Chicken DT40 cell line lacking DJ-1, the gene responsible for familial Parkinson's disease, displays mitochondrial dysfunction

• Published in: Neuroscience research Vol. 77; no. 4; pp. 228 - 233
• Main Authors: Minakawa, Eiko N., Yamakado, Hodaka, Tanaka, Atsushi, Uemura, Kengo, Takeda, Shunichi, Takahashi, Ryosuke
• Format: Journal Article
• Language: English
• Published: Ireland Elsevier Ireland Ltd 01-12-2013
• Subjects: Animals; Avian Proteins - genetics; Cell Line; Chickens; DJ-1; DT40; Gene Knockout Techniques; Mitochondria; Mitochondria - metabolism; Mitochondria - pathology; Mitochondria - physiology; Oncogene Proteins - genetics; Oxidative stress; Oxidative Stress - genetics; Parkinson's disease; Parkinsonian Disorders - genetics; Parkinsonian Disorders - metabolism; Parkinsonian Disorders - physiopathology; Oxidative stress; Parkinson's disease; DJ-1; TMRE; H2O2; DT40; MMP; CCCP; Mitochondria; PD; AV; ROS; PI; carboxy-H2DCFDA; HO; hydrogen peroxide; mitochondrial membrane potential; carbonyl cyanide m-chlorophenylhydrazone; tetramethylrhodamine ethyl ester; propium iodide; 5-(and-6)-carboxy-2′,7′-dichloro-dihydrofluorescein diacetate; Annexin-V; reactive oxygen species; carboxy-HDCFDA
• ISSN: 0168-0102; 1872-8111
• DOI: 10.1016/j.neures.2013.09.006

•A novel model of Parkinson's disease was created using DT40, a chicken B cell line.•Targeted gene disruption of DJ-1 in DT40 was utilized to generate this model.F•These cells exhibited phenotypes compatible with DJ-1-deficient mammalian cells.•Gene-modified DT40 cells would be a relevant model of Parkinson's disease.
Parkinson's disease (PD) is the most common neurodegenerative movement disorder mainly due to gradual loss of dopaminergic neurons in the substantia nigra. Although the causative genes for autosomal recessive PD, Parkin, PINK1 and DJ-1, share a common pathway, at least in part, in mitochondrial quality control and protein quality control, their precise relationship remains elusive. Previous studies suggested the limitation of gene-modified mice model to solve this problem. DT40 is an avian leukosis virus-induced chicken B cell line with an exceptionally high ratio of targeted to random DNA integration, which enables efficient targeted disruption of multiple genes of interest. We generated DJ-1-deficient DT40 cells and analyzed PD-related phenotypes. These cells exhibited vulnerability to oxidative stress, mitochondrial dysfunction and fragmentation. Importantly, we showed that mitochondrial membrane potential and morphology are available for the phenotype analysis in DT40. These results suggest that genetically engineered DT40 cells would serve as a relevant model of PD, and help understand the genetic and functional relationship among multiple causative genes. Furthermore, in line with the recent concept of PD as a systemic disorder, elucidating the pathomechanism of PD using DT40 would lead to the development of noninvasive diagnostic tools and drug screening assays using patient-derived lymphocytes.