Probing Ca2+ release mechanisms using sea urchin egg homogenates

Methods Cell Biol. 2019:151:445-458. doi: 10.1016/bs.mcb.2018.10.007. Epub 2019 Feb 21.

Abstract

Sea urchin eggs have been extensively used to study Ca2+ release through intracellular Ca2+-permeable channels. Their amenability to homogenization yields a robust, cell-free preparation that was central to establishing the Ca2+ mobilizing actions of cyclic ADP-ribose and NAADP. Egg homogenates have continued to provide insight into the basic properties and pharmacology of intracellular Ca2+ release channels. In this chapter, we describe methods for the preparation of egg homogenates and monitoring Ca2+ release using fluorimetry and radiotracer flux.

Keywords: (45)Ca flux; Ca(2+); Cyclic-ADP ribose; Fluorimetry; Inositol trisphosphate; NAADP.

Publication types

  • Review

MeSH terms

  • Animals
  • Calcium / chemistry
  • Calcium / metabolism*
  • Calcium Signaling / drug effects
  • Cell-Free System
  • Cyclic ADP-Ribose / chemistry
  • Cyclic ADP-Ribose / metabolism*
  • Kinetics
  • NADP / analogs & derivatives*
  • NADP / chemistry
  • NADP / metabolism
  • Ovum / chemistry
  • Proteins / chemistry
  • Proteins / pharmacology
  • Sea Urchins / growth & development
  • Sea Urchins / metabolism*

Substances

  • Proteins
  • Cyclic ADP-Ribose
  • NADP
  • NAADP
  • Calcium