Xenbase Image ID: 159097

Figure 4. Abnormal touch response and primary neurogenesis in n1-src-knockdown embryos. A, Diagram showing the sequences and corresponding RNA target sequences of the splice acceptor (AMO a) and donor (AMO d) splice-blocking AMOs. B, RT-PCR analysis of c-src and n1-src mRNA expression at Stage 16 in control uninjected embryos and embryos injected with a total of 20 ng AMO a, AMO d, or AMO a+d. rpl8 is used as a ubiquitously expressed loading control. −rt, No reverse transcriptase control; water, no template control. C, Representative phenotypes of embryos at larval Stage 41 bilaterally injected at the two-cell or four-cell stage with 10 ng total of a standard control MO or the AMO a+d combination. Embryos were coinjected with 100 pg of nuclear β-galactosidase and subsequently stained with X-gal (light blue) to demonstrate successful injection targeting. D, Diagram of embryo touch reflex. Touching the skin stimulates Rohon–Beard sensory neurons (s), which synapse onto commissural interneurons (i) that activate contralateral motor neurons (m), leading to muscle contraction. E, Quantitation of touch-response phenotype of the same embryos at larval Stage 28 and 41 bilaterally injected at the two-cell or four-cell stage with 10 ng total of a standard control MO or the AMO a+d combination. Data are plotted as mean ± SEM (n = 3 independent experiments, analyzed by 1-way ANOVA with Tukey's post hoc test, ***p < 0.001, control MO vs n1-Src AMO). F, In situ hybridization analysis of tubb2b expression in differentiating primary neurons of open neural plate Stage 14 embryos unilaterally injected with 5 ng total of a standard control MO or the AMO a+d combination. The injected side shows faint blue nuclear staining with the β-galactosidase lineage tracer, and is indicated with a black asterisk. Anterior is to the left. m, Motor neurons; i, interneurons; s, sensory neurons. Image published in: Lewis PA et al. (2017) Copyright © 2017 Lewis, Bradley et al. 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 Gene Synonyms Species Stage(s) Tissue tubb2b.L N-Tub, n-tubulin, NBT, neural beta-tubulin, NST, ntubulin, Xn-tubulin X. laevis Throughout NF stage 18 neuron Rohon-Beard neuron interneuron Image source: Published Larger Image Printer Friendly View

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