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Real time RT-PCR analysis of housekeeping genes in human skeletal muscle following acute exercise Mahoney, Douglas J., Carey, Kate, Fu, Ming-Hua, Snow, Rodney, Cameron-Smith, David, Parise, Gianni and Tarnopolsky, Mark A. 2004, Real time RT-PCR analysis of housekeeping genes in human skeletal muscle following acute exercise, Physiological genomics, vol. 18, pp. 226-231. Title Real time RT-PCR analysis of housekeeping genes in human skeletal muscle following acute exercise Author(s) Mahoney, Douglas J. Carey, Kate Fu, Ming-Hua Snow, Rodney Cameron-Smith, David Parise, Gianni Tarnopolsky, Mark A. Journal name Physiological genomics Volume number 18 Start page 226 End page 231 Publisher American Physiological Society Place of publication Bethesda, Md. Publication date 2004 ISSN 1094-8341 1531-2267 Keyword(s) gene expression endogenous controls validation Summary Studies examining gene expression with RT-PCR typically normalize their mRNA data to a constitutively expressed housekeeping gene. The validity of a particular housekeeping gene must be determined for each experimental intervention. We examined the expression of various housekeeping genes following an acute bout of endurance (END) or resistance (RES) exercise. Twenty-four healthy subjects performed either a interval-type cycle ergometry workout to exhaustion (~75 min; END) or 300 single-leg eccentric contractions (RES). Muscle biopsies were taken before exercise and 3 h and 48 h following exercise. Real-time RT-PCR was performed on ß-actin, cyclophilin (CYC), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and ß2-microglobulin (ß2M). In a second study, 10 healthy subjects performed 90 min of cycle ergometry at ~65% of O2 max, and we examined a fifth housekeeping gene, 28S rRNA, and reexamined ß2M, from muscle biopsy samples taken immediately postexercise. We showed that CYC increased 48 h following both END and RES exercise (3- and 5-fold, respectively; P < 0.01), and 28S rRNA increased immediately following END exercise (2-fold; P = 0.02). ß-Actin trended toward an increase following END exercise (1.85-fold collapsed across time; P = 0.13), and GAPDH trended toward a small yet robust increase at 3 h following RES exercise (1.4-fold; P = 0.067). In contrast, ß2M was not altered at any time point postexercise. We conclude that ß2M and ß-actin are the most stably expressed housekeeping genes in skeletal muscle following RES exercise, whereas ß2M and GAPDH are the most stably expressed following END exercise. Language eng Field of Research 110602 Exercise Physiology HERDC Research category C1 Refereed article in a scholarly journal Copyright notice ©2004, American Physiological Society Persistent URL http://hdl.handle.net/10536/DRO/DU:30008698 Document type: Journal Article Collections: School of Exercise and Nutrition Sciences Centre for Physical Activity and Nutrition Research Related Links Link Description Connected to published version (restricted access) Go to link with your DU access privileges     Unless expressly stated otherwise, the copyright for items in DRO is owned by the author, with all rights reserved. Versions Version Filter Type Sat, 14 Dec 2013, 13:08:25 EST Mon, 29 Apr 2013, 14:28:03 EST Filtered Full Citation counts:   Cited 107 times in TR Web of Science Cited 109 times in Scopus Search Google Scholar Access Statistics: 457 Abstract Views, 0 File Downloads  -  Detailed Statistics Created: Mon, 13 Oct 2008, 15:39:26 EST

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