Investigation of PAR-1-type thrombin receptors in rat glioma C6 cells with a novel monoclonal anti-PAR-1 antibody (Mab COR7-6H9)

J Neurocytol. 1998 Sep;27(9):661-6. doi: 10.1023/a:1006912114880.

Abstract

Rat glioma C6 cells have been demonstrated to be a suitable model in the investigation of PAR-1-type thrombin receptors in brain. However, anti-PAR-1 antibodies, which should be very helpful tools in studying PAR-1 in rat cells, have not been available up until now. Therefore, we prepared a monoclonal anti-thrombin receptor antibody (Mab COR7-6H9) directed against the peptide sequence GRAVYLNKSRFPPMPPPPFISEDASG in the N-terminus below the thrombin cleavage site of the rat PAR-1-type thrombin receptor. Using this antibody, we demonstrated the presence of PAR-1 binding sites on the plasma membrane of rat glioma C6 cells both with confocal laser fluorescence and with scanning electron microscopy. In addition, Mab COR7-6H9 was shown to block PAR-1-mediated transmembranal signaling as demonstrated by measurement of free intracellular calcium and cyclic AMP. This novel anti-PAR-1 antibody is therefore likely to be a very helpful tool in studying PAR-1-type thrombin receptors in rat brain.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Monoclonal*
  • Fluorescent Antibody Technique
  • Glioma
  • Microscopy, Confocal
  • Microscopy, Electron, Scanning
  • Molecular Sequence Data
  • Rats
  • Receptor, PAR-1
  • Receptors, Thrombin / immunology
  • Receptors, Thrombin / metabolism*
  • Tumor Cells, Cultured

Substances

  • Antibodies, Monoclonal
  • Receptor, PAR-1
  • Receptors, Thrombin