FIGURE

Figure 4

ID

ZDB-FIG-220801-19

Publication

Gessler et al., 2022 - Control of Directed Cell Migration after Tubular Cell Injury by Nucleotide Signaling

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Figure 4

The involvement of purinergic P2ry2 receptors in zebrafish pronephros repair. (A) Single-cell RNA sequencing, performed with two-day-old zebrafish embryos, revealed expression of p2ry2 along the zebrafish embryo, while other P2 family members were expressed at low levels. (B–D) In situ hybridization revealed upregulation of all three zebrafish p2ry2 variants two hours after a laser-induced injury. (E) Depletion of zebrafish p2ry2.1, p2ry2.2 and p2ry2.3 by translation- (TBM) blocking morpholino oligonucleotides (MO) (each MO, 0.2 mM) significantly delayed the repair process. (F) Depletion of zebrafish p2ry2.1, p2ry2.2 and p2ry2.3 by CRISPR/Cas9 in combination with 8 sgRNAs delayed the repair process; however, the difference was statistically not significant (mean ± SEM; 2-way Anova). (G,H) High-resolution video microscopy revealed that p2ry2 depletion reduced track speed and cell displacement particularly in the proximal parts of the pronephros (mean ± SD; Mann–Whitney test).

Expression Data

Genes:	p2ry2.1 p2ry2.2 p2ry2.3
Fish:	WT
Condition:	ablation: pronephros
Anatomical Term:	pronephros
Stage:	Long-pec

Expression Detail

Gene	Fish	Conditions	Stage	Anatomy	Assay
p2ry2.1	WT	ablation: pronephros	Long-pec	pronephros	ISH
p2ry2.2	WT	ablation: pronephros	Long-pec	pronephros	ISH
p2ry2.3	WT	ablation: pronephros	Long-pec	pronephros	ISH

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Antibody Labeling

Phenotype Data

Fish:	WT WT + CRISPR1-p2ry2.1 + CRISPR1-p2ry2.2 + CRISPR1-p2ry2.3 + CRISPR2-p2ry2.1 + CRISPR2-p2ry2.2 + CRISPR2-p2ry2.3 + CRISPR3-p2ry2.1 + CRISPR4-p2ry2.1 WT + MO1-p2ry2.1 + MO1-p2ry2.2 + MO1-p2ry2.3
Condition:	ablation: pronephros
Knockdown Reagents:	CRISPR1-p2ry2.1 CRISPR1-p2ry2.2 CRISPR1-p2ry2.3 CRISPR2-p2ry2.1 CRISPR2-p2ry2.2 CRISPR2-p2ry2.3 CRISPR3-p2ry2.1 CRISPR4-p2ry2.1 MO1-p2ry2.1 MO1-p2ry2.2 MO1-p2ry2.3 (all 11)
Observed In:	pronephros pronephros regeneration
Stage:	Long-pec

Phenotype Detail

Fish	Conditions	Stage	Phenotype
WT	ablation: pronephros	Long-pec	pronephros p2ry2.1 expression increased amount, abnormal
WT	ablation: pronephros	Long-pec	pronephros p2ry2.2 expression increased amount, abnormal
WT	ablation: pronephros	Long-pec	pronephros p2ry2.3 expression increased amount, abnormal
WT + CRISPR1-p2ry2.1 + CRISPR1-p2ry2.2 + CRISPR1-p2ry2.3 + CRISPR2-p2ry2.1 + CRISPR2-p2ry2.2 + CRISPR2-p2ry2.3 + CRISPR3-p2ry2.1 + CRISPR4-p2ry2.1	ablation: pronephros	Long-pec	pronephros regeneration occurrence, normal
WT + MO1-p2ry2.1 + MO1-p2ry2.2 + MO1-p2ry2.3	ablation: pronephros	Long-pec	pronephros damaged, abnormal
WT + MO1-p2ry2.1 + MO1-p2ry2.2 + MO1-p2ry2.3	ablation: pronephros	Long-pec	pronephros regeneration decreased occurrence, abnormal
WT + MO1-p2ry2.1 + MO1-p2ry2.2 + MO1-p2ry2.3	ablation: pronephros	Long-pec	pronephros regeneration decreased rate, abnormal
WT + MO1-p2ry2.1 + MO1-p2ry2.2 + MO1-p2ry2.3	ablation: pronephros	Long-pec	pronephros regeneration delayed, abnormal

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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 @ Int. J. Mol. Sci.