title: PDH activation during in vitro muscle contractions in PDH kinase 2 knockout mice: effect of PDH kinase 1 compensation.
abstract: Pyruvate dehydrogenase (PDH) plays an important role in regulating carbohydrate oxidation in skeletal muscle. PDH is deactivated by a set of PDH kinases (PDK1, PDK2, PDK3, PDK4), with PDK2 and PDK4 being the most predominant isoforms in skeletal muscle. Although PDK2 is the most abundant isoform, few studies have examined its physiological role. The role of PDK2 on PDH activation (PDHa) at rest and during muscle stimulation at 10 and 40 Hz (eliciting low- and moderate-intensity muscle contractions, respectively) in isolated extensor digitorum longus muscles was studied in PDK2 knockout (PDK2KO) and wild-type (WT) mice (n = 5 per group). PDHa activity was unexpectedly 35 and 77% lower in PDK2KO than WT muscle (P = 0.043), while total PDK activity was nearly fourfold lower in PDK2KO muscle (P = 0.006). During 40-Hz contractions, initial force was lower in PDK2KO than WT muscle (P < 0.001) but fatigued similarly to ∼75% of initial force by 3 min. There were no differences in initial force or rate of fatigue during 10-Hz contractions. PDK1 compensated for the lack of PDK2 and was 1.8-fold higher in PDK2KO than WT muscle (P = 0.019). This likely contributed to ensuring that resting PDHa activity was similar between the groups and accounts for the lower PDH activation during muscle contraction, as PDK1 is a very potent inhibitor of the PDH complex. Increased PDK1 expression appears to be regulated by hypoxia inducible factor-1α, which was 3.5-fold higher in PDK2KO muscle. It is clear that PDK2 activity is essential, even at rest, in regulation of carbohydrate oxidation and production of reducing equivalents for the electron transport chain. In addition, these results underscore the importance of the overall kinetics of the PDK isoform population, rather than total PDK activity, in determining transformation of the PDH complex and PDHa activity during muscle contraction.
authors.affiliation: Centre for Muscle Metabolism and Biophysics, Brock University, St. Catharines, Ontario, Canada.
authors.forename: Emily C
authors.initials: EC
authors.lastname: Dunford
authors.forename: Eric A
authors.initials: EA
authors.lastname: Herbst
authors.forename: Nam Ho
authors.initials: NH
authors.lastname: Jeoung
authors.forename: William
authors.initials: W
authors.lastname: Gittings
authors.forename: J Greig
authors.initials: JG
authors.lastname: Inglis
authors.forename: Rene
authors.initials: R
authors.lastname: Vandenboom
authors.forename: Paul J
authors.initials: PJ
authors.lastname: LeBlanc
authors.forename: Robert A
authors.initials: RA
authors.lastname: Harris
authors.forename: Sandra J
authors.initials: SJ
authors.lastname: Peters
mesh_terms: D000818:Animals; D005260:Female; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000502:physiology; D066298:In Vitro Techniques; D007658:Ketone Oxidoreductases / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D023421:Models, Animal; D009119:Muscle Contraction / Q000502:physiology*; D018482:Muscle, Skeletal / Q000502:physiology*; D020033:Protein Isoforms / Q000502:physiology; D017346:Protein Serine-Threonine Kinases / Q000172:deficiency* / Q000235:genetics / Q000502:physiology*; D000081382:Pyruvate Dehydrogenase Acetyl-Transferring Kinase
chemical_list: C497443:Hif1a protein, mouse; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; C000631433:Pdk1 protein, mouse; C000631435:Pdk2 protein, mouse; C000631437:Pdk3 protein, mouse; C000631439:Pdk4 protein, mouse; D020033:Protein Isoforms; D000081382:Pyruvate Dehydrogenase Acetyl-Transferring Kinase; D007658:Ketone Oxidoreductases; C053164:pyruvate dehydrogenase (NADP+); D017346:Protein Serine-Threonine Kinases