siRNA-mediated gene targeting in Aedes aegypti embryos reveals that frazzled regulates vector mosquito CNS development

Anthony Clemons, Morgan Haugen, Christy Le, Akio Mori, Michael Tomchaney, David W. Severson, Molly Scheel

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Although mosquito genome projects uncovered orthologues of many known developmental regulatory genes, extremely little is known about the development of vector mosquitoes. Here, we investigate the role of the Netrin receptor frazzled (fra) during embryonic nerve cord development of two vector mosquito species. Fra expression is detected in neurons just prior to and during axonogenesis in the embryonic ventral nerve cord of Aedes aegypti (dengue vector) and Anopheles gambiae (malaria vector). Analysis of fra function was investigated through siRNA-mediated knockdown in Ae. aegypti embryos. Confirmation of fra knockdown, which was maintained throughout embryogenesis, indicated that microinjection of siRNA is an effective method for studying gene function in Ae. aegypti embryos. Loss of fra during Ae. aegypti development results in thin and missing commissural axons. These defects are qualitatively similar to those observed in Dr. melanogaster fra null mutants. However, the Aa. aegypti knockdown phenotype is stronger and bears resemblance to the Drosophila commissureless mutant phenotype. The results of this investigation, the first targeted knockdown of a gene during vector mosquito embryogenesis, suggest that although Fra plays a critical role during development of the Ae. aegypti ventral nerve cord, mechanisms regulating embryonic commissural axon guidance have evolved in distantly related insects.

Original languageEnglish
Article numbere16730
JournalPLoS One
Volume6
Issue number1
DOIs
StatePublished - 2011

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Gene Targeting
Aedes
gene targeting
small interfering RNA
Aedes aegypti
Small Interfering RNA
Culicidae
embryo (animal)
Embryonic Structures
Genes
Embryonic Development
ventral nerve cord
Gene Knockdown Techniques
Developmental Genes
Anopheles gambiae
Phenotype
Dengue
axons
Microinjections
Regulator Genes

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

siRNA-mediated gene targeting in Aedes aegypti embryos reveals that frazzled regulates vector mosquito CNS development. / Clemons, Anthony; Haugen, Morgan; Le, Christy; Mori, Akio; Tomchaney, Michael; Severson, David W.; Scheel, Molly.

In: PLoS One, Vol. 6, No. 1, e16730, 2011.

Research output: Contribution to journalArticle

Clemons, Anthony ; Haugen, Morgan ; Le, Christy ; Mori, Akio ; Tomchaney, Michael ; Severson, David W. ; Scheel, Molly. / siRNA-mediated gene targeting in Aedes aegypti embryos reveals that frazzled regulates vector mosquito CNS development. In: PLoS One. 2011 ; Vol. 6, No. 1.
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