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Originally published as Genetics Published Articles Ahead of Print on January 16, 2006.
Genetics, Vol. 172, 2325-2335, April 2006, Copyright © 2006
doi:10.1534/genetics.105.051276
A Novel Forward Genetic Screen for Identifying Mutations Affecting Larval Neuronal Dendrite Development in Drosophila melanogaster
Paul Mark B. Medina1, Lance L. Swick1, Ryan Andersen, Zachary Blalock and Jay E. Brenman2
Department of Cell and Developmental Biology and Neuroscience Center, University of North Carolina–Chapel Hill School of Medicine, Chapel Hill, North Carolina 27599
2 Corresponding author: Neuroscience Research Bldg., Room 8109A, UNC School of Medicine, Box 7250, 103 Mason Farm Rd., Chapel Hill, NC 27599-7250.
E-mail: brenman{at}med.unc.edu
Vertebrate and invertebrate dendrites are information-processing compartments that can be found on both central and peripheral neurons. Elucidating the molecular underpinnings of information processing in the nervous system ultimately requires an understanding of the genetic pathways that regulate dendrite formation and maintenance. Despite the importance of dendrite development, few forward genetic approaches have been used to analyze the latest stages of dendrite development, including the formation of F-actin-rich dendritic filopodia or dendritic spines. We developed a forward genetic screen utilizing transgenic Drosophila second instar larvae expressing an actin, green fluorescent protein (GFP) fusion protein (actin::GFP) in subsets of sensory neurons. Utilizing this fluorescent transgenic reporter, we conducted a forward genetic screen of >4000 mutagenized chromosomes bearing lethal mutations that affected multiple aspects of larval dendrite development. We isolated 13 mutations on the X and second chromosomes composing 11 complementation groups affecting dendrite outgrowth/branching, dendritic filopodia formation, or actin::GFP localization within dendrites in vivo. In a fortuitous observation, we observed that the structure of dendritic arborization (da) neuron dendritic filopodia changes in response to a changing environment.
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