Journal
PHYSIOLOGICAL GENOMICS
Volume 18, Issue 2, Pages 226-231Publisher
AMER PHYSIOLOGICAL SOC
DOI: 10.1152/physiolgenomics.00067.2004
Keywords
gene expression; endogenous controls; validation
Categories
Ask authors/readers for more resources
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 (similar to75 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 beta-actin, cyclophilin (CYC), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and beta2-microglobulin (beta2M). In a second study, 10 healthy subjects performed 90 min of cycle ergometry at similar to65% of Vo(2) (max), and we examined a fifth housekeeping gene, 28S rRNA, and reexamined beta2M, 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). beta-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, beta 2M was not altered at any time point postexercise. We conclude that beta 2M and beta-actin are the most stably expressed housekeeping genes in skeletal muscle following RES exercise, whereas beta 2M and GAPDH are the most stably expressed following END exercise.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available