Abstract
BackgroundBergström needle biopsy is widely used to sample skeletal muscle in order to study cell signaling directly in human tissue. Consequences of the biopsy protocol design on muscle protein quantity and quality remain unclear. The aim of the present study was to assess the impact of different events surrounding biopsy protocol on the stability of the Western blot signal of eukaryotic translation initiation factor 4E binding protein 1 (4E-BP1), Akt, glycogen synthase kinase-3β (GSK-3β), muscle RING finger protein 1 (MuRF1) and p70 S6 kinase (p70 S6K). Six healthy subjects underwent four biopsies of the vastus lateralis, distributed into two distinct visits spaced by 48 hrs. At visit 1, a basal biopsy in the right leg was performed in the morning (R1) followed by a second in the left leg in the afternoon (AF). At visit 2, a second basal biopsy (R2) was collected from the right leg. Low intensity mobilization (3 × 20 right leg extensions) was performed and a final biopsy (Mob) was collected using the same incision site as R2.ResultsAkt and p70 S6K phosphorylation levels were increased by 83% when AF biopsy was compared to R1. Mob condition induced important phosphorylation of p70 S6K when compared to R2. Comparison of R1 and R2 biopsies revealed a relative stability of the signal for both total and phosphorylated proteins.ConclusionsThis study highlights the importance to standardize muscle biopsy protocols in order to minimize the method-induced variation when analyzing Western blot signals.
Highlights
Bergström needle biopsy is widely used to sample skeletal muscle in order to study cell signaling directly in human tissue
Muscle atrophy has a wide spectrum of consequences for chronically ill patients, ranging from an aggravated morbidity to a seriously impaired survival prognostic [4,5]
Total protein amount variation The impact of the conditions surrounding biopsy procedures on total protein amount was evaluated by measuring the intra-subject variation of the Western blot signal obtained with each sampling conditions (R1, AF, rested and fasted-2” (R2), mobilization and fasted” (Mob))
Summary
Bergström needle biopsy is widely used to sample skeletal muscle in order to study cell signaling directly in human tissue. Consequences of the biopsy protocol design on muscle protein quantity and quality remain unclear. At visit 2, a second basal biopsy (R2) was collected from the right leg. The phosphatidylinositol-3 kinase/Akt. Human muscle samples required for biochemical studies have been obtained repeatedly with a Bergström biopsy needle [12]. Protein levels and/or their associated activities (kinase or ligase) in muscle tissue are affected by stimuli such as exercise [13] and nutritional state [14]. In the context of studies aimed at investigating cell signaling in muscle tissue, it cannot be excluded that sampling site variation combined to unstandardized nutritional and physical activity status could impact data analysis
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
