Figure 3.

Strategy for isolation of Trpv1-expressing neurons in mouse DRG. A–F, Neurons isolated from lumbosacral adult DRG of TrpV1^PLAP-nLacZ mice were incubated with the fluorescent galactosidase substrate, DDAOG (excitation/emission maxima ∼460/610 nm). A, Five neurons that have taken up DDAOG. B, Three neurons (arrows) are visible by their fluorescent hydrolysis product (excitation/emission maxima ∼645/660), also evident in the nucleus, so deduced to express galactosidase, i.e., Trpv1 neurons. C, Merge of panels A, B. D, Neurons expressing β-galactosidase visualized using immunohistochemistry; expression is restricted to the nucleus. E, Each neuron in the field has converted DDAOG to DDAO. F, Merge of panels D, E. Bar in A applies to all micrographs and represents 50 μm. G–I, Representative outputs from flow cytometry of mouse lumbosacral DRG. G, Neurons isolated from wild-type mice were not treated with DDAOG. H, Neurons isolated from wild-type mice were treated with DDAOG, but without expressing β-galactosidase cannot hydrolyze this to form DDAO. I, Neurons isolated from of TrpV1^PLAP-nLacZ mice were treated with DDAOG; many cells have hydrolyzed DDAOG to DDAO (i.e., Trpv1 neurons) and others have taken up DDAOG but not hydrolyzed this to DDAO (i.e., Trpv1-negative neurons).