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ZDB-IMAGE-220725-29

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Ab7-mapk

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Osorio-Méndez et al., 2022 - Voltage-gated sodium channel scn8a is required for innervation and regeneration of amputated adult zebrafish fins

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Fig. 4

scn8ab Regulates neuronal activity in the brain. (A) Representative section in situ hybridization of scn8ab transcripts in brain and 2 dpa fins of wild-type fish. Black arrows indicate scn8ab expression. Black arrowhead marks amputation plane. (B) RT-PCR showing scn8ab expression in the zebrafish brain but not in uninjured or regenerating (2 dpa) fins. huC/elavl3 Is used as a brain-specific gene; and4 is a representative gene for regenerating fins; and actb2 is used as a loading control. (C) RNA-seq genome-browser tracks of fin-specific genes (and4 and cldn1), brain-specific genes (elavl3 and elavl4), and scn8ab. Brain and uninjured and 36 h postamputation (hpa) fin profiles of controls were used. (D and E) Confocal, whole-mount, immuno-stained image against pan-Nav antibody of uninjured and 2 dpa wild-type fins. Dashed white lines mark amputation plane. (F) Representative images of brain phosphorylated ERK (p-ERK; red) and HuC/D (green) immunostaining in control (heterozygous siblings) and temca placed at 33 °C for 2 h prior to dissection. White arrows indicate p-ERK⁺/HuC/D⁺ double-stained neurons. (G) Quantification of p-ERK⁺ neurons in 300 × 300 µm² dorsal telencephalon region (n = 6). Data are presented as mean ± SD. Student’s unpaired two-tailed t test. (H) RNA-seq volcano plot displaying significantly reduced genes in temca placed at 33 °C for 1 d before brain collection. (I) Top Gene Ontology terms for genes up-regulated and down-regulated in temca brains. (D–F) Scale bars, 50 µm. ac-Tub, acetylated α-tubulin; Adjust, adjusted.

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This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Proc. Natl. Acad. Sci. USA