ANO1 (TMEM16A) is a widely expressed Ca2+ activated Cl- channel (CaCC) with functions ranging from epithelial transport to sensory transmission.  ANO1 is expressed in sensory neurons and involved in the process of detecting and relaying signals that arise after painful (noxious) stimulation. These neurons also express the Ca2+ permeable heat sensor TRPV1 with previous research suggesting that ANO1 and TRPV1 exist in a nanodomain arrangement where Ca2+ entering through TRPV1 is able to directly activate ANO1 due to their close proximity in dorsal root ganglion (DRG) neurons. We have previously shown that ANO1 activation is functionally coupled to Ca2+ release through inositol trisphosphate receptors (IP3R) in DRG neurons, thus allowing the low Ca2+ sensitivity of ANO1 to be overcome. Interestingly, due to the ability of TRPV1 to induce Ca2+ release from the ER by activating phospholipase C (PLC), we hypothesised that TRPV1 may be able to activate ANO1 through IP3R Ca2+ release as well. To this end, we developed a multi-wavelength live cell imaging approach to allow us to simultaneously monitor CaCC activity and Ca2+ dynamics in DRG and revealed that capsaicin activation of TRPV1 was able to induce CaCC activity. Furthermore, CaCC activity produced by capsaicin application was attenuated after depletion of the endoplasmic reticulum (ER) Ca2+ load, suggesting that ER Ca2+ release contributed to TRPV1-induced CaCC activation. To confirm that this effect was induced by plasmalemmal and not ER-localised TRPV1 channels, we used a cell impermeable TRPV1 activator- a derivative of double knot spider toxin- with an ER-Ca2+ sensor to demonstrate that ER depletion only occurs when membrane-localised TRPV1 are activated. To understand if there was a structural arrangement of channels that allowed this coupling to be facilitated in DRG neurons, we used in situ proximity ligation assay (PLA) to show that ANO1, TRPV1 and IP3R receptors were often found in close proximity. This was also confirmed using superresolution stochastic optical reconstruction microscopy (STORM) which revealed that all 3 proteins were indeed found in close proximity. In summary, our findings demonstrate that functional coupling between ANO1 and TRPV1 in sensory neurons is facilitated by Ca2+ release through IP3R with the channels found in a nanodomain structure to enable efficient ANO1 activation.