Endogenous phosphoinositide precursors of inositol phosphates in rat brain cortical membranes

Enrique Claro, Elisabet Sarri, Fernando Picatoste

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Abstract

The appearance of Ins1P as an index of direct PtdIns breakdown by phospholipase C was examined in rat brain cortical membranes using either exogenous [3H]PtdIns substrate or [3H]inositol-prelabeled endogenous phosphoinositide substrates. Production of [3H]Ins1P was observed using exogenous [3H]PtdIns but not with endogenous substrate over the physiological range of calcium concentrations. [3H]Ins1,4P2 and [3H]Ins4P, derived from phospholipase C breakdown of polyphosphoinositides, were formed by membranes from both exogenous [3H]PtdIns and endogenous 3H-phosphoinositides, in the presence of ATP. The contribution of endogenous PtdInsP2 and PtdInsP to the generation of inositol phosphates was examined in membranes from [3H]inositol-prelabeled brain slices by adding unlabeled Ins1,4,5P3 to trap [3H]Ins1,4,5P3 generated upon breakdown of [3H]PtdInsP2. The maximal rate of [3H]Ins1,4,5P3 appearance was attained in the presence of 150-200 μM added Ins1,4,5P3 and represented 12.5% of the combined rates of formation of [3H]Ins1,4,5P3 and [3H]Ins1,4P2, similar to the content of [3H]PtdInsP2 relative to total 3H-polyphosphoinositides. The results show that, while endogenous PtdIns is not degraded by phospholipase C, the enzyme appears to be equally effective to cleave endogenous PtdInsP2 and PtdInsP. © 1993 Academic Press, Inc.
Original languageEnglish
Pages (from-to)1061-1067
JournalBiochemical and Biophysical Research Communications
Volume193
DOIs
Publication statusPublished - 1 Jan 1993

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    Claro, E., Sarri, E., & Picatoste, F. (1993). Endogenous phosphoinositide precursors of inositol phosphates in rat brain cortical membranes. Biochemical and Biophysical Research Communications, 193, 1061-1067. https://doi.org/10.1006/bbrc.1993.1733