ZFIN ID: ZDB-FIG-161104-1
Saera-Vila et al., 2016 - Autophagy Regulates Cytoplasmic Remodeling During Cell Reprogramming in a Zebrafish Model of Muscle Regeneration. Autophagy   12(10):1864-1875 Full text @ Autophagy
Anatomical Term:
Stage: Adult
Observed In:
Stage: Adult

Fig. 1

Autophagy in the regenerating EOMs. (A to L) LysoTracker Red was used to label autophagy in the regenerating LR (left side), after staining fish heads were processed for cryosectioning ((A to D). (A, B) Sections containing the regenerating LR were imaged at lower magnification. (C, D) Higher-resolution detail of the boxes in (A)and (B), respectively. ((E)to L) Craniectomy was also used to visualize LysoTracker Red labeling in situ. At 16 hpi, zebrafish heads were mounted in agarose (E), and the top of the skulls was removed (F). Brain was extracted to expose the skull base (*) where the pituitary is located and both LR muscles attach to the bone (G). Then the lateral bones of the skull were removed to allow complete visualization of the LR muscles (H). (I) Fluorescent visualization of H. LysoTracker Red (J) and GFP-Lc3 (K) clearly accumulate and colocalize (L) in the regenerating LR. (M) Fluorescence intensity of GFP signal was therefore higher in the injured muscle; in fact, the ratio of the intensities of injured vs control was higher than 1 in every fish, meaning a net increase of GFP-Lc3 content of the injured muscle (Student t test, **, P < 0.01, n = 5). (N) Western blot of Lc3 showed an increase of Lc3-II in the injured muscle, indicating again autophagy activation (Mann-Whitney test, *, P < 0.05, n = 4). Protein loading was assayed with an anti-Tubg1/tubulin antibody. +, positive control (rat protein); U, uninjured; I, injured. (O to Q) Electron microscopy showed that double-membrane organelles were easily detected in the injured muscle (P, scale bar: 500 nm; Q, scale bar: 100 nm) while virtually undetectable in the control muscle (O, scale bar: 500 nm). Check Figure 30 for a diagram of a craniectomyzed zebrafish head.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP zf155Tg resection: lateral rectus Adult lateral rectus regenerating tissue IHC
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab1-map1lc3 WB WT resection: lateral rectus Adult lateral rectus regenerating tissue
Phenotype Details
Fish Conditions Stage Phenotype
WT resection: lateral rectus Adult lateral rectus regenerating tissue ab1-map1lc3 labeling increased amount, abnormal
zf155Tg resection: lateral rectus Adult autophagy increased occurrence, abnormal
Adult muscle autophagosome increased amount, abnormal
Adult regenerating tissue autophagosome increased amount, abnormal
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