The expression of Toll-like receptor 4, 7 and co-receptors in neurochemical sub-populations of rat trigeminal ganglion sensory neurons.

The recent discovery that mammalian nociceptors express Toll-like receptors (TLRs) has raised the possibility that these cells directly detect and respond to pathogens with implications for either direct nociceptor activation or sensitization. A range of neuronal TLRs have been identified, however a detailed description regarding the distribution of expression of these receptors within sub-populations of sensory neurons is lacking. There is also some debate as to the composition of the TLR4 receptor complex on sensory neurons. Here we use a range of techniques to quantify the expression of TLR4, TLR7 and some associated molecules within neurochemically-identified sub-populations of trigeminal (TG) and dorsal root (DRG) ganglion sensory neurons. We also detail the pattern of expression and co-expression of two isoforms of lysophosphatidylcholine acyltransferase (LPCAT), a phospholipid remodeling enzyme previously shown to be involved in the lipopolysaccharide-dependent TLR4 response in monocytes, within sensory ganglia. Immunohistochemistry shows that both TLR4 and TLR7 preferentially co-localize with transient receptor potential vallinoid 1 (TRPV1) and purinergic receptor P2X ligand-gated ion channel 3 (P2X3), markers of nociceptor populations, within both TG and DRG. A gene expression profile shows that TG sensory neurons express a range of TLR-associated molecules. LPCAT1 is expressed by a proportion of both nociceptors and non-nociceptive neurons. LPCAT2 immunostaining is absent from neuronal profiles within both TG and DRG and is confined to non-neuronal cell types under naïve conditions. Together, our results show that nociceptors express the molecular machinery required to directly respond to pathogenic challenge independently from the innate immune system

Endogenous galanin potentiates spinal nociceptive processing following inflammation

We have undertaken a series of experiments using galanin null mutant mice to better define the role of endogenous galanin in spinal excitability following inflammation and in response to centrally sensitizing stimuli. We have employed a behavioural paradigm, the formalin test, as a model of tonic nociception in both galanin knock-out (gal-/-) and wild-type (gal+/+) mice. In this model, we find that gal-/- mice are markedly hypo-responsive, especially in the second phase response. Additionally, we have examined the thermal hyperalgesia which develops following peripheral injection of carrageenan into the plantar surface of one hindpaw. In this inflammatory paradigm, thermal hyperalgesia is markedly attenuated in gal-/- mice. These behavioural findings suggest that endogenous galanin contributes to nociceptive processing. We have tested this hypothesis further by employing an electrophysiological measure of spinal excitability, the flexor withdrawal reflex in gal-/- and gal+/+ mice. We found no differences in acute reflex responses to single stimuli at C-fibre strength or in the time course and magnitude of wind-up induced by a short conditioning train between non-inflamed gal+/+ and gal-/- mice. However, the Iona-lasting post-conditioning enhancement of reflex excitability was only seen in gal+/+ mice. Moreover, following carrageenan inflammation, there was a marked increase in spinal nociceptive reflex excitability in the inflamed gal+/+ mice, but this enhanced excitability was absent in gal-/- animals. These findings illustrate that endogenous galanin is necessary for the full expression of central sensitization, and as such, plays a critical role in the development of hyperalgesia following peripheral tissue injury. Copyright (C) 2001 International Association for the Study of Pain. Published by Elsevier Science B.V. All rights reserved

Functional regeneration of sensory axons into the adult spinal cord

The arrest of dorsal root axonal regeneration at the transitional zone between the peripheral and central nervous system has been repeatedly described since the early twentieth century(1). Here we show that, with trophic support to damaged sensory axons, this regenerative barrier is surmountable. In adult rats with injured dorsal roots, treatment with nerve growth factor (NGF), neurotrophin-3 (NT3) and glial-cell-line-derived neurotrophic factor (GDNF), but not brain-derived neurotrophic factor (BDNF), resulted in selective regrowth of damaged axons across the dorsal root entry zone and into the spinal cord. Dorsal horn neurons were found to be synaptically driven by peripheral nerve stimulation in rats treated with NGF, NT3 and GDNF, demonstrating functional reconnection. In behavioural studies, rats treated with NGF and GDNF recovered sensitivity to noxious heat and pressure. The observed effects of neurotrophic factors corresponded to their known actions on distinct subpopulations of sensory neurons. Neurotrophic factor treatment may thus serve as a viable treatment in promoting recovery from root avulsion injuries