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. 2014 Oct 1;124(11):4829–4842. doi: 10.1172/JCI68506

Figure 3. Effects of cholestenoic acids on zebrafish motor neurons.

Figure 3

Tg[isl1:GFP] embryos were incubated with 10 μM test compound or vehicle added to medium, and the medium was replaced every 12 hours with fresh solution (containing test compound or vehicle). Immunocytochemistry was performed using an anti-GFP antibody at 48 hpf. (A) Dorsal (top) and dorsolateral (bottom) views of the head/upper back region of embryos treated with vehicle, 3β,7α-diHCA, or 3βH,7O-CA. Arrows indicate loci III, IV, V, VII, and X (37), which are evolutionarily homologous to the cranial nerves in humans (40). Locus III contains the oculomotor neurons, locus IV the trochlear neurons, locus V the trigeminal motor neurons, locus VII the facial motor neurons, and locus X the cell bodies of the vagus nerve. Scale bar: 50 μm. (B) Quantification of islet-1–GFP signal intensity in the different loci (n = 4). 24,25-EC was used as a positive control. (C and D) mRNA levels of (C) isl1 and (D) abca1 after treatment with the indicated compounds. Data are mean ± SEM (n = 3). *P < 0.05, **P < 0.01 vs. respective vehicle, Mann-Whitney test.