Tags

Type your tag names separated by a space and hit enter

Stim1 and Orai1 mediate CRAC currents and store-operated calcium entry important for endothelial cell proliferation.
Circ Res. 2008 Nov 21; 103(11):1289-99.CircR

Abstract

Recent breakthroughs in the store-operated calcium (Ca(2+)) entry (SOCE) pathway have identified Stim1 as the endoplasmic reticulum Ca(2+) sensor and Orai1 as the pore forming subunit of the highly Ca(2+)-selective CRAC channel expressed in hematopoietic cells. Previous studies, however, have suggested that endothelial cell (EC) SOCE is mediated by the nonselective canonical transient receptor potential channel (TRPC) family, TRPC1 or TRPC4. Here, we show that passive store depletion by thapsigargin or receptor activation by either thrombin or the vascular endothelial growth factor activates the same pathway in primary ECs with classical SOCE pharmacological features. ECs possess the archetypical Ca(2+) release-activated Ca(2+) current (I(CRAC)), albeit of a very small amplitude. Using a maneuver that amplifies currents in divalent-free bath solutions, we show that EC CRAC has similar characteristics to that recorded from rat basophilic leukemia cells, namely a similar time course of activation, sensitivity to 2-aminoethoxydiphenyl borate, and low concentrations of lanthanides, and large Na(+) currents displaying the typical depotentiation. RNA silencing of either Stim1 or Orai1 essentially abolished SOCE and I(CRAC) in ECs, which were rescued by ectopic expression of either Stim1 or Orai1, respectively. Surprisingly, knockdown of either TRPC1 or TRPC4 proteins had no effect on SOCE and I(CRAC). Ectopic expression of Stim1 in ECs increased their I(CRAC) to a size comparable to that in rat basophilic leukemia cells. Knockdown of Stim1, Stim2, or Orai1 inhibited EC proliferation and caused cell cycle arrest at S and G2/M phase, although Orai1 knockdown was more efficient than that of Stim proteins. These results are first to our knowledge to establish the requirement of Stim1/Orai1 in the endothelial SOCE pathway.

Authors+Show Affiliations

Cardiovascular Sciences, MC8, Albany Medical College, 47 New Scotland Avenue, MC-8, Albany, NY 12208, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural

Language

eng

PubMed ID

18845811

Citation

Abdullaev, Iskandar F., et al. "Stim1 and Orai1 Mediate CRAC Currents and Store-operated Calcium Entry Important for Endothelial Cell Proliferation." Circulation Research, vol. 103, no. 11, 2008, pp. 1289-99.
Abdullaev IF, Bisaillon JM, Potier M, et al. Stim1 and Orai1 mediate CRAC currents and store-operated calcium entry important for endothelial cell proliferation. Circ Res. 2008;103(11):1289-99.
Abdullaev, I. F., Bisaillon, J. M., Potier, M., Gonzalez, J. C., Motiani, R. K., & Trebak, M. (2008). Stim1 and Orai1 mediate CRAC currents and store-operated calcium entry important for endothelial cell proliferation. Circulation Research, 103(11), 1289-99. https://doi.org/10.1161/01.RES.0000338496.95579.56
Abdullaev IF, et al. Stim1 and Orai1 Mediate CRAC Currents and Store-operated Calcium Entry Important for Endothelial Cell Proliferation. Circ Res. 2008 Nov 21;103(11):1289-99. PubMed PMID: 18845811.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Stim1 and Orai1 mediate CRAC currents and store-operated calcium entry important for endothelial cell proliferation. AU - Abdullaev,Iskandar F, AU - Bisaillon,Jonathan M, AU - Potier,Marie, AU - Gonzalez,Jose C, AU - Motiani,Rajender K, AU - Trebak,Mohamed, Y1 - 2008/10/09/ PY - 2008/10/11/pubmed PY - 2009/1/1/medline PY - 2008/10/11/entrez SP - 1289 EP - 99 JF - Circulation research JO - Circ. Res. VL - 103 IS - 11 N2 - Recent breakthroughs in the store-operated calcium (Ca(2+)) entry (SOCE) pathway have identified Stim1 as the endoplasmic reticulum Ca(2+) sensor and Orai1 as the pore forming subunit of the highly Ca(2+)-selective CRAC channel expressed in hematopoietic cells. Previous studies, however, have suggested that endothelial cell (EC) SOCE is mediated by the nonselective canonical transient receptor potential channel (TRPC) family, TRPC1 or TRPC4. Here, we show that passive store depletion by thapsigargin or receptor activation by either thrombin or the vascular endothelial growth factor activates the same pathway in primary ECs with classical SOCE pharmacological features. ECs possess the archetypical Ca(2+) release-activated Ca(2+) current (I(CRAC)), albeit of a very small amplitude. Using a maneuver that amplifies currents in divalent-free bath solutions, we show that EC CRAC has similar characteristics to that recorded from rat basophilic leukemia cells, namely a similar time course of activation, sensitivity to 2-aminoethoxydiphenyl borate, and low concentrations of lanthanides, and large Na(+) currents displaying the typical depotentiation. RNA silencing of either Stim1 or Orai1 essentially abolished SOCE and I(CRAC) in ECs, which were rescued by ectopic expression of either Stim1 or Orai1, respectively. Surprisingly, knockdown of either TRPC1 or TRPC4 proteins had no effect on SOCE and I(CRAC). Ectopic expression of Stim1 in ECs increased their I(CRAC) to a size comparable to that in rat basophilic leukemia cells. Knockdown of Stim1, Stim2, or Orai1 inhibited EC proliferation and caused cell cycle arrest at S and G2/M phase, although Orai1 knockdown was more efficient than that of Stim proteins. These results are first to our knowledge to establish the requirement of Stim1/Orai1 in the endothelial SOCE pathway. SN - 1524-4571 UR - https://www.unboundmedicine.com/medline/citation/18845811/Stim1_and_Orai1_mediate_CRAC_currents_and_store_operated_calcium_entry_important_for_endothelial_cell_proliferation_ L2 - http://www.ahajournals.org/doi/full/10.1161/01.RES.0000338496.95579.56?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub=pubmed DB - PRIME DP - Unbound Medicine ER -