Depletion of 14-3-3 gamma reduces the surface expression of Transient Receptor Potential Melastatin 4b (TRPM4b) Channels and attenuates TRPM4b-mediated glutamate-induced neuronal cell death

Authors
Cho, Chang-HoonKim, EunjuLee, Young-SunYarishkin, OlegYoo, Jae ChealPark, Jae-YongHong, Seong-GeunHwang, Eun Mi
Issue Date
2014-07-22
Publisher
BMC
Citation
MOLECULAR BRAIN, v.7
Abstract
Background: TRPM4 channels are Ca2+ activated nonselective cation channels which are deeply involved in physiological and pathological conditions. However, their trafficking mechanism and binding partners are still elusive. Results: We have found the 14-3-3 gamma as a binding partner for TRPM4b using its N-terminal fragment from the yeast-two hybrid screening. Ser88 at the N-terminus of TRPM4b is critical for 14-3-3 gamma binding by showing GST pull-down and co-immunoprecipitation. Heterologous overexpression of 14-3-3 gamma in HEK293T cells increased TRPM4b expression on the plasma membrane which was measured by whole-cell recordings and cell surface biotinylation experiment. Surface expression of TRPM4b was greatly reduced by short hairpin RNA (shRNA) against 14-3-3 gamma. Next, endogenous TRPM4b-mediated currents were electrophysiologically characterized by application of glutamate and 9-phenanthrol, a TRPM4b specific antagonist in HT-22 cells which originated from mouse hippocampal neurons. Glutamate-induced TRPM4b currents were significantly attenuated by shRNAs against 14-3-3 gamma or TRPM4b in these cells. Finally, glutamate-induced cell death was greatly prevented by treatment of 9-phenanthrol or 14-3-3 gamma shRNA. Conclusion: These results showed that the cell surface expression of TRPM4 channels is mediated by 14-3-3 gamma binding, and the specific inhibition of this trafficking process can be a potential therapeutic target for glutamate-induced neuronal cell death.
Keywords
CATION CHANNEL; INTRACELLULAR CALCIUM; KAINATE RECEPTORS; SINGLE-CHANNEL; PROTEINS; 14-3-3-PROTEINS; TRAFFICKING; SENSITIVITY; MODULATION; ISOFORMS; CATION CHANNEL; INTRACELLULAR CALCIUM; KAINATE RECEPTORS; SINGLE-CHANNEL; PROTEINS; 14-3-3-PROTEINS; TRAFFICKING; SENSITIVITY; MODULATION; ISOFORMS; TRPM4b; 14-3-3; Non-selective cation channels; 9-phenanthrol; HT-22; Protein-protein interaction; Surface expression; Hippocampal neurons; Calcium activated cation channels; MTT assay
ISSN
1756-6606
URI
https://pubs.kist.re.kr/handle/201004/126580
DOI
10.1186/s13041-014-0052-3
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KIST Article > 2014
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