Figure 3. Characterization of CQ effects in zebrafish ALD mutant and identification of potential for mechanism via increased scd expression.

(A) No effect of bafilomycin A1 (BMA1) on the motor behavior of WT or abcd1^sa509 mutant larvae. Distance moved after 24 hours of treatment (5 dpf) or 72 hours of treatment (7 dpf) with bafilomycin A1 at the indicated dose. (B) Effect of CQ (10 μm) on the number of Olig2⁺ cells in the spinal cord of 5 dpf abcd1^sa509 mutant larvae. Olig2⁺ cells were counted in spinal cord (region for quantification indicated by dotted line) (Z-stack confocal images of spinal cord: dorsal, top; rostral, left) using olig2:dsRed transgenic line in the abcd1^sa509 mutant background. Scale bar: 25 μM. (C) The effect of CQ (10 μm) on the expression of genes involved in fatty acid metabolism and myelin synthesis was analyzed by qRT-PCR at 7 dpf in WT or abcd1^sa509 mutant larvae (fold change of mRNA levels compared with WT control normalized to β-actin expression). abcd2, ATP-binding cassette transporter D2; mpz, myelin protein zero; plp1a, proteolipid protein 1a; mbp, myelin basic protein. (D) Loss of scd phenocopies does not worsen ALD mutant zebrafish behavior. Motor behavior of abcd1^sa509, scd/scdb, and abcd1^sa509/scd/scdb mutant 6 dpf larvae. Statistical significance by 1-way ANOVA, followed by Bonferroni’s multiple comparison test. *P < 0.05; **P < 0.01; ***P < 0.001.