Systemic RNA-interference in the honeybee Apis mellifera: Tissue dependent uptake of fluorescent siRNA after intra-abdominal application observed by laser-scanning microscopy

RNA interference has been successfully used in adult honeybees, but there are only few reports about abdominal application of dsRNA/siRNA which have reached more distant tissues than the fat body. We studied systemic RNAi in honeybees by injecting fluorescent siRNA of the ubiquitously expressed honeybee homologue of the Glycerol-3-Phosphate Dehydrogenase (amGpdh) into the abdomens of adult bees and followed them by laser scanning microscopy and qPCR. The fat body was the sole tissue emitting fluorescence and showing a decreased gene expression, whereas the siRNA had apparently not reached the other tissues. Therefore, we conclude that certain genes in other tissues than the fat body cannot be easily reached by injecting siRNA into the body cavity. In particular, the lack of amGpdh knock down in ovaries after amGpdh dsRNA injection, supports that in some cases it may be particularly difficult to interfere with gene expression in ovaries by intra-abdominal injection. In these cases alternative inhibition techniques may be required to achieve an organismic non-lethal disruption of transcription.

Highlights► First methods paper on systemic RNAi in honeybees. ► Detection of fluorescent siRNA solely in fat body after abdominal injection. ► No RNAi effect in ovaries after usage of dsRNA and siRNA. ► tissue crucial for successful RNAi when using abdominal injection.