Apical localization of a functional TRPC3/TRPC6-Ca2+-signaling complex in polarized epithelial cells: Role in apical Ca2+ influx

Bidhan C. Bandyopadhyay, William D. Swaim, Xibao Liu, Robert S. Redman, Randen L. Patterson, Indu S. Ambudkar

Research output: Contribution to journalArticle

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Abstract

Receptor-coupled [Ca2+]i increase is initiated in the apical region of epithelial cells and has been associated with apically localized Ca2+-signaling proteins. However, localization of Ca 2+ channels that are regulated by such Ca2+-signaling events has not yet been established. This study examines the localization of TRPC channels in polarized epithelial cells and demonstrates a role for TRPC3 in apical Ca2+ uptake. Endogenously and exogenously expressed TRPC3 was localized apically in polarized Madin-Darby canine kidney cells (MDCK) and salivary gland epithelial cells. In contrast, TRPC1 was localized basolaterally, whereas TRPC6 was detected in both locations. Localization of Gαq/11, inositol 1,4,5-trisphosphate receptor-3, and phospholipase Cβ1 and -β2 was also predominantly apical. TRPC3 co-immunoprecipitated with endogenous TRPC6, phospholipase Cβs, Gαq/11, inositol 1,4,5-trisphosphate receptor-3, and syntaxin 3 but not with TRPC1. Furthermore, 1-oleoyl-2-acetyl-sn-glycerol (OAG)-stimulated apical 45Ca2+ uptake was higher in TRPC3-MDCK cells compared with control (MDCK) cells. Bradykinin-stimulated apical 45Ca2+ uptake and transepithelial 45Ca 2+ flux were also higher in TRPC3-expressing cells. Consistent with this, OAG induced [Ca2+]i increase in the apical, but not basal, region of TRPC3-MDCK cells that was blocked by EGTA addition to the apical medium. Most importantly, (i) TRPC3 was detected in the apical region of rat submandibular gland ducts, whereas TRPC6 was present in apical as well as basolateral regions of ducts and acini; and (ii) OAG stimulated Ca2+ influx into dispersed ductal cells. These data demonstrate functional localization of TRPC3/TRPC6 channels in the apical region of polarized epithelial cells. In salivary gland ducts this could contribute to the regulation of salivary [Ca2+] and secretion.

Original languageEnglish (US)
Pages (from-to)12908-12916
Number of pages9
JournalJournal of Biological Chemistry
Volume280
Issue number13
DOIs
StatePublished - Apr 1 2005
Externally publishedYes

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Madin Darby Canine Kidney Cells
Epithelial Cells
Ducts
Inositol 1,4,5-Trisphosphate Receptors
Glycerol
Salivary Glands
Qa-SNARE Proteins
Phospholipases
Egtazic Acid
Salivary Ducts
Bradykinin
Submandibular Gland
Rats
Fluxes
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Apical localization of a functional TRPC3/TRPC6-Ca2+-signaling complex in polarized epithelial cells : Role in apical Ca2+ influx. / Bandyopadhyay, Bidhan C.; Swaim, William D.; Liu, Xibao; Redman, Robert S.; Patterson, Randen L.; Ambudkar, Indu S.

In: Journal of Biological Chemistry, Vol. 280, No. 13, 01.04.2005, p. 12908-12916.

Research output: Contribution to journalArticle

Bandyopadhyay, Bidhan C. ; Swaim, William D. ; Liu, Xibao ; Redman, Robert S. ; Patterson, Randen L. ; Ambudkar, Indu S. / Apical localization of a functional TRPC3/TRPC6-Ca2+-signaling complex in polarized epithelial cells : Role in apical Ca2+ influx. In: Journal of Biological Chemistry. 2005 ; Vol. 280, No. 13. pp. 12908-12916.
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