Journal: Neuroscience Bulletin

Article Title: Fibroblast Growth Factor 8 Suppresses Neurotoxic Astrocytes and Alleviates Neuropathic Pain via Spinal FGFR3 Signaling

doi: 10.1007/s12264-025-01533-x

Figure Lengend Snippet: FGF8 alleviates SNI-induced mechanical allodynia through FGFR3. A , B Paw withdrawal threshold ( A ) and von Frey–evoked response frequency ( B ) in SNI mice following a single intrathecal injection of FGF8 (1 μg in 5 μL) on day 14 post-SNI. C Time course of mechanical allodynia response over five consecutive days following FGF8 administration on day 14 post-SNI. D , E qRT-PCR analysis showed changes of Fgfr2 ( D ) and Fgfr3 ( E ) in spinal cord tissue on day 14 post-SNI. F , G Representative images showing co-localization of FGFR3 with GFAP in the SDH of sham and SNI mice ( F ) and quantification ( G ) on day 14 post-SNI. H , I Representative images and quantification of FGFR3 knockdown efficiency in spinal astrocytes using FGFR3-targeting shRNA. Arrowheads indicate the typical cell. J , K Selective knockdown of FGFR3 attenuated the analgesic effects of intrathecal FGF8 injection on day 14 post-SNI. n = 9 mice ( A , C) ; n = 4–6 mice ( B–E , J , K ); n = 10-12 slices ( G , I ) from 3 mice. Scale bar: 25 μm ( F , H ). Statistics by two-way ANOVA followed by post hoc Holm-Sidak test ( A–C , J , K ) and two-tailed Student’s t -test ( D–I ). * P < 0.05, ** P < 0.01. Data are presented as mean ± SEM. I.t, intrathecal injection.

Article Snippet: Recombinant FGF8 protein (0.5 μg or 1 μg in 5 μL PBS; R&D Systems, 423-F8-025/CF) was administered via intrathecal (i.t.) lumbar puncture between L5–L6 vertebrae under isoflurane anesthesia using a 31G needle-equipped Hamilton syringe (25 μL volume).

Techniques: Injection, Quantitative RT-PCR, Knockdown, shRNA, Two Tailed Test

Journal: Neuroscience Bulletin

Article Title: Fibroblast Growth Factor 8 Suppresses Neurotoxic Astrocytes and Alleviates Neuropathic Pain via Spinal FGFR3 Signaling

doi: 10.1007/s12264-025-01533-x

Figure Lengend Snippet: FGF8 alleviates inflammatory induction of neurotoxic astrocyte states in vitro. A Relative mRNA expression of C3 in isolated astrocytes treated with normal culture medium, LPS, or MCM, as measured by qRT-PCR. B – I Expression of reactive astrocyte markers in astrocytes treated with IL-1α, TNF-α, and C1q, with or without FGF8 co-treatment. A1-specific transcripts: C3 ( B ), H2-T23 ( D ), and Serping1 ( E ); A2-specific transcripts: S100a10 ( C ), Ptx3 ( F ), and Ptgs ( G ); pan-reactive transcripts: Lcn2 ( H ) and Cxcl10 ( I ). J–L Relative mRNA expression of synaptogenic genes Gpc4 ( J ), Gpc6 ( K ), Tsp1, and Tsp2 ( L ) in astrocytes under the same treatment conditions. M , N Relative mRNA expression of Fgfr2 ( M ) and Fgfr3 ( N ) in isolated astrocytes treated with normal culture medium, LPS, or MCM, as measured by qRT-PCR. n = 6–12 petri dishes ( A–C ) and n = 3–7 petri dishes ( M , N ) derived from pooled spinal cords of 6 mice per preparation; n = 3–6 petri dishes ( D–I ) and n = 4–6 petri dishes ( J–L ) from pooled spinal cords of 12 mice per preparation. Statistics by one-way ANOVA by post hoc Holm-Sidak test ( A–N ). * P < 0.05, ** P < 0.01. # P < 0.05, ## P < 0.01. Data are presented as mean ± SEM. LPS, lipopolysaccharide. MCM, microglia-conditioned media.

Article Snippet: Recombinant FGF8 protein (0.5 μg or 1 μg in 5 μL PBS; R&D Systems, 423-F8-025/CF) was administered via intrathecal (i.t.) lumbar puncture between L5–L6 vertebrae under isoflurane anesthesia using a 31G needle-equipped Hamilton syringe (25 μL volume).

Techniques: In Vitro, Expressing, Isolation, Quantitative RT-PCR, Derivative Assay

Journal: Neuroscience Bulletin

Article Title: Fibroblast Growth Factor 8 Suppresses Neurotoxic Astrocytes and Alleviates Neuropathic Pain via Spinal FGFR3 Signaling

doi: 10.1007/s12264-025-01533-x

Figure Lengend Snippet: Central action of FGF8 enhances inhibitory synaptic transmission in the spinal cord. A – D Representative firing traces ( A ) and quantitative statistics showing no significant changes in firing frequencies ( B ), RMP ( C ), or rheobase current ( D ) in lamina II neurons following perfusion with FGF8 (500 ng/mL). E – G Representative traces of sEPSCs of lamina II neurons ( E ) and quantitative statistics of sEPSC frequency ( F ) and amplitude ( G ). H – J Representative traces of sIPSCs of lamina II neurons ( H ) and quantitative statistics of sIPSC frequency ( I ) and amplitude ( J ). n = 18–19 neurons ( B – D ) and n = 15–19 neurons ( F , G , I , J ) from 3 mice. Statistics by two-way ANOVA followed by post hoc Holm-Sidak test ( B ), two-tailed Student’s t -test ( C , D ), and Mann-Whitney U test ( F , G , I , J ).** P < 0.01. Data are presented as mean ± SEM. AP, action potential. RMP, resting membrane potential. sEPSC, spontaneous excitatory postsynaptic current. sIPSC, spontaneous inhibitory postsynaptic current.

Article Snippet: Recombinant FGF8 protein (0.5 μg or 1 μg in 5 μL PBS; R&D Systems, 423-F8-025/CF) was administered via intrathecal (i.t.) lumbar puncture between L5–L6 vertebrae under isoflurane anesthesia using a 31G needle-equipped Hamilton syringe (25 μL volume).

Techniques: Transmission Assay, Two Tailed Test, MANN-WHITNEY, Membrane

Journal: Neuroscience Bulletin

Article Title: Fibroblast Growth Factor 8 Suppresses Neurotoxic Astrocytes and Alleviates Neuropathic Pain via Spinal FGFR3 Signaling

doi: 10.1007/s12264-025-01533-x

Figure Lengend Snippet: SNI induces persistent microglial activation in the SDH. A , B Representative immunofluorescence images ( A ) and quantification ( B ) of IBA-1 staining in the ipsilateral SDH from day 1 to day 14 post-SNI. C , D Representative Western blot images ( C ) and quantification ( D ) of IBA-1 protein levels in the L2–L5 spinal cord segments from day 4 to day 14 post-SNI. E Representative skeletonized images of individual IBA-1⁺ microglia from sham and SNI mice. F , G Quantification of microglial morphology on days 4, 7, and 14 post-SNI, showing a significant increase in soma area ( F ) and a decrease in process length ( G ) compared to sham controls. H – J Sholl analysis demonstrates reduced microglial process complexity in SNI mice compared to sham controls. n = 9 slices. K qRT-PCR analysis of Tnf-α , Il-1α , and C1q in the ipsilateral SDH from sham and SNI mice on day 14 ( n = 5–6 mice). L qRT-PCR analysis of Tnf-α , Il-1α , and C1q in spinal cord tissue from SNI mice treated with FGF8 or vehicle on day 14 ( n = 6 per group). M Representative images of C3 and GFAP co-localization and quantification of C3 and GFAP in the SDH of control and FGFR3 shRNA mice following i.t. LPS administration. n = 9 slices ( B ), n = 8–11 cells ( F – J ) from 3 mice; n = 4–6 mice ( D , K , L ); n = 12-13 slices ( M ) from 3 mice. Statistics by two-way ANOVA followed by post hoc Holm-Sidak test ( B , D , F , G ). Scale bar: 25 μm ( A , E , M ). Statistics by two-way ANOVA ( H-I ), two-tailed Student’s t -test ( K , L ), and Mann-Whitney U test ( M ). * P < 0.05, ** P < 0.01. Data are presented as mean ± SEM.

Article Snippet: Recombinant FGF8 protein (0.5 μg or 1 μg in 5 μL PBS; R&D Systems, 423-F8-025/CF) was administered via intrathecal (i.t.) lumbar puncture between L5–L6 vertebrae under isoflurane anesthesia using a 31G needle-equipped Hamilton syringe (25 μL volume).

Techniques: Activation Assay, Immunofluorescence, Staining, Western Blot, Quantitative RT-PCR, Control, shRNA, Two Tailed Test, MANN-WHITNEY

Journal: The Journal of Infectious Diseases

Article Title: FGF8 Protects Against Polymicrobial Sepsis by Enhancing the Host's Anti-infective Immunity

doi: 10.1093/infdis/jiae559

Figure Lengend Snippet: Kinetics of FGF8 production in polymicrobial sepsis. A , Male C57BL/6 mice (n = 5 per group) were subjected to sham or CLP using a 24-gauge needle. PLF, serum, lung, spleen and kidney were collected at the indicated times (6, 24, 48 hours) after CLP. FGF8 concentrations were measured by ELISA. B , Representative fluorescence images of FGF8 expression in kidney, spleen, and lung after CLP. Scale bar = 50 μm. Quantitative results are shown (n = 3 per group). C , FGF8 concentrations in serum and PLF of TLR2 −/− , TLR4 −/− , TLR2/4 −/− , and WT mice 24 hours (n = 3–4 per group) after CLP. D , Supernatant of heat-killed Pseudomonas aeruginosa (MOI = 1:100) challenged macrophages was collected at the indicated times (6, 12, 24 hours). FGF8 levels were measured by ELISA (n = 4 per group). A–D , Data are representative of 3 independent experiments; Kruskal-Wallis test followed by Dunn multiple comparisons posttest. * P < .05, ** P < .01, *** P < .001, **** P < .0001 compared within 2 groups. Abbreviations: CLP, cecal ligation and puncture; Ctrl, control; ELISA, enzyme-linked immunosorbent assay; FGF, fibroblast growth factor; MOI, multiplicity of infection; ns, not significant; PLF, peritoneal lavage fluid; TLR, Toll-like receptor; WT, wild type.

Article Snippet: Recombinant murine FGF8 (rFGF8; 423-F8; R&D Systems) was administered IP to mice at 5 and 12.5 μg/kg at the indicated times following CLP and phosphate-buffered saline (PBS) was administered to controls in the same manner.

Techniques: Enzyme-linked Immunosorbent Assay, Fluorescence, Expressing, Ligation, Control, Infection

Journal: The Journal of Infectious Diseases

Article Title: FGF8 Protects Against Polymicrobial Sepsis by Enhancing the Host's Anti-infective Immunity

doi: 10.1093/infdis/jiae559

Figure Lengend Snippet: Inhibition of endogenous FGF8 exacerbated polymicrobial sepsis. A , Survival of septic mice (n = 20 per group) with or without FGF8 neutralization after CLP using a 24-gauge needle. B , Dilutions of PLF, blood, liver, lung, and spleen tissues were obtained from septic mice (n = 10–11 per group) treated with or without monoclonal anti-FGF8 antibody at 24 hours after CLP; samples were cultured on blood agar plates and the numbers of bacterial colonies were then determined. C , Representative examples of hematoxylin and eosin-stained tissues as indicated from mice (n = 5 per group) treated or not treated with monoclonal anti-FGF8 antibody at 24 hours after CLP. The pathology scores are shown on the right side of histological images. D , Serological markers of organ injury including ALT, AST, LDH, creatinine, and urea in septic mice (n = 11–12 per group) treated with or without monoclonal anti-FGF8 antibody at 24 hours after CLP. Each dot represents an individual mouse. B–D , Nonparametric Mann-Whitney U test; ( A ) Kaplan-Meier analysis followed by log-rank test. * P < .05, ** P < .01, *** P < .001 compared within 2 groups. Abbreviations: ALT, alanine transaminase; AST, aspartate transaminase; CLP, cecal ligation and puncture; FGF, fibroblast growth factor; IgG, immunoglobulin G; LDH, lactate dehydrogenase; ns, not significant; PLF, peritoneal lavage fluid.

Article Snippet: Recombinant murine FGF8 (rFGF8; 423-F8; R&D Systems) was administered IP to mice at 5 and 12.5 μg/kg at the indicated times following CLP and phosphate-buffered saline (PBS) was administered to controls in the same manner.

Techniques: Inhibition, Neutralization, Cell Culture, Staining, MANN-WHITNEY, Ligation

Journal: The Journal of Infectious Diseases

Article Title: FGF8 Protects Against Polymicrobial Sepsis by Enhancing the Host's Anti-infective Immunity

doi: 10.1093/infdis/jiae559

Figure Lengend Snippet: FGF8 treatment improves outcomes in a murine sepsis model. A , Different levels of rFGF8 (0, 5, 12.5 μg/kg) were injected intraperitoneally into mice (n = 20 per group) immediately after CLP and survival was monitored for 14 days. B , Dilutions of blood, lungs, PLF, and spleens obtained from septic mice (n = 9–10 per group) treated with or without rFGF8 (12.5 μg/kg) 48 hours after CLP. C , Representative examples of hematoxylin and eosin-stained lung, liver, spleen, and kidney tissues from CLP mice treated with or without rFGF8 (12.5 μg/kg) after CLP. Scale bars = 400 μm. Histological scores (n = 5 per group) are shown. D , Serological markers of organ injury in septic mice (n = 13 per group) treated with or without rFGF8 (12.5 μg/kg) at 48 hours after CLP. B–D , Data are representatives of 3 independent experiments; nonparametric Mann-Whitney U test; ( A ) Kaplan-Meier analysis followed by log-rank test. * P < .05, ** P < .01 compared within 2 groups. Abbreviations: ALT, alanine transaminase; AST, aspartate transaminase; CLP, cecal ligation and puncture; FGF, fibroblast growth factor; LDH, lactate dehydrogenase; ns, not significant; PBS, phosphate-buffered saline; PLF, peritoneal lavage fluid.

Article Snippet: Recombinant murine FGF8 (rFGF8; 423-F8; R&D Systems) was administered IP to mice at 5 and 12.5 μg/kg at the indicated times following CLP and phosphate-buffered saline (PBS) was administered to controls in the same manner.

Techniques: Injection, Staining, MANN-WHITNEY, Ligation, Saline

Journal: The Journal of Infectious Diseases

Article Title: FGF8 Protects Against Polymicrobial Sepsis by Enhancing the Host's Anti-infective Immunity

doi: 10.1093/infdis/jiae559

Figure Lengend Snippet: FGF8 directly enhances bacterial phagocytosis and killing by macrophages. A , Peritoneal macrophages (1 × 10 6 cells) were stimulated with or without rFGF8 (200 ng/mL) for 6 hours and then challenged with FITC-labeled Pseudomonas aeruginosa (MOI = 1:100) for 30 minutes at 37°C. Representative images from 3 independent experiments are shown. Dot plot depicts macrophage phagocytosis levels (n = 5 per group). B , Bacterial killing of P. aeruginosa in peritoneal macrophages (5 × 10 5 cells) treated with PBS or the indicated doses of rFGF8. Dot plot depicts macrophage killing (n = 4 per group). C , Mortality of rFGF8-treated (12.5 μg/kg) septic mice in the presence or absence of macrophage depletion after CLP (n = 10 per group). D , Bacterial loads in PLF and blood of rFGF8-treated (12.5 μg/kg) septic mice in the presence or absence of macrophage depletion after CLP (n = 5 per group). E , Survival after transfer of rFGF8- or PBS-treated peritoneal macrophages in mice (n = 12 per group) after CLP. A, B, and D , Data are representatives of 3 independent experiments; ( A ) nonparametric Mann-Whitney U test; ( B and D ) Kruskal-Wallis test followed by Dunn multiple comparisons posttest; ( C and E ) Kaplan-Meier analysis followed by log-rank test. * P < .05, ** P < .01, *** P < .001, **** P < .0001 compared within 2 groups. Abbreviations: CLP, cecal ligation and puncture; CFU, colony-forming unit; DAPI, 4′,6-diamidino-2-phenylindole; FITC, fluorescein isothiocyanate; MOI, multiplicity of infection; ns, not significant; PBS, phosphate-buffered saline; rFGF, recombinant fibroblast growth factor; TRITC, tetraethyl rhodamine isothiocyanate.

Article Snippet: Recombinant murine FGF8 (rFGF8; 423-F8; R&D Systems) was administered IP to mice at 5 and 12.5 μg/kg at the indicated times following CLP and phosphate-buffered saline (PBS) was administered to controls in the same manner.

Techniques: Labeling, MANN-WHITNEY, Ligation, Infection, Saline, Recombinant

Journal: The Journal of Infectious Diseases

Article Title: FGF8 Protects Against Polymicrobial Sepsis by Enhancing the Host's Anti-infective Immunity

doi: 10.1093/infdis/jiae559

Figure Lengend Snippet: FGFR1 plays a critical role in FGF8-induced protection against experimental sepsis. A , Representative confocal images of the colocalization of FGF8 (Cy3) and FGFR (FITC) in peritoneal macrophages treated with rFGF8 (200 ng/mL). Scale bar = 20 μm. B , Peritoneal macrophages were pretreated with or without rFGF8 (200 ng/mL) followed by incubation with or without heat-inactivated Pseudomonas aeruginosa. Representative fluorescence images of phospho-FGFR1 expression are shown. Scale bar = 25 μm. C , Peritoneal macrophages (n = 4 per group) were pretreated with or without the FGFR1 inhibitor PD173074 (100 nM) for 1 hour followed by incubation with or without rFGF8 (200 ng/mL). In vitro bacterial phagocytosis and killing of P. aeruginosa . D , Mortality after blocking FGFR1 with PD173074 (1 mg/kg) and subsequent treatment with rFGF8 (12.5 μg/kg) or PBS control after CLP (n = 15 per group). Except for survival ( D ), data are presented as means and are representative of 3 independent experiments; ( C ) Kruskal-Wallis test followed by Dunn multiple comparisons posttest; ( D ) Kaplan-Meier analysis followed by log-rank test. * P < .05, *** P < .001, **** P < .0001 compared within 2 groups. Abbreviations: CLP, cecal ligation and puncture; FGFR, FGF receptor; ns, not significant; rFGF, recombinant fibroblast growth factor; FITC, Fluorescein Isothiocyanate; Cy3, Cyanine 3.

Article Snippet: Recombinant murine FGF8 (rFGF8; 423-F8; R&D Systems) was administered IP to mice at 5 and 12.5 μg/kg at the indicated times following CLP and phosphate-buffered saline (PBS) was administered to controls in the same manner.

Techniques: Incubation, Fluorescence, Expressing, In Vitro, Blocking Assay, Control, Ligation, Recombinant

Journal: The Journal of Infectious Diseases

Article Title: FGF8 Protects Against Polymicrobial Sepsis by Enhancing the Host's Anti-infective Immunity

doi: 10.1093/infdis/jiae559

Figure Lengend Snippet: FGF8-enhanced antibacterial functions of macrophages are regulated by ERK1/2 signaling pathways. A , Peritoneal murine macrophages were treated with or without rFGF8 (200 ng/mL) for the indicated times and examined for the presence of phosphorylated ERK1/2, P38, STAT1, and Akt. Three independent experiments were performed with comparable results and representative blots are shown. B , Effects of rFGF8 on the activation of MAPK, STAT, PI3 K signaling pathways in murine macrophages. Peritoneal macrophages were treated with or without rFGF8 (200 ng/mL) and then incubated with heat-inactivated Pseudomonas aeruginosa . Total cellular proteins were extracted from murine macrophages for the detection of phosphorylated ERK1/2, P38, STAT1, and Akt with the indicated antibodies by western blot analysis. Experiments were performed in 3 independent experiments with consistent results and representative blots are shown. Peritoneal macrophages (n = 4 per group) were pretreated with the ERK1/2 inhibitor U0126 (20 μM) for 1 hour followed by incubation with or without rFGF8 (200 ng/mL). C , Analysis of in vitro bacterial phagocytosis and killing of P. aeruginosa . D , Mortality after blocking ERK1/2 signaling pathway with U0126 (10 mg/kg) and subsequent treatment with rFGF8 (12.5 μg/kg) or PBS control after CLP (n = 20 per group). Except for survival rate ( D ), data are presented as means and are representative of 3 independent experiments; ( C ) Kruskal-Wallis test followed by Dunn multiple comparisons posttest; ( D ) Kaplan-Meier analysis followed by log-rank test. * P < .05, ** P < .01, **** P < .0001 compared within 2 groups. Abbreviations: CFU, colony-forming unit; CLP, cecal ligation and puncture; DMSO, dimethyl sulfoxide; ns, not significant; PBS, phosphate-buffered saline; rFGF, recombinant fibroblast growth factor.

Article Snippet: Recombinant murine FGF8 (rFGF8; 423-F8; R&D Systems) was administered IP to mice at 5 and 12.5 μg/kg at the indicated times following CLP and phosphate-buffered saline (PBS) was administered to controls in the same manner.

Techniques: Protein-Protein interactions, Activation Assay, Incubation, Western Blot, In Vitro, Blocking Assay, Control, Ligation, Saline, Recombinant

Journal: The Journal of Infectious Diseases

Article Title: FGF8 Protects Against Polymicrobial Sepsis by Enhancing the Host's Anti-infective Immunity

doi: 10.1093/infdis/jiae559

Figure Lengend Snippet: FGF8 is a candidate biomarker for sepsis. A , Levels of FGF8 at admission measured by ELISA in serum samples collected from 73 adult patients with sepsis, 96 child patients with sepsis, and corresponding healthy controls. B , Receiver operating characteristic analysis of FGF8 for diagnosis of sepsis (AUC = 0.89 for adult and AUC = 0.81 for children). C , Comparison of serum FGF8 levels between male and female patients with sepsis and healthy controls. D , Comparison of serum FGF8 levels between adults and children in healthy controls and patients with sepsis. A , C , and D , Nonparametric Mann-Whitney U test. **** P < .0001 compared within 2 groups. Abbreviations: AUC, area under the curve; CI, confidence interval; ELISA, enzyme-linked immunosorbent assay; FGF, fibroblast growth factor; ns, not significant.

Article Snippet: Recombinant murine FGF8 (rFGF8; 423-F8; R&D Systems) was administered IP to mice at 5 and 12.5 μg/kg at the indicated times following CLP and phosphate-buffered saline (PBS) was administered to controls in the same manner.

Techniques: Biomarker Discovery, Enzyme-linked Immunosorbent Assay, Comparison, MANN-WHITNEY

Journal: Communications Biology

Article Title: Sonic hedgehog and fibroblast growth factor 8 regulate the evolution of amniote facial proportions

doi: 10.1038/s42003-025-07522-0

Figure Lengend Snippet: Fgf8 expression (orange) in whole mount in situ hybridization in Anolis embryos at developmental Stage 3 ( A ), Stage 4 ( B ), Stage 5 ( C ), and Stage 6 ( D ). Fn frontal process, Fnp frontonasal process, Mxp maxillary process, Mp mandibular process. Scale bar = 200 μm. Stage 18 embryos treated at oviposition with DMSO as control ( E ) and treated with 100 μM Infigratinib ( F ). Scale bar = 1 mm. Skull of Anolis Stage 18 in DMSO ( G ) and 100 μM Infigratinib ( H ) rendered from µCT, premaxilla in red, maxilla in blue, nasal in yellow, not to scale.

Article Snippet: Expression of Fgf8 (blue) and Shh (red) in early stages of head development in lizard ( Anolis sagrei ), chicken ( Gallus gallus ) , – and mouse ( Mus musculus ) , , , .

Techniques: Expressing, In Situ Hybridization, Control

Journal: Communications Biology

Article Title: Sonic hedgehog and fibroblast growth factor 8 regulate the evolution of amniote facial proportions

doi: 10.1038/s42003-025-07522-0

Figure Lengend Snippet: Expression of Fgf8 (blue) and Shh (red) in early stages of head development in lizard ( Anolis sagrei ), chicken ( Gallus gallus ) , – and mouse ( Mus musculus ) , , , . Images not to scale. N nasal pit, Mxp maxillary process, Mp Mandibular process.

Article Snippet: Expression of Fgf8 (blue) and Shh (red) in early stages of head development in lizard ( Anolis sagrei ), chicken ( Gallus gallus ) , – and mouse ( Mus musculus ) , , , .

Techniques: Expressing