Slepecky N, Ulfendahl M
Department of Physiology II, Karolinska Institute, Stockholm, Sweden.
J Submicrosc Cytol Pathol. 1988 Jan;20(1):37-45.
Individual isolated outer hair cells (OHCs) from the cochlea were maintained in a collagen gel and viewed in the light microscope. They were observed during fixation and processing for transmission electron microscopy and individual cells were selected for observation in the electron microscope. Application of glutaraldehyde at several concentrations caused OHCs to become shorter. Shrinkage occurred during dehydration but there was no further change during infiltration with the epoxy resin. Ultrastructural analysis of isolated cells fixed with glutaraldehyde and postfixed with osmium tetroxide showed that these cells were similar to cells fixed in the intact cochlea. The glutaraldehyde-induced cell shape change is similar to the shortening seen in intact OHCs in response to the application of solutions containing high potassium or caffeine. Application of glutaraldehyde to cells pretreated with potassium or caffeine caused further shortening. Glutaraldehyde-induced cell shape change was not blocked by the application of tetracaine, which did prevent potassium-induced and caffeine-induced shortening. Glutaraldehyde-induced cell shape change was not stopped by short treatment with N-ethylmaleimide, which did inhibit potassium-induced shortening. Results from these experiments suggest that the glutaraldehyde-induced OHC shape change is not caused by an effect on the membrane or by calcium activation of a contractile response. Shortening may be caused by shrinkage due to cross-linking of proteins.
从耳蜗分离出的单个外毛细胞(OHCs)被置于胶原凝胶中,并在光学显微镜下观察。在固定和处理以进行透射电子显微镜观察的过程中对它们进行观察,并挑选单个细胞在电子显微镜下观察。应用几种浓度的戊二醛会使OHCs变短。脱水过程中会发生收缩,但在用环氧树脂渗透时没有进一步变化。对用戊二醛固定并用四氧化锇后固定的分离细胞进行超微结构分析表明,这些细胞与在完整耳蜗中固定的细胞相似。戊二醛诱导的细胞形状变化类似于完整OHCs在应用含高钾或咖啡因的溶液时所见到的缩短。将戊二醛应用于用钾或咖啡因预处理过的细胞会导致进一步缩短。戊二醛诱导的细胞形状变化不会被应用丁卡因所阻断,而丁卡因确实能防止钾诱导和咖啡因诱导的缩短。戊二醛诱导的细胞形状变化不会因用N - 乙基马来酰亚胺短时间处理而停止,而N - 乙基马来酰亚胺确实能抑制钾诱导的缩短。这些实验结果表明,戊二醛诱导的OHC形状变化不是由对膜的影响或收缩反应的钙激活引起的。缩短可能是由于蛋白质交联导致的收缩引起的。