RNAi-mediated double gene knockdown and gustatory perception measurement in honey bees (Apis mellifera)

Ying Wang, Nicholas Baker, Gro Amdam

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

This video demonstrates novel techniques of RNA interference (RNAi) which downregulate two genes simultaneously in honey bees using double-stranded RNA (dsRNA) injections. It also presents a protocol of proboscis extension response (PER) assay for measuring gustatory perception. RNAi-mediated gene knockdown is an effective technique downregulating target gene expression. This technique is usually used for single gene manipulation, but it has limitations to detect interactions and joint effects between genes. In the first part of this video, we present two strategies to simultaneously knock down two genes (called double gene knockdown). We show both strategies are able to effectively suppress two genes, vitellogenin (vg) and ultraspiracle (usp), which are in a regulatory feedback loop. This double gene knockdown approach can be used to dissect interrelationships between genes and can be readily applied in different insect species. The second part of this video is a demonstration of proboscis extension response (PER) assay in honey bees after the treatment of double gene knockdown. The PER assay is a standard test for measuring gustatory perception in honey bees, which is a key predictor for how fast a honey bee's behavioral maturation is. Greater gustatory perception of nest bees indicates increased behavioral development which is often associated with an earlier age at onset of foraging and foraging specialization in pollen. In addition, PER assay can be applied to identify metabolic states of satiation or hunger in honey bees. Finally, PER assay combined with pairing different odor stimuli for conditioning the bees is also widely used for learning and memory studies in honey bees.

Original languageEnglish (US)
JournalJournal of visualized experiments : JoVE
Issue number77
DOIs
StatePublished - Jan 1 2013

Fingerprint

Taste Perception
Gene Knockdown Techniques
Honey
Bees
RNA Interference
RNA
Genes
Assays
Down-Regulation
Satiation
Vitellogenins
Hunger
Double-Stranded RNA
Pollen
Age of Onset
Insects
Odors
Gene expression
Learning
Gene Expression

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

RNAi-mediated double gene knockdown and gustatory perception measurement in honey bees (Apis mellifera). / Wang, Ying; Baker, Nicholas; Amdam, Gro.

In: Journal of visualized experiments : JoVE, No. 77, 01.01.2013.

Research output: Contribution to journalArticle

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