Optical investigations reveal the effects of 2-aminoethyldiphenyl borate on STIM1 puncta formation
2-Aminoethyldiphenyl borate (2-APB) is the most commonly used pharmacological agent in the study of calcium release-activated channels (CRACs); however, its inhibitory mechanism to CRACs remains unclear. To address this issue, we systematically employed confocal imaging, dual-wavelength excitation p...
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doaj-8f59dbf248be4f1089efcc352f144ba52020-11-25T01:35:16ZengWorld Scientific PublishingJournal of Innovative Optical Health Sciences1793-54581793-72052018-03-011121850003-11850003-1410.1142/S179354581850003710.1142/S1793545818500037Optical investigations reveal the effects of 2-aminoethyldiphenyl borate on STIM1 puncta formationTao Yu0Shangbin Chen1Jingying Pan2Conglin Su3Jun He4Wuhan Children’s Hospital (Wuhan Maternal and Child Healthcare Hospital), Tongji Medical College, Huazhong University of Science and Technology, Wuhan, P. R. ChinaBritton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics — Huazhong University of Science and Technology, Wuhan, P. R. ChinaDivision of Histology and Embryology, School of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, P. R. ChinaDivision of Histology and Embryology, School of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, P. R. ChinaDivision of Histology and Embryology, School of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, P. R. China2-Aminoethyldiphenyl borate (2-APB) is the most commonly used pharmacological agent in the study of calcium release-activated channels (CRACs); 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 Orai1, two key components of CRACs. Imaging results support that there are two signaling pathways (Orai1-independent and Orai1-dependent) for the formation of STIM1 puncta. 2-APB could dose dependently block Orai1-independent but not Orai1-dependent STIM1 puncta formation, despite its obvious inhibition effect on store-operated Ca2+ 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 Ca2+ entry and promote the interaction between Orai1 and CAD by FRET measurements. Therefore, we proposed that inhibitory action of 2-APB on SOCE might attribute to its direct inhibitory effects on Orai1 channel itself, but not the interference on puncta formation between STIM1 and Orai1.http://www.worldscientific.com/doi/pdf/10.1142/S1793545818500037SOCECRAC2-aminoethyldiphenyl borateSTIM1Orai1confocal imagingFRET |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Tao Yu Shangbin Chen Jingying Pan Conglin Su Jun He |
spellingShingle |
Tao Yu Shangbin Chen Jingying Pan Conglin Su Jun He Optical investigations reveal the effects of 2-aminoethyldiphenyl borate on STIM1 puncta formation Journal of Innovative Optical Health Sciences SOCE CRAC 2-aminoethyldiphenyl borate STIM1 Orai1 confocal imaging FRET |
author_facet |
Tao Yu Shangbin Chen Jingying Pan Conglin Su Jun He |
author_sort |
Tao Yu |
title |
Optical investigations reveal the effects of 2-aminoethyldiphenyl borate on STIM1 puncta formation |
title_short |
Optical investigations reveal the effects of 2-aminoethyldiphenyl borate on STIM1 puncta formation |
title_full |
Optical investigations reveal the effects of 2-aminoethyldiphenyl borate on STIM1 puncta formation |
title_fullStr |
Optical investigations reveal the effects of 2-aminoethyldiphenyl borate on STIM1 puncta formation |
title_full_unstemmed |
Optical investigations reveal the effects of 2-aminoethyldiphenyl borate on STIM1 puncta formation |
title_sort |
optical investigations reveal the effects of 2-aminoethyldiphenyl borate on stim1 puncta formation |
publisher |
World Scientific Publishing |
series |
Journal of Innovative Optical Health Sciences |
issn |
1793-5458 1793-7205 |
publishDate |
2018-03-01 |
description |
2-Aminoethyldiphenyl borate (2-APB) is the most commonly used pharmacological agent in the study of calcium release-activated channels (CRACs); 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 Orai1, two key components of CRACs. Imaging results support that there are two signaling pathways (Orai1-independent and Orai1-dependent) for the formation of STIM1 puncta. 2-APB could dose dependently block Orai1-independent but not Orai1-dependent STIM1 puncta formation, despite its obvious inhibition effect on store-operated Ca2+ 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 Ca2+ entry and promote the interaction between Orai1 and CAD by FRET measurements. Therefore, we proposed that inhibitory action of 2-APB on SOCE might attribute to its direct inhibitory effects on Orai1 channel itself, but not the interference on puncta formation between STIM1 and Orai1. |
topic |
SOCE CRAC 2-aminoethyldiphenyl borate STIM1 Orai1 confocal imaging FRET |
url |
http://www.worldscientific.com/doi/pdf/10.1142/S1793545818500037 |
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