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1 Department of Physiology and Pharmacology, Karolinska Institute, Stockholm, Sweden; Department of Laboratory Medicine, Division of Clinical Physiology, Karolinska Institute, Stockholm, Sweden
2 Department of Laboratory Medicine, Division of Clinical Physiology, Karolinska Institute, Stockholm, Sweden
3 Department of Laboratory Medicine, Division of Clinical Physiology, Karolinska Institute, Stockholm, Sweden; Center for Genomics and Bioinformatics, Karolinska Institute, Stockholm, Sweden
4 Department of Physiology and Pharmacology, Karolinska Institute, Stockholm, Sweden
* To whom correspondence should be addressed. E-mail: Jamie.Timmons{at}cgb.ki.se.
Vascular endothelial growth factor A (VEGF-A) contributes to muscle tissue angiogenesis following aerobic exercise training. The temporal response of the VEGF-A isoforms and their target receptors has not been comprehensively profiled in human skeletal muscle. We combined submaximal exercise with and without reduced leg blood flow, to establish if ischemia induced metabolic stress was an important physiological stimuli responsible for regulating the VEGF-A system in humans. Nine healthy men performed two 45-min bouts of one-leg knee extension exercise, with and without blood flow restriction. Muscle biopsies were obtained at rest and 2 h and 6 h after exercise. Expression (mRNA) of the VEGF-A splice variants and related receptors (VEGFR1, VEGFR2 and neuropilin-1 (NP-1)) were determined using qPCR. VEGF-Atotal expression increased more robustly after exercise with reduced blood flow and initially this principally reflected an increase in VEGF-A165. Six hr after exercise there was a relatively greater increase in VEGF-A189 and this response was not influenced by blood flow conditions. VEGFR1 mRNA expression increased 2 h after exercise, NP-1 expression was transiently reduced; while all 3 receptors increased by 6 h. There was no evidence for the expression of the inhibitory VEGF-A165B variant in human skeletal muscle. Our study, reflecting both VEGF-A ligand and receptors, implicates metabolic perturbation as a regulator of human muscle angiogenesis and demonstrates that VEGF-A splice variants are distinctly regulated. Our findings also indicate that all three receptor genes exhibit different pre-translational regulation, in response to exercise in humans.
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