Pae S S, Saunders J C
Department of Otorhinolaryngology, Head and Neck Surgery, University of Pennsylvania, Philadelphia 19104.
Proc Natl Acad Sci U S A. 1994 Feb 1;91(3):1153-7. doi: 10.1073/pnas.91.3.1153.
Segments of the chick basilar papilla were isolated and maintained in culture medium. The sensory hair bundle of individual hair cells was observed with light microscopy and stimulated with a water microjet at 600 Hz. Hair bundle motion was slowed by illuminating the microscope with stroboscopic light, and water jet intensity was systematically varied in decibel (dB) steps until a visual detection level (VDL) threshold of hair bundle motion was achieved. The VDL threshold of many hair cells was measured in each isolated papilla. However, only one of eight extracellular calcium concentrations (0.0, 0.0001, 0.001, 0.01, 0.1, 1.25, 6.0, and 12.0 mM) was used with each papilla. In a second series, a calcium ionophore (ionomycin) was added to the culture medium, and VDL thresholds were again measured at seven of these extracellular calcium concentrations. With extracellular calcium alone, the stimulus level needed to achieve threshold was reduced by 2.73 dB between 0.1 and 0.01 mM. This change in threshold represented a 1.37-fold decrease in hair bundle stiffness. When ionomycin was added to the culture medium, a progressively greater stimulus intensity was needed to achieve threshold as calcium concentration increased. The 11.7-dB increase in threshold, with the addition of ionomycin, between 0.0001 and 6.0 mM extracellular calcium was equivalent to a 3.85-fold increase in bundle stiffness. These large changes in hair-bundle stiffness, as a function of the extra- or intracellular calcium environment, may play an important role in the micromechanical behavior of the hair cell during sound simulation.
分离出鸡基底乳头的片段并在培养基中培养。用光学显微镜观察单个毛细胞的感觉毛束,并用频率为600Hz的水微射流进行刺激。用频闪灯照射显微镜可减慢毛束运动,水射流强度以分贝(dB)步长系统地变化,直到达到毛束运动的视觉检测水平(VDL)阈值。在每个分离的乳头中测量许多毛细胞的VDL阈值。然而,每个乳头仅使用八种细胞外钙浓度(0.0、0.0001、0.001、0.01、0.1、1.25、6.0和12.0mM)中的一种。在第二个系列中,向培养基中添加钙离子载体(离子霉素),并再次在这些细胞外钙浓度中的七种浓度下测量VDL阈值。仅使用细胞外钙时,在0.1和0.01mM之间达到阈值所需的刺激水平降低了2.73dB。阈值的这种变化表示毛束刚度降低了1.37倍。当向培养基中添加离子霉素时,随着钙浓度的增加,达到阈值需要逐渐增加的刺激强度。在细胞外钙浓度为0.0001至6.0mM之间添加离子霉素后,阈值增加11.7dB相当于束刚度增加3.85倍。作为细胞外或细胞内钙环境的函数,毛束刚度的这些大变化可能在声音模拟过程中毛细胞的微机械行为中起重要作用。