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. 2018 Jun 19;7:e34519. doi: 10.7554/eLife.34519

Figure 4. Pancreatic islet parasympathetic innervation is derived from the neural crest.

(A) Lineage tracing of neural crest cells in Tg(sox10:CreERT2, myl7:GFP); Tg(ubb:loxP-CFP-loxP-nuc-mCherry) zebrafish following 5 µM tamoxifen treatment from 16 to 24 hpf and staining at 90 and 120 hpf. (B) Whole mount immunostaining at 90 hpf for GFP (elavl3-positive cells), mCherry (neural-crest-derived cells), Somatostatin (delta cells), Insulin (beta cells), and Glucagon (alpha cells). (C–E) Whole mount immunostaining at 90 (C, D) and 120 (E) hpf for acetylated Tubulin (nerves), Insulin (beta cells), mCherry (neural-crest-derived cells), and DAPI (DNA). Yellow arrows point to neural-crest-derived cells on the periphery of the pancreatic islet; blue arrowheads point to neural-crest-derived cells projecting neural extensions toward the pancreatic islet, and some of these cells are adjacent to the vagus nerve (V).

Figure 4.

Figure 4—figure supplement 1. Neural crest cells are essential for the establishment of islet parasympathetic innervation.

Figure 4—figure supplement 1.

(A) Immunostaining analysis of innervation density in wild-type and sox10 mutants at 80 hpf. (B) Wild-type and sox10-/- zebrafish at 80 hpf. (C) Body length measurements at 80 hpf, mean ± SEM, n = 26–27 animals, p-values from t tests are presented. (D) Whole larva free glucose level measurements at 80 hpf, mean ± SEM, n = 21–22 batches of five larvae per replicate. (E–F) Whole mount immunostaining at 80 hpf for acetylated Tubulin (nerves), Insulin (beta cells), Somatostatin (delta cells), and DAPI (DNA) of wild-type and sox10 mutants. sox10 mutants display a severe decline in enteric nervous density and a complete loss of pancreatic innervation, n = 8 animals.