Increasing the expression of calcium-permeable TRPC3 and TRPC7 channels enhances constitutive secretion.
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.
Lavender
Verna
V
Department of Biology and Biochemistry, University of Bath, Bath BA2 7AY, UK.
Chong
Setareh
S
Ralphs
Katherine
K
Wolstenholme
Adrian J
AJ
Reaves
Barbara J
BJ
D000818:Animals; D019556:COS Cells; D002118:Calcium / Q000378:metabolism*; D002460:Cell Line; D002522:Chlorocebus aethiops; D006056:Golgi Apparatus / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D008856:Microscopy, Fluorescence; D051381:Rats; D050052:TRPC Cation Channels / Q000235:genetics / Q000378:metabolism*
D050052:TRPC Cation Channels; C104603:TRPC3 cation channel; C498501:TRPC7 protein, human; C096351:transient receptor potential cation channel, subfamily C, member 1; D002118:Calcium