Journal: Journal of Pain Research

Article Title: Wrist-Ankle Acupuncture Alleviates Paclitaxel-Induced Neuropathic Pain in Mice by Neuronal α7 Nicotinic Acetylcholine Receptor–Dependent Modulation of Spinal Glutamatergic/NMDAR Signaling

doi: 10.2147/JPR.S604818

Figure Lengend Snippet: WAA alleviates paclitaxel-induced mechanical allodynia and thermal hyperalgesia in a neuronal α7nAChR–dependent manner. Data are presented as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001. ( a ) Experimental design and timeline for the Chrna7 flox/flox mouse cohort; detailed procedures are provided in the Methods. ( b ) Representative Western blot bands of α7nAChR in the spinal dorsal horn. ( c ) Quantification of α7nAChR protein levels 21 days after lumbar dorsal horn microinjection of control virus or rAAV-hSyn-SV40-NLS-Cre (N=3, t = 3.2, P = 0.033). ( d ) Representative immunofluorescence images showing colocalization of α7nAChR (red) and NeuN (green); scale bar: 100 μm. ( e ) Quantification of neuronal association/targeting using Manders’ overlap coefficient (fraction of α7nAChR overlapping NeuN, N=3, t = 41.833, P = 0.001). ( f ) Behavioral assessments of mechanical (paw withdrawal threshold, PWT) and thermal sensitivity (paw withdrawal latency, PWL) at days 0, 7, and 14; WAA-induced analgesia was attenuated after dorsal horn neuronal α7nAChR knockdown (N=9,day 14: F PWT =342.009, P PWT < 0.001; F PWL =333.245, P PWL < 0.001). ( g ) Representative Western blot bands of α7nAChR in the spinal dorsal horn across indicated groups following paclitaxel administration and WAA treatment. ( h ) Densitometric quantification of α7nAChR protein levels corresponding to panel ( g ) (N=3, F =46.729, P < 0.001). ( i ) Representative immunofluorescence images of α7nAChR (red) and NeuN (green) in the spinal cord dorsal horn on day 14 after model induction; scale bar: 100 μm. ( j ) Quantification of α7nAChR immunofluorescence intensity (mean intensity), corresponding to panel ( i ) (N=3, F =81.123, P < 0.001).

Article Snippet: Except for the saline group, paclitaxel (PTX)-induced CIPN was established as previously described by Toma et al Briefly, a 5 mg/mL PTX (MedChemExpress) stock solution was prepared in 10% DMSO and 90% (20% sulfobutylether-β-cyclodextrin [SBE-β-CD] in saline; MedChemExpress).

Techniques: Western Blot, Microinjection, Control, Virus, Immunofluorescence, Knockdown

Journal: Journal of Pain Research

Article Title: Wrist-Ankle Acupuncture Alleviates Paclitaxel-Induced Neuropathic Pain in Mice by Neuronal α7 Nicotinic Acetylcholine Receptor–Dependent Modulation of Spinal Glutamatergic/NMDAR Signaling

doi: 10.2147/JPR.S604818

Figure Lengend Snippet: Intrathecal NMDAR activation attenuates the analgesic and spinal molecular effects of WAA in paclitaxel-induced CIPN. Data are presented as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001. ( a ) Experimental design and timeline for the C57BL/6 cohort; detailed procedures are provided in the Methods. ( b ) Behavioral assessment of PWT and PWL at days 0, 7, and 14. Intrathecal administration of an NMDAR agonist prior to WAA reduced the behavioral analgesic benefit of WAA. (N=9; two-way repeated-measures ANOVA; day 14: F PWT =281.188, P PWT < 0.001; F PWL =461.233, P PWL < 0.001). ( c ) Glutamate (glutamic acid) content in the spinal dorsal horn. (N=3, F =54.286, P < 0.001). ( d ) Representative Western blot bands of proteins associated with glutamatergic/NMDAR pathway activity in the spinal dorsal horn. ( e – i ) Densitometric quantification of NR2B ( e ) VGluT2 ( f ) p-JNK ( g ) p-CREB ( h ) and CGRP ( i ) in the spinal cord dorsal horn.(N=3, NR2B: F =11.877, P < 0.01; VGluT2: F =36.553, P < 0.001; p-JNK: F =8.216, P < 0.01; p-CREB: F =16.911, P < 0.01; CGRP: F =25.487, P < 0.001).

Article Snippet: Except for the saline group, paclitaxel (PTX)-induced CIPN was established as previously described by Toma et al Briefly, a 5 mg/mL PTX (MedChemExpress) stock solution was prepared in 10% DMSO and 90% (20% sulfobutylether-β-cyclodextrin [SBE-β-CD] in saline; MedChemExpress).

Techniques: Activation Assay, Western Blot, Activity Assay

Journal: Journal of Pain Research

Article Title: Wrist-Ankle Acupuncture Alleviates Paclitaxel-Induced Neuropathic Pain in Mice by Neuronal α7 Nicotinic Acetylcholine Receptor–Dependent Modulation of Spinal Glutamatergic/NMDAR Signaling

doi: 10.2147/JPR.S604818

Figure Lengend Snippet: α7nAChR in VGluT2-expressing glutamatergic neurons is required for the analgesic and spinal glutamatergic/NMDAR-modulatory effects of WAA in paclitaxel-induced CIPN. Data are presented as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001. ( a ) Experimental design and timeline for VGluT2 neuron–specific Chrna7 conditional knockout mice ( Chrna7 flox/flox ;Slc17a6 -Cre + , α7-cKO(VGluT2)) and littermate Cre-negative controls ( Chrna7 flox/flox ;Slc17a6 -Cre − ); detailed procedures are provided in the Methods. ( b ) Representative Western blot bands of α7nAChR in the spinal dorsal horn. ( c ) Densitometric quantification of α7nAChR protein levels in α7-cKO(VGluT2) mice and controls (N=3, t =9.954, P =0.001). ( d ) Representative immunofluorescence images of the spinal dorsal horn showing α7nAChR (red) and VGluT2 (green); scale bar, 100 μm. ( e ) Quantification of α7nAChR immunofluorescence intensity (mean intensity) corresponding to panel ( d ). (N=3, t =33.319, P < 0.001). ( f ) Behavioral assessment of PWT and PWL at days 0, 7, and 14. VGluT2 neuron–specific α7nAChR deletion attenuated the analgesic effects of WAA (N=9; two-way repeated-measures ANOVA; day 14: F PWT =308.427, P PWT < 0.001; F PWL =301.306, P PWL < 0.001). ( g ) Glutamate (glutamic acid) content in the spinal dorsal horn. (N=3, F =37.871, P <0.001). ( h )Representative Western blot bands of proteins associated with glutamatergic/NMDAR pathway activity in the spinal dorsal horn. ( i – m ) Densitometric quantification of NR2B ( i ) VGluT2 ( j ) p-JNK ( k ) p-CREB ( l ) and CGRP ( m ) in the spinal dorsal horn (N=3, NR2B: F =36.601, P < 0.001; VGluT2: F =11.265, P < 0.001; p-JNK: F =21.336, P < 0.001; p-CREB: F =13.144, P < 0.01; CGRP: F =15.896, P < 0.001).

Article Snippet: Except for the saline group, paclitaxel (PTX)-induced CIPN was established as previously described by Toma et al Briefly, a 5 mg/mL PTX (MedChemExpress) stock solution was prepared in 10% DMSO and 90% (20% sulfobutylether-β-cyclodextrin [SBE-β-CD] in saline; MedChemExpress).

Techniques: Expressing, Knock-Out, Western Blot, Immunofluorescence, Activity Assay

Journal: bioRxiv

Article Title: Live cell imaging reveals paclitaxel-induced lysosome motility and function disruption in DRG neurons

doi: 10.64898/2026.05.19.726221

Figure Lengend Snippet: A . Representative western images showing p62 bands in blot DRGs of PTX-injected mice or vehicle, plus eFT508. B . Western blot analy is of p62 in DRGs of WT mice (n = 3-7 per group) treated with PTX and vehicle, plus eFT508, or eFT508 only. Data are presented as mean ± SEM ** p < 0.01 as determined by two-way ANOVA followed by Bonferroni’s multiple comparisons test.

Article Snippet: On DIV7, mDRG cultures were treated with PTX (Selleck Chemicals, Cat# S1150) or PTX + eFT508 (MedChem Express, Cat#HY-100022) for 24 hrs.

Techniques: Western Blot, Injection

Journal: bioRxiv

Article Title: Live cell imaging reveals paclitaxel-induced lysosome motility and function disruption in DRG neurons

doi: 10.64898/2026.05.19.726221

Figure Lengend Snippet: A . Representative confocal images of mouse DRG neurons under vehicle (0.002% DMSO), 100 nM PTX, and 100 nM PTX + 100 nM eFT508 conditions showing TFEB expression in gray, peripherin (magenta) to identify neurons, and DAPI (blue) to stain nuclei. Images were captured using a 40x objective lens with an additional 8x digital zoom to highlight the nucleus and cytosol. Contrast has been uniformly adjusted for presentation purposes only. B . Normalized mean gray intensities reveals a significant increase in nuclear TFEB expression after 24 hours of PTX treatment, which was attenuated by concurrent administration of eFT508. Interestingly, there is also a marked decrease in cytosolic TFEB expression, as shown in C . Mouse DRG cultures were prepared independently from two mice. We analyzed 58, 60, and 74 neurons for vehicle, 100 nM PTX, and 100 nM PTX + eFT508 conditions, respectively. Data are presented as violin plots showing the distribution of individual measurements. Image scale bar = 2 μm. One-way ANOVA with Tukey’s multiple comparisons test *p < 0.05, ** p < 0.01, **** p < 0.0001.

Article Snippet: On DIV7, mDRG cultures were treated with PTX (Selleck Chemicals, Cat# S1150) or PTX + eFT508 (MedChem Express, Cat#HY-100022) for 24 hrs.

Techniques: Expressing, Staining

Journal: bioRxiv

Article Title: Live cell imaging reveals paclitaxel-induced lysosome motility and function disruption in DRG neurons

doi: 10.64898/2026.05.19.726221

Figure Lengend Snippet: A . Schematic diagram of a human spinal cord section with DRGs. Lines indicate where the nerves are cut to isolate the DRG. Details on the number of neurons assessed and the number of biological replicates are included. B . Representative 10X images of human DRG neurons under various experimental conditions showing p-eIF4E expression in magenta, peripherin (green) to identify neurons, and DAPI (blue) to stain nuclei. Contrast has been uniformly adjusted for presentation purposes only. C . hDRG neurons treated with 100 nM PTX in vitro for 24 hours showed significant upregulation of p-eIF4E, and concurrent treatment with 100 nM eFT508 demonstrated robust inhibition of p-eIF4E after 24 hours. DRG cultures were independently prepared from 4 organ donors. N = 13, 14, 13 technical replicates for Vehicle (0.002% DMSO), 100 nM PTX, and 100 nM PTX + eFT508 conditions, respectively. Data are presented as mean ± SEM. Image scale bar = 100 μm. One-way ANOVA with Tukey’s multiple comparisons test ** p < 0.01, **** p < 0.0001.

Article Snippet: On DIV7, mDRG cultures were treated with PTX (Selleck Chemicals, Cat# S1150) or PTX + eFT508 (MedChem Express, Cat#HY-100022) for 24 hrs.

Techniques: Expressing, Staining, In Vitro, Inhibition