Quantitative rt-PCR analysis of uncoupling protein isoforms in mouse brain cortex: methodological optimization and comparison of expression with brown adipose tissue and skeletal muscle

Journal of Cerebral Blood Flow and Metabolism : Official Journal of the International Society of Cerebral Blood Flow and Metabolism
Sylvain LengacherLuc Pellerin

Abstract

Uncoupling proteins (UCPs) present in the inner mitochondrial membrane are involved in uncoupling respiration from ATP synthesis. Five UCP isoforms have been identified but information about their presence and level of expression in the central nervous system remains incomplete. To determine the nature and proportion of UCP isoform mRNAs present in brain cortex, we developed and optimized a specific quantitative reverse-transcription polymerase chain reaction procedure. Optimal range of RNA concentrations to be used in the reverse-transcriptase reaction was determined. Primer design and concentration were optimized for each target gene while polymerase chain reaction efficiency was assessed for a range of reverse-transcriptase dilutions. Genomic contribution to the quantitative signal was evaluated for each isoform and minimized. Three reference genes were tested for normalization, and beta-actin was found to be the most stable among tissues. Results indicate that brain cortex contains significant amounts of all UCP mRNAs, with UCP5 and UCP4 being the most abundant, as opposed to brown adipose tissue and skeletal muscle, which predominantly express UCP1 and UCP3, respectively. These data provide a first quantitative assessment ...Continue Reading

References

Jun 1, 1991·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·D RicquierF Bouillaud
Apr 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·W R Pearson, D J Lipman
Jan 14, 1967·Nature·P Mitchell, J Moyle
Jan 1, 1984·Physiological Reviews·D G Nicholls, R M Locke
Oct 1, 1996·Genome Research·U E GibsonP M Williams
Sep 1, 1997·Nucleic Acids Research·S F AltschulD J Lipman
Jan 5, 2000·Journal of Biotechnology·O ThellinE Heinen
Jan 6, 2000·The Biochemical Journal·D Ricquier, F Bouillaud
Aug 6, 2000·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·X X YuG Pan
May 30, 2001·Brain Research. Molecular Brain Research·R MacdonaldK L Philpott
Sep 27, 2001·The Journal of Biological Chemistry·M NibbelinkL Casteilla
Feb 16, 2002·Methods : a Companion to Methods in Enzymology·K J Livak, T D Schmittgen
Jul 17, 2003·Acta Physiologica Scandinavica·C Erlanson-Albertsson
Sep 27, 2003·Journal of Neurochemistry·Shigeru YamadaKatsuya Nagai

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Citations

Aug 24, 2005·Antioxidants & Redox Signaling·Jeong Sook Kim-Han, Laura L Dugan
May 9, 2009·Journal of Neuroinflammation·Antoinette DefauxFlorianne Monnet-Tschudi
Jan 1, 2008·Advances and Applications in Bioinformatics and Chemistry : AABC·Hyeon O KimPamela J Skinner
Mar 9, 2010·FEBS Letters·Petr JezekKeith D Garlid
Jan 22, 2015·Proceedings of the National Academy of Sciences of the United States of America·Willem J LaursenElena O Gracheva
Apr 21, 2012·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Malgorzata SlocinskaWieslawa Jarmuszkiewicz
Feb 29, 2008·The Journal of Investigative Dermatology·Shinobu MoriMasayuki Saito
Nov 11, 2008·Free Radical Biology & Medicine·Rocío GuevaraPilar Roca
Dec 18, 2007·Toxicology and Applied Pharmacology·Florianne Monnet-TschudiPaul Honegger
Nov 12, 2005·Bioscience Reports·Fábio T S NogueiraPaulo Arruda
May 22, 2013·Comparative Biochemistry and Physiology. Part A, Molecular & Integrative Physiology·Malgorzata SlocinskaWieslawa Jarmuszkiewicz
Feb 19, 2013·Developmental Biology·Petra KrausThomas Lufkin
Jul 8, 2016·IUBMB Life·Malgorzata SlocinskaWieslawa Jarmuszkiewicz
Aug 16, 2017·Veterinary Pathology·Elise E B LaDouceurChristopher M Reilly
May 26, 2005·Physiological Genomics·Patrick A KellerNatalia Giovannini
Apr 25, 2012·Neuromolecular Medicine·Nirinjini Naidoo
Sep 23, 2014·The Journal of Biological Chemistry·Hélène Perreten LambertSylvain Lengacher
Jan 10, 2020·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Kristen P TolsonAlexander S Kauffman
Oct 16, 2020·The Journal of Veterinary Medical Science·Andhika Yudha PrawiraYoshinao Z Hosaka

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Methods Mentioned

BETA
PCR

Software Mentioned

BLAST
Primer Express
ABI 7000
ClustalW

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