation of KiSS1 gene expression by estradiol (E2) has been reported in the hypothalamus. Estrogen receptor α (ERα level is recognized as a marker of breast cancer, raising the question of whether expression of KiSS1 and its G-protein-coupled receptor (GPR54) is down- or upregulated by estrogens in breast cancer cells. KiSS1 was found to be expressed in MDA-MB-231, MCF7, and T47D cell lines, but not in ZR75-1, L56Br, and MDA-MB-435 cells. KiSS1 mRNA levels decreased significantly in ERα-negative MDA-MB-231 cells expressing recombinant ERα. In contrast, tamoxifen (TAM) treatment of ERα-positive MCF7 and T47D cells increased KiSS1 and GPR54 levels. The clinical relevance of this negative regulation of KiSS1 and GPR54 by E2 was then studied in postmenopausal breast cancers. KiSS1 mRNA increased with the grade of the breast tumors. ERα-positive invasive primary tumors expressed sevenfold lower KiSS1 levels than ERα-negative tumors. Among ERα-positive breast tumors from postmenopausal women treated with TAM, high KiSS1 combined with high GPR54 mRNA tumoral levels was unexpectedly associated with shorter relapse-free survival (RFS) relative to tumors expressing low tumoral mRNA levels of both genes. The contradictory observation of putative metastasis inhibitor role of kisspeptins and RFS to TAM treatment suggests that evaluation of KiSS1 and its receptor tumoral mRNA levels could be new interesting markers of the tumoral resistance to anti-estrogen treatment.

Sign in / Sign up

Export Citation Format

Share Document