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为研究听毛细胞的钙信号转导 ,观察了钙离子载体 ionomycin引起毛细胞胞内游离钙变化的过程。分离的豚鼠耳蜗外毛细胞经钙敏荧光染料 fluo-3和 fura-red染色后 ,用激光扫描共聚焦显微镜进行检测 ,以 fluo-3 /fura-red荧光比值指示胞内游离钙浓度高低。 5～ 10μmol/L ionomycin作用后 ,外毛细胞胞内游离钙由静态比值 1.69± 0 .5 9增至峰值时的 2 .78± 1.43 (x± s,n=10 ,P<0 .0 5 )。外毛细胞胞内游离钙增高的时程有两种模式 ,一种为胞内游离钙在药物作用后 10～ 3 5 s即有明显增加 (n=6) ,部分呈双相升高 ;另一种在药物作用后经过约 2 0 0～ 3 0 0 s的潜伏期 ,胞内游离钙方出现明显增加 (n=4)。一个外毛细胞在ionomycin作用后有钙波形成。ionomycin作用后外毛细胞胞内游离钙动态变化的多种模式显示了其作用机制和内耳毛细胞钙调控过程的复杂性 ,保证了钙在复杂的听觉外周过程中起重要的调节作用
To study calcium signaling in hair cells, we observed the intracellular free calcium changes induced by ionomycin. The isolated cochlear outer hair cells were stained with calcium-sensitive fluorescent dyes fluo-3 and fura-red and detected by laser scanning confocal microscopy. Fluo-3 / fura-red fluorescence ratio indicated the intracellular free calcium concentration. After 5 ~ 10μmol / L ionomycin, the intracellular free calcium in outer hair cells increased from 1.69 ± 0.59 to 2.78 ± 1.43 (x ± s, n = 10, P <0. 0 5). There are two modes of intracellular free calcium elevation in epidermal cells. One is that intracellular free calcium is significantly increased (n = 6) 10 to 35 s after drug action, and some are in biphasic elevation. After a period of incubation of about 200 ~ 300 s after drug administration, there was a significant increase in intracellular free calcium (n = 4). An outer hair cell forms calcium waves after the action of ionomycin. Various patterns of the intracellular free calcium dynamics of the outer hair cells after ionomycin exposure demonstrate their mechanism of action and the complexity of calcium regulation processes in the inner ear hair cells, thus ensuring that calcium plays an important regulatory role in complex auditory peripheral processes