Nrps are upregulated during zebrafish heart regeneration. (A) Absolute qPCR analysis of nrp family genes at 1, 3, 7, 14, 30 and 60 days following cryoinjury or sham surgery. Basal expression was evaluated in uninjured hearts of age-matched wild-type fish. Bars represent normalized copy number per reaction. Data are mean±s.e.m. **P<0.01, ***P<0.005, ****P<0.001 (one-way ANOVA with Sidak's post hoc test for multiple comparisons of n=4 or 5 with each n being a pool of five ventricles). (B) Adult zebrafish ventricle lysates 1, 3, 7, 14 and 30 days following surgery, immunoblotted for Nrp1 and Gapdh (left); western blot quantification of Nrp1 protein in sham and cryoinjured ventricles 1, 3, 7, 14 and 30 days following surgery (right) (n=4 or 5, with each nbeing a pool of three ventricles). (C) In situ hybridization with digoxigenin-labeled antisense riboprobes were used to detect nrp family isoforms in sham-operated and cryoinjured adult zebrafish hearts 1, 3 and 14 dpci (n≥3). Arrowheads indicate gene expression within the epicardium. CI, cryoinjured; epi, epicardium; IA, injured area. Scale bars: 250 µm.
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