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anti cgrp  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti cgrp
    Anti Cgrp, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 144 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/anti+cgrp/CGRP+Rabbit+mAb/pmc13675831-108-38-42
    Average 96 stars, based on 144 article reviews
    anti cgrp - by Bioz Stars, 2026-11
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    Article Title: Morphine Tolerance Gated through EZH2-Mediated Suppression of Trpc5 in Spinal GABAergic Interneurons in Male Mice.
    Article Snippet: www.advancedsciencenews.com www.advancedscience.com with 10% normal donkey serum (Jackson ImmunoResearch, 017-000- 121, West Grove, Pennsylvania, USA) and 0.3% triton X 100 in PBS for 2 h at room temperature

    Article Title: Spinal Cord Injury Versus Sciatic Nerve Injury: Divergent Neuromuscular Pathologies and Neural Circuitry Remodeling.
    Article Snippet: Spinal cord injury (SCI) and sciatic nerve injury (SNI) represent distinct neurotraumamodelswith different pathological outcomes.. Although both injuries result in severe motor and sensory dysfunction in the lower limbs, their divergent mechanisms of muscle atrophy and neural circuit remodeling remain poorly understood.. This study systematically compared electrophysiological and morphological alterations at multiple time points post-injury.

    Article Title: AhR inhibition promotes axon regeneration via a stress-growth switch.
    Article Snippet: Primary antibodies for IF were as follows: anti-AHR (rabbit, 1:300, Enzo, BML-SA210, AB_10540536), anti-ATF3 (rabbit, 1:300, Santa Cruz, sc-188, AB_2258513), anti-tubulin β3 (TUJ1, mouse, 1:1,000, BioLegend, 801201, AB_2313773), anti-tubulin β3 (D71G9, rabbit, 1:300, Cell Signaling, 5568S, AB_10694505), anti-SCG10/STMN2 (rabbit, 1:1,000, Novus, NBP1-49461, AB_10011569), anti-GFP (chicken, 1:1,000, Aves Lab, GFP-1020, AB_10000240), anti-IBA1 (rabbit, 1:1,000, Wako, 019-19741, AB_839504), anti-pS6 ribosomal protein-S235/236 (rabbit, 1:300, Cell Signaling, 2211, AB_331679), anti-5hmC (rabbit, 1:500, Active Motif, 39769, AB_10013602), anti-CD8a (rat, 1:100, Invitrogen, 14-0081-82, AB_467087), anti-CD4 (rat, 1:100, Invitrogen, 14-0041-82, AB_467063), anti-CD68 (rat, 1:100, BioLegend, 137002, AB_2044004), anti-CD45 (rat, 1:100, BD Pharmingen, 550539, AB_2174426), anti-PU1 (E.388.3) (rabbit, 1:300, Thermo Fisher Scientific, MA5-15064, AB_10986949), anti-F4/80 (rat, 1:300, Thermo Fisher Scientific, 14-4801-81, AB_467557), antiCD206 (goat, 1:200, R&D systems, AF2535, AB_2063012), anti-SOX10 (goat, 1:50, R&D systems, AF2864, AB_442208), anti-PGP9.5 (rabbit, 1:800, Neuromics, RA12103, AB_2315126), anti-NF-H (chicken, EMD Millipore, AB5539, 1:1,000, AB_11212161), anti-CSPG (CS-56) (mouse, Sigma-Aldrich, C8035, 1:100, AB_476879), anti-GFAP (chicken, Aves Labs, GFAP, 1:500, AB_2858088), anti-puromycin (mouse, DSHB, PMY-2A4, 1:100, AB_2619605), anti-p-eIF2α (S51) (D9G8) (rabbit, Cell Signaling, 3398, 1:300, AB_2096481), anti-E-cadherin (24E10) (rabbit, Cell Signaling, 3195, 1:300, AB_2291471), anti-Ly6G (rat, BioLegend, 127601, 1:100, AB_1089179), anti-HIF1α (rabbit, Novus, NB100-479, 1:300, AB_10000633), anti-CGRP (rabbit, Cell Signaling, 14959, 1:300, AB_2798662).

    Article Title: Remote Activation of Spinal TRPV1 by Magnetic Nanocubes Confers Cardioprotection Against Myocardial Ischemia‐Reperfusion Injury
    Article Snippet: The primary antibodies used were rabbit anti‐CGRP (14959, CST) and guinea pig anti‐SP (GP14110, Neuromics).

    Article Title: Transcranial Photobiomodulation Attenuates Neurogenic Inflammation and Sensory Hypersensitivity in a Nitroglycerin‐Induced Migraine Model: Toward a Nonpharmacological Therapy
    Article Snippet: After the membranes were blocked with 5% milk, they were treated with the primary antibodies listed below: anti‐PACAP (Cat No.: ab181205, Abcam), anti‐c‐Fos (Cat No.: ab222699, Abcam), anti‐TRPA1 (Cat No.: NB110‐40763SS, Novus Biologicals), anti‐TRPV1 (Cat No.: 22686‐1‐AP, PTGLAB), anti‐CGRP (Cat No.: 14959T, Cell Signal), anti‐IL‐1 β (Cat No.: ab254360, Abcam), anti‐NF‐κB p65 phospho S536 (Cat No.: ab76302, Abcam), and anti‐ β ‐actin (Cat No: ab8227, Abcam).

    Article Title: Morphine Tolerance Gated through EZH2‐Mediated Suppression of Trpc5 in Spinal GABAergic Interneurons in Male Mice
    Article Snippet: Cells wells without cells were washed by calcium‐free kreb's buffer comprising sulfinpyrazone three times and incubated with sulfinpyrazone in calcium‐free kreb's buffer for 30 min. After 30 min of incubation, 96‐well plates were transferred to a fluorescence laser plate reader (FLIPR Tetra; Molecular Devices, Sunnyvale, CA, USA) incubation chamber and the emitted fluorescence were recorded at 521 nm after excitation at 488 nm (F base and F 0 ).

    Staining:

    Article Title: Beyond polarization: single-cell atlas unveils macrophage heterogeneity and neuro-immune crosstalk in bone regeneration.
    Article Snippet: Bone regeneration is a complex and carefully orchestrated physiological process of bone formation.. Immune cells, especially macrophages, play a key role in MSC recruitment, osteoblast differentiation and osteoclast formation.. However, the traditional concept of macrophage polarization is inadequate to match its functional diversity and mechanistic complexity.

    Cell-Signaling:

    Article Title: Semaphorin 7A regulates axon outgrowth in subcutaneous white adipose tissue
    Article Snippet: .. Primary antibodies: anti-TH (1:500, Millipore, Cat# ab152), anti-CGRP (1:500, Cell Signaling, Cat# D5R8F), and anti-plexin C1 (1:500, R&D Systems, Cat# AF5375). .. Secondary antibodies: Goat anti-rabbit IgG (H + L) Alexa Fluor Plus 594 (1:1000, Invitrogen, Cat# A32740) and donkey anti-sheep IgG (H + L) Alexa Fluor 594 (1:1000, Invitrogen, Cat# A-11016).



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    Generation of iPSC line BO-VC1 -derived nociceptive neurons. Differentiation of ( A ) human iPSC line BO-VC1 into nociceptive neurons was done by dual SMAD inhibition, suppressing TGF-ß and BMP4 signaling, in knockout serum replacement (KSR) medium (day 0–4). This was followed by an overlapping inhibition of glycogen synthase kinase-3ß (GSK-3ß), vascular endothelial growth factor (VEGF), and Notch signaling (day 2–12), with KSR medium incremented with N2/B27 medium every second day by 25% from day 4 on. After reseeding of the ( B ) immature sensory neurons, final maturation was performed in N2/B27 medium infused with the neurotrophic factors glial cell line-derived neurotrophic factor (GDNF), nerve growth factor (NGF), and brain-derived neurotrophic factor (BDNF) (day 13 - approx. 60) according to the protocols of Chambers and Schoepf leading to the differentiation of ( C ) intermediate and finally ( D ) mature nociceptors (Scale bar: 500 μm). Mature nociceptive neurons express ( E ) the neuron specific microtubule element βIII-tubulin (βIIITUB) and vGLUT2, the sensory neuron markers ( F ) insulin gene enhancer protein 1 (ISL1) as well as the brain-specific homeobox/POU domain protein 3 A (BRN3A). Additionally, iPSC-derived differentiated cells express the nociceptive markers ( G ) transient receptor potential cation channel subfamily V member 1(TRPV1), H voltage-gated sodium ion channel 1.9 (Na V 1.9) and I TWIK-related spinal cord potassium channel (TRESK). Moreover, co-expression of ( J ) the calcitonin gene-related peptide <t>(CGRP),</t> the nociceptive marker high affinity nerve growth factor <t>receptor</t> <t>(TRKA)</t> and the estrogen receptor G-protein coupled estrogen receptor 1 (GPER1) was confirmed (Scale bars: 50 μm) ( K ) Quantification of the percentage of positive cells for the respective markers was 95.96 ± 2,495% for CGRP, 89.68 ± 6,137% for GPER1, 91.67 ± 5.270% for Na V 1.9, 95.83 ± 4.167% for TRKA, 80.38 ± 7.143% for TRPV1 and 98.55 ± 0.2899% for TRESK. L Realtime-PCR analysis in order to quantify nociceptive neuron marker and GPER1 gene expression of HDFa-derived iPSCs (line BO-VC1) and corresponding nociceptive neurons relative to HDFa cell samples mean value. Means ± SEM (standard error of the mean) from three independent passages were statistically analyzed by one-way ANOVA with Turkey’s multiple comparison test (L). n = 3 (* p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001)
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    Generation of iPSC line BO-VC1 -derived nociceptive neurons. Differentiation of ( A ) human iPSC line BO-VC1 into nociceptive neurons was done by dual SMAD inhibition, suppressing TGF-ß and BMP4 signaling, in knockout serum replacement (KSR) medium (day 0–4). This was followed by an overlapping inhibition of glycogen synthase kinase-3ß (GSK-3ß), vascular endothelial growth factor (VEGF), and Notch signaling (day 2–12), with KSR medium incremented with N2/B27 medium every second day by 25% from day 4 on. After reseeding of the ( B ) immature sensory neurons, final maturation was performed in N2/B27 medium infused with the neurotrophic factors glial cell line-derived neurotrophic factor (GDNF), nerve growth factor (NGF), and brain-derived neurotrophic factor (BDNF) (day 13 - approx. 60) according to the protocols of Chambers and Schoepf leading to the differentiation of ( C ) intermediate and finally ( D ) mature nociceptors (Scale bar: 500 μm). Mature nociceptive neurons express ( E ) the neuron specific microtubule element βIII-tubulin (βIIITUB) and vGLUT2, the sensory neuron markers ( F ) insulin gene enhancer protein 1 (ISL1) as well as the brain-specific homeobox/POU domain protein 3 A (BRN3A). Additionally, iPSC-derived differentiated cells express the nociceptive markers ( G ) transient receptor potential cation channel subfamily V member 1(TRPV1), H voltage-gated sodium ion channel 1.9 (Na V 1.9) and I TWIK-related spinal cord potassium channel (TRESK). Moreover, co-expression of ( J ) the calcitonin gene-related peptide <t>(CGRP),</t> the nociceptive marker high affinity nerve growth factor <t>receptor</t> <t>(TRKA)</t> and the estrogen receptor G-protein coupled estrogen receptor 1 (GPER1) was confirmed (Scale bars: 50 μm) ( K ) Quantification of the percentage of positive cells for the respective markers was 95.96 ± 2,495% for CGRP, 89.68 ± 6,137% for GPER1, 91.67 ± 5.270% for Na V 1.9, 95.83 ± 4.167% for TRKA, 80.38 ± 7.143% for TRPV1 and 98.55 ± 0.2899% for TRESK. L Realtime-PCR analysis in order to quantify nociceptive neuron marker and GPER1 gene expression of HDFa-derived iPSCs (line BO-VC1) and corresponding nociceptive neurons relative to HDFa cell samples mean value. Means ± SEM (standard error of the mean) from three independent passages were statistically analyzed by one-way ANOVA with Turkey’s multiple comparison test (L). n = 3 (* p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001)
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    Generation of iPSC line BO-VC1 -derived nociceptive neurons. Differentiation of ( A ) human iPSC line BO-VC1 into nociceptive neurons was done by dual SMAD inhibition, suppressing TGF-ß and BMP4 signaling, in knockout serum replacement (KSR) medium (day 0–4). This was followed by an overlapping inhibition of glycogen synthase kinase-3ß (GSK-3ß), vascular endothelial growth factor (VEGF), and Notch signaling (day 2–12), with KSR medium incremented with N2/B27 medium every second day by 25% from day 4 on. After reseeding of the ( B ) immature sensory neurons, final maturation was performed in N2/B27 medium infused with the neurotrophic factors glial cell line-derived neurotrophic factor (GDNF), nerve growth factor (NGF), and brain-derived neurotrophic factor (BDNF) (day 13 - approx. 60) according to the protocols of Chambers and Schoepf leading to the differentiation of ( C ) intermediate and finally ( D ) mature nociceptors (Scale bar: 500 μm). Mature nociceptive neurons express ( E ) the neuron specific microtubule element βIII-tubulin (βIIITUB) and vGLUT2, the sensory neuron markers ( F ) insulin gene enhancer protein 1 (ISL1) as well as the brain-specific homeobox/POU domain protein 3 A (BRN3A). Additionally, iPSC-derived differentiated cells express the nociceptive markers ( G ) transient receptor potential cation channel subfamily V member 1(TRPV1), H voltage-gated sodium ion channel 1.9 (Na V 1.9) and I TWIK-related spinal cord potassium channel (TRESK). Moreover, co-expression of ( J ) the calcitonin gene-related peptide <t>(CGRP),</t> the nociceptive marker high affinity nerve growth factor <t>receptor</t> <t>(TRKA)</t> and the estrogen receptor G-protein coupled estrogen receptor 1 (GPER1) was confirmed (Scale bars: 50 μm) ( K ) Quantification of the percentage of positive cells for the respective markers was 95.96 ± 2,495% for CGRP, 89.68 ± 6,137% for GPER1, 91.67 ± 5.270% for Na V 1.9, 95.83 ± 4.167% for TRKA, 80.38 ± 7.143% for TRPV1 and 98.55 ± 0.2899% for TRESK. L Realtime-PCR analysis in order to quantify nociceptive neuron marker and GPER1 gene expression of HDFa-derived iPSCs (line BO-VC1) and corresponding nociceptive neurons relative to HDFa cell samples mean value. Means ± SEM (standard error of the mean) from three independent passages were statistically analyzed by one-way ANOVA with Turkey’s multiple comparison test (L). n = 3 (* p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001)
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    Generation of iPSC line BO-VC1 -derived nociceptive neurons. Differentiation of ( A ) human iPSC line BO-VC1 into nociceptive neurons was done by dual SMAD inhibition, suppressing TGF-ß and BMP4 signaling, in knockout serum replacement (KSR) medium (day 0–4). This was followed by an overlapping inhibition of glycogen synthase kinase-3ß (GSK-3ß), vascular endothelial growth factor (VEGF), and Notch signaling (day 2–12), with KSR medium incremented with N2/B27 medium every second day by 25% from day 4 on. After reseeding of the ( B ) immature sensory neurons, final maturation was performed in N2/B27 medium infused with the neurotrophic factors glial cell line-derived neurotrophic factor (GDNF), nerve growth factor (NGF), and brain-derived neurotrophic factor (BDNF) (day 13 - approx. 60) according to the protocols of Chambers and Schoepf leading to the differentiation of ( C ) intermediate and finally ( D ) mature nociceptors (Scale bar: 500 μm). Mature nociceptive neurons express ( E ) the neuron specific microtubule element βIII-tubulin (βIIITUB) and vGLUT2, the sensory neuron markers ( F ) insulin gene enhancer protein 1 (ISL1) as well as the brain-specific homeobox/POU domain protein 3 A (BRN3A). Additionally, iPSC-derived differentiated cells express the nociceptive markers ( G ) transient receptor potential cation channel subfamily V member 1(TRPV1), H voltage-gated sodium ion channel 1.9 (Na V 1.9) and I TWIK-related spinal cord potassium channel (TRESK). Moreover, co-expression of ( J ) the calcitonin gene-related peptide <t>(CGRP),</t> the nociceptive marker high affinity nerve growth factor <t>receptor</t> <t>(TRKA)</t> and the estrogen receptor G-protein coupled estrogen receptor 1 (GPER1) was confirmed (Scale bars: 50 μm) ( K ) Quantification of the percentage of positive cells for the respective markers was 95.96 ± 2,495% for CGRP, 89.68 ± 6,137% for GPER1, 91.67 ± 5.270% for Na V 1.9, 95.83 ± 4.167% for TRKA, 80.38 ± 7.143% for TRPV1 and 98.55 ± 0.2899% for TRESK. L Realtime-PCR analysis in order to quantify nociceptive neuron marker and GPER1 gene expression of HDFa-derived iPSCs (line BO-VC1) and corresponding nociceptive neurons relative to HDFa cell samples mean value. Means ± SEM (standard error of the mean) from three independent passages were statistically analyzed by one-way ANOVA with Turkey’s multiple comparison test (L). n = 3 (* p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001)
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    Generation of iPSC line BO-VC1 -derived nociceptive neurons. Differentiation of ( A ) human iPSC line BO-VC1 into nociceptive neurons was done by dual SMAD inhibition, suppressing TGF-ß and BMP4 signaling, in knockout serum replacement (KSR) medium (day 0–4). This was followed by an overlapping inhibition of glycogen synthase kinase-3ß (GSK-3ß), vascular endothelial growth factor (VEGF), and Notch signaling (day 2–12), with KSR medium incremented with N2/B27 medium every second day by 25% from day 4 on. After reseeding of the ( B ) immature sensory neurons, final maturation was performed in N2/B27 medium infused with the neurotrophic factors glial cell line-derived neurotrophic factor (GDNF), nerve growth factor (NGF), and brain-derived neurotrophic factor (BDNF) (day 13 - approx. 60) according to the protocols of Chambers and Schoepf leading to the differentiation of ( C ) intermediate and finally ( D ) mature nociceptors (Scale bar: 500 μm). Mature nociceptive neurons express ( E ) the neuron specific microtubule element βIII-tubulin (βIIITUB) and vGLUT2, the sensory neuron markers ( F ) insulin gene enhancer protein 1 (ISL1) as well as the brain-specific homeobox/POU domain protein 3 A (BRN3A). Additionally, iPSC-derived differentiated cells express the nociceptive markers ( G ) transient receptor potential cation channel subfamily V member 1(TRPV1), H voltage-gated sodium ion channel 1.9 (Na V 1.9) and I TWIK-related spinal cord potassium channel (TRESK). Moreover, co-expression of ( J ) the calcitonin gene-related peptide <t>(CGRP),</t> the nociceptive marker high affinity nerve growth factor <t>receptor</t> <t>(TRKA)</t> and the estrogen receptor G-protein coupled estrogen receptor 1 (GPER1) was confirmed (Scale bars: 50 μm) ( K ) Quantification of the percentage of positive cells for the respective markers was 95.96 ± 2,495% for CGRP, 89.68 ± 6,137% for GPER1, 91.67 ± 5.270% for Na V 1.9, 95.83 ± 4.167% for TRKA, 80.38 ± 7.143% for TRPV1 and 98.55 ± 0.2899% for TRESK. L Realtime-PCR analysis in order to quantify nociceptive neuron marker and GPER1 gene expression of HDFa-derived iPSCs (line BO-VC1) and corresponding nociceptive neurons relative to HDFa cell samples mean value. Means ± SEM (standard error of the mean) from three independent passages were statistically analyzed by one-way ANOVA with Turkey’s multiple comparison test (L). n = 3 (* p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001)
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    NM-LP TK /RSV-MnCDs alleviates discogenic pain in IVDD. (A) Experimental timeline schematic. (B–C) Open-field behavioral analysis showing locomotor activity parameters (active time, central mean speed, total distance, and mean speed) across groups (n = 5). (D–E) PWL and PWT test showing alleviation of thermal and mechanical allodynia by NM-LP TK /RSV-MnCDs in LSI mice (n = 5). (F–G) IHC staining and quantification analyses of DRG sections showing expression of sensory neuropeptides <t>CGRP</t> and SP (n = 5). Scale bar, 100 μm. (H–I) IF co-labeling <t>of</t> <t>PGP9.5</t> and CGRP/SP in DRG, and corresponding quantification analyses (n = 5). Scale bar, 100 μm. Data are presented as mean ± SD. For comparisons among multiple groups at a single time point, one-way ANOVA followed was employed to determine statistical significance. For datasets involving multiple groups evaluated over time, including PWL and PWT assessments, a two-way repeated measures ANOVA was utilized. ∗, P < 0.05; ∗∗, P < 0.01; ∗∗∗, P < 0.001; ∗∗∗∗, P < 0.0001; ns = not significant.
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    Image Search Results


    Journal: Cell Reports Medicine

    Article Title: A biomimetic hybrid membrane vesicle nanoplatform attenuates tendinopathy through neuroinflammation modulation and tendon regeneration

    doi: 10.1016/j.xcrm.2026.102937

    Figure Lengend Snippet:

    Article Snippet: Rabbit Polyclonal anti-CGRP , ABclonal , Cat# A5542; RRID: AB_2766323.

    Techniques: Recombinant, Lysis, Hydroxyproline Assay, Enzyme-linked Immunosorbent Assay, Isolation, SYBR Green Assay, Bicinchoninic Acid Protein Assay, Staining, Software

    Generation of iPSC line BO-VC1 -derived nociceptive neurons. Differentiation of ( A ) human iPSC line BO-VC1 into nociceptive neurons was done by dual SMAD inhibition, suppressing TGF-ß and BMP4 signaling, in knockout serum replacement (KSR) medium (day 0–4). This was followed by an overlapping inhibition of glycogen synthase kinase-3ß (GSK-3ß), vascular endothelial growth factor (VEGF), and Notch signaling (day 2–12), with KSR medium incremented with N2/B27 medium every second day by 25% from day 4 on. After reseeding of the ( B ) immature sensory neurons, final maturation was performed in N2/B27 medium infused with the neurotrophic factors glial cell line-derived neurotrophic factor (GDNF), nerve growth factor (NGF), and brain-derived neurotrophic factor (BDNF) (day 13 - approx. 60) according to the protocols of Chambers and Schoepf leading to the differentiation of ( C ) intermediate and finally ( D ) mature nociceptors (Scale bar: 500 μm). Mature nociceptive neurons express ( E ) the neuron specific microtubule element βIII-tubulin (βIIITUB) and vGLUT2, the sensory neuron markers ( F ) insulin gene enhancer protein 1 (ISL1) as well as the brain-specific homeobox/POU domain protein 3 A (BRN3A). Additionally, iPSC-derived differentiated cells express the nociceptive markers ( G ) transient receptor potential cation channel subfamily V member 1(TRPV1), H voltage-gated sodium ion channel 1.9 (Na V 1.9) and I TWIK-related spinal cord potassium channel (TRESK). Moreover, co-expression of ( J ) the calcitonin gene-related peptide (CGRP), the nociceptive marker high affinity nerve growth factor receptor (TRKA) and the estrogen receptor G-protein coupled estrogen receptor 1 (GPER1) was confirmed (Scale bars: 50 μm) ( K ) Quantification of the percentage of positive cells for the respective markers was 95.96 ± 2,495% for CGRP, 89.68 ± 6,137% for GPER1, 91.67 ± 5.270% for Na V 1.9, 95.83 ± 4.167% for TRKA, 80.38 ± 7.143% for TRPV1 and 98.55 ± 0.2899% for TRESK. L Realtime-PCR analysis in order to quantify nociceptive neuron marker and GPER1 gene expression of HDFa-derived iPSCs (line BO-VC1) and corresponding nociceptive neurons relative to HDFa cell samples mean value. Means ± SEM (standard error of the mean) from three independent passages were statistically analyzed by one-way ANOVA with Turkey’s multiple comparison test (L). n = 3 (* p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001)

    Journal: Stem Cell Research & Therapy

    Article Title: Activation of the G-protein coupled estrogen receptor 1 (GPER1) reduces transient receptor potential vanilloid 1 (TRPV1) activity and human iPSC-derived nociceptive neuron firing

    doi: 10.1186/s13287-026-05174-3

    Figure Lengend Snippet: Generation of iPSC line BO-VC1 -derived nociceptive neurons. Differentiation of ( A ) human iPSC line BO-VC1 into nociceptive neurons was done by dual SMAD inhibition, suppressing TGF-ß and BMP4 signaling, in knockout serum replacement (KSR) medium (day 0–4). This was followed by an overlapping inhibition of glycogen synthase kinase-3ß (GSK-3ß), vascular endothelial growth factor (VEGF), and Notch signaling (day 2–12), with KSR medium incremented with N2/B27 medium every second day by 25% from day 4 on. After reseeding of the ( B ) immature sensory neurons, final maturation was performed in N2/B27 medium infused with the neurotrophic factors glial cell line-derived neurotrophic factor (GDNF), nerve growth factor (NGF), and brain-derived neurotrophic factor (BDNF) (day 13 - approx. 60) according to the protocols of Chambers and Schoepf leading to the differentiation of ( C ) intermediate and finally ( D ) mature nociceptors (Scale bar: 500 μm). Mature nociceptive neurons express ( E ) the neuron specific microtubule element βIII-tubulin (βIIITUB) and vGLUT2, the sensory neuron markers ( F ) insulin gene enhancer protein 1 (ISL1) as well as the brain-specific homeobox/POU domain protein 3 A (BRN3A). Additionally, iPSC-derived differentiated cells express the nociceptive markers ( G ) transient receptor potential cation channel subfamily V member 1(TRPV1), H voltage-gated sodium ion channel 1.9 (Na V 1.9) and I TWIK-related spinal cord potassium channel (TRESK). Moreover, co-expression of ( J ) the calcitonin gene-related peptide (CGRP), the nociceptive marker high affinity nerve growth factor receptor (TRKA) and the estrogen receptor G-protein coupled estrogen receptor 1 (GPER1) was confirmed (Scale bars: 50 μm) ( K ) Quantification of the percentage of positive cells for the respective markers was 95.96 ± 2,495% for CGRP, 89.68 ± 6,137% for GPER1, 91.67 ± 5.270% for Na V 1.9, 95.83 ± 4.167% for TRKA, 80.38 ± 7.143% for TRPV1 and 98.55 ± 0.2899% for TRESK. L Realtime-PCR analysis in order to quantify nociceptive neuron marker and GPER1 gene expression of HDFa-derived iPSCs (line BO-VC1) and corresponding nociceptive neurons relative to HDFa cell samples mean value. Means ± SEM (standard error of the mean) from three independent passages were statistically analyzed by one-way ANOVA with Turkey’s multiple comparison test (L). n = 3 (* p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001)

    Article Snippet: Nevertheless, incubation with the first antibodies CGRP (1:500; # 414 004, Synaptic Systems GmbH, Göttingen, Germany), TRKA (1:500; # MA515509 , Thermo Fisher Scientific, Darmstadt, Germany), GPER1 (1:500; # ab39742, Abcam, ), BRN3A (1:100, # ab245230, Abcam, Cambridge, UK), ISL1 (1:100, #H00003670-M05, Abnova, Taipeh, Taiwan), TRPV1 (1:1000, # SAB5700857, Sigma-Aldrich, Merck, Darmstadt, Germany), vGlut2 (1:400, # MAB5504, Sigma-Aldrich, Merck, Darmstadt, Germany), ßIIITUB (1:300, # PA1-41331, Thermo Fisher Scientific, Darmstadt, Germany or 1:250, #302 304, Synaptic Systems GmbH, Göttingen, Germany) and Na V 1.9 (1:100, # ab65160, Abcam, Cambridge, UK) was done at 4 °C overnight.

    Techniques: Derivative Assay, Inhibition, Knock-Out, Expressing, Marker, Gene Expression, Comparison

    NM-LP TK /RSV-MnCDs alleviates discogenic pain in IVDD. (A) Experimental timeline schematic. (B–C) Open-field behavioral analysis showing locomotor activity parameters (active time, central mean speed, total distance, and mean speed) across groups (n = 5). (D–E) PWL and PWT test showing alleviation of thermal and mechanical allodynia by NM-LP TK /RSV-MnCDs in LSI mice (n = 5). (F–G) IHC staining and quantification analyses of DRG sections showing expression of sensory neuropeptides CGRP and SP (n = 5). Scale bar, 100 μm. (H–I) IF co-labeling of PGP9.5 and CGRP/SP in DRG, and corresponding quantification analyses (n = 5). Scale bar, 100 μm. Data are presented as mean ± SD. For comparisons among multiple groups at a single time point, one-way ANOVA followed was employed to determine statistical significance. For datasets involving multiple groups evaluated over time, including PWL and PWT assessments, a two-way repeated measures ANOVA was utilized. ∗, P < 0.05; ∗∗, P < 0.01; ∗∗∗, P < 0.001; ∗∗∗∗, P < 0.0001; ns = not significant.

    Journal: Bioactive Materials

    Article Title: An intelligent nanoliposome alleviates disc degeneration and discogenic pain by inhibiting neurovascular ingrowth via a “Soil-conditioning, seed-modulating, and weeds-suppressing” strategy

    doi: 10.1016/j.bioactmat.2025.11.048

    Figure Lengend Snippet: NM-LP TK /RSV-MnCDs alleviates discogenic pain in IVDD. (A) Experimental timeline schematic. (B–C) Open-field behavioral analysis showing locomotor activity parameters (active time, central mean speed, total distance, and mean speed) across groups (n = 5). (D–E) PWL and PWT test showing alleviation of thermal and mechanical allodynia by NM-LP TK /RSV-MnCDs in LSI mice (n = 5). (F–G) IHC staining and quantification analyses of DRG sections showing expression of sensory neuropeptides CGRP and SP (n = 5). Scale bar, 100 μm. (H–I) IF co-labeling of PGP9.5 and CGRP/SP in DRG, and corresponding quantification analyses (n = 5). Scale bar, 100 μm. Data are presented as mean ± SD. For comparisons among multiple groups at a single time point, one-way ANOVA followed was employed to determine statistical significance. For datasets involving multiple groups evaluated over time, including PWL and PWT assessments, a two-way repeated measures ANOVA was utilized. ∗, P < 0.05; ∗∗, P < 0.01; ∗∗∗, P < 0.001; ∗∗∗∗, P < 0.0001; ns = not significant.

    Article Snippet: For immunofluorescence (IF) staining, antigen retrieval was conducted using citrate buffer (0.1 M, pH 6.0), followed by blocking in 10 % normal goat serum (Solarbio, Cat. No. SL038) at room temperature for 1 h. Sections were incubated overnight at 4 °C with primary antibodies targeting ACAN (Cat. No. DF7561, Affinity), COL2A1 (Cat. No. AF0135, Affinity), MMP3 (Cat. No. AF0217, Affinity), MMP13 (Cat. No. 18165-1-AP, Proteintech), CD31 (Cat. No. AF0077, Affinity), p-LATS1 (Cat. No. AF7170, Affinity), p-YAP (Cat. No. AF3328, Affinity), Tuj1 (Cat. No. 66375, Proteintech), CGRP (Cat. No. DF7386, Affinity), SP (Cat. No. DF7522, Affinity), and PGP9.5 (Cat. No. AF5490, Affinity) followed by incubation with appropriate fluorescent secondary antibodies for 2 h at room temperature and counterstaining with DAPI (Beyotime, Cat. No. C1006).

    Techniques: Activity Assay, Immunohistochemistry, Expressing, Labeling