Characterisation of Na-v types endogenously expressed in human SH-SY5Y neuroblastoma cells

Vetter, Irina, Mozar, Christine A., Durek, Thomas, Wingerd, Joshua S., Alewood, Paul F., Christie, Macdonald J. and Lewis, Richard J. (2012) Characterisation of Na-v types endogenously expressed in human SH-SY5Y neuroblastoma cells. Biochemical Pharmacology, 83 11: 1562-1571. doi:10.1016/j.bcp.2012.02.022


Author Vetter, Irina
Mozar, Christine A.
Durek, Thomas
Wingerd, Joshua S.
Alewood, Paul F.
Christie, Macdonald J.
Lewis, Richard J.
Title Characterisation of Na-v types endogenously expressed in human SH-SY5Y neuroblastoma cells
Journal name Biochemical Pharmacology   Check publisher's open access policy
ISSN 0006-2952
1873-2968
Publication date 2012-06
Sub-type Article (original research)
DOI 10.1016/j.bcp.2012.02.022
Volume 83
Issue 11
Start page 1562
End page 1571
Total pages 10
Place of publication Philadelphia PA, United States
Publisher Elsevier
Collection year 2013
Language eng
Formatted abstract
The human neuroblastoma cell line SH-SY5Y is a potentially useful model for the identification and characterisation of Nav modulators, but little is known about the pharmacology of their endogenously expressed Navs. The aim of this study was to determine the expression of endogenous Nav α and β subunits in SH-SY5Y cells using PCR and immunohistochemical approaches, and pharmacologically characterise the Nav isoforms endogenously expressed in this cell line using electrophysiological and fluorescence approaches. SH-SY5Y human neuroblastoma cells were found to endogenously express several Nav isoforms including Nav1.2 and Nav1.7. Activation of endogenously expressed Navs with veratridine or the scorpion toxin OD1 caused membrane depolarisation and subsequent Ca2+ influx through voltage-gated L- and N-type calcium channels, allowing Nav activation to be detected with membrane potential and fluorescent Ca2 dyes. μ-Conotoxin TIIIA and ProTxII identified Nav1.2 and Nav1.7 as the major contributors of this response. The Nav1.7-selective scorpion toxin OD1 in combination with veratridine produced a Nav1.7-selective response, confirming that endogenously expressed human Nav1.7 in SH-SY5Y cells is functional and can be synergistically activated, providing a new assay format for ligand screening.
Keyword SH-SY5Y
Ca2+
Na(v)1.7
FLIPR
ProTxII
OD1
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2013 Collection
Institute for Molecular Bioscience - Publications
 
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