Fig. 5. Axon outgrowth was suppressed by injection into embryos of a non-coding RNA bearing the triple tripartite hnRNP K RNA-binding motif. (Aa-Cb) Parasagittal optical sections of whole-mount immunostained stage 37/38 animals. (Aa-Ad) Animals receiving the triple tripartite binding motif. Aa shows the uninjected (Uninj.) side with arrowheads indicating motor axons. Animals were immunostained for the indicated axonal markers (NF-M; peripherin; neuronal (N) β-tubulin). Arrowheads in Ab-Ad point to severely atrophied fibers. (Ba-Cb) Two control RNAs yielded normal axon outgrowth and expression of NF-M on the injected side. Scale bars: 100 μm. (Da-Dd) In dissociated embryonic spinal cord cultures, the triple tripartite binding motif RNA yielded neurons (positively stained for N-β tubulin) with marked defects in microtubule organization and that either lacked neurites (Da) or had only short, immature neurites (Db). Neurons from indicated control RNA cultures (Dc,Dd) were normal. Scale bars: 20 μm. (E) Top: Mean (± s.d.) fraction of N-β tubulin-positive cells that had neurites. Bottom left: Mean (± s.d.) total length of neuritic arbor per cell. Bottom right: Mean (± s.d.) number of higher order branches (secondary and higher) per cell. 1, uninjected; 2, single tripartite binding motif-injected; 3, triple tripartite binding motif-injected; 4, triple mutated tripartite motif-injected; 5, triple scrambled tripartite motif-injected cultures. Because embryos were unilaterally injected, only the half derived from the uninjected side was expected to be normal (indicated by dashed line). **P<0.01, t-test. Numbers of neurons tabulated were: upper graph (four cultures each), bars 1-5: 610, 625, 631, 591, 632, respectively; lower graphs (three cultures each), bars 1-5: 169, 158, 155, 142, 184, respectively.
Image published in: Liu Y and Szaro BG (2011)
Copyright © 2011. 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.
| Gene | Synonyms | Species | Stage(s) | Tissue |
|---|---|---|---|---|
| nefm.L | nef-m, nef3, nefm-a, nefm-b, nf-m | X. laevis | Throughout NF stage 37 and 38 | spinal cord |
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