Abstract
TRPV4 is a non-selective cation channel that tunes the function of different tissues including the vascular endothelium, lung, chondrocytes, and neurons. GSK1016790A is the selective and potent agonist of TRPV4 and a pharmacological tool that is used to study the TRPV4 physiological function in vitro and in vivo. It remains unknown how the sensitivity of TRPV4 to this agonist is regulated. The spatial and temporal dynamics of receptors are the major determinants of cellular responses to stimuli. Membrane translocation has been shown to control the response of several members of the transient receptor potential (TRP) family of ion channels to different stimuli. Here, we show that TRPV4 stimulation with GSK1016790A caused an increase in [Ca2+]i that is stable for a few minutes. Single molecule analysis of TRPV4 channels showed that the density of TRPV4 at the plasma membrane is controlled through two modes of membrane trafficking, complete, and partial vesicular fusion. Further, we show that the density of TRPV4 at the plasma membrane decreased within 20 min, as they translocate to the recycling endosomes and that the surface density is dependent on the release of calcium from the intracellular stores and is controlled via a PI3K, PKC, and RhoA signaling pathway.
Highlights
Transient receptor potential vanilloid sub type 4 (TRPV4) is a non-selective multifunctional cation channel that is involved in different physiological function and pathologies (Darby et al, 2016; White et al, 2016; Rajasekhar et al, 2017)
We found that treatment of Human Umbilical Vein Endothelial Cells (HUVECs) and TRPV4-HEK293 cells with GSK1016790A lead to the activation of the TRPV4 and an increase in [Ca2+]i within the first-minute post-treatment (Figure 1B, Supplementary Image 1, Supplementary Video 1)
We found that GSK1016790A caused the concentration-dependent cytoplasmic aggregation of TRPV4 with an EC50 value of 31 nM
Summary
Transient receptor potential vanilloid sub type 4 (TRPV4) is a non-selective multifunctional cation channel that is involved in different physiological function and pathologies (Darby et al, 2016; White et al, 2016; Rajasekhar et al, 2017). Little information is available on the trafficking of transient receptor potential (TRP) channels to and from the plasma membrane, this is important for controlling channel function (Baratchi et al, 2017a). Several studies on the TRPV family members have shown that trafficking into to the plasma membrane plays an important role in the channel responses to different stimuli (Baratchi et al, 2016). Insulin-like growth factor (IGF) stimulation is reported to induce the insertion of TRPV2 into the plasma membrane (Boels et al, 2001), inflammatory mediators control the regulated membrane trafficking of TRPV1(Van Buren et al, 2005), and shear stress mediates membrane trafficking of TRPV4 channels (Baratchi et al, 2016)
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