The shaker rat is an X-linked recessive spontaneous model of progressive Purkinje cell (PC) degeneration exhibiting a shaking ataxia and wide stance. Generation of Wistar Furth (WF) / Brown Norwegian (BN) F1 hybrids and genetic mapping of F2 sib-sib offspring using polymorphic markers narrowed the candidate gene region to 26Mbp denoted by the last recombinant genetic marker DXRat21 at 133 Mbp to qter. In contrast to complete penetrance, stereotypic phenotype and a narrow window for age of onset in the WF background, the F2 hybrid phenotype was more varied and included later age of onset and likely non-penetrance of the mutation. By deep RNA-sequencing, 5 variants were found in the candidate region, 4 were novel without known annotation. One of the variants caused an arginine (R) to cysteine (C) change at codon 35 of the ATPase, Ca2+ transporting, plasma membrane 3 (Atp2b3) gene encoding PMCA3 that has high expression in the cerebellum. The variant was well supported by hundreds of overlapping reads, found in 100% of all affected replicas; and 0% of the wild-type (WT) replicas. The mutation segregated with disease in all affected animals and the amino acid change was found in an evolutionarily conserved region of PMCA3. Despite strong genetic evidence for pathogenicity, in vitro analyses of PMCA3R35C function did not show any differences to WT PMCA3. As Atp2b3 mutation leads to congenital ataxia in humans, the identified Atp2b3 missense change in the shaker rat presents a good candidate for the shaker rat phenotype based on genetic criteria, but cannot yet be considered a definite pathogenic variant owing to lack of functional changes.
- Received August 17, 2015.
- Accepted February 29, 2016.
- © 2016. Published by The Company of Biologists Ltd
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