A mouse model of autosomal dominant spastic ataxia and myopathy caused by a mutation in Tuba4a

ABSTRACT Hereditary ataxias are a heterogeneous group of neurodegenerative disorders characterized by impaired balance and coordination, often due to cerebellar dysfunction. Despite advances in identifying genetic causes, animal models remain essential for dissecting underlying mechanisms and testing therapeutic strategies. Here we describe a mouse model of spastic ataxia and myopathy caused by a missense mutation in Tuba4a (n.A626C, p.Gln176Pro). In an ENU mutagenesis screen, a male C57BL/6J mouse exhibiting muscle wasting and an intention tremor starting at approximately 4 weeks-of-age was identified. The male was bred by in vitro fertilization to BALB/cByJ oocyte donors. Genetic mapping determined dominant inheritance and localized the mutation to Chromosome 1. Genome sequencing revealed single nucleotide polymorphisms (SNPs) in serine threonine kinase 36 (Stk36Y1003N) and alpha-tubulin 4A (Tuba4aQ176P) in the mapping interval. These SNPs were CRISPR-engineered into C57BL/6J mice, which confirmed the Tuba4aQ176P variant as the causative mutation. Mutant mice are normal at 3 weeks, except for decrement in muscle response following repetitive nerve stimulation. However, by 30 days these mice have ataxia, Purkinje neuron degeneration, and extensive skeletal muscle defects, which contribute to a decreased lifespan. Dominant TUBA4A mutations in humans are associated with spastic ataxia type 11 (SPAX11), congenital myopathy type 26 (CMYO26), and frontotemporal dementia/amyotrophic lateral sclerosis type 9 (FTDALS9). Our mice exhibit hallmark features of SPAX11 and CMYO26, but do not show motor neuron degeneration. This specificity makes this model a valuable tool for studying cell-type selective effects of TUBA4A mutations in neurodegeneration and myopathy. Competing Interest Statement The authors have declared no competing interest. Abbreviations - ENU - N-ethyl-N-nitrosourea - SNP - single nucleotide polymorphism - CMYO - congenital myopathy - SPAX - spastic ataxia - FTD - frontotemporal dementia - ALS - amyotrophic lateral sclerosis - CRISPR - clustered regularly interspaced short palindromic repeats - Bp - base pairs - WES - whole exome sequencing - WGS - whole genome sequencing - EMG - electromyography - CMAP - compound muscle action potential - NCV - nerve conduction velocity - RNS - repetitive nerve stimulation - WT - wild-type - NMJ - neuromuscular junction - CV/LFB - cresyl violet luxol fast blue - PN - Purkinje neuron - TEM - transmission electron microscop - IVF - in vitro fertilization - MRI - magnetic resonance imaging - PD - Parkinson’s disease - PCR - polymerase chain reaction - DNA - deoxyribonucleic acid - PVDF - polyvinylidene difluoride - PBS-T - phosphate buffered saline with Tween 20 - HRP - horseradish peroxidase - NBF - neutral buffered formaline - PFA - paraformaldehyde - BSA - bovine serum albumin