Polarized expression of GABA transporters in Madin-Darby canine kidney cells and cultured hippocampal neurons

J Biol Chem. 1996 Mar 22;271(12):6917-24. doi: 10.1074/jbc.271.12.6917.

Abstract

At least three high affinity Na+- and Cl--dependent gamma-aminobutyric acid (GABA) transporters are known to exist in the rat and mouse brain. These transporters share 50-65% amino acid sequence identity with the kidney betaine transporter which also transports GABA but with lower affinity. The betaine transporter (BGT) is expressed on the basolateral surface of polarized Madin-Darby canine kidney (MDCK) cells. Recent evidence suggests that the signals and mechanisms involved in membrane protein sorting share many functional characteristics in polarized neurons and epithelial cells. It was previously shown that the rat GABA transporter GAT-1 is located in the presynaptic membrane of axons where it plays a role in terminating GABAergic neurotransmission. When expressed in MDCK cells by transfection, GAT-1 was sorted to the apical membrane. In this report, we have localized the other two GABA transporters, GAT-2 and GAT-3, in transfected MDCK cells by GABA uptake, immunofluorescence, and cell surface biotinylation. GAT-3, like GAT-1, localized to the apical membrane of MDCK cells while GAT-2, like BGT, localized to the basolateral membrane. We have also expressed BGT in low density cultures of hippocampal neurons by microinjection and immunolocalized it to the dendrites. The distribution of GAT-3 in these neurons after transfection was axonal as well as somatodendritic. These results indicate that highly homologous subtypes of GABA transporters are sorted differently when expressed in epithelial cells or neurons and suggest that these two cell types share the capacity to distinguish among these isoforms and target them to distinct destinations.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Axons / metabolism
  • Base Sequence
  • Carrier Proteins / genetics*
  • Cell Line
  • DNA Primers
  • DNA, Complementary
  • Dogs
  • GABA Plasma Membrane Transport Proteins
  • Hippocampus / cytology
  • Hippocampus / metabolism*
  • Kidney / cytology
  • Kidney / metabolism*
  • Membrane Proteins / genetics*
  • Membrane Transport Proteins*
  • Microinjections
  • Molecular Sequence Data
  • Neurons / metabolism*
  • Organic Anion Transporters*
  • gamma-Aminobutyric Acid / metabolism

Substances

  • Carrier Proteins
  • DNA Primers
  • DNA, Complementary
  • GABA Plasma Membrane Transport Proteins
  • Membrane Proteins
  • Membrane Transport Proteins
  • Organic Anion Transporters
  • Slc6a1 protein, mouse
  • Slc6a1 protein, rat
  • Slc6a11 protein, mouse
  • Slc6a12 protein, mouse
  • betaine plasma membrane transport proteins
  • gamma-Aminobutyric Acid