Journal: Development (Cambridge, England)

Article Title: N-cadherin facilitates trigeminal sensory neuron outgrowth and target tissue innervation

doi: 10.1242/dev.204369

Figure Lengend Snippet: Blocking N-cadherin-mediated adhesion reduces trigeminal neuron axon outgrowth. (A) Schematic of a N-cadherin function-blocking assay. Created in BioRender. McIntosh, A. (2025) https://BioRender.com/4yud5sc . This figure was sublicensed under CC-BY 4.0 terms. (B-D″) Workflow for in vitro analyses. (B,C,D) Representative images of wild-type trigeminal ganglion explants after 24 h of incubation and fixation (pre-treatment control explants, n =31), or post-treatment with either anti-N-cadherin (C, n =9) or rat IgG (D, n =7) antibody, followed by Tubb3 immunostaining. Scale bar: 1 mm. (B′,C′,D′) Binary images generated after ridge detection processing on Fiji for pre-treatment (B′), anti-N-cadherin-treated (C′) or rat IgG-treated (D′) explants. Dotted lines indicate perimeter of explanted tissue cropped during processing. (B″,C″,D″) Processed images overlaid with concentric circle function to count the number of axonal intersections at increasing intervals of 175 µm. (E) Mean signal intensity for explant outgrowth±s.e.m. (F) Mean number of axonal intersections±s.e.m. Significant differences between pre-treatment control explants and anti-N-cadherin-treated explants are observed at 525 µm ( P <0.05), and 175, 350, 875, 1225 and 1400 µm ( P <0.01). Significant differences between pre-treatment controls and rat IgG-treated explants are observed at 1750 µm ( P <0.05), 1575 µm ( P <0.01), and 175-1400 µm ( P <0.0001). Significant differences between anti-N-cadherin- and rat IgG-treated explants are observed at 175, 350 and 700 µm ( P <0.05), and 525 µm ( P <0.01). (G) Mean segment length±s.e.m. for pre-treatment, anti-N-cadherin-treated and rat IgG-treated explants. (H) Violin plots of segment lengths for pre-treatment, anti-N-cadherin-treated and rat IgG-treated explants. Solid black lines indicate median; dotted black lines indicate the interquartile range. Statistical significance was determined using unpaired t -tests and Mann–Whitney tests (E-G) with adjustments made for multiple comparisons (F). * P <0.05, ** P <0.01.

Article Snippet: After 24 h of incubation, fresh media containing either an antibody to block N-cadherin function, MNCD2 (anti-N-cadherin, Developmental Studies Hybridoma Bank, 0.1 μg/ml) or a normal rat IgG control (Santa Cruz Biotechnology, 1.6 μg/ml) replaced the initial explant plating media, and explants were incubated overnight.

Techniques: Blocking Assay, In Vitro, Incubation, Control, Immunostaining, Generated, MANN-WHITNEY

Journal: Development (Cambridge, England)

Article Title: N-cadherin facilitates trigeminal sensory neuron outgrowth and target tissue innervation

doi: 10.1242/dev.204369

Figure Lengend Snippet: The majority of placodal neurons rely on N-cadherin for axon outgrowth. (A-D″) Representative images of trigeminal explant cultures with RFP-labeled placodal neurons after rat IgG (A-B″, n =8) or anti-N-cadherin (C-D″, n =8) treatment, and RFP and Tubb3 immunostaining. Areas outlined in A and C are shown at higher magnification in B-B″ and D-D″, respectively. Dotted lines in A and C indicate the perimeter of the explanted tissue. Scale bars: in A, 500 µm for A and C; in B, 100 µm for B-B″,D-D″. (E) Mean signal intensity for Tubb3 and RFP±s.e.m. (F) Quantitative PCR for RFP normalized against chick 18s rRNA of rat IgG- and anti-N-cadherin-treated explants ( n =3). (G) Trigeminal ganglion explant culture schematic, with cores (inside the explant) and axons (outside the explant) comprising the explant culture. Created in BioRender. McIntosh, A. (2025) https://BioRender.com/gg9u4t2 . This figure was sublicensed under CC-BY 4.0 terms. (H) Ratio of RFP signal found within axons and cores±s.e.m. Statistical significance was determined using unpaired t -tests and Mann–Whitney tests * P <0.05, *** P <0.001.

Article Snippet: After 24 h of incubation, fresh media containing either an antibody to block N-cadherin function, MNCD2 (anti-N-cadherin, Developmental Studies Hybridoma Bank, 0.1 μg/ml) or a normal rat IgG control (Santa Cruz Biotechnology, 1.6 μg/ml) replaced the initial explant plating media, and explants were incubated overnight.

Techniques: Labeling, Immunostaining, Real-time Polymerase Chain Reaction, MANN-WHITNEY

Journal: Development (Cambridge, England)

Article Title: N-cadherin facilitates trigeminal sensory neuron outgrowth and target tissue innervation

doi: 10.1242/dev.204369

Figure Lengend Snippet: Blocking N-cadherin-mediated adhesion reduces neurite complexity in neural crest-derived neurons. (A) Schematic of N-cadherin function blocking assay after labeling neural crest-derived neurons with PiggyBacGFP. Created in BioRender. McIntosh, A. (2025) https://https://BioRender.com/cobg1l4 . This figure was sublicensed under CC-BY 4.0 terms. (B,C) Representative images of trigeminal explant cultures with GFP-labeled neural crest-derived neurons after control rat IgG (B, n =7) or anti-N-cadherin (C, n =7) treatment. (B′,C′) GFP immunostaining and neurite traced images. Arrowheads and asterisks indicate examples of primary and secondary neurites, respectively. Scale bars: in B, 50 µm for B,C; in B′, 25 µm for B′,C′. (D,E) Number (D) and length (E) of GFP-positive neural crest-derived neurites after rat IgG or anti-N-cadherin treatment. Data are mean ±s.e.m. Statistical significance was determined using unpaired t -tests and Mann–Whitney tests. * P <0.05, ** P <0.01; ns, not significant.

Article Snippet: After 24 h of incubation, fresh media containing either an antibody to block N-cadherin function, MNCD2 (anti-N-cadherin, Developmental Studies Hybridoma Bank, 0.1 μg/ml) or a normal rat IgG control (Santa Cruz Biotechnology, 1.6 μg/ml) replaced the initial explant plating media, and explants were incubated overnight.

Techniques: Blocking Assay, Derivative Assay, Labeling, Control, Immunostaining, MANN-WHITNEY