Figure 7.

CLN3 induced degeneration is suppressed by the downregulation of ROCK2 genetically and pharmacologically (A) ROCK2 protein/gene interactome represented with IPA showing that some of the interactors of ROCK2 are also altered in “crude” thalamic synaptosomes. (B) QWB bands and quantification of control and Cln3 ^−/− synaptic and “non synaptic” fraction. Upregulation of ROCK2 is higher in synapses than in the non-synaptic fraction indicating a potential synaptic specific response. *P < 0.05 (T-student test). (C) “Signaling by Rho family GTPases” canonical pathway represented by IPA. Upregulation/activation of ROCK2 impacts in actin nucleation and polymerization that may disrupt actin dynamics in the synapse. (A&C) Coloured nodes illustrate proteins present in the thalamic dataset in Cln3 ^−/− with respect WT. Nodes in grey represent proteins changed <20%, down-/up-regulated proteins by >20% are represented in green and red respectively, orange indicates predicted activation and blue; predicted inhibition. (D) ROCK2 upregulation at early stage of disease is a converved event across animal models of injury, two adult-onset neurodegenerative diseases (Huntington and Spinocerebelar ataxia)² and two childhood neurodegenerative conditions (SMA and the lysosomal storage disorders-NCLs). (E and H) Representative light microscope images of Drosophila eyes and corresponding 200x zoom of eye structure of (A) Disease Model (DM) (B) DM + 1.5 mM Fasudil. (C) DM + Rok RNAi TRiP and (D) Control fly (Canton S). (I) Bar chart representing the % of normalized suppression calculated from the average eye surface areas of three independent experiments as “(x- average eye surface area of the disease model)/(average of eye surface area of the control - average eye surface area of disease model) × 100” for each of the three independent experiments. Scale bar = 100um. ***P < 0.001, *P < 0.05 (One-way ANOVA and Tukey’s multiple comparison test as a post-hoc).