Johnson Stuart L, Forge Andrew, Knipper Marlies, Münkner Stefan, Marcotti Walter
Department of Biomedical Science, University of Sheffield, Sheffield, United Kingdom.
J Neurosci. 2008 Jul 23;28(30):7670-8. doi: 10.1523/JNEUROSCI.0785-08.2008.
The mammalian cochlea is specialized to recognize and process complex auditory signals with remarkable acuity and temporal precision over a wide frequency range. The quality of the information relayed to the auditory afferent fibers mainly depends on the transfer characteristics of inner hair cell (IHC) ribbon synapses. To investigate the biophysical properties of the synaptic machinery, we measured changes in membrane capacitance (DeltaC(m)) in low-frequency (apical region, approximately 300 Hz) and high-frequency (basal, approximately 30 kHz) gerbil IHCs maintained in near physiological conditions (1.3 mm extracellular Ca(2+) and body temperature). With maturation, the Ca(2+) efficiency of exocytosis improved in both apical and basal IHCs and was more pronounced in the latter. Prehearing IHCs showed a similar Ca(2+) cooperativity of exocytosis despite the smaller DeltaC(m) in apical cells. After maturation, DeltaC(m) in high-frequency IHCs increased linearly with the Ca(2+) current, whereas, somewhat surprisingly, the relationship was significantly more nonlinear in low-frequency cells. This tonotopic difference seemed to be correlated with ribbon synapse morphology (spherical in apical and ellipsoid in basal IHCs) but not with the expression level of the proposed Ca(2+) sensor otoferlin or the spatial coupling between Ca(2+) channels and active zones. Repetitive stimulation of adult IHCs showed that vesicle pool refilling could become rate limiting for vesicle release, with high-frequency IHCs able to sustain greater release rates. Together, our findings provide the first evidence for a tonotopic difference in the properties of the synaptic machinery in mammalian IHCs, which could be essential for fine-tuning their receptor characteristics during sound stimulation.
哺乳动物的耳蜗专门用于在很宽的频率范围内以极高的敏锐度和时间精度识别和处理复杂的听觉信号。传递给听觉传入纤维的信息质量主要取决于内毛细胞(IHC)带状突触的传递特性。为了研究突触机制的生物物理特性,我们测量了处于接近生理条件(细胞外钙离子浓度1.3 mM,体温)下的低频(顶端区域,约300 Hz)和高频(基部,约30 kHz)沙鼠IHC的膜电容变化(ΔCm)。随着成熟,顶端和基部IHC的胞吐作用的钙离子效率均有所提高,且在后者中更为明显。尽管顶端细胞的ΔCm较小,但听力前的IHC显示出类似的胞吐作用的钙离子协同性。成熟后,高频IHC中的ΔCm随钙离子电流呈线性增加,而令人惊讶的是,低频细胞中的这种关系明显更非线性。这种音调拓扑差异似乎与带状突触形态(顶端为球形,基部为椭圆形)相关,但与所提出的钙离子传感器 otoferlin的表达水平或钙离子通道与活性区之间的空间耦合无关。对成年IHC的重复刺激表明,囊泡池的重新填充可能成为囊泡释放的速率限制因素,高频IHC能够维持更高的释放速率。总之,我们的研究结果首次证明了哺乳动物IHC中突触机制特性存在音调拓扑差异,这对于在声音刺激过程中微调其受体特性可能至关重要。