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Proceedings of the International Workshop On INTERNATIONAL WORKSHOP 115 PROCEEDINGS OF THE INTERNATIONAL WORKSHOP ON “CALCIUM RELEASE AND CELLULAR CALCIUM SIGNALLING DOMAINS” MARBELLA, CHILE, SEPTEMBER 28 - OCTOBER 2, 2003 Organized by FONDAP Center for Molecular Studies of the Cell, Facultad de Medicina. Universidad de Chile CECILIA HIDALGO, EDITOR 116 INTERNATIONAL WORKSHOP INTERNATIONAL WORKSHOP 117 Biol Res 37: ???-???, 2004 BR INTERNATIONAL WORKSHOP “CALCIUM RELEASE AND CELLULAR CALCIUM SIGNALLING DOMAINS” MARBELLA, CHILE, SEPTEMBER 28-OCTOBER 2, 2003 PROGRAM Sunday, Sept. 28 Chair: Susan Hamilton Paul Allen (USA). Structure Function Studies of Opening lecture: Ernesto Carafoli (Italy) Calcium RyR1 signalling, a historical perspective Colin Taylor (UK). Structure and regulation of InsP3 receptors Chair: Cecilia Hidalgo Roberto Coronado (USA). Functional interactions between dihydropyridine and Monday, Sept. 29 - FIRST TOPIC: CALCIUM ryanodine receptors in skeletal muscle RELEASE CHANNELS Kurt Beam (USA). Arrangement of proteins involved in excitation-contraction coupling as Session 1: Calcium Release Channels (RyR and analyzed by FRET InsP3-gated channels). Structural aspects and Hector Valdivia (USA). Modulation of single channel studies cardiac ryanodine receptors by sorcin, a novel regulator of excitation-contraction coupling Chair: Barbara Ehrlich Humbert De Smedt (Belgium). InsP3- Clara Franzini-Armstrong (USA). The induced Ca2+ release and Calmodulin junctional supramolecular complex Susan Hamilton (USA) RYR1 Phosphorylation POSTER SESSION 2 by PKA and the role of FKBP12 Gregory Mignery (USA) Structure and Session 4: Regulation of Calcium Release Channels function of InsP3 Receptor (RyR and InsP3-gated channels), continued Ricardo Bull (Chile) Regulation of single RyR channels from brain by endogenous modulators Chair: Enrique Jaimovich Allan Williams (UK) The interaction of Sandor Gyorke (USA). SR intra-luminal Ca ryanoids with single RyR channels signalling in the heart: the role of Kevin Foskett (USA) Novel model of calcium calsequestrin and inositol 1,4,5-trisphosphate regulation of Cecilia Hidalgo (Chile) Redox regulation of InsP3 receptor channel gating in native calcium release through RyR channels endoplasmic reticulum David Yule (USA). Regulation of Ca2+ release by InsP3 receptor phosphorylation POSTER SESSION 1 Plenary lecture: Ole Petersen (UK) Local and Session 2: Calcium Release Channels: RyR, global Ca2+ signals: physiology and pathophysiology NAADP- and InsP3-gated channels. Chair: Enrique Jaimovich Chair: Kevin Foskett Michael Fill (USA) Calcium regulation of Wednesday, Oct. 1st. SECOND TOPIC: CELLULAR single ryanodine receptor channels RESPONSES TRIGGERED BY CALCIUM Barbara Ehrlich (USA). Regulation of the RELEASE InsP3 receptor from both sides Antony Galione (UK). NAADP signalling Session 5: Cellular Calcium Signals generated by mechanisms Calcium Release through RyR-channels and Eduardo Chini (USA). NAADP a new InsP3-gated channels second messenger? Chair: Kevin Foskett Tuesday Sept. 30 - Session 3: Regulation of Calcium Martin Schneider (USA). Local Ca2+ release Release Channels (RyR and InsP3-gated channels) events in skeletal muscle. 118 INTERNATIONAL WORKSHOP Eduardo Ríos (USA). Control of dual Thursday, Oct. 2nd THIRD TOPIC: CELLULAR isoforms of Ca release channels in muscle CALCIUM SIGNALLING David Eisner (UK). Control and miscontrol of the cardiac calcium transient Session 7: Cellular Calcium Signalling Domains Martin Bootman (UK) Spatial and temporal properties of calcium transients in cardiac Chair: Cecilia Hidalgo myocytes Pedro Verdugo (USA). Calcium/Proton Enrique Jaimovich (Chile). InsP3 dependent signalling in secretion: crosstalk, redundancy, calcium signals in skeletal muscle cells or complementation? Susan Wray (UK). Calcium signals and Patricia Camacho (USA). ER chaperones function in smooth muscle modulate intracellular calcium oscillations Shmuel Muallen (USA). Homer proteins in Jordi Molgó (France). Localization of InsP3 Ca2+ signaling receptors in muscle satellite cells, and components of the mouse neuromuscular junction POSTER SESSION 3 Patrick Delmas (France). Polycystin ion channel signalling complex Ian Parker (USA) Imaging single-channel Session 6: Mitochondrial and Store-Operated Ca2+ microdomains; and effects of buffers on Calcium Signals interactions between InsP3 receptors Alex Verkhratsky (UK) ER Ca2+ homeostasis Chair: Ernesto Carafoli 2+ and neuronal signalling Rosario Rizzuto (Italy). Mitochondrial Ca David Friel (USA) Interplay between ER homeostasis: molecular determinants and Ca2+ uptake and release fluxes in neurons and functional role its impact on [Ca2+] dynamics in neurons Javier Garcia-Sancho (Spain). The contribution of mitochondria to the genesis of the calcium Session 8: Nuclear Calcium Signals and Calcium- signal in excitable cells dependent Gene Expression James Putney (USA). TRPCs and mechanisms of calcium entry Chair: Ole Petersen Ricardo Boland (Argentina). Modulation of M. Angélica Carrasco (Chile) Signal transduction store operated calcium channels by the and gene expression regulated by calcium release α steroid hormone 1 ,25-dihydroxy-vitamin D3 from internal stores in excitable cells Isaac Pessah (USA). Excitation-SOC entry James Lechleiter (USA) The role of calcium coupling in skeletal myotubes in cell survival and cell death INTERNATIONAL WORKSHOP 119 ABSTRACTS AND SESSION SUMMARIES 120 INTERNATIONAL WORKSHOP OPENING LECTURE: receptors in the plasma membrane. In sarcoplasmic reticulum Ca2+ is released through a channel which is sensitive to the alkaloid ryanodine (the ryanodine receptor), CALCIUM SIGNALLING, A but probably operates also in non-muscle cells, where it is HISTORICAL PERSPECTIVE activated by another ligand, cyclic ADP ribose. The nicotinic acid derivative of NADP (NAADP) has also been 2+ ERNESTO CARAFOLI shown to discharge Ca from a store which is different 2+ Department of Biochemistry, University of Padova, from the stores sensitive to InsP3 or to cADPr. Ca 35121 Padova, Italy, and Venetian Institute of released from the reticulum creates micro domains of high Molecular Medicine (VIMM), 35129 Padova, Italy concentration, which activate the electrophoretic uptake uniporter. The uniporter is essentially inactive at the ionic 2+ When the foundations of physiology were established, Ca2+ Ca concentrations of the cytosol at rest. When a vicinal 2+ was recognised as a structural element, essentially for the Ca hotspot is generated the uniporter rapidly dissipates it. 2+ stability of bones. Unexpectedly, however, a stunning Mitochondrial Ca uptake, long considered unimportant, experiment performed by Ringer 120 years ago showed that has now come back as a powerful means to regulate cell 2+ 2+ Ca2+, was also a carrier of biological information. In the Ca , and, especially, Ca -sensitive intramitochondrial case of Ringer, the information was carried to the dehydrogenases. The system, however, also has another myofibrils of heart cells, eliciting their contraction. In the important role. Since mitochondria can also accumulate 2+ decades to follow, Ca2+ was found to transmit signals to an phosphate, they store large amounts of Ca as insoluble 2+ increasing number of cellular functions: Ca2+ is now phosphate in situations of persistent cellular Ca overload, 2+ universally recognised as the most important transducer of temporarily freeing the cytosol of excess Ca . biological signals. Mitochondria are thus vital in the protection of cells in 2+ The signalling function demands that the concentration and situations of Ca emergency, a function that stresses the 2+ the movements of free Ca2+ inside cells be carefully ambivalent role of the Ca signal, which is essential to the controlled. Ca2+ inside cells is indeed maintained at a proper functioning of cells when correctly controlled, but concentration oscillating around 10-7 M, since most targets becomes a conveyor of doom when control fails. respond to the calcium signal in this concentration range. Deviations of Ca2+ to concentrations much above this level will lead to the permanent activation of all Ca2+ targets, a situation incompatible with cell life. The maintenance of the concentration of Ca2+ in the sub-µM range is achieved through the reversible complexation to binding proteins, the best known of which are the EF-hand proteins: These proteins buffer Ca2+, however, their most important task is the processing of the information it carries. They decode the Ca2+ signal by undergoing conformational changes, at the end of which Ca2+ information is transferred to the targets. Other cellular proteins only buffer Ca2+ without processing its signal. They are intrinsic to membranes, and transport it across them. They are located in the plasma membrane and in the membrane of organelles, and belong to several classes: Channels, ATPases (pumps), Na+/ Ca2+ (H+/Ca2+) exchangers and an electrophoretic Ca2+ uniporter in the inner mitochondrial membrane. These transporters function with different Ca2+ affinities and total transport capacities, thus satisfying the various Ca2+ homeostasis demands of cells. Channels, mediate the influx of Ca2+ into cells, and are gated by voltage changes, by specific ligands, or by the emptying of cellular Ca2+ stores. Pumps operate with high Ca2+ affinity in the plasma membrane (PMCA), in the endo(sarco)plasmic reticulum(SERCA) and in the Golgi membranes (HMCA). The pumps are the products of multigene families: Of the four basic isoforms of the PMCA, two are ubiquitously distributed, two are restricted to
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