[3H]PtdIns hydrolysis in postmortem human brain membranes is mediated by the G-proteins G(q/11) and phospholipase C-β

R. S. Jope, L. Song, R. Powers

Research output: Contribution to journalArticle

31 Scopus citations

Abstract

A method utilizing exogenously added [3H]PtdIns incubated with membranes prepared from postmortem human brain has been shown to provide a means of measuring agonist-induced, guanosine 5'-O-(thiotriphosphate) (GTP[S])-dependent hydrolysis of [3H]PtdIns, thus allowing investigations of the activity of the phosphoinositide second-messenger system in accessible human brain tissue. Agonists inducing [3H]PtdIns hydrolysis include carbachol, trans-1-aminocyclopentyl-1,3-dicarboxylate (ACPD; a glutamatergic metabotropic receptor agonist), serotonin and ATP, with the latter two agonists producing the largest responses. In addition to ATP, [3H]PtdIns hydrolysis was induced by ADP and by 2-methylthio-ATP, indicating that P2-purinergic receptors mediate this process. Subtype-selective antibodies were used to identify G(q/11) and phospholipase C-β as the G-protein and phospholipase C subtypes that mediated GTP[S]-induced and agonist-induced [3H]PtdIns hydrolysis. These results demonstrate that this method reveals that agonist-induced, GTP[S]dependent [3H]PtdIns hydrolysis is retained in postmortem human brain membranes with properties similar to rat brain. This method should allow studies of the modulation of phosphoinositide hydrolysis in human brain and investigations of potential alterations in postmortem brain from subjects with neurological and psychiatric diseases.

Original languageEnglish (US)
Pages (from-to)655-659
Number of pages5
JournalBiochemical Journal
Volume304
Issue number2
DOIs
StatePublished - Jan 1 1994
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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