Norepinephrinergic projection from locus coeruleus to parafascicular nucleus promotes pain and anxiety-like behaviors in mice

Chronic neuropathic pain is frequently comorbid with anxiety disorders, yet the neural circuits underlying this interaction remain poorly defined. The parafascicular nucleus of the thalamus (PF) integrates nociceptive and affective signals, but its specific regulatory mechanisms in pain-anxiety comorbidity are not well known. Using spared nerve injury (SNI) model mice, we combined viral neural tracing, chemogenetics, pharmacology, and electrophysiology to dissect the locus coeruleus (LC)-PF neural pathway. Viral tracing revealed monosynaptic projections from norepinephrinergic (NEergic) neurons in the dorsal LC to calcium/calmodulin dependent protein kinase IIα (CaMKIIα)- immunopositive neurons within the PF. Chemogenetic inhibition/activation of this pathway were performed in naïve and SNI mice, alongside intra-PF microinjection of the alpha-2 adrenergic receptor (ADRA2) antagonist yohimbine. Behavioral tests assessed mechanical/thermal hypersensitivity and anxiety-like behaviors. Results showed that 92.1% of PF-projecting LC neurons were NEergic, with 70.1% localized dorsally. Chemogenetic inhibition of LCNE-PFCaMKIIα neural pathway significantly alleviated both acute-phase mechanical hypersensitivity (< 7 days post-surgery) and chronic-phase anxiety-like behaviors in SNI mice, while activation of this pathway induced pain sensitization and anxiety-like behaviors in naïve mice. Intra-PF yohimbine reversed SNI-induced allodynia and anxiety-like behaviors. Electrophysiology confirmed yohimbine increased PF neuronal intrinsic excitability. These results suggest that the LCNE-PFCaMKIIα neural pathway promotes neuropathic pain and comorbid anxiety via ADRA2-mediated suppression of PF neuronal activity. Targeted inhibition of this circuit may represent a therapeutic strategy for pain-related affective disorders.