5-HT2A Receptor-Induced Morphological Reorganization of PKC{gamma}-Expressing Interneurons Gates Inflammatory Mechanical Allodynia in Rat

Mechanical allodynia, a widespread pain symptom that still lacks effective therapy, is associated with the activation of a dorsally directed polysynaptic circuit within the spinal dorsal horn (SDH) or medullary dorsal horn (MDH), whereby tactile inputs into deep SDH/MDH can gain access to superficial SDH/MDH, eliciting pain. Inner lamina II (II_i) interneurons expressing the isoform of protein kinase C (PKC⁺) are key elements for allodynia circuits, but how they operate is still unclear. Combining behavioral, ex vivo electrophysiological, and morphological approaches in an adult rat model of facial inflammatory pain (complete Freund's adjuvant, CFA), we show that the mechanical allodynia observed 1 h after CFA injection is associated with the following (1) sensitization (using ERK1/2 phosphorylation as a marker) and (2) reduced dendritic arborizations and enhanced spine density in exclusively PKC⁺ interneurons, but (3) depolarized resting membrane potential (RMP) in all lamina II_i PKC⁺/PKC^– interneurons. Blocking MDH 5HT_2A receptors (5-HT_2AR) prevents facial mechanical allodynia and associated changes in the morphology of PKC⁺ interneurons, but not depolarized RMP in lamina II_i interneurons. Finally, activation of MDH 5-HT_2AR in naive animals is enough to reproduce the behavioral allodynia and morphological changes in PKC⁺ interneurons, but not the electrophysiological changes in lamina II_i interneurons, induced by facial inflammation. This suggests that inflammation-induced mechanical allodynia involves strong morphological reorganization of PKC⁺ interneurons via 5-HT_2AR activation that contributes to open the gate for transmission of innocuous mechanical inputs to superficial SDH/MDH pain circuitry. Preventing 5-HT_2AR-induced structural plasticity in PKC⁺ interneurons might represent new avenues for the specific treatment of inflammation-induced mechanical hypersensitivity.

SIGNIFICANCE STATEMENT Inflammatory or neuropathic pain syndromes are characterized by pain hypersensitivity such as mechanical allodynia (pain induced by innocuous mechanical stimuli). It is generally assumed that mechanisms underlying mechanical allodynia, because they are rapid, must operate at only the level of functional reorganization of spinal or medullary dorsal horn (MDH) circuits. We discovered that facial inflammation-induced mechanical allodynia is associated with rapid and strong structural remodeling of specifically interneurons expressing the isoform of protein kinase C (PKC) within MDH inner lamina II. Moreover, we elucidated a 5-HT_2A receptor to PKC/ERK1/2 pathway leading to the behavioral allodynia and correlated morphological changes in PKC interneurons. Therefore, descending 5-HT sensitize PKC interneurons, a putative "gate" in allodynia circuits, via 5-HT_2A receptor-induced structural reorganization.

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