Identification and characterization of the gene associated with the spontaneous autosomal recessive Spinning mice

本实验室发现了一种新的自发性常染色体隐性合并全身性肌张力障碍模型

A new spontaneously autosomal recessive model of combined generalized dystonia was identified in our lab after outbreeding of BALB/cAnNCrl mouse line obtained from Charles River Laboratories with C57BL/6J from The Jackson Laboratory. Phenotypes maintained after backcross breeding to BALB/cAnNCrl and inbreeding with C57BL/6J. Homozygote mice display active movement in a circular direction (Spinning) and dystonia (jerky head movements, head tossing with the neck twisted), but without hearing loss. Candidate genes including Cacng2 (Stargazer mice), Cacna2d2 (Ducky mice), Cacnb4 (Lethargic mice), Cacna1a (Tottering mice), and Myo7a (Tonado rat) were excluded by DNA and RNA Sanger sequencing and QRT-PCR. RNA sequencing (RNA-seq) was performed in 6 samples (3 normal controls and 3 Spinning mice) and a candidate gene list was generated which includes Slc16a14, Col6a5, Lrrc66, Marveld3, Armc3, Tc2n, Zfp979, and other 5 candidate genes. Our central hypothesis is that the circling behavior in the Spinning mice is most likely due to a loss of function mutation of a novel gene which may be related to the dopamine signaling pathway. The major goal of this application is to identify the casual gene of the Spinning mice, and characterize its function and reveal if its human homologue gene are related with dystonia patients. In Aim #1, we will identify the disease gene using RNA-seq and or whole genome sequencing, and genetic mapping strategies. In Aim #2, we will characterize the disease gene by QRT-PCR, Western blot, and immunohistochemistry to identify the expression pattern of the disease gene in brain regions, among normal controls and the Spinning mice; and identify the molecular functional pathway of the disease gene with RNA-seq and other in-silico analysis. We will also screen all exons of the disease gene in at least 2000 of our 3155 dystonia patients to identify if the gene is also associated with human dystonia. Completion of these objectives will improve and expand our understanding of dystonia and movement disorders.