title: Effects of glucose and insulin on glucokinase activity in rat hypothalamus.
abstract: In an attempt to study the role of glucokinase (GK) and the effects of glucose and peptides on GK gene expression and on the activity of this enzyme in the hypothalamus, we used two kinds of biological models: hypothalamic GT1-7 cells and rat hypothalamic slices. The expression of the GK gene in GT1-7 cells was reduced by insulin (INS) and was not modified by different glucose concentrations, while GK enzyme activities were significantly reduced by the different peptides. Interestingly, a distinctive pattern of GK activities between the ventromedial hypothalamus (VMH) and lateral hypothalamus (LH) were found, with higher enzyme activities in the VMH as the glucose concentrations rose, while LH enzyme activities decreased at 2.8 and 20 mM glucose, the latter effect being prevented by incubation with INS. These effects were produced only by d-glucose and the modifications found were due to GK, but not to other hexokinases. In addition, GK activities in the VMH and the LH were reduced by glucagon-like peptide 1, leptin, orexin B, INS, and neuropeptide Y (NPY), but this effect was only statistically significant for NPY in LH. Our results indicate that the effects of both glucose and peptides occur on GK enzyme activities rather than on GK gene transcription. Moreover, the effects of glucose and INS on GK activity suggest that in the brain GK behaves in a manner opposite to that in the liver, which might facilitate its role in glucose sensing. Finally, hypothalamic slices seem to offer a good physiological model to discriminate the effects between different areas.
authors.affiliation: Department of Biochemistry and Molecular Biology, Faculty of Medicine, Complutense University, Madrid, Spain. mcsanz@med.ucm.es
authors.forename: Carmen
authors.initials: C
authors.lastname: Sanz
authors.forename: Isabel
authors.initials: I
authors.lastname: Roncero
authors.forename: Patricia
authors.initials: P
authors.lastname: Vázquez
authors.forename: M Angeles
authors.initials: MA
authors.lastname: Navas
authors.forename: Enrique
authors.initials: E
authors.lastname: Blázquez
mesh_terms: D000818:Animals; D015153:Blotting, Western / Q000379:methods; D002460:Cell Line; D004789:Enzyme Activation / Q000187:drug effects; D015870:Gene Expression / Q000187:drug effects; D052216:Glucagon-Like Peptide 1 / Q000494:pharmacology; D005941:Glucokinase / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D005947:Glucose / Q000494:pharmacology*; D007004:Hypoglycemic Agents / Q000494:pharmacology*; D007026:Hypothalamic Area, Lateral / Q000187:drug effects / Q000201:enzymology; D007031:Hypothalamus / Q000187:drug effects / Q000201:enzymology*; D007328:Insulin / Q000494:pharmacology*; D047908:Intracellular Signaling Peptides and Proteins / Q000494:pharmacology; D020738:Leptin / Q000494:pharmacology; D008297:Male; D009478:Neuropeptide Y / Q000494:pharmacology; D009479:Neuropeptides / Q000494:pharmacology; D000068797:Orexins; D011401:Promoter Regions, Genetic; D051381:Rats; D017208:Rats, Wistar; D020133:Reverse Transcriptase Polymerase Chain Reaction; D046509:Tissue Culture Techniques; D014697:Ventromedial Hypothalamic Nucleus / Q000187:drug effects / Q000201:enzymology
chemical_list: D007004:Hypoglycemic Agents; D007328:Insulin; D047908:Intracellular Signaling Peptides and Proteins; D020738:Leptin; D009478:Neuropeptide Y; D009479:Neuropeptides; D000068797:Orexins; D052216:Glucagon-Like Peptide 1; D005941:Glucokinase; D005947:Glucose