Safarian et al., 2020 - Visuomotor deficiency in panx1a knockout zebrafish is linked to dopaminergic signaling. Scientific Reports   10:9538 Full text @ Sci. Rep.

Fig. 3 ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

Fig. 4 ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

PHENOTYPE:
Fish:
Conditions:
Observed In:
Stage: Day 6

Fig. 5 ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

PHENOTYPE:
Fish:
Observed In:
Stage: Day 6

Fig. 6 ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

EXPRESSION / LABELING:
Genes:
Fish:
Anatomical Terms:
Stage: Day 6
PHENOTYPE:
Fish:
Observed In:
Stage: Day 6

Fig. 7 ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

Fig. 8 ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

Figure 1

Generating panx1a−/− fish using TALENs – (a) The zebrafish panx1a gene structure with six coding exons. The position of the left and right TALENs sequence with the spacer sequence and AfeI restriction site in blue and red is highlighted. (b) Larval survival rates (in %) one day after (1 dpf) microinjection. A dose of 30 pg TALENs pair resulted in more than 50% survival rate and was selected for the experiments generating the panx1a−/− fish line. (c) The RFLP-assay shows the loss of the AfeI recognition sequence (indicated by uncut) in four out of ten F0 larvae tested. (d) A sequencing alignment is demonstrating small deletions causing frameshift mutations found in three different F0 larvae. (e) The predicted sequence features of the mutated panx1a were visualized using the Protter open-source tool (wlab.ethz.ch/protter). A 4 bp deletion in Panx1a exon 4 resulted in a frameshift causing a premature stop codon at amino acid 195. (f) Localization of the truncated Panx1a protein – Confocal images of transiently transfected proteins Panx1awt-EGFP (left panel) and Panx1aΔ4-EGFP (right panel) in Neuro2a cells. Nuclei were stained with DAPI. Scale bar: 10 μm.

Figure 2

Phenotypic characterization of panx1a−/− larvae – (a) Age-matched wild type TL and panx1a−/− larvae (6 dpf) showing regular morphology. (b) RT-qPCR analysis of pannexin expression in 6 dpf larva. (c) The panx1a expression is reduced in the adult panx1a−/− zebrafish. The expression was determined using an affinity purified rabbit anti-panx1a antibody directed against the carboxy-terminal 129 amino acids of the Panx1a protein. Detection of the PSD-95 protein in cone terminals served as an internal control. Cell nuclei were stained with DAPI. In wild type TL fish arrows indicate Panx1a immunoreactivity found in the horizontal cell layer as reported previously19,20. The immunoreactivity was significantly reduced in panx1a−/− fish. Residual staining was overlapping with DAPI stained nuclei and considered as unspecific. Abbreviations: PL, photoreceptor layer; ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer; GCL, ganglion cell layer. Scale bar 100 µm. n.d., Not detected. Significance: ***p-value < 0.001.

EXPRESSION / LABELING:
Genes:
Antibodies:
Fish:
Anatomical Terms:
Stage Range: Day 6 to Adult
PHENOTYPE:
Fish:
Observed In:
Stage Range: Day 6 to Adult
Acknowledgments:
ZFIN wishes to thank the journal Scientific Reports for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Sci. Rep.