Peter M N, Paasche G, Reich U, Lenarz T, Warnecke A
Klinik für Hals‑, Nasen- und Ohrenheilkunde, Medizinische Hochschule Hannover, Carl-Neuberg-Str. 1, 30625, Hannover, Deutschland.
Exzellenz Cluster "Hearing4all", Medizinische Hochschule Hannover, Hannover, Deutschland.
HNO. 2019 Apr;67(4):251-257. doi: 10.1007/s00106-019-0641-7.
Increasing numbers of cochlear implant patients have residual hearing. Despite surgical and pharmacological efforts to preserve residual hearing, a significant number of these patients suffer a late, unexplained loss of residual hearing. Surgical trauma can be excluded as the cause. To investigate this phenomenon and because cells in their native environment react differently to stimuli (such as electrical current) than isolated cells, whole-organ explants from cochleae may be a better model. For early detection of synaptic changes in the organ of Corti, a high-resolution microscopic technique such as stimulated emission depletion (StED) can be used. The aim of this study was establishment of a qualitative and quantitative technique to determinate changes in the organ of Corti and its synapses after electrical stimulation. Explanted organs of Corti from postnatal rats (P2-4) were cultured on a coverslip for 24 h and subsequently exposed to biphasic pulsed electrical stimulation (amplitude 0.44-2.0 mA, pulse width 400 μs, interpulse delay 120 μs, repetition 1 kHz) for another 24 h. For visualization, the cytoskeleton and the ribbon synapses were stained immunocytochemically. For an early detectable response to electrical stimulation, the number of synapses was quantified. Organs of Corti without electrical stimulation served as a reference. Initial research has shown that electrical stimulation can cause changes in ribbon synapses and that StED can detect these alterations. The herein established model could be of great importance for identification of molecular changes in the organ of Corti in response to electrical or other stimuli.
越来越多的人工耳蜗植入患者有残余听力。尽管在手术和药物方面努力保留残余听力,但仍有相当数量的患者后期出现不明原因的残余听力丧失。手术创伤可排除为病因。为了研究这一现象,并且由于细胞在其原生环境中对刺激(如电流)的反应与分离细胞不同,来自耳蜗的全器官外植体可能是一个更好的模型。为了早期检测柯蒂氏器中的突触变化,可以使用高分辨率显微镜技术,如受激发射损耗显微镜(StED)。本研究的目的是建立一种定性和定量技术,以确定电刺激后柯蒂氏器及其突触的变化。将出生后大鼠(P2 - 4)的柯蒂氏器外植体在盖玻片上培养24小时,随后再暴露于双相脉冲电刺激(幅度0.44 - 2.0 mA,脉冲宽度400 μs,脉冲间隔120 μs,重复频率1 kHz)24小时。为了可视化,对细胞骨架和带状突触进行免疫细胞化学染色。为了对电刺激有早期可检测的反应,对突触数量进行定量。未接受电刺激的柯蒂氏器作为对照。初步研究表明,电刺激可导致带状突触发生变化,并且StED可以检测到这些改变。本文建立的模型对于识别柯蒂氏器中响应电刺激或其他刺激的分子变化可能具有重要意义。