Improvement in Conditions for Solubilisation and Characterisation of Brain D2 Dopamine Receptors Using Various Detergents

Mark Wheatley, Jean M. Hall, Patricia A. Frankham, Philip G. Strange

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Abstract: A series of detergents of varying chemical properties has been tested for solubilisation of bovine caudate nucleus D2 dopamine receptors using [3H]spiperone binding to assay the solubilised sites. The properties of the lysophosphatidylcholine (LPC)‐ and 3‐[(3‐cholamidopropyl)dimethylammonio]‐1‐propane‐sulphonate (CHAPS)‐solubilised preparations are described in detail. The preparations are truly solubilised, and sucrose density gradient and gel filtration data are reported. Specific [3H]spiperone binding in the LPC‐solubilised preparation assayed at 4°C is solely to D2 dopamine re‐ceptors. If the assay temperature is raised to 25°C, the amount of specific [3H]spiperone binding is largely unchanged, but it forms a greater proportion of the total [3H]spiperone binding owing to a reduction in nonstereospecific (spirodecanone) [3H]spiperone binding at the higher temperature. The effect of raising the assay temperature is important as it enables more precise deter‐minations of specific [3H]spiperone binding to be made. Part of the specific [3H]spiperone binding at 25°C is to solubilised S2 serotonin receptors in addition to D2 dopamine receptors. Good correlations are observed between the affinities for binding of ligands to the solubilised D2 receptors and corresponding data obtained on membrane‐bound receptors. Agonist binding in LPC‐solubilised preparations is insensitive to guanine nucleotides. It is speculated that the spirodecanone sites represent, in part, proteolysed or damaged D2 dopamine, or S2 serotonin, receptors. In the CHAPS‐solubilised preparation the pharmacological profile of [3H]spiperone binding is unclear when assayed at 4°C, but in assays at 25°C a clear serotonin S2 receptor component of specific [3H]spiperone binding can be discerned. No clear D2 dopamine receptor component can be detected in this preparation under these conditions.

Original languageEnglish
Pages (from-to)926-934
Number of pages9
JournalJournal of Neurochemistry
Volume43
Issue number4
DOIs
Publication statusPublished - Oct 1984
Externally publishedYes

Keywords

  • Bovine caudate nucleus
  • D dopamine receptors
  • Detergents
  • Solubilisation
  • [H]Spiperone binding

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

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