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, doi: 10.1152/physiolgenomics.00067.2004.

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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, RodneyORCID iD for 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
Keyword(s) gene expression
endogenous controls
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
DOI 10.1152/physiolgenomics.00067.2004
Field of Research 110602 Exercise Physiology
HERDC Research category C1 Refereed article in a scholarly journal
Copyright notice ©2004, American Physiological Society
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