Fig. 8

Overexpression of pmpcb in glial cell effectively rescues the neurogenic defects and promotes stress resistance in zebrafish. A, B Construction of zebrafish transgenic Tg(gfap:pmpcb) line. Schematic diagram of the construction of Tg(gfap:pmpcb) transgenic line (A), and this transgenic zebrafish highly expresses pmpcb in glial cell at 24, 48 and 72 hpf (B). C, D WB analysis of Pmpcb (C) and GFP (D) at 72 hpf in WTs and pmpcb−/− larvae at 72 hpf, and the calculation of the relatively translational levels of Pmpcb and GFP. E WB analysis of Sox2 and Gfap in the WT siblings, pmpcb−/−, and pmpcb−/− larvae rescued with Tg(gfap:pmpcb) at 72 hpf, and the calculation of the relatively translational levels of Sox2 and Gfap. F Transcriptional expression of gfap and vimentin at 72 hpf (F, left) in the WT siblings, pmpcb−/−, and pmpcb−/− larvae rescued with Tg(gfap:pmpcb), and the calculation of the relative expression levels of gfap and vimentin (F, right). G WB analysis of Hsp60 and Lias proteins in the WT siblings, pmpcb−/−, and pmpcb−/− larvae rescued with Tg(gfap:pmpcb) at 72 hpf (G, left), and the calculation of the relatively translational levels of Hsp60 and Lias (G, right). H WB analysis of β-Catenin and β-Catenin-pS552 in the WT siblings, pmpcb−/−, and pmpcb−/− larvae rescued with Tg(gfap:pmpcb) at 72 hpf (H, left), and the calculation of the relatively translational levels of β-Catenin and β-Catenin-pS552 (H, left). I Transcriptional expression of axin2 at 72 hpf in the WT siblings, pmpcb−/−, and pmpcb−/− larvae rescued with Tg(gfap:pmpcb) (I, left), and the calculation of the relative expression levels of axin2 (I, right). J Cumulative mortality of WT siblings and Tg(gfap:pmpcb) transgenic larvae after microinjection with 104 CFU/mL Aeromonas hydrophila (Ah) at 72 hpf. hpi, hours post infection. K Cumulative mortality of WT siblings and Tg(gfap:pmpcb) embryos exposed to 7.8 μM Cu.2+ stress before sphere stage. hpf: hours post fertilization. B, F, I, lateral view, anterior to the left, and dorsal to the up. Each experiment was repeated at least three times, and a representative result is shown. Data are analyzed using GraphPad Prism 9.5 for t-test, and are presented as mean ± SD. ***P < 0.001, **P < 0.01, *P < 0.05, ns, not significant. Scale bar, 200 μm (B, F, I)