Two-photon permeabilization and calcium measurements in cellular organelles

Gerasimenko, Oleg Vsevolodovich ORCID: https://orcid.org/0000-0003-2573-8258 and Gerasimenko, Julia Vladimirovna ORCID: https://orcid.org/0000-0002-2262-2543 2010. Two-photon permeabilization and calcium measurements in cellular organelles. Live Cell Imaging: Methods and Protocols, Vol. 591. Methods in Molecular Biology, vol. 2. Springer, pp. 201-210. (10.1007/978-1-60761-404-3_12)

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Abstract

Inositol trisphosphate and cyclic ADP-ribose, main intracellular Ca2+ messengers, induce release from the intracellular Ca2+ stores via inositol trisphosphate and ryanodine receptors, respectively. Recently, studies using novel messenger nicotinic acid adenine dinucleotide phosphate (NAADP) releasing Ca2+ from calcium stores in organelles other than endoplasmic reticulum (ER) have been conducted. However, technical difficulties of Ca2+ measurements in relatively small Ca2+ stores prompted us to develop a new, more sensitive, and less damaging two-photon permeabilization technique. Applied to pancreatic acinar cells, this technique allowed us to show that all three messengers – IP3, cADPR, and NAADP – release Ca2+ from two intracellular stores: the endoplasmic reticulum and an acidic store in the granular region. This chapter describes a detailed procedure of using this technique with pancreatic acinar cells.

Item Type:	Book Section
Date Type:	Publication
Status:	Published
Schools:	Biosciences
Subjects:	Q Science > Q Science (General)
Publisher:	Springer
ISBN:	978160761403-6
ISSN:	1064-3745
Last Modified:	20 Oct 2022 07:50
URI:	https://orca.cardiff.ac.uk/id/eprint/26568

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