Expression and regulation of nampt in human islets

PLoS One. 2013;8(3):e58767. doi: 10.1371/journal.pone.0058767. Epub 2013 Mar 11.

Abstract

Nicotinamide phosphoribosyltransferase (Nampt) is a rate-limiting enzyme in the mammalian NAD+ biosynthesis of a salvage pathway and exists in 2 known forms, intracellular Nampt (iNampt) and a secreted form, extracellular Nampt (eNampt). eNampt can generate an intermediate product, nicotinamide mononucleotide (NMN), which has been reported to support insulin secretion in pancreatic islets. Nampt has been reported to be expressed in the pancreas but islet specific expression has not been adequately defined. The aim of this study was to characterize Nampt expression, secretion and regulation by glucose in human islets. Gene and protein expression of Nampt was assessed in human pancreatic tissue and isolated islets by qRT-PCR and immunofluorescence/confocal imaging respectively. Variable amounts of Nampt mRNA were detected in pancreatic tissue and isolated islets. Immunofluorescence staining for Nampt was found in the exocrine and endocrine tissue of fetal pancreas. However, in adulthood, Nampt expression was localized predominantly in beta cells. Isolated human islets secreted increasing amounts of eNampt in response to high glucose (20 mM) in a static glucose-stimulated insulin secretion assay (GSIS). In addition to an increase in eNampt secretion, exposure to 20 mM glucose also increased Nampt mRNA levels but not protein content. The secretion of eNampt was attenuated by the addition of membrane depolarization inhibitors, diazoxide and nifedipine. Islet-secreted eNampt showed enzymatic activity in a reaction with increasing production of NAD+/NADH over time. In summary, we show that Nampt is expressed in both exocrine and endocrine tissue early in life but in adulthood expression is localized to endocrine tissue. Enzymatically active eNampt is secreted by human islets, is regulated by glucose and requires membrane depolarization.

MeSH terms

  • Adolescent
  • Adult
  • Age Factors
  • Aged
  • Aged, 80 and over
  • Child
  • Child, Preschool
  • Enzyme Activation
  • Female
  • Gene Expression Regulation* / drug effects
  • Glucagon / metabolism
  • Glucose / metabolism
  • Glucose / pharmacology
  • Humans
  • Infant
  • Infant, Newborn
  • Insulin / metabolism
  • Islets of Langerhans / drug effects
  • Islets of Langerhans / metabolism*
  • Male
  • Middle Aged
  • Nicotinamide Phosphoribosyltransferase / genetics*
  • Nicotinamide Phosphoribosyltransferase / metabolism
  • Pancreas / drug effects
  • Pancreas / metabolism
  • Protein Binding
  • Proteomics
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Young Adult

Substances

  • Insulin
  • RNA, Messenger
  • Glucagon
  • Nicotinamide Phosphoribosyltransferase
  • Glucose

Grant support

The authors have no support or funding to report.