Xenbase Image ID: 138166

Fig. 5. Tau knockdown selectively disrupted the structural integrity of microtubules containing a neuronal class II b-tubulin. Newly differentiating, embryonic cultured spinal cord neurons containing tau antisense morpholino oligonucleotide (tau MO; B1–3, D2, E2, F2, G2) or control MO (A1–2, D1, E1, F1, G1) were stained for the neuron-specific b-tubulin II isotype (Nb-tubulin; Xenopus gene symbol, tubb2b), ubiquitous a- and b-tubulins, tyrosinated (Tyr-), and acetylated (Ac-) a-tubulins, respectively. (A1–B3) In contrast to the generally filamentous structures seen in the axons and growth cones (A2) of control MO neurons stained for Nb-tubulin, significantly more short, discontinuous fragments (arrowheads in B1 and 2) were seen in the axons and growth cones (B3) of tau MO neurons. (C) Quantitation of the number ( SEM) of discontinuous, fragmented microtubules (puncta) stained by the Nb-tubulin antibody along the neuritic shaft (left; /100 lm) and within growth cones (right; /100 lm2) of control and tau MO cultured neurons (three cultures each). **Significantly different from control, as determined by t-test; P < 0.01. (D1–G2) As determined by immunostaining, neither the levels of expression nor distributions of ubiquitous b-tubulins (D1 and 2), ubiquitous a-tubulins, Tyr-tubulin (F1 and 2) and Ac-tubulin (G1 and 2) differed between tau MO and control MO cultured neurons. Scale bars (20 lm in all panels) in (A2), (B1), (E1), (F1) and (G1) apply to (B3), (B2), (E2), (F2) and (G2), respectively. Neurons were imaged by conventional epifluorescence with a 1009 objective (NA 1.4). (H) Quantitation of the relative fluorescence intensity (RFI; see Statistical analyses in Materials and methods for details) within neurites for each of the tubulin antibodies in (D1–G2) revealed no significant differences in tau MO neurons relative to that of control MO neurons (RFI = 1.0; t-test, three cultures each). Image published in: Liu Y et al. (2015) Copyright © 2015. Image reproduced with permission of the Publisher, John Wiley & Sons. Gene Synonyms Species Stage(s) Tissue tubb2b.S N-Tub, n-tubulin, NBT, neural beta-tubulin, NST, ntubulin, Xn-tubulin X. laevis Throughout NF stage 25 spinal neuron axon tuba4b.L alpha tubulin, alpha-tubulin, tuba4, tuba4a X. laevis Throughout NF stage 25 axon spinal neuron Image source: Published Experiment + Assay Source Phenotypes and Disease Xla Wt + Mapt MO + neural tube explant + NF37/38 (immunohistochemistry) fig.5.b1-b3, c Anatomical Phenotype abnormal axon microtubule bundle morphology decreased size of the axon microtubule bundle Xla Wt + Mapt MO + neural tube explant + NF37/38 (immunohistochemistry) fig.5.b3, c Anatomical Phenotype abnormal growth cone morphology abnormal microtubule morphology decreased size of the microtubule Larger Image Printer Friendly View

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