title: Changes in glucose transport and protein kinase Cbeta(2) in rat skeletal muscle induced by hyperglycaemia.
abstract: OBJECTIVE
  We have previously reported that hyperglycaemia activates glucose transport in skeletal muscle by a Ca(2+)-dependent pathway, which is distinct from the insulin-signalling pathway. The aim of this study was to explain the signalling mechanism by which hyperglycaemia autoregulates glucose transport in skeletal muscle.
  
  
  METHODS
  Isolated rat soleus muscle was incubated in the presence of various concentrations of glucose or 3-O-methylglucose and protein kinase C and phospholipase C inhibitors. Glucose transport activity, cell surface glucose transporter 1 and glucose transporter 4 content and protein kinase C translocation was determined.
  
  
  RESULTS
  High concentrations of 3-O-methylglucose led to a concentration-dependent increase in [(3)H]-3-O-methylglucose transport in soleus muscle. Dantrolene, an inhibitor of Ca(2+) released from the sarcoplasmic reticulum, decreased the V(max) and the K(m) of the concentration-response curve. Protein kinase C inhibitors (H-7 and GF109203X) inhibited the stimulatory effect of high glucose concentrations on hexose transport, whereas glucose transport stimulated by insulin was unchanged. Incubation of muscle with glucose (25 mmol/l) and 3-O-methylglucose (25 mmol/l) led to a three fold gain in protein kinase Cbeta(2) in the total membrane fraction, whereas membrane content of protein kinase Calpha, beta(1), delta, epsilon and theta were unchanged. A short-term increase in the extracellular glucose concentration did not change cell surface recruitment of glucose transporter 1 or glucose transporter 4, as assessed by exofacial photolabelling with [(3)H]-ATB-BMPA bis-mannose.
  
  
  CONCLUSIONS
  Protein kinase Cbeta(2) is involved in a glucose-sensitive, Ca(2+)-dependent signalling pathway, which is possibly involved in the regulation of glucose transport in skeletal muscle. This glucose-dependent increase in 3-0-methylglucose transport is independent of glucose transporter 4 and glucose transporter 1 translocation to the plasma membrane and may involve modifications of cell surface glucose transporter activity.
authors.affiliation: Departments of Clinical Physiology, and Physiology and Pharmacology, Karolinska Institute, Karolinska Hospital, Stockholm, Sweden.
authors.forename: Y
authors.initials: Y
authors.lastname: Kawano
authors.forename: J
authors.initials: J
authors.lastname: Rincon
authors.forename: A
authors.initials: A
authors.lastname: Soler
authors.forename: J W
authors.initials: JW
authors.lastname: Ryder
authors.forename: L A
authors.initials: LA
authors.lastname: Nolte
authors.forename: J R
authors.initials: JR
authors.lastname: Zierath
authors.forename: H
authors.initials: H
authors.lastname: Wallberg-Henriksson
mesh_terms: D019325:3-O-Methylglucose / Q000378:metabolism*; D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D005947:Glucose / Q000378:metabolism* / Q000494:pharmacology; D051272:Glucose Transporter Type 1; D051275:Glucose Transporter Type 4; D006943:Hyperglycemia / Q000378:metabolism*; D066298:In Vitro Techniques; D007328:Insulin / Q000494:pharmacology; D007527:Isoenzymes / Q000378:metabolism*; D007700:Kinetics; D008297:Male; D009004:Monosaccharide Transport Proteins / Q000378:metabolism*; D009124:Muscle Proteins; D018482:Muscle, Skeletal / Q000187:drug effects / Q000378:metabolism*; D011493:Protein Kinase C / Q000378:metabolism*; D064546:Protein Kinase C beta; D051381:Rats; D017208:Rats, Wistar
chemical_list: D051272:Glucose Transporter Type 1; D051275:Glucose Transporter Type 4; D007328:Insulin; D007527:Isoenzymes; D009004:Monosaccharide Transport Proteins; D009124:Muscle Proteins; C495679:Slc2a1 protein, rat; C495694:Slc2a4 protein, rat; D019325:3-O-Methylglucose; D011493:Protein Kinase C; D064546:Protein Kinase C beta; D005947:Glucose