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However, in neurons,<jats:styled-content style=\"fixed-case\">STIM<\/jats:styled-content>2 over\u2010expression appears to play a role in calcium homeostasis that is different from<jats:styled-content style=\"fixed-case\">STIM<\/jats:styled-content>1 over\u2010expression. The aim of this study was to establish the role and localization of native<jats:styled-content style=\"fixed-case\">STIM<\/jats:styled-content>2 in the neuronal cell. Co\u2010immunoprecipitation experiments revealed that the interaction between endogenous<jats:styled-content style=\"fixed-case\">STIM<\/jats:styled-content>2 and<jats:styled-content style=\"fixed-case\">ORAI<\/jats:styled-content>1 was greater in a low\u2010calcium medium than in a high\u2010calcium medium. Using a Proximity Ligation Assay (<jats:styled-content style=\"fixed-case\">PLA<\/jats:styled-content>), the number of apparent complexes of endogenous<jats:styled-content style=\"fixed-case\">STIM<\/jats:styled-content>2 with<jats:styled-content style=\"fixed-case\">ORAI<\/jats:styled-content>1 was quantified. No change in the number of<jats:styled-content style=\"fixed-case\">PLA<\/jats:styled-content>signals was observed in the presence of thapsigargin, which depletes calcium from the endoplasmic reticulum (<jats:styled-content style=\"fixed-case\">ER<\/jats:styled-content>). However, the number of apparent<jats:styled-content style=\"fixed-case\">STIM<\/jats:styled-content>2\u2010<jats:styled-content style=\"fixed-case\">ORAI<\/jats:styled-content>1 complexes increased when intracellular and subsequently<jats:styled-content style=\"fixed-case\">ER<\/jats:styled-content>calcium concentrations were decreased by<jats:styled-content style=\"fixed-case\">BAPTA<\/jats:styled-content>\u2010<jats:styled-content style=\"fixed-case\">AM<\/jats:styled-content>or a low\u2010calcium medium. Both Fura\u20102 acetoxymethyl ester calcium imaging and<jats:styled-content style=\"fixed-case\">PLA<\/jats:styled-content>in the same neuronal cell indicated that the calcium responses correlated strongly with the number of endogenous<jats:styled-content style=\"fixed-case\">STIM<\/jats:styled-content>2\u2010<jats:styled-content style=\"fixed-case\">ORAI<\/jats:styled-content>1 complexes. The small drop in calcium levels in the<jats:styled-content style=\"fixed-case\">ER<\/jats:styled-content>caused by decreased intracellular calcium levels appeared to initiate the calcium\u2010sensitive and thapsigargin\u2010insensitive interaction between<jats:styled-content style=\"fixed-case\">STIM<\/jats:styled-content>2 and<jats:styled-content style=\"fixed-case\">ORAI<\/jats:styled-content>1.<\/jats:p><jats:p><jats:boxed-text content-type=\"graphic\" position=\"anchor\"><jats:graphic xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" mimetype=\"image\/png\" position=\"anchor\" specific-use=\"enlarged-web-image\" xlink:href=\"graphic\/jnc12320-fig-0007-m.png\"><jats:alt-text>image<\/jats:alt-text><\/jats:graphic><\/jats:boxed-text><\/jats:p><jats:p>We show in neuronal somata the formation of endogenous complexes of stromal interaction molecule 2 (STIM2) with ORAI1 calcium channels. Their number increased when intracellular Ca<jats:sup>2+<\/jats:sup>concentrations were decreased by the Ca<jats:sup>2+<\/jats:sup>chelator BAPTA\u2010AM or a low\u2010calcium medium (EGTA), but did not in the presence of thapsigargin (TG). We conclude that the small drop of Ca<jats:sup>2+<\/jats:sup>level in endoplasmic reticulum, due to the decreased level of intracellular Ca<jats:sup>2+<\/jats:sup>, is sufficient to trigger STIM2\u2010ORAI1 complex formation in a thapsigargin\u2010insensitive manner.<\/jats:p>","DOI":"10.1111\/jnc.12320","type":"journal-article","created":{"date-parts":[[2013,5,25]],"date-time":"2013-05-25T12:12:36Z","timestamp":1369483956000},"page":"727-738","source":"Crossref","is-referenced-by-count":56,"title":["Native<scp>STIM<\/scp>2 and<scp>ORAI<\/scp>1 proteins form a calcium\u2010sensitive and thapsigargin\u2010insensitive complex in cortical neurons"],"prefix":"10.1111","volume":"126","author":[{"given":"Joanna","family":"Gruszczynska\u2010Biegala","sequence":"first","affiliation":[{"name":"International Institute of Molecular and Cell Biology Laboratory of Neurodegeneration Warsaw Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jacek","family":"Kuznicki","sequence":"additional","affiliation":[{"name":"International Institute of Molecular and Cell Biology Laboratory of Neurodegeneration Warsaw Poland"},{"name":"Nencki Institute of Experimental Biology Warsaw Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2013,6,12]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1189\/jlb.0610381"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1126\/scisignal.2000522"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymeth.2008.09.009"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0166-2236(00)01891-9"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbamcr.2008.11.008"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2007.11.039"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M110.118984"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ceca.2010.01.006"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1096\/fj.10-178210"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1523\/JNEUROSCI.5053-10.2011"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1093\/molehr\/gar071"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M110.206946"},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0019285"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M609630200"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1038\/355353a0"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1228757"},{"key":"e_1_2_8_18_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M802904200"},{"key":"e_1_2_8_19_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.neuint.2008.10.005"},{"key":"e_1_2_8_20_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cub.2005.05.055"},{"key":"e_1_2_8_21_1","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.200604015"},{"key":"e_1_2_8_22_1","doi-asserted-by":"publisher","DOI":"10.1152\/ajpcell.1999.276.1.C221"},{"key":"e_1_2_8_23_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M604589200"},{"key":"e_1_2_8_24_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1471-4159.2012.07840.x"},{"key":"e_1_2_8_25_1","doi-asserted-by":"publisher","DOI":"10.1096\/fj.07-9449com"},{"key":"e_1_2_8_26_1","doi-asserted-by":"publisher","DOI":"10.1038\/ncb1435"},{"key":"e_1_2_8_27_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0143-4160(03)00143-X"},{"key":"e_1_2_8_28_1","doi-asserted-by":"crossref","first-page":"413","DOI":"10.55782\/ane-2009-1753","article-title":"Immunolocalization of STIM1 in the mouse brain","volume":"69","author":"Skibinska\u2010Kijek A.","year":"2009","journal-title":"Acta Neurobiol. 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