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galanin receptor 2 mediates galanin-induced activation of large conductance Ca2+-dependent K+ channels(BK) in human embryonic kidney(HEK293) cells

Galanin receptor 2 mediates galanin-induced activation of la...

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中国神经科学学会第十届全国学术会议

作者： Yun-Fei BAI Na Clara PAN Zhi-Qing David XU Beijing Center of Neural Regeneration & Repair Department of NeurobiologyBeijing Key Laboratory of Brain Major DisordersCapital Medical University

Objective The large conductance Ca-activated K(BK) channels,which are widely distributed in the central nervous system(CNS),have been considered as intracellular calcium sensors in neuron and play an important feedback role in controlling Ca flux and subsequent Ca-dependent processes including neurotransmitter release and cellular *** The effects of neuropeptide galanin on BK channels in human embryonic kidney(HEK293) cells co-expressing both GalR2 and BK alpha subunit were examined by determining the whole-cell currents and single-channel ***(1) The application of galanin enhanced the currents of BK channels,in a PTX-independent and concentration-dependent *** ED50 value of dose-dependent curve was 71.8+16.9 nM.(2) galanin-induced activation of BK was mediated by GalR2,but not GalR1 nor GalR3 as galanin did not facilitate BK current in the cells co-expressed cDNAs of BK with GalR1 or GalR3.(3) Chelating intracellular Ca by either the slow Ca buffer EGTA or the fast Ca buffer BAPTA in the pipette solution abolished galanin-mediated activation of BK channels,indicating the important role of Ca in galanin-induced BK current.(4) The galanin-induce BK current persisted when the bath solution was replaced by Ca-free solution,suggesting the extracellular Ca in not required to lead activation of BK channels.(5) The galanin-induced BK channels activation mediated by the Ca efflux from the sarcoplasmic reticulum /endoplasmic reticulum(SR/ER) was confirmed by application of thapsigargin,an irreversible inhibitor that emptied Ca pool from SR/ER.(6) Inositol-1,4,5-triphosphate receptor(IP3R) on ER was identified as the mediator responsible for increased intracellular Ca that activated BK *** Activation of GalR2,by its ligand galanin,led to elevation of intracellular Ca level due to Ca efflux through IP3R from ER and open BK channels.

关键词： BK channel Ca2+ efflux galanin receptor 2 IP3R

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galanin and its receptor system promote the repair of injured sciatic nerves in diabetic rats

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Neural Regeneration Research 2016年第9期11卷 1517-1526页

作者： Xiao- feng Xu Dan-dan Zhang Jin-chi Liao Li Xiao Qing Wang Wei Qiu The Third Affiliated Hospital of Sun Yat-sen University Guangzhou Guangdong Province China South China Botanical Garden Chinese Academy of Sciences Guangzhou Guangdong Province China

Various studies have reported that galanin can promote axonal regeneration of dorsal root ganglion neurons in vitro and inhibit neuropathic pain. However, little is known about its effects on diabetic peripheral neuropathy, and in vivo experimental data are lacking. We hypothesized that repeated applications of exogenous galanin over an extended time frame may also repair nerve damage in diabetic peripheral neuropathy, and relieve pain in vivo. We found that neuropathic pain occurred in streptozotocin-induced diabetic rats and was more severe after sciatic nerve pinch injury at 14 and 28 days than in diabetic sham-operated rats. Treatment with exogenous galanin alleviated the neuropathic pain and promoted sciatic nerve regeneration more effectively in diabetic rats than in non-diabetic rats after sciatic nerve pinch injury. This was accompanied by changes in the levels of endogenous galanin, and its receptors galanin receptor 1 and galanin receptor 2 in the dorsal root ganglia and the spinal dorsal horn when compared with nerve pinch normal rats. Our results show that application of exogenous galanin daily for 28 days can promote the regeneration of injured sciatic nerves, and alleviate neuropathic pain in diabetic rats.

关键词： nerve regeneration peripheral nerve injury diabetes sciatic nerve galanin galanin receptor 1 galanin receptor 2 neuropathicpain dorsal root ganglion spinal dorsal horn neural regeneration

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Ubiquitination regulates Ga1R2 trafficking and degradation

Ubiquitination regulates Ga1R2 trafficking and degradation

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中国神经科学学会第十届全国学术会议

作者： Xia-Ni GONG Yu-Tao YANG Zhi-Qing David XU Beijing Center of Neural Regeneration ＆ Repair Department of NeurobiologyBeijing Key Laboratory of Brain Major DisordersCapital Medical University

Objective Trafficking and degradation are important for regulation of G protein-coupled receptors（GPCRs）sensitization as well as temporal and spatial aspects of ***,one of posttranslational modifications,is known to regulate GPCRs trafficking and *** present study is to investigate the role of galanin receptor2（GalR2） ubiquitinaition and identify the critical *** The ubiquitination of GalR2 was identified by immunoprecipitation *** accumulation of GalR2 was measured with Western blot *** internalization of GalR2 was observed by antibody-feeding assay together with confocal microscope,and analyzed with ***（1） GalR2 was constitutively ubiquitinated.（2） Ubiquitinated GalR2 accumulated when proteasome was inhibited.（3）GalR2 lysin mutantion significantly enhanced constitutive internalization of the *** Ubiquitination regulates GalR2 cell surface retention and proteasome-related degradation.

关键词： galanin receptor 2 G protein coupled receptor trafficking ubiquitination

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