Whole-mount identification of gene transcripts in aphids: Protocols and evaluation of probe accessibility

Chun Che Chang, Ting Yu Huang, Chun Liang Shih, Gee Way Lin, Te Pin Chang, Hui Chiu, Wen Chang Chang

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12 引文 斯高帕斯(Scopus)

摘要

In situ hybridization has become a powerful tool for detecting the temporal and spatial distribution of gene transcripts in prokaryotes and eukaryotes. We report an efficient protocol for whole-mount identification of the expression of mRNAs in the parthenogenetic pea aphid Acyrthosiphon pisum, an emerging model organism with a growing accumulation of genome sequencing data. In addition to steps common for most animal in situ hybridization protocols, we describe processing methods specific to aphids, the accessibility of antisense riboprobes of different lengths in whole-mounted aphids, and signal intensity versus probe lengths. To find substrate combinations that clearly contrast single and double in situ signals in A. pisum, we tested our protocols using riboprobes constructed from two conserved germline markers, Apvasa ana Apnanos, and examined colocalized signals in the germaria and developing oocytes. Finally, we propose conditions for stringent permeabilization that may be applied to tissues deep within the aphid embryo.
原文英語
頁(從 - 到)186-196
頁數11
期刊Archives of Insect Biochemistry and Physiology
68
發行號4
DOIs
出版狀態已發佈 - 八月 1 2008
對外發佈

ASJC Scopus subject areas

  • 生理學
  • 生物化學
  • 昆蟲科學

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