title: Preparation and use of sea urchin egg homogenates for studying NAADP-mediated Ca²⁺ release.
abstract: NAADP and other Ca(2+)-mobilizing messengers are membrane impermeant and thus must be added directly to cell-free or broken-cell preparations to effect Ca(2+) release. The sea urchin egg homogenate, where the biological activity of NAADP was first reported, remains the gold standard cell-free system for studying NAADP-mediated Ca(2+) release. Here we describe how to prepare sea urchin egg homogenate and use it to measure NAADP-mediated Ca(2+) release.
authors.affiliation: Department of Pharmacology, University of Oxford, Oxford OX1 3QT, United Kingdom.
authors.forename: Antony
authors.initials: A
authors.lastname: Galione
authors.affiliation: Department of Pharmacology, University of Oxford, Oxford OX1 3QT, United Kingdom.
authors.forename: Kai-Ting
authors.initials: KT
authors.lastname: Chuang
authors.affiliation: Department of Pharmacology, University of Oxford, Oxford OX1 3QT, United Kingdom.
authors.forename: Tim M
authors.initials: TM
authors.lastname: Funnell
authors.affiliation: Department of Pharmacology, University of Oxford, Oxford OX1 3QT, United Kingdom.
authors.forename: Lianne C
authors.initials: LC
authors.lastname: Davis
authors.affiliation: Department of Pharmacology, University of Oxford, Oxford OX1 3QT, United Kingdom.
authors.forename: Anthony J
authors.initials: AJ
authors.lastname: Morgan
authors.affiliation: Department of Pharmacology, University of Oxford, Oxford OX1 3QT, United Kingdom.
authors.forename: Margarida
authors.initials: M
authors.lastname: Ruas
authors.affiliation: Department of Pharmacology, University of Oxford, Oxford OX1 3QT, United Kingdom.
authors.forename: John
authors.initials: J
authors.lastname: Parrington
authors.affiliation: Department of Pharmacology, University of Oxford, Oxford OX1 3QT, United Kingdom.
authors.forename: Grant C
authors.initials: GC
authors.lastname: Churchill
mesh_terms: D000255:Adenosine Triphosphate / Q000494:pharmacology; D000814:Aniline Compounds; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D020013:Calcium Signaling / Q000187:drug effects / Q000502:physiology; D002458:Cell Fractionation / Q000379:methods*; D005260:Female; D015544:Inositol 1,4,5-Trisphosphate / Q000494:pharmacology; D008297:Male; D009249:NADP / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology; D010063:Ovum / Q000187:drug effects* / Q000378:metabolism* / Q000648:ultrastructure; D012617:Sea Urchins; D014966:Xanthenes
chemical_list: D000814:Aniline Compounds; D014966:Xanthenes; C059715:Fluo-3; D009249:NADP; C024376:NAADP; D015544:Inositol 1,4,5-Trisphosphate; D000255:Adenosine Triphosphate; D002118:Calcium