Canonical transient receptor potential channel 4 (TRPC4) co-localizes with the scaffolding protein ZO-1 in human fetal astrocytes in culture

Xianyuan Song, Yongmei Zhao, Leontine Narcisse, Heather Duffy, Yvonne Kress, Sunhee Lee, Celia F. Brosnan

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

Members of the mammalian transient receptor potential (TRP) family form cation-permeable channels at the plasma membrane implicated in capacitative calcium influx after activation by either second-messenger-mediated pathways or store depletion, or both. This study shows that with the use of RT-PCR, Western blotting, and immunohistochemistry, resting astrocytes express TRPC4 at the cell membrane, particularly at sites of cell-to-cell contact. By confocal imaging and immunoelectron microscopy, we detected co-localization of TRPC4 with the scaffolding protein zonula occludens 1 (ZO-1), and demonstrated that immunoprecipitation with antibodies to ZO-1 brought down TRPC4, and vice-versa. It has been proposed that the targeting of TRPC4 to the cell membrane is dependent on the interaction of the C-terminal TRL motif with PDZ domains. Using transfection of astrocytes with myc-tagged TRPC4 or TRL-motif truncated TRPC4 (δTRL), we found that δTRL localized predominantly to a juxtanuclear compartment, whereas the wild-type protein showed cell surface distribution. Deletion of the TRL motif also reduced plasma membrane expression as assessed by cell surface biptinylation experiments. Using GST fusion proteins, we found that TRPC4 interacted with the PDZ1 domain of ZO-1 and that this was also dependent on the TRL motif. Thus, our data demonstrate that the PDZ-interacting domain of TRPC4 controls its cell surface localization. These data implicate TRPC4 in the regulation of calcium homeostasis in astrocytes, particularly as part of a signaling complex that forms at junctional sites between astrocytes.

Original languageEnglish (US)
Pages (from-to)418-429
Number of pages12
JournalGLIA
Volume49
Issue number3
DOIs
StatePublished - Feb 2005

Fingerprint

Zonula Occludens-1 Protein
Transient Receptor Potential Channels
Astrocytes
Cell Membrane
PDZ Domains
Tight Junctions
Calcium
Immunoelectron Microscopy
Second Messenger Systems
Immunoprecipitation
Confocal Microscopy
Transfection
Cations
Membrane Proteins
Homeostasis
Western Blotting
Immunohistochemistry

Keywords

  • Astrocytes
  • Calcium
  • Gap junctions
  • PDZ domains
  • Transient receptor potential channels
  • ZO-1

ASJC Scopus subject areas

  • Immunology

Cite this

Canonical transient receptor potential channel 4 (TRPC4) co-localizes with the scaffolding protein ZO-1 in human fetal astrocytes in culture. / Song, Xianyuan; Zhao, Yongmei; Narcisse, Leontine; Duffy, Heather; Kress, Yvonne; Lee, Sunhee; Brosnan, Celia F.

In: GLIA, Vol. 49, No. 3, 02.2005, p. 418-429.

Research output: Contribution to journalArticle

Song, Xianyuan ; Zhao, Yongmei ; Narcisse, Leontine ; Duffy, Heather ; Kress, Yvonne ; Lee, Sunhee ; Brosnan, Celia F. / Canonical transient receptor potential channel 4 (TRPC4) co-localizes with the scaffolding protein ZO-1 in human fetal astrocytes in culture. In: GLIA. 2005 ; Vol. 49, No. 3. pp. 418-429.
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