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document
Fine needle aspiration coupled with real-time PCR : a painless methodology to study adaptive functional changes in skeletal muscle . BACKGROUND AND AIM 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 . METHODS AND 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 . CONCLUSION 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 .
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Intervention_Surgical
Fine needle aspiration coupled with real-time PCR
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Intervention_Physical
methodology
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Intervention_Surgical
fine needle aspiration technique
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Intervention_Physical
Multiplex tandem RT-PCR
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Intervention_Physical
painless methodology
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Outcome_Physical
PGC-1alpha and COX5B
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Outcome_Physical
PGC-1alpha and MT-CO2
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Participant_Sample-size
21
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Participant_Condition
healthy
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Participant_Condition
different fitness levels