ZFIN ID: ZDB-FIG-160824-20
Huang et al., 2016 - The circadian clock regulates autophagy directly through the nuclear hormone receptor Nr1d1/Rev-erbα and indirectly via Cebpb/(C/ebpβ) in zebrafish. Autophagy   12(8):1292-309 Full text @ Autophagy
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Condition:
Anatomical Term:
Stage Range: Day 4 to Day 5
PHENOTYPE:
Fish:
Condition:
Observed In:
Stage Range: Long-pec to Adult

Fig. 4

Disrupted autophagy activities and altered expression of autophagy genes in nr1d1 mutant zebrafish. (A) Zebrafish larval autolysosomes were estimated by expression of mCherry-Map1lc3b at 56 hpf. The scale bar in the lower left of the wild-type panel represents 100 microns. The right upper panels show high magnification highlighting a hindbrain region in both wild-type and nr1d1 mutant larva labeled with mCherry-Map1lc3b. (B) Transmission electron micrograph of wild-type and nr1d1 mutant liver sections at ZT 0. The scale bar in the lower left of the wild-type panel represents one micron. (C) Quantification of autolysosomes in (A), calculated manually. (D) Quantification of autophagosome abundance in (B) with software ImageJ. The figure represents one of the 3 independent sets of samples. Data represent mean ± s.d.(**, Pd0 .01). The Student t test was conducted, (**, P ≤ 0 .01). (E) RT-PCR analysis of autophagy genes in nr1d1 (7bp or +124,-2bp) mutant zebrafish at different time points under DD condition. Zebrafish larvae were raised under light/dark (14 h/10 h) condition for the first 3 dpf, and then placed under constant darkness condition. Approximately 50 zebrafish larvae were pooled for each time point. The mRNA expression levels were analyzed by the JTK-CYCLE method. ADJ.P for adjusted minimal p-values (*, P ≤ 0 .05; ***, P ≤ 0 .001), AMP for amplitude. Two-way ANOVA with the Tukey post hoc test was conducted (*, P ≤ 0 .05; **, P ≤ 0 .01).

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
atg4a nr1d1sus010/sus010 constant dark Day 4 to Day 5 whole organism RTPCR
nr1d1sus012/sus012 constant dark Day 4 whole organism RTPCR
Day 5 whole organism RTPCR
atg4da nr1d1sus010/sus010 constant dark Day 4 to Day 5 whole organism RTPCR
atp6v1d nr1d1sus010/sus010 constant dark Day 4 to Day 5 whole organism RTPCR
becn1 nr1d1sus010/sus010 constant dark Day 4 to Day 5 whole organism RTPCR
bnip3 nr1d1sus010/sus010 constant dark Day 4 to Day 5 whole organism RTPCR
gabarapa nr1d1sus010/sus010 constant dark Day 4 to Day 5 whole organism RTPCR
map1lc3b nr1d1sus010/sus010 constant dark Day 4 to Day 5 whole organism RTPCR
nr1d1sus012/sus012 constant dark Day 4 whole organism RTPCR
Day 5 whole organism RTPCR
ulk1a nr1d1sus010/sus010 constant dark Day 4 to Day 5 whole organism RTPCR
nr1d1sus012/sus012 constant dark Day 4 whole organism RTPCR
Day 5 whole organism RTPCR
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
nr1d1sus010/sus010 control Long-pec hindbrain autolysosome increased amount, abnormal
Adult liver autophagosome increased amount, abnormal
nr1d1sus010/sus010 constant dark Day 4 - Day 5 whole organism atg4a expression increased amount, abnormal
Day 4 - Day 5 whole organism atg4da expression increased amount, abnormal
Day 4 - Day 5 whole organism atp6v1d expression increased amount, abnormal
Day 4 - Day 5 whole organism becn1 expression increased amount, abnormal
Day 4 - Day 5 whole organism bnip3 expression increased amount, abnormal
Day 4 - Day 5 whole organism gabarapa expression increased amount, abnormal
Day 4 - Day 5 whole organism map1lc3b expression increased amount, abnormal
Day 4 - Day 5 whole organism ulk1a expression increased amount, abnormal
nr1d1sus012/sus012 constant dark Day 4 whole organism atg4a expression increased amount, abnormal
Day 4 whole organism map1lc3b expression increased amount, abnormal
Day 4 whole organism ulk1a expression increased amount, abnormal
Day 5 whole organism atg4a expression increased amount, abnormal
Day 5 whole organism map1lc3b expression increased amount, abnormal
Day 5 whole organism ulk1a expression increased amount, abnormal
Acknowledgments:
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 @ Autophagy