The University of St Andrews

Research@StAndrews:FullText >
Biology (School of) >
Biology >
Biology Theses >

Please use this identifier to cite or link to this item:
This item has been viewed 8 times in the last year. View Statistics

Files in This Item:

File Description SizeFormat
Peter James Calcraft PhD thesis.PDF34.61 MBAdobe PDFView/Open
Title: Two-pore channels and NAADP-dependent calcium signalling
Authors: Calcraft, Peter James
Supervisors: Evans, A. Mark
Keywords: Nicotinic acid adenine dinucleotide phosphate
Two-pore channel
Issue Date: 23-Jun-2010
Abstract: Nicotinic acid adenine dinucleotide phosphate (NAADP) is a potent Ca²⁺ mobilising messenger in mammalian and non-mammalian cells. Studies on a variety of cell types suggest that NAADP evokes Ca²⁺ release from a lysosome-related store and via activation of a receptor distinct from either ryanodine receptors (RyR) or inositol 1,4,5-trisphosphate (IP₃) receptors (IP₃R). However, the identity of the NAADP receptor has, until now, remained elusive. In this thesis I have shown that NAADP-evoked Ca²⁺ release from lysosomes is underpinned by two-pore channels (TPCs), of which there are 3 subtypes, TPC1, TPC2 and TPC3. When stably over-expressed in HEK293 cells, TPC2 was found to be specifically targeted to lysosomes, while TPC1 and TPC3 were targeted to endosomes. Initial Ca²⁺ signals via TPC2, but not those via TPC1, were amplified into global Ca²⁺ waves by Ca²⁺-induced Ca²⁺ release (CICR) from the endoplasmic reticulum (ER) via IP₃Rs. I have shown that, consistent with a role for TPCs in NAADP-mediated Ca²⁺ release, TPC2 is expressed in pulmonary arterial smooth muscle cells (PASMCs), is likely targeted to lysosomal membranes, and that TPCs also underpin NAADP-evoked Ca²⁺ signalling in this cell type. However, and in contrast to HEK293 cells, in PASMCs NAADP evokes spatially restricted Ca²⁺ bursts that are amplified into global Ca²⁺ waves by CICR from the sarcoplasmic reticulum (SR) via a subpopulation of RyRs, but not via IP₃Rs. I have demonstrated that lysosomes preferentially co-localise with RyR subtype 3 (RyR3) in the perinuclear region of PASMCs to comprise a “trigger zone” for Ca²⁺ signalling by NAADP, away from which a propagating Ca²⁺ wave may be carried by subsequent recruitment of RyR2. The identification of TPCs as a family of NAADP receptors may further our understanding of the mechanisms that confer the versatility of Ca²⁺ signalling which is required to regulate such diverse cellular functions as gene expression, fertilization, cell growth, and ultimately cell death.
Other Identifiers:
Type: Thesis
Publisher: University of St Andrews
Appears in Collections:Biology Theses

This item is protected by original copyright

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.


DSpace Software Copyright © 2002-2012  Duraspace - Feedback
For help contact: | Copyright for this page belongs to St Andrews University Library | Terms and Conditions (Cookies)