Acute sensitization by NGF of the response of small-diameter sensory neurons to capsaicin

Abstract

We investigated acute sensitization by nerve growth factor (NGF) of the response of small-diameter (>30 μm) dissociated dorsal root ganglion (DRG) cells to brief repeated puffs of capsaicin as a model for thermal hyperalgesia induced by NGF. We have previously shown that placing NGF in the bath after an initial puff of capsaicin can completely overcome the tachyphylaxis normally observed in response to a second puff 10 min later, and this response is often substantially larger than the first. If tachyphylaxis is abolished by carrying out the experiment in Ca2+-free solution, NGF still elicits potentiation of the second puff. However, the amount of potentiation is considerably less than that observed when tachyphylaxis also takes place. Thus it is concluded that NGF has two effects: overcoming tachyphylaxis and potentiation. With three puffs of capsaicin separated by 10 min, we have found that the potentiation established after 10 min exposure to NGF is no longer evident 10 min after removal of NGF. In Ca2+-free solution the potentiation can last up to 1 h after removal of NGF. These results suggest that the initial behavioral sensitization elicited by NGF could result from a direct effect on the sensory neuron but that its later components most likely involve other mechanisms. We have also investigated the contribution of various second-messenger pathways in these actions of NGF by treating the cells with blockers of MAP kinase (PD98059), protein kinase A (PA; PKAI14-22, H89), and PKC (Bisindolylmaleimide I). Surprisingly, PD98059, which previously has been shown to diminish the enhancement of capsaicin responses of dissociated neurons when exposed to NGF for several days, had no effect on the acute response to NGF; nor did the PKC inhibitor. However, PKA inhibitors reduced the capsaicin response of the cells to NGF (as determined from the NGF effect on tachyphylaxis). Consistent with these findings we confirmed that forskolin, a PKA activator, enhances the effect of NGF on the capsaicin response. The percentage of small cells sensitized by NGF under these conditions, as determined by its ability to reduce tachyphylaxis, was 64%. This suggests that about two-thirds of DRG cells <30 μm and sensitive to capsaicin express a functional trkA receptor.