Calcium Signalling Laboratory

Confocal images of the plasma membrane of liver cells transfected with a GFP-tagged plasma membrane reporter

Signals conveyed from one part of the body to another, require the release of chemical messengers (hormones and neurotransmitters) from one set of cells and their action on target cells. The processes by which hormones and neurotransmitters act on target cells require increases in the concentration of calcium ions in the target cell. This is often achieved by calcium entering the cell through channels in the plasma membrane. The aims of this research are to study the physiological functions of these calcium channels, their structures and mechanisms of opening, and how these channels are altered in diseased states.

Investigators

Greg Barritt, BSc(Hons), PhD

Visiting Research Scholars

Carlijn Buis (University Medical Centre, Groningen, Netherlands), Visiting Research Scholar
Alwine Kist (University Medical Centre, Groningen, Netherlands), Visiting Research Scholar
Marije van der Steen (University Medical Centre, Groningen, Netherlands), Visiting Research Scholar
Joris Wakkie (University Medical Centre, Groningen, Netherlands), Visiting Research Scholar
Susanne Zeile ( University of Mannheim, Germany), Visiting Research Scholar

Support Staff

Claire Wilson, BSc(Hons)

Students

Yit-Lung Khung, PhD student
Brian Tam, PhD student
Yabin Zhou, PhD student

Contacts

Prof Greg Barritt

Ph: (08) 8204 4260 or (08) 8204 4419 Int Ph: +61 8 8204 4260 or +61 8 8204 4419 Fax: (08) 8374 0139 Int Fax: +61 8 8374 0139 Email: greg.barritt@flinders.edu.au

Research Projects

• Receptor-activated calcium channels
  There is now considerable evidence that most cells possess receptor-activated Ca 2+ channels as well as voltage-operated Ca 2+ channels. However, the structures and mechanisms of activation of receptor-activated Ca 2+ channels are not well understood. Recently it has been shown that store-operated Ca 2+ channels (STIM1 and Orai1) and members of the TRP (transient receptor potential) channel family are important receptor-activated Ca 2+ channels in animal cells. Mutations in TRP channels may be responsible for some forms of photoreceptor cell and neuronal cell degeneration. The aims of this research are to (i) study the roles of STIM and Orai proteins in store-operated Ca 2+ channels and (ii) to identify TRP channels in non-excitable and excitable cells. Messenger RNA encoding subtypes of TRP channels have been identified in using RT-PCR techniques, and some of the TRP proteins have been detected by (a) using specific antibodies directed against the proteins and (b) functionally. Experiments are presently under-way to further elucidate the physiological functions of these TRP channels.

Collaborative Research

Dr Andy Miller and Dr Sarah Ho, The Hong Kong University of Science & Technology: Calcium signalling in zebra fish liver.

Dr Grigori Rychkov, Dr Michael Roberts and Dr Tom Litjens, University of Adelaide: Store-operated calcium channels.

Prof Natalia Prevarskaya, INSERM, Cedex, France: TRP Channels.

Dr Vincent Nieuwenhuijs and Prof Robert Porte, University Medical Centre, Groningen, Holland: Ischemic reperfusion injury.

Media Appearances

• ABC Local Radio, (2005) Radio interview
• 5AA, (2005) Radio interview
• Channel 7, (2005) Television interview

Publications

Barritt GJ, Chen J and Rychkov GY (2008) Ca 2+-permeable channels in the hepatocyte plasma membrane and their roles in hepatocyte physiology. Biochem Biophys Acta - Mol Cell Res, 1783, 651-672

Aromataris EC, Castro J, Rychkov GY and Barritt GJ (2008) Store-operated Ca 2+ channels and stromal interaction molecular 1 (STIM1) are targets for the actions of bile acids on liver cells. Biochem Biophys Acta - Mol Cell Res, 1783, 874-885

Khung YL, Barritt G and Voelcker NH (2008) Using continuous porous silicon gradients to study the influence of surface topography on the behaviour of neuroblastoma cells. Exper Cell Res, 314, 789-800

Beck B, Bavencoffe A, Bidaux G, Lemonnier L, Thebault S, Shuba Y, Barritt G, Skryma R and Prevarskaya N (2007) Prospects for prostate cancer imaging and therapy using high-affinity TRPM8 activators. Cell Calcium 41:285-94

Nieuwenhuijs VB, de Bruijn MT, Schiesser M, Morphett A, Padbury RT, Barritt GJ (2007) Ischemic preconditioning and intermittent ischemia preserve bile flow in a rat model of ischemia reperfusion injury. Dig Dis Sci. 52(5):1159-67

Litjens T, Nguyen T, Castro J, Aromataris EC, Jones L, Barritt GJ, Rychkov GY (2007) Phospholipase C-gamma1 is required for the activation of store-operated Ca2+ channels in liver cells. Biochem J. 405(2):269-76

Rychkov G, Barritt GJ (2007) TRPC1 Ca(2+)-permeable channels in animal cells. Handb Exp Pharmacol. (179):23-52. Review

Prevarskaya N, Zhang L, Barritt G (2007) TRP channels in cancer. Biochem Biophys Acta 1772:937-46

Aromartis EC, Roberts ML, Barritt GJ and Rychkov GY (2006) Glucagon activates Ca 2+ and CI - channels in rat hepatocytes. J Physiol, 573, 611-625

Nieuwenhuijs VB, de Bruijn MT, Padbury RTA and Barritt GJ (2006) Hepatic ischaemia reperfusion injury: roles of Ca 2+ and other intracellular mediators of impaired bile flow and hepatocyte damage. Digest Dis Sci, 51, 1087-1102

de Bruijn MT, Nieuwenhuijs VB, Padbury RTA, Hughes R, Stahl J and Barritt GJ (2006) The effects of ischaemic preconditioning and intermittent ischaemia on bile flow in rats. ANZ J Surg. 75(Suppl), A73

Zhang L and Barritt GJ (2006) TRPM8 in prostate cancer cells: a potential diagnostic and prognostic marker with a secretory function? Endocrine-Related Cancer, 13, 27-38

Rychkov GY, Litjens T, Roberts ML, Barritt GJ (2005) Arachidonic acid inhibits the store-operated Ca2+ current in rat liver cells. Biochem J. 385(Pt 2):551-6

Rychkov GY, Litjens T, Roberts ML, Barritt GJ (2005) ATP and vasopressin activate a single type of store-operated Ca2+ channel, identified by patch-clamp recording, in rat hepatocytes. Cell Calcium. 37(2):183-91

de Bruijn MT, Nieuwenhuijs VB, Padbury RTA, Hughes R, Stahl J and Barritt GJ (2005) The effects of ischaemic preconditioning and intermittent ischaemia on bile flow in rats. ANZ J. Surg. 75(Suppl), A73

Ong HL, Barritt GJ (2005) Transient receptor potential and other ion channels as pharmaceutical targets in airway smooth muscle cells. Respirology. 9(4):448-57. Review

Barritt G and Rychkov G (2005) TRPs as mechanosensitive channels. Nature Cell Biology 7, 105-7

Ruengrairatanaroje P, Hahn M, Brereton HM, Zhang L, Froscio M, Petronijevic T and Barritt GJ (2004) Development of a tetracycline-inducible system for expression of the Ca2+ permeable TRPL channel and the killing of prostate cancer cells. Cancer Therapy 2, 389-402

Chan C, Harland ML, Webb SE, Chen J, Miller AL and Barritt GJ (2004) Evaluation, using targeted aequorins, of the roles of the endoplasmic reticulum and its (Ca2+ + Mg2+) ADP-ases in the activation of store-operated Ca2+ channels in liver cells. Cell Calcium 35, 317-331

Zhang L. and Barritt GJ (2004) Evidence that TRPM8 is an androgen-dependent Ca2+ channel required for the survival of prostate cancer cells. Cancer Res. 64, 8365-8373

Litjens T, Harland ML, Roberts ML, Barritt GJ and Rychkov G (2004) Fast Ca2+-dependent inactivation of the store-operated Ca2+ current (ISOC) in liver cells: a role for calmodulin. J. Physiol. 558.1, 85-97

Gregory RB, Hughes R and Barritt GJ (2004) Induction of cholestasis in the perfused rat liver by 2-aminoethyl diphenylborate, an inhibitor of the hepatocyte plasma membrane Ca2+ channels. J. Gastroenterol. Hepatol. 19, 1128-1134

Gregory RB, Hughes R, Riley AM, Potter BVL., Wilcox RA and Barritt GJ (2004) Inositol trisphosphate analogues selective for types I and II inositol trisphosphate receptors exert differential effects on vasopressin-stimulated Ca2+ inflow and Ca2+ release from intracellular stores in rat hepatocytes. Biochem. J. 381, 519-526

Chong MP, Barritt GJ and Crouch MF (2004) Insulin potentiates EGFR activation and signaling in fibroblasts. Biochem. Biophys. Res. Commun. 322, 535-541

Ong HL and Barritt GJ (2004) Transient receptor potential (TRP) and other ion channels as pharmaceutical targets in airway smooth muscle cells. Respirology 9, 448-457

Ong HL, Brereton HM, Harland ML, Barritt GJ (2003) Evidence for the expression of transient receptor potential proteins in guinea pig airway smooth muscle cells. Respirology 8(1):23-32

Gregory RB, Barritt GJ (2003) Evidence that Ca2+-release-activated Ca2+ channels in rat hepatocytes are required for the maintenance of hormone-induced Ca2+ oscillations. Biochem J. 369: 1-7

Gregory RB, Sykiotis D, Barritt GJ (2003) Evidence that store-operated Ca2+ channels are more effective than intracellular messenger-activated non-selective cation channels in refilling rat hepatocyte intracellular Ca2+ stores. Cell Calcium 34(3):241-51

Chen J, Barritt GJ (2003) Evidence that TRPC1(transient receptor potential canonical 1) forms a Ca(2+)-permeable channel linked to the regulation of cell volume in liver cells obtained using small interfering RNA targeted against TRPC1. Biochem J 373: 327-36

Zhang L, Brereton HM, Hahn M, Froscio M, Tilley WD, Brown MP, Barritt GJ (2003) Expression of Drosophila Ca2+ permeable transient receptor potential-like channel protein in a prostate cancer cell line decreases cell survival. Cancer Gene Ther 10(8):611-25

Wang Y-J, Gregory RB and Barritt GJ (2002) Maintenance of the filamentious actin cytoskeleton is necessary for the activation of store-operated Ca2+ channels but not other types of plasma membrane Ca2+ channels in rat hepatocytes. Biochem. J. 363, 117-126

Ong HL, Chen J, Chataway T, Brereton H, Zhang L, Downs T, Tsiokas L and Barritt GJ (2002) Specific detection of the endogenous transient receptor potential (TRP)-1 protein in liver and airway smooth muscle cells using immunoprecipitation and Western blot analysis. Biochem. J. 364, 641-648

Tesfai Y, Brereton HM and Barritt GJ (2001) A diacylglycerol-activated Ca2+ channel in PC-12 cells (an adrenal chromaffin cell line) correlates with expression of the TRP-6 (transient receptor potential) protein. Biochem. J. 358, 717-726

Gregory RB, Rychkov G, Barritt GJ (2001) Evidence that 2-aminoethyl diphenylborate is a novel inhibitor of store operated Ca2+ channels in liver cells, and acts through a mechanism which does not involve inositol trisphosphate receptors. Biochem J 1;354(Pt2): 285-90

Brereton HM, Chen J, Rychkov G, Harland ML and Barritt GJ (2001) Maitotoxin activates an endogenous non-selective cation channel and is an effective initiator of the activation of the heterologously expressed hTRPC-1 (transient receptor potential) non-selective cation channel in H4-IIE liver cells. Biochim. Biophys. Acta. 1540, 107-126

Rychkov G, Brereton HM, Harland ML, Barritt GJ (2001) Plasma membrane Ca2+ release-activated Ca2+ channels with a high selectivity for Ca2+ identified by patch-clamp recording in rat liver cells. Hepatology 33, 938-947

Barritt GJ (2000) Calcium signalling in liver cells. Calcium: The Molecular Basis of Calcium Action in Biology and Medicine (Pochet, R., Donato, R., Haiech, J., Heizmann, CW, Gerke, V., eds.), Kluwer Academic Publishers, Netherlands, pp. 73-94

Brereton HM, Harland ML, Auld A and Barritt GJ (2000) Evidence that the TRP-1 protein is unlikely to account for store-operated Ca2+ inflow in Xenopus laevis oocytes. Molecular and Cellular Biochemistry 214: 63-74

Wang Y, Gregory RB and Barritt GJ (2000) Regulation of F-actin and endoplasmic reticulum organisation by the trimeric G-protein Gi2 in rat hepatocytes: Implication for the activation of store-operated Ca2+ inflow. Journal of Biological Chemistry 275: 22229-22237

Auld A, Chen J, Brereton HM, Wang Y-J, Gregory RB and Barritt GJ (2000) Store-operated Ca2+ inflow in Reuber hepatoma cells is inhibited by voltage-operated Ca2+ channel antagonists, and in contrast to freshly-isolated hepatocytes does not require a pertussis toxin-sensitive trimeric GTP-binding protein. Biochimica et Biophysica Acta 1497: 11-26

Updated December 15, 2009