RNAi-mediated knockdown showing impaired cell survival in Drosophila wing Imaginal disc

Makoto Umemori, Okiko Habara, Tatsunori Iwata, Kousuke Maeda, Kana Nishinoue, Atsushi Okabe, Masahiko Takemura, Kuniaki Takahashi, Kaoru Saigo, Ryu Ueda, Takashi Adachi-Yamada

Research output: Contribution to journalArticlepeer-review

17 Scopus citations

Abstract

The genetically amenable organism Drosophila melanogaster has been estimated to have 14,076 protein coding genes in the genome, according to the flybase release note R5.13 (http://flybase.bio.indiana.edu/ static_pages/docs/release_notes.html). Recent application of RNA interference (RNAi) to the study of developmental biology in Drosophila has enabled us to carry out a systematic investigation of genes affecting various specific phenotypes. In order to search for genes supporting cell survival, we conducted an immunohistochemical examination in which the RNAi of 2,497 genes was independently induced within the dorsal compartment of the wing imaginal disc. Under these conditions, the activities of a stress-activated protein kinase JNK (c-Jun N-terminal kinase) and apoptosis-executing factor Caspase-3 were monitored. Approximately half of the genes displayed a strong JNK or Caspase-3 activation when their RNAi was induced. Most of the JNK activation accompanied Caspase-3 activation, while the opposite did not hold true. Interestingly, the area activating Caspase-3 was more broadly seen than that activating JNK, suggesting that JNK is crucial for induction of non-autonomous apoptosis in many cases. Furthermore, the RNAi of essential factors commonly regulating transcription and translation showed a severe and cell-autonomous apoptosis but also elicited another apoptosis at an adjacent area in a non-autonomous way. We also found that the frequency of apoptosis varies depending on the tissues.

Original languageEnglish (US)
Pages (from-to)11-20
Number of pages10
JournalGene Regulation and Systems Biology
Volume2009
Issue number3
DOIs
StatePublished - 2009

Keywords

  • Drosophila RNAi apoptosis JNK caspase-3

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