Itch is relayed to higher centers by projection neurons in the spinal and medullary dorsal horn. We employed a double-label method to map the ascending projections of pruriceptive and nociceptive trigeminal and spinal neurons. The retrograde tracer fluorogold (FG) was stereotaxically injected into the right thalamus or lateral parabrachial area (LPb) in mice. Seven days later, mice received intradermal (id) microinjection of histamine, chloroquine, capsaicin, or vehicle into the left cheek. Id histamine, chloroquine and capsaicin elicited similar distributions of Fos-positive neurons in the medial aspect of the superficial medullary and spinal dorsal horn from the trigeminal subnucleus caudalis to C2. Of neurons retrogradely labeled from the thalamus, 43, 8 and 22% were Fos-positive following id histamine, chloroquine or capsaicin. Of the Fos-positive neurons following pruritic or capsaicin stimuli, ∼1–2% were retrogradely labeled with FG. Trigeminoparabrachial projection neurons exhibited a higher incidence of double-labeling in the superficial dorsal horn. Of the neurons retrogradely labeled from LPb, 36, 29 and 33% were Fos-positive following id injection of histamine, chloroquine or capsaicin, respectively. Of Fos-positiveneurons elicited by id histamine, chloroquine and capsaicin, respectively, 3.7,4.3 and 4.1% were retrogradely labeled from LPb. The present resultsindicate that, overall, relatively small subpopulations of pruriceptive and/ornociceptive neurons innervating the cheek project to thalamus or LPb. Theseresults imply that the vast majority of pruritogen- and algogen-responsivespinal neurons are likely to function as interneurons relaying information toprojection neurons and/or participating in segmental nocifensive circuits.
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