Effect of Insulin Deprivation on Muscle Mitochondrial ATP Production and Gene Transcript Levels in Type 1 Diabetic Subjects

• Published in: Diabetes (New York, N.Y.) Vol. 56; no. 11; pp. 2683 - 2689
• Main Authors: Karakelides, Helen, Asmann, Yan W., Bigelow, Maureen L., Short, Kevin R., Dhatariya, Ketan, Coenen-Schimke, Jill, Kahl, Jane, Mukhopadhyay, Debabrata, Nair, K. Sreekumaran
• Format: Journal Article
• Language: English
• Published: Alexandria, VA American Diabetes Association 01.11.2007
• Subjects: Adenosine Triphosphate - metabolism; Adult; Amino acids; Biological and medical sciences; Blood Glucose - metabolism; Body Mass Index; Cardiovascular disease; Catheters; Cytochrome; Diabetes; Diabetes Mellitus, Type 1 - blood; Diabetes Mellitus, Type 1 - genetics; Diabetes Mellitus, Type 1 - metabolism; Diabetes. Impaired glucose tolerance; Diabetics; DNA Primers; Dosage and administration; Drug therapy; Endocrine pancreas. Apud cells (diseases); Endocrinopathies; Etiopathogenesis. Screening. Investigations. Target tissue resistance; Evaluation; Fatty Acids, Nonesterified - blood; Genes; Genetic aspects; Genetic transcription; Glucose; Health aspects; Humans; Hypotheses; Insulin; Insulin - deficiency; Insulin resistance; Medical sciences; Mitochondria, Muscle - metabolism; Mitochondrial DNA; Muscle, Skeletal - metabolism; Oligonucleotide Array Sequence Analysis; Oxidative Phosphorylation; Peptides; Phosphorylation; Plasma; Polymerase Chain Reaction; Protons; Research design; Transcription (Genetics); Transcription, Genetic; Type 1 diabetes; Vascular endothelial growth factor; United States; Endocrinopathy; Human; Immunopathology; Pancreatic hormone; Mitochondria; Gene; Type 1 diabetes; Autoimmune disease; Muscle; ATP; Insulin
• ISSN: 0012-1797; 1939-327X; 1939-327X
• DOI: 10.2337/db07-0378

Effect of Insulin Deprivation on Muscle Mitochondrial ATP Production and Gene Transcript Levels in Type 1 Diabetic Subjects Helen Karakelides 1 , Yan W. Asmann 1 , Maureen L. Bigelow 1 , Kevin R. Short 1 , Ketan Dhatariya 1 , Jill Coenen-Schimke 1 , Jane Kahl 1 , Debabrata Mukhopadhyay 2 and K. Sreekumaran Nair 1 1 Division of Endocrinology and Metabolism and Endocrine Research Unit, Mayo Clinic, Rochester, Minnesota 2 Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, Minnesota Address correspondence and reprint requests to K. Sreekumaran Nair, MD, PhD, Mayo Clinic, 200 First St. SW, Joseph 5-194, Rochester, MN 55905. E-mail: nair.sree{at}mayo.edu Abstract OBJECTIVE—Muscle mitochondrial dysfunction occurs in many insulin-resistant states, such as type 2 diabetes, prompting a hypothesis that mitochondrial dysfunction may cause insulin resistance. We determined the impact of insulin deficiency on muscle mitochondrial ATP production by temporarily depriving type 1 diabetic patients of insulin treatment. RESEARCH DESIGN AND METHODS—We withdrew insulin for 8.6 ± 0.6 h in nine C-peptide–negative type 1 diabetic subjects and measured muscle mitochondrial ATP production and gene transcript levels (gene array and real-time quantitative PCR) and compared with insulin-treated state. We also measured oxygen consumption (indirect calorimetry); plasma levels of glucagon, bicarbonate, and other substrates; and urinary nitrogen. RESULTS—Withdrawal of insulin resulted in increased plasma glucose, branched chain amino acids, nonesterified fatty acids, β-hydroxybutyrate, and urinary nitrogen but no change in bicarbonate. Insulin deprivation decreased muscle mitochondrial ATP production rate (MAPR) despite an increase in whole-body oxygen consumption and altered expression of many muscle mitochondrial gene transcripts. Transcript levels of genes involved in oxidative phosphorylation were decreased, whereas those involved in vascular endothelial growth factor (VEGF) signaling, inflammation, cytoskeleton signaling, and integrin signaling pathways were increased. CONCLUSIONS—Insulin deficiency and associated metabolic changes reduce muscle MAPR and expression of oxidative phosphorylation genes in type 1 diabetes despite an increase in whole-body oxygen consumption. Increase in transcript levels of genes involved in VEGF, inflammation, cytoskeleton, and integrin signaling pathways suggest that vascular factors and cell proliferation that may interact with mitochondrial changes occurred. COX5B, cytochrome c oxidase subunit 5 CRU, Clinical Research Unit HIF, hypoxia-inducible factor IPA, ingenuity pathway analysis MAPR, mitochondrial ATP production rate NIH, National Institutes of Health SDH, succinate dehydrogenase TCA, tricarboxylic acid TFAM, mitochondrial transcription factor A UCP, uncoupling protein UQCR, ubiquinol cytochrome c reductase VEGF, vascular endothelial growth factor Footnotes Published ahead of print at http://diabetes.diabetesjournals.org on 27 July 2007. DOI: 10.2337/db07-0378. Additional information for this article can be found in an online appendix at http://dx.doi.org/10.2337/db07-0378 . The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. Received March 20, 2007. Accepted July 23, 2007. DIABETES