XB-IMG-77381
Xenbase Image ID: 77381
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Fig. 3. Dominant-negative KATP channel subunits cause defects in endogenous eye development and induce a variety of ectopic eye tissues. (A) Stage 42 tadpoles showed a significantly high incidence of eye phenotypes (both malformed eyes and ectopic eye tissue) upon microinjection of dominant-negative KATP mRNAs (DNKir6.1p and DNKir6.1-ER), in comparison to control injections, in all four cells of four-cell embryos. Values are mean ± s.e.m. (n=5). *, P<0.05; **, P<0.01; one-way ANOVA, Tukey's post test. (B-H) Stage 42 control tadpoles (B,C) and those microinjected with DNKir6.1p mRNA (D-H) as detailed in A. Green arrow and arrowhead mark endogenous eyes. Red arrow and arrowheads mark ectopic eye tissues. E shows multiple ectopic patches of RPE. Note whole ectopic eyes with lens (F,G) and ectopic eye tissue in tail (H). (I,J) β-crystallin immunostaining (red) in stage 42 whole tadpole (I) and lateral sections through tadpoles (J) injected with DNKir6.1p mRNA in all four cells at the four-cell stage. Blue arrowheads mark distinct ectopic lens tissue in head and tail region, with green arrowheads marking the endogenous eye lens tissue. (K,L) Transverse sections of stage 42 tadpoles microinjected with DNKir6.1p mRNA in all four cells at the four-cell stage. Green arrows marks endogenous eyes and red arrowheads mark ectopic eye tissues. Note that the ectopic eye is present on the ventral side of the tadpole in K and along lateral mesoderm in L. (M) Hematoxylin and Eosin staining of the endogenous eye (i) and ectopic eye tissue (ii) show a similar histological organization of cell layers. (N) A narrow range of membrane voltage induces ectopic eye tissue. Quantification of tadpoles with ectopic eye tissues at stage 42 upon microinjection of neonatal Nav1.5 channel mRNA in all four cells at the four-cell stage. To vary the Vmem in injected cells by control of influx of positive ions through this channel, embryos were incubated in different concentrations of sodium gluconate (10, 15, 20 and 40 mM); 20 mM is isomolar with intracellular sodium (equilibrium). Ectopic eyes were induced only in the hyperpolarizing condition created by 15 mM sodium gluconate. See also supplementary material Fig. S3, Table S2. Image published in: Pai VP et al. (2012) Copyright © 2012. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License.
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