Alternative splicing in Acad8 resulting a mitochondrial defect and progressive hepatic steatosis in mice.
Journal Article (Journal Article)
Using a combination of N-ethyl-N-nitrosourea-mediated mutagenesis and metabolomics-guided screening, we identified mice with elevated blood levels of short-chain C4-acylcarnitine and increased urine isobutyryl-glycine. Genome-wide homozygosity screening, followed by fine mapping, located the disease gene to 15-25 Mb of mouse chromosome 9 where a candidate gene, Acad8, encoding mitochondrial isobutyryl-CoA dehydrogenase was located. Genomic DNA sequencing revealed a single-nucleotide mutation at -17 of the first intron of Acad8 in affected mice. cDNA sequencing revealed an intronic 28-bp insertion at the site of the mutation, which caused a frame shift with a premature stop codon. In vitro splicing assay confirmed that the mutation was sufficient to activate an upstream, aberrant 3' splice site. There was a reduction in the expression of Acad8 at both the mRNA and protein levels. The mutant mice grew normally but demonstrated cold intolerance at young age with a progressive hepatic steatosis. Homozygous mutant mice hepatocytes had abnormal mitochondria with crystalline inclusions, suggestive of mitochondriopathy. This mouse model of isobutyryl-CoA dehydrogenase deficiency could provide us a better understanding of the possible role of IBD deficiency in mitochondriopathy and fatty liver.
Full Text
Duke Authors
Cited Authors
- Sabbagha, NGAA-A; Kao, H-J; Yang, C-F; Huang, C-C; Lin, W-D; Tsai, F-J; Chen, T-H; Tarn, W-Y; Wu, J-Y; Chen, Y-T
Published Date
- July 2011
Published In
Volume / Issue
- 70 / 1
Start / End Page
- 31 - 36
PubMed ID
- 21659959
Electronic International Standard Serial Number (EISSN)
- 1530-0447
Digital Object Identifier (DOI)
- 10.1203/PDR.0b013e31821b89ee
Language
- eng
Conference Location
- United States