Metabotropic glutamate receptor 3 (mGlu3; mGluR3; GRM3) in schizophrenia: Antibody characterisation and a semi-quantitative western blot study

Aintzane García-Bea, Mary A. Walker, Thomas M. Hyde, Joel E. Kleinman, Paul J. Harrison, Tracy A. Lane

Research output: Contribution to journalArticle

Abstract

Background Metabotropic glutamate receptor 3 (mGlu3, mGluR3), encoded by GRM3, is a risk gene for schizophrenia and a therapeutic target. It is unclear whether expression of the receptor is altered in the disorder or related to GRM3 risk genotype. Antibodies used to date to assess mGlu3 in schizophrenia have not been well validated. Objective To characterise six commercially available anti-mGlu3 antibodies for use in human brain, and then conduct a semi-quantitative study of mGlu3 immunoreactivity in schizophrenia. Methods Antibodies tested using Grm3−/− and Grm2−/−/3−/− mice and transfected HEK293T/17 cells. Western blotting on membrane protein isolated from superior temporal cortex of 70 patients with schizophrenia and 87 healthy comparison subjects, genotyped for GRM3 SNP rs10234440. Results One (out of six) anti-mGlu3 antibodies was fully validated, a C-terminal antibody which detected monomeric (~ 100 kDa) and dimeric (~ 200 kDa) mGlu3. A second, N-terminal, antibody detected the 200 kDa band but also produced non-specific bands. Using the C-terminal antibody for western blotting in human brain, mGlu3 immunoreactivity was found to decline with age, and was affected by pH and post mortem interval. There were no differences in monomeric or dimeric mGlu3 immunoreactivity in schizophrenia or in relation to GRM3 genotype. The antibody was not suitable for immunohistochemistry. Interpretation These data highlight the value of knockout mouse tissue for antibody validation, and the need for careful antibody characterisation. The schizophrenia data show that involvement of GRM3 in the disorder and its genetic risk architecture is not reflected in total membrane mGlu3 immunoreactivity in superior temporal cortex.

Original languageEnglish (US)
Pages (from-to)18-27
Number of pages10
JournalSchizophrenia Research
Volume177
Issue number1-3
DOIs
StatePublished - Nov 1 2016

Keywords

  • Antibody validation
  • Group II metabotropic glutamate receptor
  • Human brain
  • Methods
  • Psychosis

ASJC Scopus subject areas

  • Psychiatry and Mental health
  • Biological Psychiatry

Fingerprint Dive into the research topics of 'Metabotropic glutamate receptor 3 (mGlu3; mGluR3; GRM3) in schizophrenia: Antibody characterisation and a semi-quantitative western blot study'. Together they form a unique fingerprint.

  • Cite this