Rbfox Splicing Factors Promote Neuronal Maturation and Axon Initial Segment Assembly

Neuron. 2018 Feb 21;97(4):853-868.e6. doi: 10.1016/j.neuron.2018.01.020. Epub 2018 Feb 1.

Abstract

Neuronal maturation requires dramatic morphological and functional changes, but the molecular mechanisms governing this process are not well understood. Here, we studied the role of Rbfox1, Rbfox2, and Rbfox3 proteins, a family of tissue-specific splicing regulators mutated in multiple neurodevelopmental disorders. We generated Rbfox triple knockout (tKO) ventral spinal neurons to define a comprehensive network of alternative exons under Rbfox regulation and to investigate their functional importance in the developing neurons. Rbfox tKO neurons exhibit defects in alternative splicing of many cytoskeletal, membrane, and synaptic proteins, and display immature electrophysiological activity. The axon initial segment (AIS), a subcellular structure important for action potential initiation, is diminished upon Rbfox depletion. We identified an Rbfox-regulated splicing switch in ankyrin G, the AIS "interaction hub" protein, that regulates ankyrin G-beta spectrin affinity and AIS assembly. Our data show that the Rbfox-regulated splicing program plays a crucial role in structural and functional maturation of postmitotic neurons.

Keywords: AnkG; Rbfox; actin cytoskeleton; alternative splicing; axon initial segment; neuronal maturation.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3T3 Cells
  • Alternative Splicing*
  • Animals
  • Ankyrins / metabolism
  • Axon Initial Segment / metabolism*
  • DNA-Binding Proteins
  • Embryonic Stem Cells / metabolism
  • Gene Expression Regulation, Developmental
  • Mice
  • Mice, Knockout
  • Nerve Tissue Proteins / metabolism
  • Nuclear Proteins / metabolism
  • RNA Splicing Factors / metabolism*
  • Spinal Cord / embryology*
  • Spinal Cord / metabolism

Substances

  • Ank3 protein, mouse
  • Ankyrins
  • DNA-Binding Proteins
  • Nerve Tissue Proteins
  • NeuN protein, mouse
  • Nuclear Proteins
  • RNA Splicing Factors
  • Rbfox1 protein, mouse
  • Rbfox2 protein, mouse