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Article: The use of cells doubly labelled with [ 14C]inositol and [ 3H]inositol to search for a hormone-sensitive inositol lipid pool with atypically rapid metabolic turnover

TitleThe use of cells doubly labelled with [ 14C]inositol and [ 3H]inositol to search for a hormone-sensitive inositol lipid pool with atypically rapid metabolic turnover
Authors
Issue Date1989
PublisherSociety for Endocrinology. The Journal's web site is located at http://joe.endocrinology-journals.org
Citation
Journal Of Endocrinology, 1989, v. 122 n. 1, p. 379-389 How to Cite?
AbstractSome, though not all, previous studies have suggested that the inositol lipid which is hydrolysed during transmembrane signalling in response to receptor activation might be drawn from a metabolically discrete and relatively small hormone-sensitive lipid pool that turns over more rapidly than the bulk of membrane inositol lipid. In order to seek evidence for the existence of this putative hormone-sensitive lipid pool, we have double-labelled cells by growing them for 3 days in a medium containing [ 14C]inositol and then supplying them with [ 3H]inositol for the final 2 h before stimulation. We anticipated that stimulation of these doubly labelled cells might provoke the formation, from the postulated hormone-sensitive pool, of small quantities of relatively 3H-enriched inositol phosphates, and that these could be harvested from cells (provided that the cytosolic inositol monophosphatase and inositol 1,4-bisphosphate/inositol 1,3,4-trisphosphate 1-phosphatase activities are first inhibited by Li +). Experiments of this type, using both vasopressin-stimulated WRK1 rat mammary tumour cells and 3T3 mouse fibroblasts stimulated by prostaglandin F(2α), have largely failed to demonstrate the formation of relatively 3H-enriched inositol phosphates. There was a tendency for phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bis-phosphate to have slightly higher 3H : 14C ratios than phosphatidylinositol, but the 3H : 14C ratios than phosphatidylinositol, but the 3H : 14C ratios of the inositol phosphates formed in stimulated cells were not substantially greater than the 3H : 14C ratios of the inositol lipids. We therefore conclude, at least for the two cell lines that we studied, that hormone-stimulated inositol lipid hydrolysis can call, either directly or indirectly, upon the majority of the inositol lipid complement of the stimulated cell.
Persistent Identifierhttp://hdl.handle.net/10722/147335
ISSN
2015 Impact Factor: 4.498
2015 SCImago Journal Rankings: 1.910
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorMaccallum, SHen_US
dc.contributor.authorBarker, CJen_US
dc.contributor.authorHunt, PAen_US
dc.contributor.authorWong, NSen_US
dc.contributor.authorKirk, CJen_US
dc.contributor.authorMichell, RHen_US
dc.date.accessioned2012-05-29T06:03:00Z-
dc.date.available2012-05-29T06:03:00Z-
dc.date.issued1989en_US
dc.identifier.citationJournal Of Endocrinology, 1989, v. 122 n. 1, p. 379-389en_US
dc.identifier.issn0022-0795en_US
dc.identifier.urihttp://hdl.handle.net/10722/147335-
dc.description.abstractSome, though not all, previous studies have suggested that the inositol lipid which is hydrolysed during transmembrane signalling in response to receptor activation might be drawn from a metabolically discrete and relatively small hormone-sensitive lipid pool that turns over more rapidly than the bulk of membrane inositol lipid. In order to seek evidence for the existence of this putative hormone-sensitive lipid pool, we have double-labelled cells by growing them for 3 days in a medium containing [ 14C]inositol and then supplying them with [ 3H]inositol for the final 2 h before stimulation. We anticipated that stimulation of these doubly labelled cells might provoke the formation, from the postulated hormone-sensitive pool, of small quantities of relatively 3H-enriched inositol phosphates, and that these could be harvested from cells (provided that the cytosolic inositol monophosphatase and inositol 1,4-bisphosphate/inositol 1,3,4-trisphosphate 1-phosphatase activities are first inhibited by Li +). Experiments of this type, using both vasopressin-stimulated WRK1 rat mammary tumour cells and 3T3 mouse fibroblasts stimulated by prostaglandin F(2α), have largely failed to demonstrate the formation of relatively 3H-enriched inositol phosphates. There was a tendency for phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bis-phosphate to have slightly higher 3H : 14C ratios than phosphatidylinositol, but the 3H : 14C ratios than phosphatidylinositol, but the 3H : 14C ratios of the inositol phosphates formed in stimulated cells were not substantially greater than the 3H : 14C ratios of the inositol lipids. We therefore conclude, at least for the two cell lines that we studied, that hormone-stimulated inositol lipid hydrolysis can call, either directly or indirectly, upon the majority of the inositol lipid complement of the stimulated cell.en_US
dc.languageengen_US
dc.publisherSociety for Endocrinology. The Journal's web site is located at http://joe.endocrinology-journals.orgen_US
dc.relation.ispartofJournal of Endocrinologyen_US
dc.subject.meshAnimalsen_US
dc.subject.meshArginine Vasopressin - Pharmacologyen_US
dc.subject.meshBradykinin - Pharmacologyen_US
dc.subject.meshCell Lineen_US
dc.subject.meshFibroblasts - Drug Effects - Metabolismen_US
dc.subject.meshInositol Phosphates - Metabolismen_US
dc.subject.meshMammary Neoplasms, Experimental - Metabolismen_US
dc.subject.meshMiceen_US
dc.subject.meshPhosphatidylinositols - Metabolismen_US
dc.subject.meshRatsen_US
dc.subject.meshTumor Cells, Cultured - Drug Effects - Metabolismen_US
dc.titleThe use of cells doubly labelled with [ 14C]inositol and [ 3H]inositol to search for a hormone-sensitive inositol lipid pool with atypically rapid metabolic turnoveren_US
dc.typeArticleen_US
dc.identifier.emailWong, NS:nswong@hkucc.hku.hken_US
dc.identifier.authorityWong, NS=rp00340en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.pmid2769159-
dc.identifier.scopuseid_2-s2.0-0024351886en_US
dc.identifier.volume122en_US
dc.identifier.issue1en_US
dc.identifier.spage379en_US
dc.identifier.epage389en_US
dc.identifier.isiWOS:A1989AE40200044-
dc.publisher.placeUnited Kingdomen_US
dc.identifier.scopusauthoridMaccallum, SH=6602909714en_US
dc.identifier.scopusauthoridBarker, CJ=7201818641en_US
dc.identifier.scopusauthoridHunt, PA=7202324475en_US
dc.identifier.scopusauthoridWong, NS=7202836641en_US
dc.identifier.scopusauthoridKirk, CJ=7102433740en_US
dc.identifier.scopusauthoridMichell, RH=7006966596en_US

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