XB-IMG-173554
Xenbase Image ID: 173554
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Rescue of the ability to regenerate tails during the refractory period in the X. laevis tadpoles. (A) Different rates of regeneration success in the refractory tadpoles with or without Ras-dva1/2 overexpression. The average percent mean values of regenerating tadpoles with different success rates marked by colors: gray - no regeneration, pink – partial regeneration, red – normal regeneration, in tadpole butches injected with fluorescein (FLD, as control) or overexpressing xRas-dva1 and xRas-dva2 mRNAs. n – number of injected tadpoles in three independent experiments. Differences of percent of normal regeneration between regenerates injected by FLD and mRNA are statistically significant, two-tailed t-test, p < 0,05. Percent differences of not regenerating tadpoles in FLD and mRNA injected tadpoles is statistically significant, two-tailed t-test, p < 0,05. (B) The measurements of the length of regenerated tail tips by 8 day after amputation of refractory tadpoles, developed from embryos injected by FLD (control) or xRas-dva1 or xRas-dva2 mRNA. n – number of injected tadpoles in three independent experiments. Two-tailed t-test, p < 0,001 (asterisk). The transmitted light (C) and fluorescent (D) images of regenerated tails of 8 dpa refractory tadpoles injected with fluorescein (FLD) or xRas-dva1 mRNA + FLD and xRas-dva2 mRNA + FLD. FLD refractory tadpoles show regeneration arrest which is common for refractory period. Refractory tadpoles with Ras-dva genes overexpression effectively regenerate their tails. (E–G) The transmitted light images of regenerating tail tips of control tadpoles (FLD) or xRas-dva1 mRNA and xRas-dva2 mRNA at 4 dpa in refractory period show regeneration arrest in control tip but normal regeneration of notochord (nt), spinal cord (sc), muscles (m), melanophores (mp) in the tadpole’s tails with xRas-dva1 or xRas-dva2 overexpression. The red dashed line indicates the amputation level. Scale bar 1 mm. (H) qRT-PCR analysis of the expression of early regeneration marker genes Fgf20 and Msx1b during the regeneration process (at 0 and 2 dpa) after amputation in refractory period in tadpoles tails injected with FLD or xRas-dva1 or xRas-dva2 mRNAs at early developmental stages. The scheme of samples harvesting is the same as described in Fig. 2A. The value of normalized PCR signal in the 0 dpa sample, harvested immediately after amputation, was taken as an arbitrary unit (a.u.) in each series. Dpa - days post amputation. Error bars indicate SD, t-test, p < 0.05 (asterisk). Image published in: Ivanova AS et al. (2018) © The Author(s) 2018. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View |