Phenotypes of Drosophila brain neurons in primary culture reveal a role for fascin in neurite shape and trajectory

Robert Kraft, Mindy M. Escobar, Martha L. Narro, Jackie L. Kurtis, Alon Efrat, Jacobus J Barnard, Linda L Restifo

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

Subtle cellular phenotypes in the CNS may evade detection by routine histopathology. Here, we demonstrate the value of primary culture for revealing genetically determined neuronal phenotypes at high resolution. Gamma neurons of Drosophila melanogaster mushroom bodies (MBs) are remodeled during metamorphosis under the control of the steroid hormone 20-hydroxyecdysone (20E). In vitro, wild-type γ neurons retain characteristic morphogenetic features, notably a single axon-like dominant primary process and an arbor of short dendrite-like processes, as determined with microtubule-polarity markers. We found three distinct genetically determined phenotypes of cultured neurons from grossly normal brains, suggesting that subtle in vivo attributes are unmasked and amplified in vitro. First, the neurite outgrowth response to 20E is sexually dimorphic, being much greater in female than in male γ neurons. Second, the γ neuron-specific "naked runt" phenotype results from transgenic insertion of an MB-specific promoter. Third, the recessive, panneuronal "filagree" phenotypemapsto singed, which encodes the actin-bundling protein fascin. Fascin deficiency does not impair the 20E response, but neurites fail to maintain their normal, nearly straight trajectory, instead forming curls and hooks. This is accompanied by abnormally distributed filamentous actin. This is the first demonstration of fascin function in neuronal morphogenesis. Our findings, along with the regulation of human Fascin1 (OMIM 602689) by CREB (cAMP response element-binding protein) binding protein, suggest FSCN1 as a candidate gene for developmental brain disorders. We developed an automated method of computing neurite curvature and classifying neurons based on curvature phenotype. This will facilitate detection of genetic and pharmacological modifiers of neuronal defects resulting from fascin deficiency.

Original languageEnglish (US)
Pages (from-to)8734-8747
Number of pages14
JournalJournal of Neuroscience
Volume26
Issue number34
DOIs
StatePublished - Aug 23 2006

Fingerprint

Neurites
Drosophila
Phenotype
Neurons
Brain
Mushroom Bodies
Actins
Ecdysterone
Genetic Databases
Developmental Genes
Cyclic AMP Response Element-Binding Protein
Brain Diseases
Dendrites
Drosophila melanogaster
Morphogenesis
Protein Binding
Microtubules
Axons
fascin
Carrier Proteins

Keywords

  • Actin
  • Cytoskeleton
  • Ecdysone
  • Mushroom body
  • Neurite curvature
  • Neurogenetics
  • Plasticity
  • Sexual dimorphism
  • Software

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Phenotypes of Drosophila brain neurons in primary culture reveal a role for fascin in neurite shape and trajectory. / Kraft, Robert; Escobar, Mindy M.; Narro, Martha L.; Kurtis, Jackie L.; Efrat, Alon; Barnard, Jacobus J; Restifo, Linda L.

In: Journal of Neuroscience, Vol. 26, No. 34, 23.08.2006, p. 8734-8747.

Research output: Contribution to journalArticle

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