Single-cell RT-PCR analysis of N-methyl-D-aspartate receptor subunit expression in rat locus coeruleus neurones

Naunyn Schmiedebergs Arch Pharmacol. 2001 Jan;363(1):120-3. doi: 10.1007/s002100000348.

Abstract

Whole-cell patch-clamp recordings were performed on 12- to 15-day-old rat locus coeruleus neurones in a midpontine slice preparation. Application of noradrenaline (100 microM) and N-methyl-D-aspartate (NMDA; 100 microM) induced a small outward current and a distinct inward current, respectively. Single-cell reverse transcriptase-polymerase chain reaction (scRT-PCR), used to analyse the expression pattern of NMDA receptor subunits 2A, 2B, and 2C (NR2A-C) subsequent to electrophysiological characterization, demonstrated differences in the capacity of individual locus coeruleus neurones to express NR2A-C mRNA. NR2C mRNA expression predominated over those of NR2A and NR2B mRNA in most neurones. In addition, in neurones containing NR2C mRNA NMDA induced significantly larger currents than in cells lacking expression of this gene. RT-PCR studies performed on tissue preparations of adult rats also revealed a distinct expression of NR2C mRNA. In conclusion, the present data demonstrate differences in the mRNA expression pattern of NR2A-C of individual locus coeruleus neurones with a predominant NR2C mRNA expression in the majority of the cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Excitatory Amino Acid Agonists / pharmacology
  • Gene Expression
  • Locus Coeruleus / cytology
  • Locus Coeruleus / metabolism*
  • Locus Coeruleus / physiology
  • Membrane Potentials / drug effects
  • N-Methylaspartate / pharmacology
  • Neurons / drug effects
  • Neurons / metabolism*
  • Neurons / physiology
  • Norepinephrine / pharmacology
  • Patch-Clamp Techniques
  • Protein Subunits
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Rats
  • Rats, Wistar
  • Receptors, N-Methyl-D-Aspartate / genetics*
  • Receptors, N-Methyl-D-Aspartate / physiology
  • Reverse Transcriptase Polymerase Chain Reaction

Substances

  • Excitatory Amino Acid Agonists
  • NR2A NMDA receptor
  • NR2B NMDA receptor
  • Protein Subunits
  • RNA, Messenger
  • Receptors, N-Methyl-D-Aspartate
  • N-Methylaspartate
  • Norepinephrine