Increasing the expression of calcium-permeable TRPC3 and TRPC7 channels enhances constitutive secretion

Autor: Katherine L. Ralphs, Adrian J. Wolstenholme, Setareh Chong, Barbara J. Reaves, Verna Lavender
Rok vydání: 2008
Předmět:
BFA
Brefeldin A
Golgi Apparatus
Biochemistry
chemistry.chemical_compound
0302 clinical medicine
TRPC3
Chlorocebus aethiops
TRPC
canonical TRP
TRPC
0303 health sciences
DMEM
Dulbecco's modified Eagle's medium
transient receptor potential (TRP)
trans-Golgi network
HRP
horseradish peroxidase
SNAP
NSF-attachment protein
Brefeldin A
EGFP
enhanced green fluorescent protein
Immunohistochemistry
Cell biology
COS Cells
symbols
TNFR
tumour necrosis factor receptor
SEAP
a secreted form of alkaline phosphatase
Intracellular
Research Article
TRP
transient receptor potential
IP3
inositol trisphosphate
NSF
N-ethylmaleimide-sensitive factor
cation channel trafficking
Immunoblotting
Biology
hTRPC
human TRPC
Cell Line
ER
endoplasmic reticulum
03 medical and health sciences
symbols.namesake
PLC
phospholipase C
Animals
Humans
Secretion
canonical TRP (TRPC)
Molecular Biology
Secretory pathway
TRPC Cation Channels
030304 developmental biology
calcium
HEK-293 cell
human embryonic kidney cell
TGN
trans-Golgi network
Endoplasmic reticulum
FCS
foetal calf serum
Cell Biology
Golgi apparatus
TRPML
mucolipin TRP
secretory pathway
Rats
TRPM8
melastatin TRP8
Microscopy
Fluorescence
chemistry
dTRP
Drosophila TRP
DAG
diacylglycerol
030217 neurology & neurosurgery
Zdroj: Biochemical Journal
ISSN: 1470-8728
0264-6021
DOI: 10.1042/bj20071488
Popis: The hTRPC [human TRPC (canonical transient receptor potential)] family of non-selective cation channels is proposed to mediate calcium influx across the plasma membrane via PLC (phospholipase C)-coupled receptors. Heterologously expressed hTRPC3 and hTRPC7 have been localized at the cell surface; however, a large intracellular component has also been noted but not characterized. In the present study, we have investigated the intracellular pool in COS-7 cells and have shown co-localization with markers for both the TGN (trans-Golgi network) and the cis-Golgi cisternae by immunofluorescence microscopy. Addition of BFA (Brefeldin A) to cells expressing hTRPC3 or hTRPC7 resulted in the redistribution of the Golgi component to the endoplasmic reticulum, indicating that this pool is present in both the Golgi stack and the TGN. Expression of either TRPC3 or TRPC7, but not TRPC1 or the cell surface marker CD8, resulted in a 2–4-fold increase in secreted alkaline phosphatase in the extracellular medium. Based on these results, we propose that an additional function of these members of the hTRPC family may be to enhance secretion either by affecting transport through the Golgi stack or by increasing fusion at the plasma membrane.
Databáze: OpenAIRE
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