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从人嵴帽分离的前庭毛细胞的电生理特性。

Electrophysiological properties of vestibular hair cells isolated from human crista.

作者信息

Mohamed Nesrien, Al-Amin Mohammad, Meredith Frances L, Kalmanson Olivia, Dondzillo Anna, Cass Stephen, Gubbels Samuel, Rennie Katherine J

机构信息

Department of Otolaryngology-Head & Neck Surgery, University of Colorado School of Medicine, Aurora, CO, United States.

出版信息

Front Neurol. 2025 Jan 22;15:1501914. doi: 10.3389/fneur.2024.1501914. eCollection 2024.

Abstract

INTRODUCTION

The vast majority of cellular studies on mammalian vestibular hair cells have been carried out in rodent models due in part to the inaccessibility of human inner ear organs and reports describing electrophysiological recordings from human inner ear sensory hair cells are scarce. Here, we obtained freshly harvested vestibular neuroepithelia from adult translabyrinthine surgical patients to obtain electrophysiological recordings from human hair cells.

METHODS

Whole cell patch clamp recordings were performed on hair cells mechanically isolated from human cristae to characterize voltage-dependent and pharmacological properties of membrane currents. Hair cells were classified as type I or type II according to morphological characteristics and/or their electrophysiological properties.

RESULTS

Type I hair cells exhibited low voltage-activated K currents (IKLV) at membrane potentials around the mean resting membrane potential (-63 mV) and large slowly activating outward K currents in response to depolarizing voltage steps. Recordings from type II hair cells revealed delayed rectifier type outward K currents that activated above the average resting potential of -55 mV and often showed some inactivation at more depolarized potentials. Perfusion with the K channel blocker 4-aminopyridine (1 mM) substantially reduced outward current in both hair cell types. Additionally, extracellular application of 8-bromo-cGMP inhibited IKLV in human crista type I hair cells suggesting modulation via a nitric oxide/cGMP mechanism. A slow hyperpolarization-activated current (Ih) was observed in some hair cells in response to membrane hyperpolarization below -100 mV.

DISCUSSION

In summary, whole cell recordings from isolated human hair cells revealed ionic currents that strongly resemble mature current phenotypes previously described in hair cells from rodent vestibular epithelia. Rapid access to surgically obtained adult human vestibular neuroepithelia allows translational studies crucial for improved understanding of human peripheral vestibular function.

摘要

引言

由于人类内耳器官难以获取,大部分关于哺乳动物前庭毛细胞的细胞研究都是在啮齿动物模型中进行的,而且描述人类内耳感觉毛细胞电生理记录的报告也很稀少。在这里,我们从成年经迷路手术患者身上获取了新鲜收获的前庭神经上皮,以记录人类毛细胞的电生理活动。

方法

对从人嵴帽机械分离的毛细胞进行全细胞膜片钳记录,以表征膜电流的电压依赖性和药理学特性。根据形态特征和/或电生理特性将毛细胞分为I型或II型。

结果

I型毛细胞在接近平均静息膜电位(-63 mV)的膜电位下表现出低电压激活的钾电流(IKLV),并且在去极化电压阶跃时表现出大的缓慢激活的外向钾电流。II型毛细胞的记录显示,延迟整流型外向钾电流在高于-55 mV的平均静息电位时激活,并且在更去极化的电位下通常表现出一些失活。用钾通道阻滞剂4-氨基吡啶(1 mM)灌注可显著降低两种毛细胞类型的外向电流。此外,细胞外应用8-溴-cGMP可抑制人嵴帽I型毛细胞中的IKLV,提示通过一氧化氮/cGMP机制进行调节。在一些毛细胞中,当膜超极化至-100 mV以下时观察到缓慢的超极化激活电流(Ih)。

讨论

总之,从分离的人类毛细胞进行的全细胞记录揭示了离子电流,这些电流与先前在啮齿动物前庭上皮的毛细胞中描述的成熟电流表型非常相似。快速获取手术获得的成年人类前庭神经上皮允许进行转化研究,这对于更好地理解人类外周前庭功能至关重要。

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