Immunolabeling of central serotonin 5-HT1D beta receptors in the rat, mouse, and guinea pig with a specific anti-peptide antiserum

J Neurochem. 1995 Dec;65(6):2671-81. doi: 10.1046/j.1471-4159.1995.65062671.x.


A synthetic peptide (25 amino acids) corresponding to a specific portion of the third intracytoplasmic loop of the rat serotonin 5-HT1B/1D beta receptor was coupled to keyhole limpet hemocyanin and injected monthly into rabbits. Anti-peptide antibodies were detected by enzyme-linked immunosorbent assay and characterized by immunoprecipitation of the 5-HT1B/1D beta receptor in CHAPS-solubilized extracts from rat striatal membranes. Up to 60% of solubilized striatal serotonin-O-carboxymethylglycyl[125I]iodotyrosinamide ([125I]GTI; a selective 5-HT1B/1D radioligand) binding sites were immunoprecipitated and subsequently pharmacologically identified as 5-HT1B receptors. The remaining 40% of [125I]GTI binding sites were shown to be 5-HT1D receptors. In addition, these antibodies were successfully used in immunofluorescence experiments to detect the 5-HT1B/1D beta, but not the 5-HT1D/1D alpha, receptor in transiently transfected LLC-PK1 cells. Immunoautoradiographic experiments performed with brain sections from the rat, mouse, and guinea pig showed that the substantia nigra and globus pallidus contained the highest densities of 5-HT1D beta receptor-like immunoreactivity. Comparison of the regional distribution of immunolabeling with that of the specific binding of [125I]GTI in the brain of these species further confirmed that the anti-peptide antibodies selectively recognized only the 5-HT1D beta component of [125I]GTI specific receptor binding sites.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Autoradiography
  • Binding Sites
  • Corpus Striatum / metabolism*
  • Dipeptides / metabolism
  • Guinea Pigs
  • Immune Sera
  • Immunologic Techniques
  • Iodine Radioisotopes
  • LLC-PK1 Cells
  • Male
  • Mice
  • Molecular Sequence Data
  • Rabbits
  • Rats
  • Rats, Sprague-Dawley
  • Receptors, Serotonin / genetics
  • Receptors, Serotonin / metabolism*
  • Serotonin / analogs & derivatives
  • Serotonin / metabolism
  • Swine
  • Transfection


  • Dipeptides
  • Immune Sera
  • Iodine Radioisotopes
  • Receptors, Serotonin
  • serotonin-O-carboxymethyl-Gly-Tyr
  • Serotonin