Sugar and protein are the major macronutrients＇ sources, and their balanced intake is important for animal＇s health. It has been observed that animals are able to change food preference in an imbalanced nutritional ...Sugar and protein are the major macronutrients＇ sources, and their balanced intake is important for animal＇s health. It has been observed that animals are able to change food preference in an imbalanced nutritional condition to selectively consume nutrients that are deficient in the body （Dethier, 1976）. Early studies in both Drosophila and mouse have demonstrated that animals exhibit food rejection to imbalanced diets lacking essential amino acids （Hao et al., 2005; Bjordal et al., 2014）. Furthermore, the food preference change upon protein depri- vation has been characterized using a two choice assay in Drosophila （Ribeiro and Dickson, 2010; Vargas et al., 2010）. Different from protein food, sugar is the main energy source, and sugar deficiency severely affects animal survival （Lee et al., 2008）. However, whether animals adopt a strategy of fast food preference switch upon sugar deprivation had not been investigated, and the neural mechanisms underlying this behavior regulation remain poorly understood.展开更多
Description Cellular membranes present natural borders for signal transduction between cells and their environment. Nature developed different strategies to enable signals to cross the membrane barrier. The goal of th...Description Cellular membranes present natural borders for signal transduction between cells and their environment. Nature developed different strategies to enable signals to cross the membrane barrier. The goal of this meeting is to discuss the molecular mechanisms of transmembrane signaling on the basis of three protein classes, i.e.展开更多
The superfamily of G protein-coupled receptors (GPCRs) contains immense structural and functional diversity and mediates a myriad of biological processes upon activation by various extracellular signals.Critical roles...The superfamily of G protein-coupled receptors (GPCRs) contains immense structural and functional diversity and mediates a myriad of biological processes upon activation by various extracellular signals.Critical roles of GPCRs have been established in bone development,remodeling,and disease.Multiple human GPCR mutations impair bone development or metabolism,resulting in osteopathologies.Here we summarize the disease phenotypes and dysfunctions caused by GPCR gene mutations in humans as well as by deletion in animals.To date,92 receptors (5 glutamate family,67 rhodopsin family,5 adhesion,4 frizzled/taste2 family,5 secretin family,and 6 other 7TM receptors) have been associated with bone diseases and dysfunctions (36 in humans and 72 in animals).By analyzing data from these 92 GPCRs,we found that mutation or deletion of different individual GPCRs could induce similar bone diseases or dysfunctions,and the same individual GPCR mutation or deletion could induce different bone diseases or dysfunctions in different populations or animal models.Data from human diseases or dysfunctions identified 19 genes whose mutation was associated with human BMD:9 genes each for human height and osteoporosis;4 genes each for human osteoarthritis (OA) and fracture risk;and 2 genes each for adolescent idiopathic scoliosis (AIS),periodontitis,osteosarcoma growth,and tooth development.Reports from gene knockout animals found 40 GPCRs whose deficiency reduced bone mass,while deficiency of 22 GPCRs increased bone mass and BMD;deficiency of 8 GPCRs reduced body length,while 5 mice had reduced femur size upon GPCR deletion.Furthermore,deficiency in 6 GPCRs induced osteoporosis;4 induced osteoarthritis;3 delayed fracture healing;3 reduced arthritis severity;and reduced bone strength,increased bone strength,and increased cortical thickness were each observed in 2 GPCR-deficiency models.The ever-expanding number of GPCR mutation-associated diseases warrants accelerated molecular analysis,population studies,and investigation of phenotype correlation with SNPs to elucidate GPCR function in human diseases.展开更多
近日,中国科学院上海药物研究所徐华强/赵丽华团队联合山东大学于晓/孙金鹏团队、浙江大学基础医学院张岩团队等,在《自然-通讯》(Nature Communications)上,在线发表了题为"Structure insights into selective coupling of G prot...近日,中国科学院上海药物研究所徐华强/赵丽华团队联合山东大学于晓/孙金鹏团队、浙江大学基础医学院张岩团队等,在《自然-通讯》(Nature Communications)上,在线发表了题为"Structure insights into selective coupling of G protein subtypes by a class B Gprotein-coupled receptor"的研究成果.科研人员在揭示B类GPCRs一个亚家族的两个受体激素识别、受体激活和G蛋白偶联的特异性机制基础上,解析了CRF2R分别与Go蛋白和G11蛋白的复合物三维结构,即B类GPCRs偶联Go和G11的首个复合物结构,系统性阐述了促肾上腺皮质激素释放因子受体偶联Gs、G11和Go蛋白的分子作用机制,为B类GPCRs的G蛋白选择性偶联激活机制奠定了研究基础.展开更多
Adipogenesis is the healthy expansion of white adipose tissue(WAT),serving as a compensatory response to maintain metabolic homeostasis in the presence of excess energy in the body.Therefore,the identification of nove...Adipogenesis is the healthy expansion of white adipose tissue(WAT),serving as a compensatory response to maintain metabolic homeostasis in the presence of excess energy in the body.Therefore,the identification of novel regulatory molecules in adipogenesis,specifically membrane receptors such as G protein-coupled receptors(GPCRs),holds significant clinical promise.These receptors can serve as viable targets for pharmaceuticals,offering potential for restoring metabolic homeostasis in individuals with obesity.We utilized trajectory inference methods to analyze three distinct single-nucleus sequencing(sNuc-seq)datasets of adipose tissue and systematically identified GPCRs with the potential to regulate adipogenesis.Through verification in primary adipose progenitor cells(APCs)of mice,we discovered that ADGRD1 promoted the differentiation of APCs,while GPR39 inhibits this process.In the obese mouse model induced by a high-fat diet(HFD),both gain-of-function and loss-of-function studies validated that ADGRD1 promoted adipogenesis,thereby improving metabolic homeostasis,while GPR39 inhibited adipogenesis,leading to metabolic dysfunction.Additionally,through the analysis of 2,400 ChIP-seq data and 1,204 bulk RNA-seq data,we found that the transcription factors(TFs)MEF2D and TCF12 regulated the expression of ADGRD1 and GPR39,respectively.Our study revealed the regulatory role of GPCRs in adipogenesis,providing novel targets for clinical intervention of metabolic dysfunction in obese patients.展开更多
G-protein coupled receptors(GPCRs)are the largest family of druggable targets.In recent years,GPCR structural biology has made great advances,revealing the three-dimensional structures of many GPCRs and their interact...G-protein coupled receptors(GPCRs)are the largest family of druggable targets.In recent years,GPCR structural biology has made great advances,revealing the three-dimensional structures of many GPCRs and their interactions with ligands,proteins,and membrane components,which also have inspired a surge of structure-based drug discovery campaigns.This article provides a comprehensive summary of the currently available structural insights into the allosteric pockets of GPCRs and their regulatory mechanisms governing GPCR conformational changes.Furthermore,this article also presents several structure-inspired studies that utilize both orthosteric and allosteric modulation to discover small molecular modulators targeting GPCRs.The article emphasizes the promising potential of drug discovery targeting GPCR allosteric sites,while acknowledging the challenges arising from the limited structural information regarding the lipids and cholesterols in the membrane.Finally,the article discusses the future prospects of using large-scale or focused compound libraries to discover novel chemotypes,as well as the application of artificial intelligence(AI)in structure-based virtual screening(SBVS)against GPCRs.展开更多
Understanding the cellular mechanisms of drug addiction remains a key task in current brain research.While neuron-based mechanisms have been extensively explored over the past three decades,recent evidence indicates a...Understanding the cellular mechanisms of drug addiction remains a key task in current brain research.While neuron-based mechanisms have been extensively explored over the past three decades,recent evidence indicates a critical involvement of astrocytes,the main type of non-neuronal cells in the brain.In response to extracellular stimuli,astrocytes modulate the activity of neurons,synaptic transmission,and neural network properties,collectively influencing brain function.G protein-coupled receptors(GPCRs)expressed on astrocyte surfaces respond to neuron-and environment-derived ligands by activating or inhibiting astrocytic signaling,which in turn regulates adjacent neurons and their circuitry.In this review,we focus on the dopamine D1 receptors(D1R)and metabotropic glutamate receptor 5(mGLUR5 or GRM5)—two GPCRs that have been critically implicated in the acquisition and maintenance of addiction-related behaviors.Positioned as an introductory-level review,this article briefly discusses astrocyte biology,outlines earlier discoveries about the role of astrocytes in substance-use disorders(SUDs),and provides detailed discussion about astrocytic D1Rs and mGLUR5s in regulating synapse and network functions in the nucleus accumbens(NAc)—a brain region that mediates addictionrelated emotional and motivational responses.This review serves as a stepping stone for readers of Engineering to explore links between astrocytic GPCRs and drug addiction and other psychiatric disorders.展开更多
Activating and inactivating mutations in numerous human G protein-coupled receptors(GPCRs)are associated with a wide range of disease phenotypes.Here we use several class A GPCRs with a particularly large set of ident...Activating and inactivating mutations in numerous human G protein-coupled receptors(GPCRs)are associated with a wide range of disease phenotypes.Here we use several class A GPCRs with a particularly large set of identified disease-associated mutations,many of which were biochemically characterized,along with known GPCR structures and current models of GPCR activation,to understand the molecular mechanisms yielding pathological phenotypes.Based on this mechanistic understanding we also propose different therapeutic approaches,both conventional,using small molecule ligands,and novel,involving gene therapy.展开更多
The brain is,after the adipose tissue,the organ with the greatest amount of lipids and diversity in their composition in the human body.In neurons,lipids are involved in signaling pathways controlling autophagy,a lyso...The brain is,after the adipose tissue,the organ with the greatest amount of lipids and diversity in their composition in the human body.In neurons,lipids are involved in signaling pathways controlling autophagy,a lysosome-dependent catabolic process essential for the maintenance of neuronal homeostasis and the function of the primary cilium,a cellular antenna that acts as a communication hub that transfers extracellular signals into intracellular responses required for neurogenesis and brain development.A crosstalk between primary cilia and autophagy has been established;however,its role in the control of neuronal activity and homeostasis is barely known.In this review,we briefly discuss the current knowledge regarding the role of autophagy and the primary cilium in neurons.Then we review the recent literature about specific lipid subclasses in the regulation of autophagy,in the control of primary cilium structure and its dependent cellular signaling in physiological and pathological conditions,specifically focusing on neurons,an area of research that could have major implications in neurodevelopment,energy homeostasis,and neurodegeneration.展开更多
G-protein-coupled receptors(GPCRs)are the largest family of transmembrane receptors and regulate various physiological and pathological processes.Despite extensive studies,the roles of GPCRs in mouse embryonic stem ce...G-protein-coupled receptors(GPCRs)are the largest family of transmembrane receptors and regulate various physiological and pathological processes.Despite extensive studies,the roles of GPCRs in mouse embryonic stem cells(mESCs)remain poorly understood.Here,we show that GPR160,a class A member of GPCRs,is dramatically downregulated concurrent with mESC differentiation into embryoid bodies in vitro.Knockdown of Gpr160 leads to downregulation of the expression of pluripotency-associated transcription factors and upregulation of the expression of lineage markers,accompanying with the ar-rest of the mESC cell-cycle in the G0/G1 phase.RNA-seq analysis shows that GPR160 participates in the JAK/STAT signaling pathway crucial formaintaining ESC stemness,and the knockdown of Gpr160 results in the downregulation of STAT3 phosphorylation level,which in turn is partially rescued by colivelin,a STAT3 activator.Consistent with these observations,GPR160 physically interacts with JAK1,and co-operates with leukemia inhibitory factor receptor(LIFR)and gp130 to activate the STAT3 pathway.In summary,our results suggest that GPR160 regulates mESC self-renewal and pluripotency by interacting with the JAK1-LIFR-gp130 complex to mediate the JAK1/STAT3 signaling pathway.展开更多
Objectives:To investigate whether the protective actions of ginsenoside Rb1(Rb1)on astrocytes are mediated through the G_(s)-type G-protein-coupled receptor(GPCR-G_(s)).Methods:Primary astrocyte cultures derived from ...Objectives:To investigate whether the protective actions of ginsenoside Rb1(Rb1)on astrocytes are mediated through the G_(s)-type G-protein-coupled receptor(GPCR-G_(s)).Methods:Primary astrocyte cultures derived from neonatal mouse brain were used.Astrocyte injury was induced via oxygen-glucose deprivation/re-oxygenation(OGD/R).Cell morphology,viability,lactate dehydrogenase(LDH)leakage,apoptosis,glutamate uptake,and brain-derived neurotrophic factor(BDNF)secretion were assessed to gauge cell survival and functionality.Western blot was used to investigate the cyclic adenosine monophosphate(cAMP)and protein kinase B(Akt)signaling pathways.GPCR-G_(s)-specific inhibitors and molecular docking were used to identify target receptors.Results:Rb1 at concentrations ranging from 0.8 to 5μM did not significantly affect the viability,glutamate uptake,or BDNF secretion in normal astrocytes.OGD/R reduced astrocyte viability,increasing their LDH leakage and apoptosis rate.It also decreased glutamate uptake and BDNF secretion by these cells.Rb1 had protective effects of astrocytes challenged by OGD/R,by improving viability,reducing apoptosis,and enhancing glutamate uptake and BDNF secretion.Additionally,Rb1 activated the cAMP and Akt pathways in these cells.When the GPCR-G_(s) inhibitor NF449 was introduced,the protective effects of Rb1 completely disappeared,and its activation of cAMP and Akt signaling pathways was significantly inhibited.Conclusion:Rb1 protects against astrocytes from OGD/R-induced injury through GPCR-G_(s) mediation.展开更多
基金supported by the grants from the National Science Foundation of China(Nos. 91132709 and 31130027)
文摘Sugar and protein are the major macronutrients＇ sources, and their balanced intake is important for animal＇s health. It has been observed that animals are able to change food preference in an imbalanced nutritional condition to selectively consume nutrients that are deficient in the body （Dethier, 1976）. Early studies in both Drosophila and mouse have demonstrated that animals exhibit food rejection to imbalanced diets lacking essential amino acids （Hao et al., 2005; Bjordal et al., 2014）. Furthermore, the food preference change upon protein depri- vation has been characterized using a two choice assay in Drosophila （Ribeiro and Dickson, 2010; Vargas et al., 2010）. Different from protein food, sugar is the main energy source, and sugar deficiency severely affects animal survival （Lee et al., 2008）. However, whether animals adopt a strategy of fast food preference switch upon sugar deprivation had not been investigated, and the neural mechanisms underlying this behavior regulation remain poorly understood.
文摘Description Cellular membranes present natural borders for signal transduction between cells and their environment. Nature developed different strategies to enable signals to cross the membrane barrier. The goal of this meeting is to discuss the molecular mechanisms of transmembrane signaling on the basis of three protein classes, i.e.
基金supported by grants from the National Key Research and Development Program of China(2018YFC1105102 to J.L.,2016YFC0902102 to J.L.and J.X.)the National Natural Science Foundation of China(81722020,91749204,81472048 to J.L.,81330049 to M.L.,81330059 and 81572640 to J.X.)+2 种基金the Innovation Program of Shanghai Municipal Education Commission(14ZZ051 to J.L.,2017ZZ01017 to J.X.)the Science and Technology Commission of Shanghai Municipality(12ZR1447900 to J.L.,17JC1400903 and 17411950300 to J.X.)the Fundamental Research Funds for the Central Universities(to J.L.)
文摘The superfamily of G protein-coupled receptors (GPCRs) contains immense structural and functional diversity and mediates a myriad of biological processes upon activation by various extracellular signals.Critical roles of GPCRs have been established in bone development,remodeling,and disease.Multiple human GPCR mutations impair bone development or metabolism,resulting in osteopathologies.Here we summarize the disease phenotypes and dysfunctions caused by GPCR gene mutations in humans as well as by deletion in animals.To date,92 receptors (5 glutamate family,67 rhodopsin family,5 adhesion,4 frizzled/taste2 family,5 secretin family,and 6 other 7TM receptors) have been associated with bone diseases and dysfunctions (36 in humans and 72 in animals).By analyzing data from these 92 GPCRs,we found that mutation or deletion of different individual GPCRs could induce similar bone diseases or dysfunctions,and the same individual GPCR mutation or deletion could induce different bone diseases or dysfunctions in different populations or animal models.Data from human diseases or dysfunctions identified 19 genes whose mutation was associated with human BMD:9 genes each for human height and osteoporosis;4 genes each for human osteoarthritis (OA) and fracture risk;and 2 genes each for adolescent idiopathic scoliosis (AIS),periodontitis,osteosarcoma growth,and tooth development.Reports from gene knockout animals found 40 GPCRs whose deficiency reduced bone mass,while deficiency of 22 GPCRs increased bone mass and BMD;deficiency of 8 GPCRs reduced body length,while 5 mice had reduced femur size upon GPCR deletion.Furthermore,deficiency in 6 GPCRs induced osteoporosis;4 induced osteoarthritis;3 delayed fracture healing;3 reduced arthritis severity;and reduced bone strength,increased bone strength,and increased cortical thickness were each observed in 2 GPCR-deficiency models.The ever-expanding number of GPCR mutation-associated diseases warrants accelerated molecular analysis,population studies,and investigation of phenotype correlation with SNPs to elucidate GPCR function in human diseases.
文摘近日,中国科学院上海药物研究所徐华强/赵丽华团队联合山东大学于晓/孙金鹏团队、浙江大学基础医学院张岩团队等,在《自然-通讯》(Nature Communications)上,在线发表了题为"Structure insights into selective coupling of G protein subtypes by a class B Gprotein-coupled receptor"的研究成果.科研人员在揭示B类GPCRs一个亚家族的两个受体激素识别、受体激活和G蛋白偶联的特异性机制基础上,解析了CRF2R分别与Go蛋白和G11蛋白的复合物三维结构,即B类GPCRs偶联Go和G11的首个复合物结构,系统性阐述了促肾上腺皮质激素释放因子受体偶联Gs、G11和Go蛋白的分子作用机制,为B类GPCRs的G蛋白选择性偶联激活机制奠定了研究基础.
基金supported by the National Natural Science Foundation of China(32270603,31925021,82130022,92357305,82341226,32271232)the National Key Research and Development Program of China(2021YFF1201100,2018YFA0800701)the Fundamental Research Funds for the Central Universities(BMU2021YJ057)。
文摘Adipogenesis is the healthy expansion of white adipose tissue(WAT),serving as a compensatory response to maintain metabolic homeostasis in the presence of excess energy in the body.Therefore,the identification of novel regulatory molecules in adipogenesis,specifically membrane receptors such as G protein-coupled receptors(GPCRs),holds significant clinical promise.These receptors can serve as viable targets for pharmaceuticals,offering potential for restoring metabolic homeostasis in individuals with obesity.We utilized trajectory inference methods to analyze three distinct single-nucleus sequencing(sNuc-seq)datasets of adipose tissue and systematically identified GPCRs with the potential to regulate adipogenesis.Through verification in primary adipose progenitor cells(APCs)of mice,we discovered that ADGRD1 promoted the differentiation of APCs,while GPR39 inhibits this process.In the obese mouse model induced by a high-fat diet(HFD),both gain-of-function and loss-of-function studies validated that ADGRD1 promoted adipogenesis,thereby improving metabolic homeostasis,while GPR39 inhibited adipogenesis,leading to metabolic dysfunction.Additionally,through the analysis of 2,400 ChIP-seq data and 1,204 bulk RNA-seq data,we found that the transcription factors(TFs)MEF2D and TCF12 regulated the expression of ADGRD1 and GPR39,respectively.Our study revealed the regulatory role of GPCRs in adipogenesis,providing novel targets for clinical intervention of metabolic dysfunction in obese patients.
基金This work is supported by Beijing Municipal Science&Technology Commission(Z201100005320012 to N.H.)and Tsinghua University.
文摘G-protein coupled receptors(GPCRs)are the largest family of druggable targets.In recent years,GPCR structural biology has made great advances,revealing the three-dimensional structures of many GPCRs and their interactions with ligands,proteins,and membrane components,which also have inspired a surge of structure-based drug discovery campaigns.This article provides a comprehensive summary of the currently available structural insights into the allosteric pockets of GPCRs and their regulatory mechanisms governing GPCR conformational changes.Furthermore,this article also presents several structure-inspired studies that utilize both orthosteric and allosteric modulation to discover small molecular modulators targeting GPCRs.The article emphasizes the promising potential of drug discovery targeting GPCR allosteric sites,while acknowledging the challenges arising from the limited structural information regarding the lipids and cholesterols in the membrane.Finally,the article discusses the future prospects of using large-scale or focused compound libraries to discover novel chemotypes,as well as the application of artificial intelligence(AI)in structure-based virtual screening(SBVS)against GPCRs.
基金partially supported by the National Institutes of Health(NIH)grants DA056213(to Alexander K.Zinsmaier),DA053388(to Eric J.Nestler),DA023206(to Yan Dong),DA060868(to Yan Dong),and DA040620(to Eric J.Nestler and Yan Dong)。
文摘Understanding the cellular mechanisms of drug addiction remains a key task in current brain research.While neuron-based mechanisms have been extensively explored over the past three decades,recent evidence indicates a critical involvement of astrocytes,the main type of non-neuronal cells in the brain.In response to extracellular stimuli,astrocytes modulate the activity of neurons,synaptic transmission,and neural network properties,collectively influencing brain function.G protein-coupled receptors(GPCRs)expressed on astrocyte surfaces respond to neuron-and environment-derived ligands by activating or inhibiting astrocytic signaling,which in turn regulates adjacent neurons and their circuitry.In this review,we focus on the dopamine D1 receptors(D1R)and metabotropic glutamate receptor 5(mGLUR5 or GRM5)—two GPCRs that have been critically implicated in the acquisition and maintenance of addiction-related behaviors.Positioned as an introductory-level review,this article briefly discusses astrocyte biology,outlines earlier discoveries about the role of astrocytes in substance-use disorders(SUDs),and provides detailed discussion about astrocytic D1Rs and mGLUR5s in regulating synapse and network functions in the nucleus accumbens(NAc)—a brain region that mediates addictionrelated emotional and motivational responses.This review serves as a stepping stone for readers of Engineering to explore links between astrocytic GPCRs and drug addiction and other psychiatric disorders.
基金This work was supported by NIH grants GM077561,EY011500,and GM109955(VVG)and DAAD RISE worldwide summer scholarship and Eberhard Karls Universitat Tubingen(Germany)Master’s program(HS).
文摘Activating and inactivating mutations in numerous human G protein-coupled receptors(GPCRs)are associated with a wide range of disease phenotypes.Here we use several class A GPCRs with a particularly large set of identified disease-associated mutations,many of which were biochemically characterized,along with known GPCR structures and current models of GPCR activation,to understand the molecular mechanisms yielding pathological phenotypes.Based on this mechanistic understanding we also propose different therapeutic approaches,both conventional,using small molecule ligands,and novel,involving gene therapy.
基金funded by grants from Fondo Nacional de Desarrollo Científico y Tecnológico,FONDECYT 1200499 to EM,11200592 to MJY,1211329 to ACby the ANID PIA ACT172066 to EM and AC+3 种基金by the ANID postdoctoral fellowship 3210630 to MPHCby the ANID doctoral fellowship 21230122 to DPNby the ANID doctoral fellowship 21211189 to PRby the ANID doctoral fellowship by the ANID doctoral fellowship 21210611 to FDC。
文摘The brain is,after the adipose tissue,the organ with the greatest amount of lipids and diversity in their composition in the human body.In neurons,lipids are involved in signaling pathways controlling autophagy,a lysosome-dependent catabolic process essential for the maintenance of neuronal homeostasis and the function of the primary cilium,a cellular antenna that acts as a communication hub that transfers extracellular signals into intracellular responses required for neurogenesis and brain development.A crosstalk between primary cilia and autophagy has been established;however,its role in the control of neuronal activity and homeostasis is barely known.In this review,we briefly discuss the current knowledge regarding the role of autophagy and the primary cilium in neurons.Then we review the recent literature about specific lipid subclasses in the regulation of autophagy,in the control of primary cilium structure and its dependent cellular signaling in physiological and pathological conditions,specifically focusing on neurons,an area of research that could have major implications in neurodevelopment,energy homeostasis,and neurodegeneration.
基金This work was funded by grants from the National Key Research and Development Program of China(2019YFA0801402)the National Natural Science Foundation of China(82271890)+3 种基金the Shanghai Key Clinical Specialty Project(shslczdzk05705)the Shanghai Top Priority Key Discipline Project(2017zz02019)Innovative Research Team of High-Level Local Universities in Shanghai(SHSMU-ZDCX20212200)the Macao Science and Technology Development fund(FDCT)(0092/2022/A2 and 003/2022/ALC).
文摘G-protein-coupled receptors(GPCRs)are the largest family of transmembrane receptors and regulate various physiological and pathological processes.Despite extensive studies,the roles of GPCRs in mouse embryonic stem cells(mESCs)remain poorly understood.Here,we show that GPR160,a class A member of GPCRs,is dramatically downregulated concurrent with mESC differentiation into embryoid bodies in vitro.Knockdown of Gpr160 leads to downregulation of the expression of pluripotency-associated transcription factors and upregulation of the expression of lineage markers,accompanying with the ar-rest of the mESC cell-cycle in the G0/G1 phase.RNA-seq analysis shows that GPR160 participates in the JAK/STAT signaling pathway crucial formaintaining ESC stemness,and the knockdown of Gpr160 results in the downregulation of STAT3 phosphorylation level,which in turn is partially rescued by colivelin,a STAT3 activator.Consistent with these observations,GPR160 physically interacts with JAK1,and co-operates with leukemia inhibitory factor receptor(LIFR)and gp130 to activate the STAT3 pathway.In summary,our results suggest that GPR160 regulates mESC self-renewal and pluripotency by interacting with the JAK1-LIFR-gp130 complex to mediate the JAK1/STAT3 signaling pathway.
基金supported by the grant International Cooperation Project of Prevention and Treatment of Major Diseases with Chinese Medicine(GZYYGJ2021047)the High-end Experts Support Program from the Ministry of Science and Technology(DL 2021110001L)the Basic Research Funds from the Ministry of Education(1000061223731).
文摘Objectives:To investigate whether the protective actions of ginsenoside Rb1(Rb1)on astrocytes are mediated through the G_(s)-type G-protein-coupled receptor(GPCR-G_(s)).Methods:Primary astrocyte cultures derived from neonatal mouse brain were used.Astrocyte injury was induced via oxygen-glucose deprivation/re-oxygenation(OGD/R).Cell morphology,viability,lactate dehydrogenase(LDH)leakage,apoptosis,glutamate uptake,and brain-derived neurotrophic factor(BDNF)secretion were assessed to gauge cell survival and functionality.Western blot was used to investigate the cyclic adenosine monophosphate(cAMP)and protein kinase B(Akt)signaling pathways.GPCR-G_(s)-specific inhibitors and molecular docking were used to identify target receptors.Results:Rb1 at concentrations ranging from 0.8 to 5μM did not significantly affect the viability,glutamate uptake,or BDNF secretion in normal astrocytes.OGD/R reduced astrocyte viability,increasing their LDH leakage and apoptosis rate.It also decreased glutamate uptake and BDNF secretion by these cells.Rb1 had protective effects of astrocytes challenged by OGD/R,by improving viability,reducing apoptosis,and enhancing glutamate uptake and BDNF secretion.Additionally,Rb1 activated the cAMP and Akt pathways in these cells.When the GPCR-G_(s) inhibitor NF449 was introduced,the protective effects of Rb1 completely disappeared,and its activation of cAMP and Akt signaling pathways was significantly inhibited.Conclusion:Rb1 protects against astrocytes from OGD/R-induced injury through GPCR-G_(s) mediation.
