Optical investigations reveal the effects of 2-aminoethyldiphenyl borate on STIM1 puncta formation

查看全文 作　　者：Tao [2]Yu;Shangbin [3]Chen;Jingying [1]Pan;Conglin [1]Su;Jun [1]He 机构地区：[1]Division of Histology and Embryology School of Basic Medicine,Tongji Medical College Huazhong University of Science and Technology Wuhan,P.R.China;[2]Wuhan Children's Hospital(Wuhan Maternal and Child Healthcare Hospital)Tongji Medical College,Huazhong University of Science and Technology,Wuhan,P.R.China;[3]Britton Chance Center for Biomedical Photonics Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology,Wuhan,P.R.China 出　　处：《Journal of Innovative Optical Health Sciences》索引2018年第2期,共14页 基　　金：supported by the National Natural Sciences Foundation of China(Grant Nos.31371217 and 30871311). 摘　　要：2-A minoethyldiphenyl borate(2-APB)is the most commonly used pharmacological agent in the study of calcium release-activated channels(CRACa);however,its inhibitory mechanism to CRACs remains unclear.To address this issue,we systematically employed confocal imaging,dual-wavelength excitation photometry and FRET to examine the effects of 2-APB on the dynamic activities and function of STIM1 and Orail,two key components of CRACs.Imaging results support that there are two signaling pathways(Orail-independent and Orail-dependent)for the formation of STM1 puncta.2 APB could dose dependently block Orail-independent but not Oril-dependent STIM1 puncta formation,despite its obvious inhibition effect on store-opented Ca^(2+)entry(SOCE).In addition,we found that although 2-APB could not visibly alter near plasma membrane CAD-eYFP localization,it could completely block CAD-YFP-induced constitutive Ca^(2+)entry and promnote the interaction between Orail and CAD by FRET mea-surements.Therefore,we proposed that inhibitory action of 2-APB on SOCE might attribute to its direct inhbitory effects on Orail channel itself,but not the interference on puncta formation between STIM1 and Orail. 关 键 词：SOCE CRAC 2-arminoethyldiphenyl bornate STIM1 Orail confocal imaging FRET