Abstract
The first morphological change after neuronal differentiation is the microtubule-dependent initiation of thin cell protrusions called neurites. Here we performed a siRNA-based morphometric screen in P19 stem cells to evaluate the role of 408 microtubule-regulating genes during this early neuromorphogenesis step. This screen uncovered several novel regulatory factors, including specific complex subunits of the microtubule motor dynein involved in neurite initiation and a novel role for the microtubule end-binding protein EB2 in attenuation of neurite outgrowth. Epistasis analysis suggests that competition between EB1 and EB2 regulates neurite length, which links its expression to neurite outgrowth. We propose a model that explains how microtubule regulators can mediate cellular morphogenesis during the early steps of neuronal development by controlling microtubule stabilization and organizing dynein-generated forces.
Highlights
Neurons grow specialized cell protrusions that form the basis of highly interconnected cellular networks in the brain [1]
To study the role of microtubule-regulating genes in neuronal development, an automated morphometric screen was performed in P19 stem cells by combining the induction of neuronal differentiation with efficient gene knockdown via co-transfection of a neurogenic transcription factor and siRNA oligonucleotides [5] (Figure 1A)
We present a comprehensive morphometric siRNA-based screen to study the role of all known microtubuleregulating genes in early neuronal development
Summary
Neurons grow specialized cell protrusions that form the basis of highly interconnected cellular networks in the brain [1]. Actin can form distinct, dynamic supramolecular structures that play multiple roles in cellular morphogenesis. We found that the microtubule motor cytoplasmic dynein can power cellular shape changes in the absence of actin dynamics [4], suggesting that microtubules might play an active role in morphogenic processes. We extended on these observations and performed a morphometric screen in P19 stem cells to analyze the role of microtubule-regulating genes in early neuronal development. Using this strategy, we identified several regulators, which influence neurite formation
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