title: Leptin downregulates expression of the gene encoding glucagon in alphaTC1-9 cells and mouse islets.
abstract: OBJECTIVE
  Leptin, released by adipocytes, can modulate glucose homeostasis through direct actions on pancreatic alpha and beta cells. Although this hormone rapidly regulates alpha cell exocytosis, its long-term effects on glucagon gene expression are currently unknown.
  
  
  METHODS
  We analysed glucagon mRNA levels and protein content in alphaTC1-9 cells and isolated mouse islets cultured with leptin, as well as in islets from mice treated in vivo with leptin. We also studied the involvement of the signal transducers and activators of transcription (STAT) pathway by western blot, immunofluorescence and interference RNA.
  
  
  RESULTS
  Leptin incubation (0.0625-18.75 nmol/l) for 24 h inhibited glucagon gene expression in alphaTC1-9 cells. This inhibitory effect was also observed in isolated mouse islets cultured with leptin, as well as in islets from mice treated with leptin for 5 days. In contrast, no changes were detected in islets from db/db mice, which lack leptin receptors. Leptin treatment also reduced the glucagon protein content in alphaTC1-9 cells and mouse islets. Moreover, leptin induced phosphorylation of STAT3 and its translocation to the nucleus, which was confirmed by western blot analysis in alphaTC1-9 cells and by immunofluorescence in isolated alpha cells. Interestingly, the effect of leptin on glucagon mRNA levels was significantly reduced by Stat3 knockdown. In contrast, pharmacological inhibition of the phosphoinositide 3-kinase pathway did not affect leptin actions.
  
  
  CONCLUSIONS
  Our results demonstrate that leptin can regulate glucagon gene expression in alpha cells via a STAT3 pathway, and are important for understanding the role of leptin in glucose homeostasis.
authors.affiliation: Instituto de Bioingeniería, Universidad Miguel Hernandez, Avenida de la Universidad, 03202 Elche, Spain.
authors.forename: L
authors.initials: L
authors.lastname: Marroquí
authors.forename: E
authors.initials: E
authors.lastname: Vieira
authors.forename: A
authors.initials: A
authors.lastname: Gonzalez
authors.forename: A
authors.initials: A
authors.lastname: Nadal
authors.forename: I
authors.initials: I
authors.lastname: Quesada
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D005260:Female; D005934:Glucagon / Q000235:genetics / Q000378:metabolism*; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D007515:Islets of Langerhans / Q000187:drug effects* / Q000378:metabolism*; D020738:Leptin / Q000494:pharmacology*; D051379:Mice; D008810:Mice, Inbred C57BL; D034741:RNA, Small Interfering; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D020738:Leptin; D034741:RNA, Small Interfering; D005934:Glucagon