| Abstract |
The biological characteristics of the globular substance, a precursor of otoconia, are unclear. In the present study, the ATP-induced internal free Ca2+ concentration ([Ca2+]i) changes of the globular substance and the ATP distribution in the vestibular organ were investigated using a Ca2+ indicator, fluo 3, and an adenine nucleotide-specific fluorochrome, quinacrine, by means of confocal laser scanning microscopy. [Ca2+]i showed a rapid and dose-dependent increase in response to ATP with a 50% effective concentration (EC50) of 16.7 microM. This reaction was independent of external Ca2+, indicating the presence of an internal Ca2+ reservoir. Neither adenosine, alpha, beta-methylene-ATP, 3'-O-(4-benzoylbenzoyl)-ATP, ADP, nor UTP evoked this reaction, whereas 2-methylthio-ATP induced an increase of [Ca2+]i with an EC50 of 14.4 microM. Moreover, P2 antagonists, reactive blue 2 and suramin, and a phospholipase C inhibitor, U-73122, inhibited the ATP-induced [Ca2+]i increase. These findings indicate the presence of a P2Y purinoceptor on the globular substance. In addition, granular fluorescence was observed in the quinacrine-stained macular sensory epithelium, indicating the presence of ATP-containing granules in this tissue. These results suggest that a paracrine mechanism involving ATP may exist in the macula and that this mechanism regulates the biological behavior of the globular substance.
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