Mechanism of ATP release from cultured marginal cells of stria vascularis in neonatal rat
10.3760/cma.j.issn.1673-0860.2012.06.008
- VernacularTitle:新生大鼠耳蜗血管纹缘细胞释放ATP的机制
- Author:
Ya-Ting PENG
1
;
Jun YANG
Author Information
1. 上海交通大学医学院附属新华医院
- Keywords:
Adenosine triphosphate;
Stria vascularis;
Ion channels;
Intracellular signaling peptides and proteins
- From:
Chinese Journal of Otorhinolaryngology Head and Neck Surgery
2012;47(6):471-475
- CountryChina
- Language:Chinese
-
Abstract:
Objective To further confirm release of adenosine triphosphate (ATP) from cultured marginal cells in vitro of stria vascularis in neonatal rat,and to explore the mechanism of ATP release from marginal cells.Methods Isolation and in vitro culture of marginal cells of neonatal rats' cochlea.ATP released by marginal cells in extracellular fluid were detected using bioluminescence assay when add regants separately as follow:bafilomycin At,didecyl adipate (DDA),extracellular K+,thapsigargin,extracellular Ca2+,U73122,and aristolochic acid.Results The concentrations of ATP in the extracellular fluid significantly and gradually decreased along with increasing concentrations of bafilornycin A1. The concentrations of ATP in the extracellular fluid were in linear increased with DDA was added to marginal cell suspensions.ATP concentrations increased as the concentration of extracellular K+ was increased,and reached the peak with a K+ concentration of 9.15 mmol/L. At higher K+ concentrations, ATP concentrations decreased. With the addition of increasing concentrations of thapsigargin to test marginal cells,ATP concentrations were significantly deereased.When extracellular Ca2 + was completely chelated,marginal cells continued to release ATP. Moreover,as extracellular Ca2 + increased,the release of ATP decreased.However,the amount of ATP releas remained to a baseline when extracellular concentration of Ca2+ reached 1.25 mmol/L or above.When concentrations of U73122 remained within the range of 0.25 to 1.25 μmol/L,as U73122 increased,the release of ATP decreased.When concentrations of aristolochic acid ranging from 12.5 to 100.0 μmol/L were added to the marginal cells suspension,the release of ATP was significantly decreased.However,with concentrations of aristolochic acid less than 100.0 μ mol/L,the release of ATP tended to be not significantly different from the amount of ATP released by control group.Conclusions ATP could be release from marginal cells cultured in vitro of vascular stria in neonatal rats.ATP release from marginal cells has relevant with calcium pump,K + channel state and signaling pathway related enzymes.
