Article
Induction of ATPs releasing from vascular stria marginal cells of cochlea in neonatal rats by change of ion concentration
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Published: | April 4, 2012 |
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Objectives: To find the evidences of adenosine triphosphate (ATPs) presenting in vascular stria marginal cells of cochlea in neonatal rats by in vitro culture, and the change of ion concentration may induce the ATPs releasing from the vascular stria marginal cells of cochlea in neonatal rats.
Methods: The day 1 to 3 aged Sprague-Dawley rats were used for in vitro culturing, purification and identification of stria vascular cells. The ATP vesicles were observed after stained with quinacrine, which is the specific marker of ATP vesicles, under fluorescence microscope. The concentration of extra-cellular ATPs released from marginal cells was measured by bioluminescence assay, as well as the ATP releasing induced by change of extra-cellular ion concentration of marginal cells.
Results: By identification of the purity of keratin and Vimentin, both are epithelial marker, the in vitro cultured vascular stria marginal cells of cochlea of neonatal rats were confirmed as marginal cells. Large amount of green-colored star-like spots were observed in the plasma of marginal cells under fluorescence microscope after quinacrine staining. The extra-cellular concentration of ATPs released from marginal cells was measured by bioluminescence assay, and the ATP concentration was calculated by value of fluorescence, and it was noticed that the Ca2+ pathway in marginal cells might be activated by some extra-cellular ion concentration changes, which in turn changed the extra-cellular ATP concentration.
Conclusion: ATP vesicles did presenting in the vascular stria marginal cells of cochlea in neonatal rats, and ATPs did secreted by activation of Ca2+ pathway in marginal cells.