Na+/Ca2+ exchanger isoform 1 (NCX1) and canonical transient receptor potential channel 6 (TRPC6) are recruited by STIM1 to mediate Store-Operated Calcium Entry in primary cortical neurons. (January 2022)

• Record Type: Journal Article
• Authors: Tedeschi, Valentina; Sisalli, Maria Josè; Pannaccione, Anna; Piccialli, Ilaria; Molinaro, Pasquale; Annunziato, Lucio; Secondo, Agnese
• Is Part Of: Cell calcium. Volume 101(2022)
• Publication Date: 2022-01
• Subjects: SOCE Store-Operated-Calcium-Entry -- STIM1 Stromal Interaction Molecule 1 -- ER Endoplasmic reticulum -- NCX1 Na+/Ca2+ Exchanger 1; SOCE -- STIM1 -- Na+/Ca2+ exchanger -- TRPC6 -- ER Ca2+ refilling -- cortical neurons; Calcium -- Metabolism -- Periodicals; Vertebrates -- Physiology -- Periodicals; Calcium -- Physiological effect -- Periodicals; Cell physiology -- Periodicals; Calcium in the body -- Periodicals; 572.516
• DOI: 10.1016/j.ceca.2021.102525
• Languages: English
• ISSNs: 0143-4160

Abstract

Highlights: Na + /Ca 2+ exchanger 1 (NCX1) activity is modulated by STIM1 in cortical neurons NCX1 is recruited by STIM1 to take part in neuronal SOCE TRPC6 is recruited by STIM1 in primary cortical neurons Thapsigargin-mediated activation of TRPC6 produces a Na + gradient useful for NCX1 reverse mode activation Abstract: Excessive calcium (Ca 2+ ) release from the endoplasmic reticulum (ER) represents an important hallmark of several neurodegenerative diseases. ER is recharged from Ca 2+ through the so-called Store-Operated Calcium Entry (SOCE) thus providing Ca 2+ signals to regulate critical cell functions. Single transmembrane-spanning domain protein stromal interacting molecule 1 (STIM1), mainly residing in the ER, and plasmalemmal channel Orai1 represent the SOCE key components at neuronal level. However, many other proteins participate to ER Ca 2+ refilling including the Na + /Ca 2+ exchanger isoform 1 (NCX1), whose regulation by ER remains unknown. In this study, we tested the possibility that neuronal NCX1 may take part to SOCE through the interaction with STIM1. In rat primary cortical neurons and in nerve growth factor (NGF)-differentiated PC12 cells NCX1 knocking down by siRNA strategy significantly prevented SOCE as well as SOCE pharmacological inhibition by SKF-96365 and 2-APB. A significant reduction of SOCE was recorded also in synaptosomes from ncx1 − / − mice brain compared with ncx1 + / + mice. Double labeling confocal experiments showed a large co-localization between NCX1 and STIM1 in rat primary cortical neurons. Accordingly, NCX1 and STIM1 co-immunoprecipitated and functionally interacted each other during ischemic preconditioning, a phenomenon inducing ischemic tolerance. However, STIM1 knocking down reduced NCX1 activity recorded by either patch-clamp electrophysiology or Fura-2 single-cell microfluorimetry. Furthermore, canonical transient receptor potential channel 6 (TRPC6) was identified as the mechanism mediating local increase of sodium (Na + ) useful to drive NCX1 reverse mode and, therefore, NCX1-mediated Ca 2+ refilling. In fact, TRPC6 not only interacted with STIM1, as shown by the co-localization and co-immunoprecipitation with the ER Ca 2+ sensor, but it also mediated 1, 3-Benzenedicarboxylic acid, 4, 4′-[1, 4, 10-trioxa-7, 13-diazacyclopentadecane-7, 13-diylbis(5-methoxy-6, 12-benzofurandiyl)]bis-, tetrakis[(acetyloxy)methyl] ester (SBFI)-monitored Na + increase elicited by thapsigargin in primary cortical neurons. Accordingly, efficient TRPC6 knockdown prevented thapsigargin-induced intracellular Na + elevation and SOCE. Collectively, we identify NCX1 as a new partner of STIM1 in mediating SOCE, whose activation in the reverse mode may be facilitated by the local increase of Na + concentration due to the interaction between STIM1 and TRPC6 in primary cortical neurons. Graphical Abstract: Image, graphical abstract