Journal: Cell Reports Methods
Article Title: Development of a 3-dimensional organotypic model with characteristics of peripheral sensory nerves
doi: 10.1016/j.crmeth.2024.100835
Figure Lengend Snippet: Structural characteristics and ganglion expression of representative ion channels, receptors, and sensory neuron markers in the sensory nerve organotypic model (A) Schematic of the organotypic model, showing its structure. (B) Electron micrographs of an axon bundle cross-section containing abundant myelinated nerve fibers 28 days after seeding. (C) Enlarged images of whole-axon-bundle coronal section (dotted red square in B). A fibers (green), myelinating Schwann cells (red), C fibers (orange) and non-myelinating Schwann cells (blue) were present in the axon bundle. The enlarged views of the A fiber (a), multi-layered myelin sheath (b), and C fiber (c) correspond to the dotted squares in (B). (D) Electron micrographs of an axon bundle sagittal section showing a node of Ranvier 28 days after seeding. (E) 3D reconstructed image of the axon bundle sagittal section in (D), showing a node of Ranvier. (F) Schematic showing sample collection of a ganglion-like structure of an organotypic culture for RT-PCR. (G–I) RT-PCR bands showing mRNA levels of (G) cation channels, (H) nerve fiber markers, (I) functional receptors and pro-nociceptive cytokines and chemokines in ganglia of organotypic cultures 28 days after seeding. GAPDH was used as an internal control. n = 3 . (J–N) Confocal images showing Nav1.8-positive (J, green), TRPV1-positive (K, green), TRPA1-positive (K, red), TrkA-positive (L, green), CGRP-positive (L, red), TrkB-positive (M, green), NF200-positive (M and N, red), and Npy2r-positive (N, green) neuronal cell bodies in the ganglion-like structure of an organotypic model 28 days after seeding. Scale bars: 10 μm (B), 500 nm (C), 1 μm (D), and 50 μm (J–N).
Article Snippet: Rabbit anti-Nav1.8 , Alomone labs , Cat#ASC-016-GP; RRID: AB_2040188.
Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Functional Assay, Control