animal position  (Med Associates Inc)

Journal: bioRxiv

Article Title: Identification of a Neuroimmune Circuit that Regulates Allergic Inflammation in the Esophagus

doi: 10.1101/2024.11.16.623883

Figure Lengend Snippet: (A) Whole esophagus of BIII-tubulin in NaV1.8-Cre R26CAG-floxStop-tdTomato mice. Shown are surfaces of NaV1.8+ nerves (red) within 200nm of basal epithelium (blue) with BIII-tubulin (white) around the perimeter. (B) ( top ) Cross-sectional slice of whole esophagus from NaV1.8 R26CAG-floxStop-tdTomato co-stained with BIII-tubulin (white). Yellow asterisk labels the lumen of the esophagus. ( bottom ) NaV1.8+ and BIII-tubulin+ signal in whole murine esophagus section demonstrating the distinct distribution of nociceptive (red) and myelinated (white) nerves. (C) Distribution of BIII-tubulin and NaV1.8tdT nerves as measured from the basal layer of the epithelium. Blue line designates the median. **p<0.005, Mann-Whittney test. (D) Immunofluorescence image of BIII-tubulin (white) in murine whole mount esophagus.

Article Snippet: Nociceptive neuron reporter animals were generated by crossing the NaV1.8-Cre positive animal (Jax Stock#: 036564) with the tdTomato (tdTom) reporter mouse (B6.Cg-Gt(ROSA)26Sortm14(CAG-tdTomato)Hze/J) purchased from The Jackson Laboratory (Stock#: 007914).

Techniques: Staining, Immunofluorescence

Journal: bioRxiv

Article Title: Identification of a Neuroimmune Circuit that Regulates Allergic Inflammation in the Esophagus

doi: 10.1101/2024.11.16.623883

Figure Lengend Snippet: Increased esophageal sensory neuron innervation density in the lamina propria of allergen-treated mice. (A) Representative immunofluorescence images of BIII-tubulin in longitudinal esophageal cuts from NaV1.8-Cre R26CAG-floxStop-tdTomato mice treated with PBS and intranasal ALT. (B) Murine Esophageal NaV1.8-tdT+ nerve area; LP tissue control vs Alternaria treated, ****p<0.0001. Data were obtained from 3 independent experiments; allergen treated (n=8) vs. Control (n=4). (C) Murine Esophageal BIII-tubulin nerve density. Data were obtained from 2 independent experiments; allergen treated (n=6) vs. Control (n=5). Individual data points represent a separate section of esophagus. Two-way ANOVA with multiple comparisons, horizontal line shows mean.

Article Snippet: Nociceptive neuron reporter animals were generated by crossing the NaV1.8-Cre positive animal (Jax Stock#: 036564) with the tdTomato (tdTom) reporter mouse (B6.Cg-Gt(ROSA)26Sortm14(CAG-tdTomato)Hze/J) purchased from The Jackson Laboratory (Stock#: 007914).

Techniques: Immunofluorescence, Control

Journal: bioRxiv

Article Title: Identification of a Neuroimmune Circuit that Regulates Allergic Inflammation in the Esophagus

doi: 10.1101/2024.11.16.623883

Figure Lengend Snippet: (A-B) 60x water Sora images showing interaction between BIII-tubulin+ nerves (white) and (A) tryptase (green) and (B) eosinophil peroxidase (EPX) (green) in active EoE patient esophageal biopsy. (C) Comparison of IL4Ra, CRLF2, IL13Ra1, IL13Ra2, IL1RL1, and IL5R, expression by qRT-PCR in whole mouse naive DRG n = 4 WT mice. (D) Representative calcium transient of whole DRG calcium response to recombinant mouse IL-4 (300nm). Graph outlines total number of KCl responsive cells to IL-4 (67/392). (E-F) (E) Capsaicin responsive and (F) ATP responsive DRG neurons as a percentage of total KCl-responsive neurons (left) and fluorescence intensity changes (right), in response to capsaicin (1 um) or ATP (150 um) stimulation after vehicle (gray) or IL-4 (300nm) (green) exposure. Each dot represents one cell. *p<0.05, ****p<0.0001, percent responders: chi squared test, fluorescent intensity changes: unpaired T-test; DRG n>100 from 3 Pirt-GCAMP6 mice. Data shown as mean +/-SEM. (G) Murine esophageal Nav1.8tdT nerve density. Data were obtained from 2 independent experiments; WT control, WT allergen treated, IL-4R-/- NaV1.8 control, and IL-4R-/- NaV1.8 allergen treated. n≥4 mice per group; individual data points represent a separate section of esophagus. Two-way ANOVA with multiple comparisons, horizontal line shows mean.

Article Snippet: Nociceptive neuron reporter animals were generated by crossing the NaV1.8-Cre positive animal (Jax Stock#: 036564) with the tdTomato (tdTom) reporter mouse (B6.Cg-Gt(ROSA)26Sortm14(CAG-tdTomato)Hze/J) purchased from The Jackson Laboratory (Stock#: 007914).

Techniques: Comparison, Expressing, Quantitative RT-PCR, Recombinant, Fluorescence, Control

Journal: bioRxiv

Article Title: Identification of a Neuroimmune Circuit that Regulates Allergic Inflammation in the Esophagus

doi: 10.1101/2024.11.16.623883

Figure Lengend Snippet: (A) Heat map based on 582 differentially expressed genes in control vs allergen challenge esophagus. Upregulated genes (red) vs downregulated (blue) of expression profiles of differentially dysregulated genes. ≥1.5-fold change; P<0.05; TPM≥1; Each column represents one mouse (B) Reactome analysis of 481 upregulated genes in the esophagus of allergen treated mice. Data shows top 7 pathways based on FDR p-value. (C) Biological STRING process analysis based on 25 upregulated genes in allergen challenged WT vs IL-4R-/- NaV1.8 mice. Colors of circles correspond with pathways of the same color. Line thickness indicates the strength of data support. (D) Representative immunohistochemistry staining of eosinophil major basic protein (MBP) staining from WT control, WT allergen treated, IL-4R-/- NaV1.8 control, and IL-4R-/- NaV1.8 allergen treated. (E) Eosinophil major basic protein (MBP) pixel quantification per esophageal section comparing WT control, WT allergen treated, IL-4R-/- NaV1.8 control, and IL-4R-/- NaV1.8 allergen treated mice. n≥2 mice per group; individual data points represent a separate section of esophagus. One-way ANOVA with multiple comparisons, horizontal line shows mean.

Article Snippet: Nociceptive neuron reporter animals were generated by crossing the NaV1.8-Cre positive animal (Jax Stock#: 036564) with the tdTomato (tdTom) reporter mouse (B6.Cg-Gt(ROSA)26Sortm14(CAG-tdTomato)Hze/J) purchased from The Jackson Laboratory (Stock#: 007914).

Techniques: Control, Expressing, Immunohistochemistry, Staining

Journal: bioRxiv

Article Title: Identification of a Neuroimmune Circuit that Regulates Allergic Inflammation in the Esophagus

doi: 10.1101/2024.11.16.623883

Figure Lengend Snippet: (A) Heat map based on 224 differentially expressed genes in DRGs from WT allergen treated mice vs, IL-4R-/- NaV1.8 allergen treated mice. Upregulated genes (red) vs downregulated (blue) of expression profiles of differentially dysregulated genes. ≥1.5-fold change; P<0.05; TPM≥1; Each column represents one mouse. (E) ( left) Reactome analysis of 215 downregulated genes in the DRG of IL-4R-/- NaV1.8 allergen treated mice compared with WT allergen treated. Data shows top 8 pathways based on FDR p-value. ( right) Genes for the selected term ( grey bar ) are listed with the corresponding fold change in expression for allergen-treated IL-4R-/- NaV1. mice compared to WT allergen treated samples. The x-axes represent the negative log p-value.

Article Snippet: Nociceptive neuron reporter animals were generated by crossing the NaV1.8-Cre positive animal (Jax Stock#: 036564) with the tdTomato (tdTom) reporter mouse (B6.Cg-Gt(ROSA)26Sortm14(CAG-tdTomato)Hze/J) purchased from The Jackson Laboratory (Stock#: 007914).

Techniques: Expressing

Journal: Cell reports

Article Title: Macrophage memories of early-life injury drive neonatal nociceptive priming

doi: 10.1016/j.celrep.2024.114129

Figure Lengend Snippet: (A) Schematic of knocking out p75NTR from macrophages following neonatal incision. (B) Representative images of p75NTR labeling (green) co-stained with F4/80 (macrophages, white) in the injured hindpaw. Yellow arrows, double-positive staining. Scale bar, 25 μM. (C) There is a significant reduction (F = 4.012) in the percentage of double-positive cells in LysM;p75fl/fl animals compared to other genotypes (* p < 0.05 vs. controls (p75+/fl and p75 fl/fl) and LysM; p75+/fl, one way ANOVA, Tukey’s, n = 3–5/group). (D) We found no effect of tamoxifen prior to the second injury (No Tam vs. BL time points) on paw guarding. Following the second incision, there was robust paw guarding (F = 26.258) with a faster recovery in the LysM;p75NTRfl/fl mice (* p < 0.05 vs. BL, # p = 0.05 vs. BL p75 fl/fl, two-way RM ANOVA, Tukey’s). (E) No effect of tamoxifen on mechanical withdrawal thresholds was found prior to injury (No Tam vs. BL time points); however, there was an effect following a second incision (F = 40.516). Both groups displayed mechanical sensitivity on days 1, 3, and 7 (* p < 0.01 vs. BL). p75fl/fl controls continued to display significant sensitivities at day 14 while LysM; p75 fl/fl animals did not (p75fl/fl: * p = 0.002 vs. BL, LysM;p75fl/fl: p = 0.063 vs. BL). One day following the second incision, LysM;p75fl/fl animals were less hypersensitive than controls ( ^ p = 0.01 vs. controls). Two-way RM ANOVA, Tukey’s, n = 7/group. Colored horizontal lines indicate the duration of significance compared to BL for each group. Mean ± SEM.

Article Snippet: Myeloid/Macrophage reporter animals were generated by crossing the LysM-Cre positive animal (see Jax Stock#: 004781) with the tdTomato (tdTom) reporter mouse (B6.Cg-Gt(ROSA)26Sor tm14(CAG-tdTomato)Hze /J) purchased from The Jackson Laboratory (Stock#: 007914).

Techniques: Labeling, Staining

Journal: Cell reports

Article Title: Macrophage memories of early-life injury drive neonatal nociceptive priming

doi: 10.1016/j.celrep.2024.114129

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Myeloid/Macrophage reporter animals were generated by crossing the LysM-Cre positive animal (see Jax Stock#: 004781) with the tdTomato (tdTom) reporter mouse (B6.Cg-Gt(ROSA)26Sor tm14(CAG-tdTomato)Hze /J) purchased from The Jackson Laboratory (Stock#: 007914).

Techniques: Recombinant

Journal: bioRxiv

Article Title: EZ-SPOTs: A simple and robust high-throughput liquid handling platform

doi: 10.1101/2024.05.13.594031

Figure Lengend Snippet: Results from AST experiments. A) Dose-response curves for a single isolate with a single antibiotic where gray points (n=3) indicate replicates for each dose and black points indicate averages with standard deviation bars. Background color is based on the MIC (defined as the lowest concentration of an antibiotic that completely inhibits growth) of each isolate and is scaled based on CLSI resistance or susceptibility breakpoints for each antibiotic; B) Dose-response curves for 12 isolates against 6 antibiotics where most are seen to be highly resistant to certain antibiotics and a range of susceptibilities to others; C) MIC comparison to commercially available Sensititre panel where most MIC’s match identically and few are only one two-fold dilution off. Additionally, more specific MIC information was obtained for most isolates and antibiotics.

Article Snippet: These strains were run on the Thermo Fisher Sensititre Companion Animal Gram Positive AST plates (COMPGP1F) according to CLSI protocols (Cornell Animal Health Diagnostic Center, Bacteriology Laboratory) and identified as methicillin resistant.

Techniques: Standard Deviation, Concentration Assay, Comparison