两种新的gjb1相关的1X型腓骨肌萎缩症小鼠模型

IF 3.9 3区 医学 Q1 CLINICAL NEUROLOGY Journal of the Peripheral Nervous System Pub Date : 2023-08-07 DOI:10.1111/jns.12588
A. L. D. Tadenev, C. L. Hatton, B. Pattavina, T. Mullins, R. Schneider, L. P. Bogdanik, Robert W. Burgess
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引用次数: 0

摘要

背景:1X型腓骨肌萎缩症是由GJB1突变引起的,GJB1是与遗传性周围神经病变相关的第二大常见基因。GJB1基因编码连接蛋白32 (CX32),这是一种在髓鞘胶质细胞中表达的间隙连接蛋白。该基因是x连锁的,突变会导致功能丧失。目的大量与疾病相关的变异已被发现,其中许多变异导致蛋白质的错误运输和错误定位。现有缺失Gjb1表达的敲除小鼠提供了一种有效的CMT1X动物模型,但完全缺乏蛋白质可能无法完全概括异常CX32蛋白引起的疾病机制。为了更好地代表人类cmt1x相关突变谱,我们建立了一个新的Gjb1敲入小鼠模型。方法采用CRISPR/Cas9基因组编辑技术,制备携带Gjb1基因R15Q突变的小鼠。此外,我们在密码子7 (del2)上发现了第二个等位基因,其早期帧移位突变。使用临床相关的分子、组织学、神经生理学和行为分析小鼠。结果两种等位基因在雪旺细胞中均产生免疫荧光可检测的蛋白,部分蛋白可定位于Ranvier淋巴结。然而,这两种等位基因也会导致周围神经病变,包括髓鞘稀疏和脱髓鞘轴突,以及轴突退化和再生,主要发生在远端运动神经。神经传导速度仅在老年时轻度降低,复合肌肉动作电位振幅未降低。两种等位基因血浆中神经丝轻链水平均升高。del2小鼠在约3个月大时发病,而R15Q小鼠在5-6个月大时发病,表明功能丧失较轻。这两个等位基因在神经肌肉性能的加速旋转和握力测试中表现与野生型幼崽相当。我们已经建立并描述了两种新的CMT1X小鼠模型,这将对未来的机制和临床前研究有用。
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Two new mouse models of Gjb1-associated Charcot–Marie–Tooth disease type 1X

Background

Charcot–Marie–Tooth disease type 1X is caused by mutations in GJB1, which is the second most common gene associated with inherited peripheral neuropathy. The GJB1 gene encodes connexin 32 (CX32), a gap junction protein expressed in myelinating glial cells. The gene is X-linked, and the mutations cause a loss of function.

Aims

A large number of disease-associated variants have been identified, and many result in mistrafficking and mislocalization of the protein. An existing knockout mouse lacking Gjb1 expression provides a valid animal model of CMT1X, but the complete lack of protein may not fully recapitulate the disease mechanisms caused by aberrant CX32 proteins. To better represent the spectrum of human CMT1X-associated mutations, we have generated a new Gjb1 knockin mouse model.

Methods

CRISPR/Cas9 genome editing was used to produce mice carrying the R15Q mutation in Gjb1. In addition, we identified a second allele with an early frame shift mutation in codon 7 (del2). Mice were analyzed using clinically relevant molecular, histological, neurophysiological, and behavioral assays.

Results

Both alleles produce protein detectable by immunofluorescence in Schwann cells, with some protein properly localizing to nodes of Ranvier. However, both alleles also result in peripheral neuropathy with thinly myelinated and demyelinated axons, as well as degenerating and regenerating axons, predominantly in distal motor nerves. Nerve conduction velocities were only mildly reduced at later ages and compound muscle action potential amplitudes were not reduced. Levels of neurofilament light chain in plasma were elevated in both alleles. The del2 mice have an onset at ~3 months of age, whereas the R15Q mice had a later onset at 5–6 months of age, suggesting a milder loss of function. Both alleles performed comparably to wild type littermates in accelerating rotarod and grip strength tests of neuromuscular performance.

Interpretation

We have generated and characterized two new mouse models of CMT1X that will be useful for future mechanistic and preclinical studies.

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来源期刊
CiteScore
6.10
自引率
7.90%
发文量
45
审稿时长
>12 weeks
期刊介绍: The Journal of the Peripheral Nervous System is the official journal of the Peripheral Nerve Society. Founded in 1996, it is the scientific journal of choice for clinicians, clinical scientists and basic neuroscientists interested in all aspects of biology and clinical research of peripheral nervous system disorders. The Journal of the Peripheral Nervous System is a peer-reviewed journal that publishes high quality articles on cell and molecular biology, genomics, neuropathic pain, clinical research, trials, and unique case reports on inherited and acquired peripheral neuropathies. Original articles are organized according to the topic in one of four specific areas: Mechanisms of Disease, Genetics, Clinical Research, and Clinical Trials. The journal also publishes regular review papers on hot topics and Special Issues on basic, clinical, or assembled research in the field of peripheral nervous system disorders. Authors interested in contributing a review-type article or a Special Issue should contact the Editorial Office to discuss the scope of the proposed article with the Editor-in-Chief.
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