Metabotropic glutamate receptors in acutely isolated hippocampal astrocytes: developmental changes of mGluR5 mRNA and functional expression

Glia. 2000 Jan 1;29(1):70-80. doi: 10.1002/(sici)1098-1136(20000101)29:1<70::aid-glia7>3.0.co;2-v.

Abstract

We previously found that 82% of glial fibrillary acidic protein (GFAP)-positive hippocampal astrocytes acutely isolated from P1-10 rats responded to glutamate (Glu) with transient intracellular calcium increases via activation of a Group I metabotropic glutamate receptor (mGluR). Fewer cells responded to ATP and none to serotonin (5-HT). In this study we asked the question whether hippocampal astrocytes in older animals retain this relative pattern of expression. We have found that 77% of GFAP (+) cells from P11-20 rats responded to 50 microM Glu, 43% to ATP, and none to 5-HT. Thirty-three percent of GFAP (+) cells from P25-35 rats responded to Glu, 12% to ATP and 3% to 5-HT. In the case of the responses to Glu, pharmacological characterization and single-cell RT-PCR data confirmed that these responses were mediated by the mGluR5 subtype of group I mGluRs. Also, fewer (36%) GFAP mRNA (+) cells from P25-35 rats expressed detectable mGluR5 mRNA than those from P11-20 rats (77%). This number essentially corresponds to the number of GFAP(+) showing a Ca(2+) response to Glu. Both mGluR5a and b were expressed with equal frequency in cells from P11-20 rats, but the b form predominated in cells from older animals. Overall, our studies show that expression of mGluR5 in hippocampal astrocytes decreases with increasing age and the "a" splice variant declines to a greater extent than the "b" splice variant, corresponding to the developmental changes shown in total tissue for mGluR5.

Publication types

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

MeSH terms

  • Aging
  • Alternative Splicing
  • Animals
  • Astrocytes / cytology
  • Astrocytes / drug effects
  • Astrocytes / metabolism*
  • Calcium / metabolism
  • Culture Media, Serum-Free / pharmacology
  • Down-Regulation
  • Gene Expression Regulation, Developmental*
  • Glial Fibrillary Acidic Protein / biosynthesis
  • Hippocampus / cytology
  • Hippocampus / drug effects
  • Hippocampus / growth & development
  • Hippocampus / metabolism*
  • Intracellular Fluid / drug effects
  • Intracellular Fluid / metabolism
  • Protein Isoforms / genetics
  • Protein Isoforms / metabolism
  • RNA, Messenger / metabolism*
  • Rats
  • Rats, Sprague-Dawley
  • Receptor, Metabotropic Glutamate 5
  • Receptors, Metabotropic Glutamate / genetics*
  • Receptors, Metabotropic Glutamate / metabolism*
  • Reverse Transcriptase Polymerase Chain Reaction

Substances

  • Culture Media, Serum-Free
  • Glial Fibrillary Acidic Protein
  • Grm5 protein, rat
  • Protein Isoforms
  • RNA, Messenger
  • Receptor, Metabotropic Glutamate 5
  • Receptors, Metabotropic Glutamate
  • Calcium