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Vol. 15, Issue 1, 111-120, January 2004

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ARNO and ARF6 Regulate Axonal Elongation and Branching through Downstream Activation of Phosphatidylinositol 4-Phosphate 5-Kinase

Delia J. Hernández-Deviez ^*, Michael G. Roth , James E. Casanova , and Jean M. Wilson ^* 

^* Department of Cell Biology and Anatomy, College of Medicine, University of Arizona, Tucson, Arizona 85724; Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, Texas 75390; and Department of Cell Biology, University of Virginia Health System, Charlottesville, Virginia 22908

Submitted June 17, 2003; Revised September 15, 2003; Accepted September 22, 2003
Monitoring Editor: Keith Mostov

In the developing nervous system, controlled neurite extension and branching are critical for the establishment of connections between neurons and their targets. Although much is known about the regulation of axonal development, many of the molecular events that regulate axonal extension remain unknown. ADP-ribosylation factor nucleotide-binding site opener (ARNO) and ADP-ribosylation factor (ARF)6 have important roles in the regulation of the cytoskeleton as well as membrane trafficking. To investigate the role of these molecules in axonogenesis, we expressed ARNO and ARF6 in cultured rat hippocampal neurons. Expression of catalytically inactive ARNO or dominant negative ARF6 resulted in enhanced axonal extension and branching and this effect was abrogated by coexpression of constitutively active ARF6. We sought to identify the downstream effectors of ARF6 during neurite extension by coexpressing phosphatidyl-inositol-4-phosphate 5-Kinase [PI(4)P 5-Kinase ] with catalytically inactive ARNO and dominant negative ARF6. We found that PI(4)P 5-Kinase plays a role in neurite extension and branching downstream of ARF6. Also, expression of inactive ARNO/ARF6 depleted the actin binding protein mammalian ena (Mena) from the growth cone leading edge, indicating that these effects on axonogenesis may be mediated by changes in cytoskeletal dynamics. These results suggest that ARNO and ARF6, through PI(4)P 5-Kinase , regulate axonal elongation and branching during neuronal development.

Corresponding author. E-mail address: jeanw{at}email.arizona.edu.

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