Alternative splicing in Acad8 resulting a mitochondrial defect and progressive hepatic steatosis in mice
| Autor: | Nagham George Abd Al-Ahad Sabbagha, Fuu Jen Tsai, Hsiao Jung Kao, Jer-Yuarn Wu, Woan-Yuh Tarn, Yuan-Tsong Chen, Chih Fu Yang, Tzu Ho Chen, Cheng Chih Huang, Wei De Lin |
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| Rok vydání: | 2011 |
| Předmět: |
Male
Oxidoreductases Acting on CH-CH Group Donors Mutant DNA Mutational Analysis Molecular Sequence Data Mutagenesis (molecular biology technique) Mitochondria Liver Biology medicine.disease_cause Acyl-CoA Dehydrogenase Gene Expression Regulation Enzymologic Frameshift mutation Mice Microscopy Electron Transmission medicine Animals Metabolomics Genetic Predisposition to Disease PPAR alpha Thermosensing RNA Messenger Amino Acid Metabolism Inborn Errors Liver X Receptors Genetics Mutation Base Sequence Alternative splicing Intron Thermogenesis medicine.disease Orphan Nuclear Receptors Molecular biology Mice Mutant Strains Cold Temperature Fatty Liver Mice Inbred C57BL PPAR gamma Alternative Splicing Disease Models Animal Phenotype Ethylnitrosourea Pediatrics Perinatology and Child Health RNA splicing Disease Progression Steatosis Mitochondrial Swelling Mutagens |
| Zdroj: | Pediatric research. 70(1) |
| ISSN: | 1530-0447 |
| Popis: | Using a combination of N-ethyl-N-nitrosourea- mediated mutagenesis and metabolomics-guided screening, we iden- tified 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 mitochondri- opathy and fatty liver. (Pediatr Res 70: 31-36, 2011) |
| Databáze: | OpenAIRE |
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