A reduction in intestinal cell pHi due to loss of the Caenorhabditis elegans Na+/H+ exchanger NHX-2 increases life span

J Biol Chem. 2003 Nov 7;278(45):44657-66. doi: 10.1074/jbc.M307351200. Epub 2003 Aug 25.

Abstract

Na+/H+ exchangers are involved in cell volume regulation, fluid secretion and absorption, and pH homeostasis. NHX-2 is a Caenorhabditis elegans Na+/H+ exchanger expressed exclusively at the apical membrane of intestinal epithelial cells. The inactivation of various intestinal nutrient transport proteins has been shown previously to influence aging via metabolic potential and a mechanism resembling caloric restriction. We report here a functional coupling of NHX-2 activity with nutrient uptake that results in long lived worms. Gene inactivation of nhx-2 by RNAi led to a loss of fat stores in the intestine and a 40% increase in longevity. The NHX-2 protein was coincidentally expressed with OPT-2, an oligopeptide transporter that is driven by a transmembrane proton gradient and that is also known to be involved in fat accumulation. Gene inactivation of opt-2 led to a phenotype resembling that of nhx-2, although not as severe. In order to explore this potential functional interaction, we combined RNA interference with a genetically encoded, fluorescence-based reagent to measure intestinal intracellular pH (pHi) in live worms under physiological conditions. Our results suggest first that OPT-2 is the main dipeptide uptake pathway in the nematode intestine, and second that dipeptide uptake results in intestinal cell acidification, and finally that recovery following dipeptide-induced acidification is normally a function of NHX-2. The loss of NHX-2 protein results in decreased steady-state intestinal cell pHi, and we hypothesize that this change perturbs proton-coupled nutrient uptake processes such as performed by OPT-2. Our data demonstrate a functional role for a Na+/H+ exchanger in nutrient absorption in vivo and lays the groundwork for examining integrated acid-base physiology in a non-mammalian model organism.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • ATP-Binding Cassette Transporters / genetics
  • ATP-Binding Cassette Transporters / physiology
  • Acid-Base Equilibrium
  • Aging
  • Animals
  • Caenorhabditis elegans / physiology*
  • Caenorhabditis elegans Proteins / genetics
  • Caenorhabditis elegans Proteins / physiology*
  • Dipeptides / metabolism
  • Fluorescent Dyes
  • Gene Expression
  • Gene Silencing
  • Green Fluorescent Proteins
  • Hydrogen-Ion Concentration
  • Indicators and Reagents
  • Intestinal Absorption
  • Intestines / chemistry
  • Kinetics
  • Longevity
  • Luminescent Proteins / genetics
  • Microscopy, Confocal
  • Sodium-Hydrogen Exchangers / genetics
  • Sodium-Hydrogen Exchangers / physiology*

Substances

  • ATP-Binding Cassette Transporters
  • Caenorhabditis elegans Proteins
  • Dipeptides
  • Fluorescent Dyes
  • Indicators and Reagents
  • Luminescent Proteins
  • NHX-2 protein, C elegans
  • Sodium-Hydrogen Exchangers
  • pept-1 protein, C elegans
  • Green Fluorescent Proteins