Development of a multiplex assay to determine the expression of mitochondrial genes in human skeletal muscle.
Tom P AirdAndrew J FarquharsonJanice E DrewBrian P CarsonPublished in: Experimental physiology (2021)
Skeletal muscle is an important endocrine tissue demonstrating plasticity in response to external stimuli, including exercise and nutrition. Mitochondrial biogenesis is a common hallmark of adaptations to aerobic exercise training. Furthermore, altered expression of several genes implicated in the regulation of mitochondrial biogenesis, substrate oxidation and nicotinamide adenine dinucleotide (NAD+ ) biosynthesis following acute exercise underpins longer-term muscle metabolic adaptations. Gene expression is typically measured using real-time quantitative PCR platforms. However, interest has developed in the design of multiplex gene expression assays (GeXP) using the GenomeLab GeXP™ genetic analysis system, which can simultaneously quantify gene expression of multiple targets, holding distinct advantages in terms of throughput, limiting technical error, cost effectiveness, and quantifying gene co-expression. This study describes the development of a custom-designed GeXP assay incorporating the measurement of proposed regulators of mitochondrial biogenesis, substrate oxidation, and NAD+ biosynthetic capacity in human skeletal muscle and characterises the resting gene expression (overnight fasted and non-exercised) signature within a group of young, healthy, recreationally active males. The design of GeXP-based assays provides the capacity to more accurately characterise the regulation of a targeted group of genes with specific regulatory functions, a potentially advantageous development for future investigations of the regulation of muscle metabolism by exercise and/or nutrition.
Keyphrases
- skeletal muscle
- gene expression
- high intensity
- high throughput
- dna methylation
- poor prognosis
- genome wide
- oxidative stress
- physical activity
- endothelial cells
- insulin resistance
- genome wide identification
- transcription factor
- resistance training
- binding protein
- liver failure
- induced pluripotent stem cells
- genome wide analysis
- preterm infants
- real time pcr
- hydrogen peroxide
- pluripotent stem cells
- type diabetes
- mass spectrometry
- heart rate
- hepatitis b virus
- high resolution
- cancer therapy
- drug induced
- heart rate variability
- adipose tissue
- drug delivery
- amino acid
- cell wall
- body composition
- electron transfer
- middle aged
- mechanical ventilation