Exogenous nerve growth factor attenuates opioid-induced inhibition of voltage-activated Ba 2+ currents in rat sensory neurons

Abstract

Nerve growth factor (NGF) promotes the survival of embryonic sensory neurons and maintains the phenotypic characteristics of primary nociceptive neurons postnatally. NGF also contributes to nociceptor activation and hyperalgesia during inflammatory pain states. The purpose of this study was to determine whether NGF might have an additional pronociceptive action by interfering with opioid-mediated analgesia in primary nociceptive neurons. Sensory neurons were isolated from the dorsal root ganglia of weanling rats and kept in standard culture conditions either with or without exogenous NGF (50 ng/ml). Currents through voltage-gated calcium channels were recorded from individual neurons using the whole cell patch clamp technique with Ba 2+ as the charge carrier (I Ba). The μ-opioid agonist fentanyl (1 μM) and the GABA B agonist baclofen (50 μM) were used to test G protein-dependent inhibition of I Ba. Fentanyl inhibited I Ba by an average of 38±4% in untreated cells vs. 25±2% in NGF-treated cells ( P<0.01). NGF had no effect on I Ba current magnitude or kinetics. The NGF-induced attenuation of opioid action was observed as early as 4 h after exposure, but was not seen when NGF was applied by bath perfusion for up to 40 min, suggesting that the effect was not mediated by a rapid phosphorylation event. The effect of NGF was prevented by K-252a (100 nM), an inhibitor of TrkA autophosphorylation. Baclofen-induced inhibition of I Ba, on the other hand, was not affected by NGF treatment, suggesting that NGF modulation of opioid-mediated inhibition occurred upstream from the G protein. This was supported by the finding that GTP-γ-S, an agonist independent G protein activator, inhibited I Ba similarly in both untreated and NGF treated cells. The results show that NGF selectively attenuated opioid-mediated inhibition of I Ba via TrkA receptor activation, possibly by altering opioid receptor function.