Journal: bioRxiv

Article Title: Age-related sensory dysfunction reconfigures the spinal circuitry for touch, itch and pain

doi: 10.64898/2025.12.18.695289

Figure Lengend Snippet: a, b, In aged mice, ablation of Gpr83 + neurons by injection of PEN-SAP reverses mechanical hyperknesis ( a ; control blank SAP, n = 7; PEN-SAP, n =6) and prevents spontaneous scratching ( b ). c, d, Ablation of Tacr1 + neurons in aged mice by injection of SSP-SAP has no effect on mechanical itch sensitivity ( c ; SAP, n = 7; SSP-SAP, n =6) or spontaneous scratching ( d ). e, f, Mechanical itch sensitivity ( e ) and spontaneous scratching ( f ) are increased by administration of CNO to P56 NPY::Cre; Lbx1 FlpO ; R26 ds-hM4D mice (hM4D + ; n = 7) as compared to control NPY::Cre; R26 ds-hM4D mice (hM4D - ; n = 7). g, h, Mechanical itch sensitivity ( g ) and spontaneous scratching ( h ) are unchanged following administration of CNO to aged NPY::Cre; Lbx1 FlpO ; R26 ds-hM4D mice (hM4D + ; n = 6) compared to aged control NPY::Cre; R26 ds-hM4D mice (hM4D - ; n = 6). i, j, Chemogenetic silencing of spinal NPY + neurons potentiates hyperknesis/alloknesis ( i ) and spontaneous scratching ( j ) in young but not aged mice treated with AEW to induce dry skin itch. k, Silencing spinal NPY + neurons exacerbates histamine alloknesis in young but not aged mice. Error bars represent SEM. *p < 0.05; **p < 0.01; ***p < 0.001; ns, no significant difference.

Article Snippet: The following primary antibodies were used in this study: rabbit α-Calcrl (1:50; Invitrogen), rabbit α-Calretinin (1:250; Swant), rabbit α-Caspase 3 (1:500; Invitrogen), rabbit α-DsRed (1:1000; Clontech), rabbit α-Galanin (1:500; Peninsula Lab), chicken α-GFP (1:000; Aves), rabbit α-Gpr83 (1:500; Alomone Labs), rat α-K8 (TROMA1, Developmental Studies Hybridoma Bank, 1:100), rabbit α-NF200 (1:1000; Sigma), mouse α-NF200 (1:500; Sigma), rabbit α-NK1R (1:500; Advanced Targeting Systems), rabbit α-NPY (1:500; Peninsula Lab), rabbit α-NPY1R (1:500; Neuromics), guinea pig α-prodynorphin (1:500; Abcam), rabbit α-Parvalbumin (1:1000; Swant), rabbit α-Pax2 (1:200; Zymed), rat α-RFP (1:1000; Chromotek), rabbit α-dsRed (1:1000 Clontech), guinea pig α-vGluT1 (1:1000; Millipore).

Techniques: Injection, Control

Journal: bioRxiv

Article Title: Age-related sensory dysfunction reconfigures the spinal circuitry for touch, itch and pain

doi: 10.64898/2025.12.18.695289

Figure Lengend Snippet: a, Representative images showing a lack of TUNEL staining in Y1-immunoreactive neurons in the dorsal horn at cervical segment C5 of young (left) and aged mice (right). b-d, Programmed cell death in populations of molecularly defined excitatory neurons identified by antibodies against Y1, Tacr1, Calcrl and Gpr83, and calretinin (CR) in aged compared to young mice: summaries of population size ( b ), the proportion of TUNEL + neurons expressing each molecular marker ( c ), and the proportion of each molecularly defined population labelled by TUNEL ( d ). e, Representative images showing a lack of TUNEL staining in Y1-immunoreactive neurons in the dorsal horn of control (left) and Merkel cell-silenced mice (right). f-h, Programmed cell death in populations of molecularly defined excitatory neurons identified by immunohistochemistry in Merkel cell-silenced mice compared to controls: summaries of population size ( f ), the proportion of TUNEL + neurons expressing each molecular marker ( g ), and the proportion of each molecularly defined population labelled by TUNEL ( h ). Scale bars, 20 µm. Error bars represent SEM. ns, no significant difference.

Article Snippet: The following primary antibodies were used in this study: rabbit α-Calcrl (1:50; Invitrogen), rabbit α-Calretinin (1:250; Swant), rabbit α-Caspase 3 (1:500; Invitrogen), rabbit α-DsRed (1:1000; Clontech), rabbit α-Galanin (1:500; Peninsula Lab), chicken α-GFP (1:000; Aves), rabbit α-Gpr83 (1:500; Alomone Labs), rat α-K8 (TROMA1, Developmental Studies Hybridoma Bank, 1:100), rabbit α-NF200 (1:1000; Sigma), mouse α-NF200 (1:500; Sigma), rabbit α-NK1R (1:500; Advanced Targeting Systems), rabbit α-NPY (1:500; Peninsula Lab), rabbit α-NPY1R (1:500; Neuromics), guinea pig α-prodynorphin (1:500; Abcam), rabbit α-Parvalbumin (1:1000; Swant), rabbit α-Pax2 (1:200; Zymed), rat α-RFP (1:1000; Chromotek), rabbit α-dsRed (1:1000 Clontech), guinea pig α-vGluT1 (1:1000; Millipore).

Techniques: TUNEL Assay, Staining, Expressing, Marker, Control, Immunohistochemistry