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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).
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