A novel forward genetic screen for identifying mutations affecting larval neuronal dendrite development in Drosophila melanogaster.
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.
Medina
Paul Mark B
PM
Department of Cell and Developmental Biology and Neuroscience Center, University of North Carolina-Chapel Hill School of Medicine, Chapel Hill, North Carolina 27599-7250, USA.
Swick
Lance L
LL
Andersen
Ryan
R
Blalock
Zachary
Z
Brenman
Jay E
JE
D000818:Animals; D003433:Crosses, Genetic; D003712:Dendrites / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000196:embryology* / Q000235:genetics*; D005020:Ethyl Methanesulfonate; D005456:Fluorescent Dyes / Q000494:pharmacology; D005821:Genetic Techniques; D049452:Green Fluorescent Proteins / Q000378:metabolism; D016296:Mutagenesis; D009153:Mutagens; D009154:Mutation; D009474:Neurons / Q000378:metabolism*; D019076:Transgenes
D005456:Fluorescent Dyes; D009153:Mutagens; D049452:Green Fluorescent Proteins; D005020:Ethyl Methanesulfonate