title: Skeletal muscle PI3K p110β regulates expression of AMP-activated protein kinase.
abstract: Skeletal muscle metabolic homeostasis is maintained through numerous biochemical and physiological processes. Two principal molecular regulators of skeletal muscle metabolism include AMP-activated protein kinase (AMPK) and phosphatidylinositol 3-kinase (PI3K); however, PI3K exists as multiple isoforms, and specific metabolic actions of each isoform have not yet been fully elucidated in skeletal muscle. Given this lack of knowledge, we performed a series of experiments to define the extent to which PI3K p110β mediated expression and (or) activation of AMPK in skeletal muscle. To determine the effect of p110β inhibition on AMPK expression and phosphorylation in cultured cells, C2C12 myoblasts were treated with a pharmacological inhibitor of p110β (TGX-221), siRNA against p110β, or overexpression of kinase-dead p110β. Expression and phosphorylation of AMPK were unaffected in myoblasts treated with TGX-221 or expressing kinase-dead p110β. However, expressions of total and phosphorylated AMPK at T172 were reduced in myoblasts treated with p110β siRNA. When normalized to expression of total AMPK, phosphorylation of AMPK S485/491 was elevated in p110β-deficient myoblasts. Similar results were observed in tibialis anterior muscle from mice with conditional deletion of p110β (p110β-mKO mice). Analysis of AMPK transcript expression revealed decreased expression of Prkaa2 in p110β-deficient myoblasts and in p110β-mKO muscle. Loss of p110β had no effect on oligomycin-stimulated phosphorylation of AMPK or phosphorylated Acetyl-CoA carboxylase (ACC), although oligomycin-induced AMPK and ACC phosphorylation were increased in p110β-deficient myoblasts compared to oligomycin-stimulated control myoblasts when normalized to levels of total AMPK or ACC. Overall, these results suggest that p110β positively regulates expression of AMPK in cultured myoblasts and in skeletal muscle in vivo; moreover, despite the reduced abundance of AMPK in p110β-deficient myoblasts, loss of p110β does not appear to impair AMPK activation following stimulus. These findings thus reveal a novel role for p110β in mediating skeletal muscle metabolic signaling.
authors.affiliation: Military Performance Division, US Army Research Institute of Environmental Medicine, 10 General Greene Ave, Building 42, Natick, MA, 01760, USA. Electronic address: ronald.w.matheny.civ@mail.mil.
authors.forename: Ronald W
authors.initials: RW
authors.lastname: Matheny
authors.affiliation: Military Performance Division, US Army Research Institute of Environmental Medicine, 10 General Greene Ave, Building 42, Natick, MA, 01760, USA.
authors.forename: Mary N
authors.initials: MN
authors.lastname: Abdalla
authors.affiliation: Military Performance Division, US Army Research Institute of Environmental Medicine, 10 General Greene Ave, Building 42, Natick, MA, 01760, USA.
authors.forename: Alyssa V
authors.initials: AV
authors.lastname: Geddis
authors.affiliation: Military Performance Division, US Army Research Institute of Environmental Medicine, 10 General Greene Ave, Building 42, Natick, MA, 01760, USA.
authors.forename: Luis A
authors.initials: LA
authors.lastname: Leandry
authors.affiliation: Military Performance Division, US Army Research Institute of Environmental Medicine, 10 General Greene Ave, Building 42, Natick, MA, 01760, USA.
authors.forename: Christine M
authors.initials: CM
authors.lastname: Lynch
mesh_terms: D055372:AMP-Activated Protein Kinases / Q000378:metabolism*; D000103:Acetyl-CoA Carboxylase / Q000378:metabolism; D000818:Animals; D002384:Catalysis; D002460:Cell Line; D058534:Class I Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D017353:Gene Deletion; D005786:Gene Expression Regulation; D051379:Mice; D018345:Mice, Knockout; D018482:Muscle, Skeletal / Q000378:metabolism; D009132:Muscles / Q000378:metabolism; D032446:Myoblasts / Q000378:metabolism; D010766:Phosphorylation; D034741:RNA, Small Interfering / Q000378:metabolism*
chemical_list: D034741:RNA, Small Interfering; C509622:1-phosphatidylinositol 3-kinase p110 subunit, mouse; D058534:Class I Phosphatidylinositol 3-Kinases; D055372:AMP-Activated Protein Kinases; D000103:Acetyl-CoA Carboxylase
keywords: AMP-activated protein kinase (AMPK); Myoblast; Phosphoinositide 3-kinase (PI3K); Skeletal muscle