{"database":"bioimages","file_versions":[],"scores":null,"additional":{"omics_type":["Unknown"],"submitter":["Mehri Moradi"],"journal":["The Journal of Cell Biology"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-JCBD-201604117"],"attach_to":["JCB"],"legend":["S4<br />(C) Western blot analysis shows that Actβ protein levels are restored to 75% after application of the rescue construct.","Figure 3<br />(F) Actβ protein is very abundant in the soma and localize to axonal filopodia and axonal growth cone filopodia.","Figure 8<br />(D) Western blot analysis of lysates from actin isoform-specific knockdown motoneuron cultures probed with pan-actin antibody.(F) Immunoblots of supernatant (G-actin) and pellet fractions (F-actin) of motoneuron lysates after ultracentrifugation probed with pan-actin antibody. (H) Supernatant and pellet fractions of motoneuron lysates were labeled with Actβ antibody.","Figure 3<br />(E) Motoneurons were stained against Tau and Actα. Actα protein is highly enriched in axonal branch points and in neurites.","Video 2<br />Motoneurons transduced with lentiviral shRNA targeting Actα and coexpressing GFP show decreased dynamics of axonal filopodia movements.Related to Fig. 6, A-G.","Video 5<br />Motoneurons were transduced with lentiviral particles which coexpress GFP and imaged for 40 min at 20 sec intervals. Filopodia in growth cones are highly dynamic and show fast growth/retraction movements. Related to Fig. 7, D-G.","Figure 3<br />(G) Actγ protein localize to axonal filopodia and axonal growth cone filopodia.","Figure 9<br />(G-I) Western blot analysis of cultured motoneuron lysates probed with specific Actα (G), Actβ (H) and Actγ (I) antibodies.","Video 4<br />Axonal filopodia show reduced growth/retraction movements after shRNA-meditated knockdown of Actγ. Related to Fig. 6, A-G.","S2<br />(A) HEK293T cells were transfected with plasmids expressing HA-tagged Actα, Actβ or Actγ. Actα antibody detects only HA-Actα and not HA-Actβ or HA-Actγ.","Figure 3<br />(A-C) Immunoblots of total lysates obtained from actin isoform-specific knockdown and control motoneurons probed with specific Actα (A), Actβ (B) and Actγ (C) antibodies. Calnexin was used as loading control. All three isoforms are detected in cultured motoneurons.","Figure 10<br />(A) Immunoblot shows reduction in Smn protein levels in motoneuron lysates after shRNA-mediated knockdown of Smn. Calnexin was used as loading control. A Western blot from two independent knockdown experiments is shown (representative for n = 3).","Video 1<br />Motoneurons transduced with lentiviral particles expressing GFP were imaged for 40 min every 20 sec. Axonal filopodia form de novo along the axon shaft and show growth/retraction movements. Related to Fig. 6, A-G.","Video 7<br />ShRNA-mediated Actβ depletion decreases growth/retraction of filopodia in axonal growth cones. Related to Fig. 7, D-G."],"repository":["bioimages"],"figure_sub":["Figure 10 - A","Image 641879 (Figure 3 - F)","Image 641913 (Video 4 - None)","Video 5","Video 2 - None","Image 641902 (Figure 8 - D, F and H)","Figure S2 - A","Video 4 - None","Video 5 - None","Video 7","Video 7 - None","Video 1 - None","Image 641886 (Figure 3 - G)","Image 641889 (Figure 3 - G)","Figure S4","Figure S2","Image 641910 (Figure 3 - A-C)","Image 641898 (Figure S4 - C)","Image 641890 (Figure 3 - G)","Video 2","Video 1","Video 4","Image 641864 (Figure 3 - E)","Image 641883 (Figure 3 - F)","Image 641894 (Figure 10 - A)","Image 641858 (Figure 3 - E)","Image 641877 (Figure 3 - F)","Image 641861 (Figure 3 - E)","Image 641900 (Figure 8 - D, F and H)","Figure S4 - C","Image 641907 (Figure 3 - A-C)","Image 641880 (Figure 3 - F)","Figure 9 - G-I","Image 641884 (Figure 3 - G)","Image 641909 (Figure 3 - A-C)","Image 641857 (Figure 3 - E)","Image 641876 (Figure 3 - F)","Image 641904 (Figure 9 - G-I)","Image 641899 (Figure 8 - D, F and H)","Image 641912 (Video 2 - None)","Image 641862 (Figure 3 - E)","Image 641911 (Video 1 - None)","Image 641895 (Figure S2 - A)","Image 641881 (Figure 3 - F)","Figure 3 - E","Figure 3 - F","Figure 8 - D, F and H","Figure 3 - G","Figure 3 - A-C","Image 641875 (Figure 3 - F)","Image 641914 (Video 5 - None)","Image 641863 (Figure 3 - E)","Image 641893 (Figure 10 - A)","Image 641882 (Figure 3 - F)","Image 641860 (Figure 3 - E)","Image 641878 (Figure 3 - F)","Image 641896 (Figure S2 - A)","Image 641915 (Video 7 - None)","Image 641859 (Figure 3 - E)","Image 641903 (Figure 8 - D, F and H)","Image 641906 (Figure 9 - G-I)","Image 641885 (Figure 3 - G)","Image 641891 (Figure 3 - G)","Image 641905 (Figure 9 - G-I)","Image 641888 (Figure 3 - G)","Image 641865 (Figure 3 - E)","Figure 10","Image 641901 (Figure 8 - D, F and H)","Figure 9","Image 641892 (Figure 3 - G)","Figure 8","Image 641887 (Figure 3 - G)","Image 641908 (Figure 3 - A-C)","Image 641897 (Figure S4 - C)","Figure 3"],"pubmed_authors":["Benjamin Dombert","Rajeeve Sivadasan","Lena Saal","Mehri Moradi","Patrick Lüningschrör","Robert Blum","Michael Sendtner","Reena Jagdish Rathod","Daniela C. Dieterich"],"additional_accession":[]},"is_claimable":false,"name":"Differential roles of α-, β-, and γ-actin in axon growth and collateral branch formation in motoneurons","description":"Axonal branching and terminal arborization are fundamental events during the establishment of synaptic connectivity. They are triggered by assembly of actin filaments along axon shafts giving rise to filopodia. The specific contribution of the three actin isoforms, Actα, Actβ, and Actγ, to filopodia stability and dynamics during this process is not well understood. Here, we report that Actα, Actβ, and Actγ isoforms are expressed in primary mouse motoneurons and their transcripts are translocated into axons. shRNA-mediated depletion of Actα reduces axonal filopodia dynamics and disturbs collateral branch formation. Knockdown of Actβ reduces dynamic movements of growth cone filopodia and impairs presynaptic differentiation. Ablation of Actβ or Actγ leads to compensatory up-regulation of the ","dates":{"release":"2017-02-28T11:29:04Z","modification":"2018-11-29T11:29:04Z","creation":"2018-11-29T11:29:04Z"},"accession":"S-JCBD-201604117","cross_references":{"doi":["10.1083/jcb.201604117"]}}