The Effect of IP3 Receptor Inhibition Using 2-APB on Gentamicin Ototoxicity in Cochlear Sensory Cell.
- Author:
Jeong Hwan MOON
1
;
Jae Yun JUNG
;
Jin Chul AHN
;
Yeon Sun SEONG
;
Eun Suk LIM
;
Chung Ku RHEE
Author Information
1. Department of Otorhinolaryngology-Head and Neck Surgery, College of Medicine, Dankook University, Cheonan, Korea. jjking@dankook.ac.kr
- Publication Type:In Vitro ; Original Article
- Keywords:
2-APB;
Caspase-3;
Gentamicin;
Calcium;
IP3 receptor
- MeSH:
Apoptosis;
Calcium;
Caspase 3;
Cell Death;
Cell Line;
Cytosol;
Fluorescence;
Gentamicins*;
Hair;
Inositol 1,4,5-Trisphosphate Receptors*;
Microscopy, Confocal
- From:Korean Journal of Otolaryngology - Head and Neck Surgery
2007;50(5):385-390
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
BACKGROUND AND OBJECTIVES: Elevated intracellular calcium level is known to play important roles in the apoptotic pathway. IP3 receptor (ligand-gated channels that release Ca2+ from intracellular stores) is emerging as a key site for regulation of apoptosis. 2-Aminoethoxydiphenyl borate (2-APB) is one of the reliable IP3 receptor antagonists. We examined the effect of 2-APB on gentamicin ototoxicity in vitro, using the HEI-OC1 cell line. MATERIALS AND METHOD: HEI-OC1 cells were trWWeated with 100micrometer gentamicin. Using a CaspACE assay, we measured the caspases-3 activity in the gentamicin treated hair cells with and without 2-APB pre-incubation. We also observed intra-cellular calcium concentrations in HEI-OC1 cells using a confocal microscopy (calcium green-1 stain). Live cell imaging was performed by using fluorescence video-time lapse system. RESULTS: Cytosolic calcium elevation by gentamicin was remarkably inhibited by 2-APB. Caspases-3 activities of gentamicin treated cells were higher than those of the control. After incubation with 2-APB, caspases-3 activities and cell death of gentamicin treated cells were shown to decrease. CONCLUSION: 2-APB reduces Caspases-3 activity in the gentamicin treated HEI-OC1 cells by inhibition of cytosolic calcium increase.
