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ZIRC
ZFIN ID: ZDB-FIG-140903-31
Heijnen et al., 2014 - Ribosomal Protein Mutations Induce Autophagy through S6 Kinase Inhibition of the Insulin Pathway. PLoS Genetics   10:e1004371 Full text @ PLoS Genet.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Antibody:
Fish:
Anatomical Term:
Stage Range: Prim-5 to Long-pec
PHENOTYPE:
Fish:
Observed In:
Stage Range: Prim-5 to Long-pec

Fig. 3

Reduction of rpS7 to haploinsufficient levels in zebrafish embryos induces autophagy in RBCs.

(A) Western blot analysis of rpS7 protein levels in zebrafish carrying viral inserts in the rpS7 gene at 1 and 2 dpf. The amount of rpS7 is calculated by densitometer analysis measuring the rpS7/actin ratio (N = 3). (B) Electron micrographs of 1 dpf embryos. Representative morphology of RBCs is shown in wild type (upper left panel) and rpS7-deficient embryos (upper right and lower panels). Arrowheads indicate double-membrane autophagosomes, arrows indicate autolysosomes. N = nucleus. Size bars upper and bottom left panels = 1 μM, upper right = 500 nM, lower right = 200 nM. (C) Quantification of double-membrane structures in the RBCs of micrographs in (B). In each case 28 cells were examined. (D) O-dianisidine staining of embryos at 2 dpf. Embryos (N>100) were scored in (E) and genotyped to confirm mutation status. (F) Representative mitochondrial morphology (m) displayed in wild type (upper left panel) and rpS7-deficient embryos at 2dpf (upper right). Typical Golgi complex (G) in wild type embryos shown (lower left panel) compared to Golgi complex in rpS7-deficient embryos (lower right). Arrows denote double membranes sequestering cytosolic material, an indication for autophagosome formation. *Denotes engulfed cytoplasmic material. Size bars = 200 nM.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
rps7 Ab1-rps7 WT control Prim-5 whole organism WB
Ab1-rps7 Long-pec whole organism WB
Ab1-rps7 rps7hi1034bTg/hi1034bTg control Prim-5 whole organism WB
Ab1-rps7 Long-pec whole organism WB
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
rps7hi1034bTg/hi1034bTg control Prim-5 nucleate erythrocyte has extra parts of type nucleate erythrocyte autolysosome, abnormal
Prim-5 nucleate erythrocyte has extra parts of type nucleate erythrocyte autophagosome, abnormal
Prim-5 nucleate erythrocyte autophagy increased occurrence, abnormal
Long-pec embryonic hemopoiesis decreased process quality, abnormal
Long-pec whole organism has fewer parts of type nucleate erythrocyte, abnormal
Long-pec whole organism autophagy increased occurrence, abnormal
Long-pec whole organism Golgi apparatus morphology, abnormal
Long-pec whole organism mitochondrion morphology, abnormal
Acknowledgments:
ZFIN wishes to thank the journal PLoS Genetics for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ PLoS Genet.