Fine needle aspiration coupled with real-time PCR: a painless methodology to study adaptive functional changes in skeletal muscle.
OBJECTIVE
In this study we developed a new methodology for obtaining human skeletal muscle samples to evaluate gene expression. This approach is based on a fine needle aspiration technique, which allows us to extract a small tissue sample in a significantly less invasive manner than with classic biopsy.


RESULTS
Multiplex tandem RT-PCR was used to determine the mRNA levels of genes involved in ATP production and mitochondrial biogenesis in muscle tissue. Samples of vastus lateralis muscle were obtained from 21 healthy subjects with different fitness levels. The principal findings in our study show a strong correlation between PGC-1alpha and COX5B (p<0.001) and between PGC-1alpha and MT-CO2 (p=0.017) expression. Furthermore, a significant positive correlation between mtDNA content and the percentage of MHCI present in the aspired samples were found (p=0.028). These data are in agreement with current knowledge on skeletal muscle physiology and show the reliability of the proposed method.


CONCLUSIONS
This painless methodology can be used to investigate, in vivo, human muscle RNA and DNA adaptations in response to either physiological and/or pharmacological stimuli. This method has major clinical relevance, such as its application in clarifying the mechanisms underling metabolic and systemic disorders.
Guescini
Michele
M
Institute of Health and Physical Exercise, University of Urbino Carlo Bo, Urbino, Italy.
Fatone
Cristina
C
Stocchi
Laura
L
Guidi
Chiara
C
Potenza
Lucia
L
Ditroilo
Massimiliano
M
Ranchelli
Anna
A
Di Loreto
Chiara
C
Sisti
Davide
D
De Feo
Pierpaolo
P
Stocchi
Vilberto
V
D000222:Adaptation, Physiological; D000328:Adult; D001483:Base Sequence; D044963:Biopsy, Fine-Needle / Q000379:methods*; D018592:Cross-Over Studies; D004272:DNA, Mitochondrial / Q000235:genetics* / Q000378:metabolism; D005784:Gene Amplification; D005786:Gene Expression Regulation / Q000502:physiology; D006801:Humans; D008297:Male; D008931:Mitochondria, Muscle / Q000235:genetics* / Q000378:metabolism; D018482:Muscle, Skeletal / Q000378:metabolism / Q000473:pathology*; D010101:Oxygen Consumption; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods
D004272:DNA, Mitochondrial; D012333:RNA, Messenger