HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Bakowski, D. Articles by Parekh, A. B. Search for Related Content PubMed PubMed Citation Articles by Bakowski, D. Articles by Parekh, A. B.

An examination of the secretion-like coupling model for the activation of the Ca²⁺ release-activated Ca²⁺ current I_CRAC in RBL-1 cells

Daniel Bakowski, Maike D. Glitsch and Anant B. Parekh

1. One popular model for the activation of store-operated Ca²⁺ influx is the secretion-like coupling mechanism, in which peripheral endoplasmic reticulum moves to the plasma membrane upon store depletion thereby enabling inositol 1,4,5-trisphosphate (InsP₃) receptors on the stores to bind to, and thus activate, store-operated Ca²⁺ channels. This movement is regulated by the underlying cytoskeleton. We have examined the validity of this mechanism for the activation of I_CRAC, the most widely distributed and best characterised store-operated Ca²⁺ current, in a model system, the RBL-1 rat basophilic cell line.

2. Stabilisation of the peripheral cytoskeleton, disassembly of actin microfilaments and disaggregation of microtubules all consistently failed to alter the rate or extent of activation of I_CRAC. Rhodamine-phalloidin labelling was used wherever possible, and revealed that the cytoskeleton had been significantly modified by drug treatment.

3. Interference with the cytoskeleton also failed to affect the intracellular calcium signal that occurred when external calcium was re-admitted to cells in which the calcium stores had been previously depleted by exposure to thapsigargin/ionomycin in calcium-free external solution.

4. Application of positive pressure through the patch pipette separated the plasma membrane from underlying structures (cell ballooning). However, I_CRAC was unaffected irrespective of whether cell ballooning occurred before or after depletion of stores.

5. Pre-treatment with the membrane-permeable InsP₃ receptor antagonist 2-APB blocked the activation of I_CRAC. However, intracellular dialysis with 2-APB failed to prevent I_CRAC from activating, even at higher concentrations than those used extracellularly to achieve full block. Local application of 2-APB, once I_CRAC had been activated, resulted in a rapid loss of the current at a rate similar to that seen with the rapid channel blocker La³⁺.

6. Studies with the more conventional InsP₃ receptor antagonist heparin revealed that occupation of the intracellular InsP₃-sensitive receptors was not necessary for the activation or maintenance of I_CRAC. Similarly, the InsP₃ receptor inhibitor caffeine failed to alter the rate or extent of activation of I_CRAC. Exposure to Li⁺, which reduces InsP₃ levels by interfering with inositol monophosphatase, also failed to alter I_CRAC. Caffeine and Li⁺ did not affect the size of the intracellular Ca²⁺ signal that arose when external Ca²⁺ was re-admitted to cells which had been pre-exposed to thapsigargin/ionomycin in Ca²⁺-free external solution.

7. Our findings demonstrate that the cytoskeleton does not seem to regulate calcium influx and that functional InsP₃ receptors are not required for activation of I_CRAC. If the secretion-like coupling model indeed accounts for the activation of I_CRAC in RBL-1 cells, then it needs to be revised significantly. Possible modifications to the model are discussed.

This article has been cited by other articles:

				W. I. DeHaven, J. T. Smyth, R. R. Boyles, G. S. Bird, and J. W. Putney Jr. Complex Actions of 2-Aminoethyldiphenyl Borate on Store-operated Calcium Entry J. Biol. Chem., July 11, 2008; 283(28): 19265 - 19273. [Abstract] [Full Text] [PDF]

				C. Peinelt, A. Lis, A. Beck, A. Fleig, and R. Penner 2-Aminoethoxydiphenyl borate directly facilitates and indirectly inhibits STIM1-dependent gating of CRAC channels J. Physiol., July 1, 2008; 586(13): 3061 - 3073. [Abstract] [Full Text] [PDF]

				V. M. Bolotina Orai, STIM1 and iPLA2{beta}: a view from a different perspective J. Physiol., July 1, 2008; 586(13): 3035 - 3042. [Abstract] [Full Text] [PDF]

				W.-C. Chang, J. Di Capite, K. Singaravelu, C. Nelson, V. Halse, and A. B. Parekh Local Ca2+ Influx through Ca2+ Release-activated Ca2+ (CRAC) Channels Stimulates Production of an Intracellular Messenger and an Intercellular Pro-inflammatory Signal J. Biol. Chem., February 22, 2008; 283(8): 4622 - 4631. [Abstract] [Full Text] [PDF]

				J. T. Smyth, W. I. DeHaven, G. S. Bird, and J. W. Putney Jr Role of the microtubule cytoskeleton in the function of the store-operated Ca2+ channel activator STIM1 J. Cell Sci., November 1, 2007; 120(21): 3762 - 3771. [Abstract] [Full Text] [PDF]

				J. W. Putney Jr New molecular players in capacitative Ca2+ entry J. Cell Sci., June 15, 2007; 120(12): 1959 - 1965. [Abstract] [Full Text] [PDF]

				Y. Baba, K. Hayashi, Y. Fujii, A. Mizushima, H. Watarai, M. Wakamori, T. Numaga, Y. Mori, M. Iino, M. Hikida, et al. Coupling of STIM1 to store-operated Ca2+ entry through its constitutive and inducible movement in the endoplasmic reticulum PNAS, November 7, 2006; 103(45): 16704 - 16709. [Abstract] [Full Text] [PDF]

				K. Singaravelu, C. Lohr, and J. W. Deitmer Regulation of store-operated calcium entry by calcium-independent phospholipase A2 in rat cerebellar astrocytes. J. Neurosci., September 13, 2006; 26(37): 9579 - 9592. [Abstract] [Full Text] [PDF]

				M. M. Wu, J. Buchanan, R. M. Luik, and R. S. Lewis Ca2+ store depletion causes STIM1 to accumulate in ER regions closely associated with the plasma membrane J. Cell Biol., September 11, 2006; 174(6): 803 - 813. [Abstract] [Full Text] [PDF]

				Y. Dobrydneva, C. J. Abelt, B. Dovel, C. M. Thadigiri, R. L. Williams, and P. F. Blackmore 2-Aminoethoxydiphenyl Borate as a Prototype Drug for a Group of Structurally Related Calcium Channel Blockers in Human Platelets Mol. Pharmacol., January 1, 2006; 69(1): 247 - 256. [Abstract] [Full Text] [PDF]

				J.-P. Lievremont, G. S. Bird, and J. W. Putney Jr. Mechanism of Inhibition of TRPC Cation Channels by 2-Aminoethoxydiphenylborane Mol. Pharmacol., September 1, 2005; 68(3): 758 - 762. [Abstract] [Full Text] [PDF]

				A. B. Parekh and J. W. Putney Jr. Store-Operated Calcium Channels Physiol Rev, April 1, 2005; 85(2): 757 - 810. [Abstract] [Full Text] [PDF]

				B. Moreau, S. Straube, R. J. Fisher, J. W. Putney Jr., and A. B. Parekh Ca2+-Calmodulin-dependent Facilitation and Ca2+ Inactivation of Ca2+ Release-activated Ca2+ Channels J. Biol. Chem., March 11, 2005; 280(10): 8776 - 8783. [Abstract] [Full Text] [PDF]

				J. A. Rosado, P. C. Redondo, G. M. Salido, S. O. Sage, and J. A. Pariente Cleavage of SNAP-25 and VAMP-2 impairs store-operated Ca2+ entry in mouse pancreatic acinar cells Am J Physiol Cell Physiol, January 1, 2005; 288(1): C214 - C221. [Abstract] [Full Text] [PDF]

				V. M. Bolotina Store-Operated Channels: Diversity and Activation Mechanisms Sci. Signal., July 27, 2004; 2004(243): pe34 - pe34. [Abstract] [Full Text] [PDF]

				W.-C. Chang and A. B. Parekh Close Functional Coupling between Ca2+ Release-activated Ca2+ Channels, Arachidonic Acid Release, and Leukotriene C4 Secretion J. Biol. Chem., July 16, 2004; 279(29): 29994 - 29999. [Abstract] [Full Text] [PDF]

				P. C. Redondo, A. G. S. Harper, G. M. Salido, J. A. Pariente, S. O. Sage, and J. A. Rosado A role for SNAP-25 but not VAMPs in store-mediated Ca2+ entry in human platelets J. Physiol., July 1, 2004; 558(1): 99 - 109. [Abstract] [Full Text] [PDF]

				N. Cheshenko, B. Del Rosario, C. Woda, D. Marcellino, L. M. Satlin, and B. C. Herold Herpes simplex virus triggers activation of calcium-signaling pathways J. Cell Biol., October 27, 2003; 163(2): 283 - 293. [Abstract] [Full Text] [PDF]

				J. V. Gerasimenko, Y. Maruyama, K. Yano, N. J. Dolman, A. V. Tepikin, O. H. Petersen, and O. V. Gerasimenko NAADP mobilizes Ca2+ from a thapsigargin-sensitive store in the nuclear envelope by activating ryanodine receptors J. Cell Biol., October 27, 2003; 163(2): 271 - 282. [Abstract] [Full Text] [PDF]

				C. C. Scott, W. Furuya, W. S. Trimble, and S. Grinstein Activation of Store-operated Calcium Channels: ASSESSMENT OF THE ROLE OF SNARE-MEDIATED VESICULAR TRANSPORT J. Biol. Chem., August 15, 2003; 278(33): 30534 - 30539. [Abstract] [Full Text] [PDF]

				F.-J. Braun, O. Aziz, and J. W. Putney Jr. 2-Aminoethoxydiphenyl Borane Activates a Novel Calcium-Permeable Cation Channel Mol. Pharmacol., June 1, 2003; 63(6): 1304 - 1311. [Abstract] [Full Text] [PDF]

				M. Trebak, G. St. J. Bird, R. R. McKay, L. Birnbaumer, and J. W. Putney Jr. Signaling Mechanism for Receptor-activated Canonical Transient Receptor Potential 3 (TRPC3) Channels J. Biol. Chem., April 25, 2003; 278(18): 16244 - 16252. [Abstract] [Full Text] [PDF]

				O. Mignen, J. L. Thompson, and T. J. Shuttleworth Ca2+ Selectivity and Fatty Acid Specificity of the Noncapacitative, Arachidonate-regulated Ca2+ (ARC) Channels J. Biol. Chem., March 14, 2003; 278(12): 10174 - 10181. [Abstract] [Full Text] [PDF]

				Z. Su, D. S. Barker, P. Csutora, T. Chang, R. L. Shoemaker, R. B. Marchase, and J. E. Blalock Regulation of Ca2+ release-activated Ca2+ channels by INAD and Ca2+ influx factor Am J Physiol Cell Physiol, February 1, 2003; 284(2): C497 - C505. [Abstract] [Full Text] [PDF]

				M. D. BOOTMAN, T. J. COLLINS, L. MACKENZIE, H. L. RODERICK, M. J. BERRIDGE, and C. M. PEPPIATT 2-Aminoethoxydiphenyl borate (2-APB) is a reliable blocker of store-operated Ca2+ entry but an inconsistent inhibitor of InsP3-induced Ca2+ release FASEB J, August 1, 2002; 16(10): 1145 - 1150. [Abstract] [Full Text] [PDF]

				M. C. Ashby, M. Craske, M. K. Park, O. V. Gerasimenko, R. D. Burgoyne, O. H. Petersen, and A. V. Tepikin Localized Ca2+ uncaging reveals polarized distribution of Ca2+-sensitive Ca2+ release sites: mechanism of unidirectional Ca2+ waves J. Cell Biol., July 22, 2002; 158(2): 283 - 292. [Abstract] [Full Text] [PDF]

				R. Schindl, H. Kahr, I. Graz, K. Groschner, and C. Romanin Store Depletion-activated CaT1 Currents in Rat Basophilic Leukemia Mast Cells Are Inhibited by 2-Aminoethoxydiphenyl Borate. EVIDENCE FOR A REGULATORY COMPONENT THAT CONTROLS ACTIVATION OF BOTH CaT1 AND CRAC (Ca2+ RELEASE-ACTIVATED Ca2+ CHANNEL) CHANNELS J. Biol. Chem., July 19, 2002; 277(30): 26950 - 26958. [Abstract] [Full Text] [PDF]

				Q. Xie, Y. Zhang, C. Zhai, and J. A. Bonanno Calcium Influx Factor from Cytochrome P-450 Metabolism and Secretion-like Coupling Mechanisms for Capacitative Calcium Entry in Corneal Endothelial Cells J. Biol. Chem., May 3, 2002; 277(19): 16559 - 16566. [Abstract] [Full Text] [PDF]

				G. Kilic and J. G. Fitz Heterotrimeric G-proteins Activate Cl- Channels through Stimulation of a Cyclooxygenase-dependent Pathway in a Model Liver Cell Line J. Biol. Chem., March 29, 2002; 277(14): 11721 - 11727. [Abstract] [Full Text] [PDF]

				T. Voets, J. Prenen, A. Fleig, R. Vennekens, H. Watanabe, J. G. J. Hoenderop, R. J. M. Bindels, G. Droogmans, R. Penner, and B. Nilius CaT1 and the Calcium Release-activated Calcium Channel Manifest Distinct Pore Properties J. Biol. Chem., December 14, 2001; 276(51): 47767 - 47770. [Abstract] [Full Text] [PDF]

				T. Lockwich, B. B. Singh, X. Liu, and I. S. Ambudkar Stabilization of Cortical Actin Induces Internalization of Transient Receptor Potential 3 (Trp3)-associated Caveolar Ca2+ Signaling Complex and Loss of Ca2+ Influx without Disruption of Trp3-Inositol Trisphosphate Receptor Association J. Biol. Chem., November 2, 2001; 276(45): 42401 - 42408. [Abstract] [Full Text] [PDF]

				M. Hershfinkel, A. Moran, N. Grossman, and I. Sekler A zinc-sensing receptor triggers the release of intracellular Ca2+ and regulates ion transport PNAS, September 25, 2001; 98(20): 11749 - 11754. [Abstract] [Full Text] [PDF]

				M. C. Hermosura, M. K. Monteilh-Zoller, A. M. Scharenberg, R. Penner, and A. Fleig Dissociation of the store-operated calcium current ICRAC and the Mg-nucleotide-regulated metal ion current MagNuM J. Physiol., March 1, 2002; 539(2): 445 - 458. [Abstract] [Full Text] [PDF]