Knockdown of ZBP1 in hippocampal neurons decreases dendritic branching

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Knockdown of ZBP1 in hippocampal neurons decreases dendritic branching. Knockdown of ZBP1 in hippocampal neurons decreases dendritic branching. A, Representative images of hippocampal neurons transfected on DIV7 or DIV14 with pSuper, pSuper-ZBP1sh#1, or pSuper-ZBP1sh#2 for 3 d. Morphology of transfected cells was visualized by staining for cotransfected β-gal. B–F, Mean number of primary dendrites (B), mean total number of dendritic tips (C), dendritic branching index (D), mean total dendritic length (E), and Sholl analysis of neurons transfected on DIV7 (F). G, Representative images of hippocampal neurons transfected on DIV7 with pSuper and scrambled scZBP1sh and fixed 3 d later. Scale bar, 50 μm. H, Comparison of morphologies of dendritic arbors of neurons transfected with pSuper or scZBP1sh. The Sholl analysis revealed no significant difference. I–M, Mean number of primary dendrites (I), mean total number of dendritic tips (J), dendritic branching index (K), mean total dendritic length (L), and Sholl analysis of neurons transfected on DIV14 (M). ***p < 0.0005, **p < 0.005, *p < 0.025, Kruskal–Wallis test followed by Mann–Whitney post hoc test with Bonferroni's adjustment. N, qRT-PCR of ZBP1 expression in hippocampal neurons. RNA was isolated from hippocampal neuron cultures collected on DIV7, DIV10, DIV14, and DIV17. The graphs show the relative quantification of ZBP1 normalized to DIV7. *p < 0.05, one-sample t test. Error bars indicate SEM. Malgorzata Perycz et al. J. Neurosci. 2011;31:5271-5285 ©2011 by Society for Neuroscience