Attention! iBeetle is a first pass screen. Hence, the documented phenotypes need to be confirmed by independent experiments using non-overlapping dsRNA fragments! Phenotypes with a penetrance > 50% are frequently reproduced. See Schmitt-Engel et al. 2015 for details on reproducibility.
Usually 10 injected animals
Terms used in the pupal injection procedure
- larval pantagmatic defects
- At least two tagmata (head, thorax, abdomen or terminus) show similar/ comparable defects.
- empty eggs
- Just empty shells are visible, no cuticle has been developed.
- strong defects
- L1 cuticle larvae which show severe defects. These strong defects are subdivided into three categories
- strongly fragmented (cuticle crumbs)
- No segments or appendages are identifiable, only residues of cuticle and bristles can be identified.
- cuticle remnants (with unidentifiable segments)
- Segments or parts of appendages are recognizable, but identity is not clear.
- cuticle remnants (with some identifiable segments)
- At least one segment or appendage is clearly regonizable.
- number of eggs on slide
- affected embryos usually (not always) do not hatch. Only the not hatched were used for cuticle analysis.
For the embryonic muscle analysis the hatched as well as the not hatched were analysed. - total number of affected eggs/embryos/larvae
- Summary of all different phenotypes/ phenotypic series which were annotated.
- inside-out
- The L1 larva shows a partial or complete inversion., e.g. bristles, appendages or parts of the abdominal segments are inverted into the interior of the cuticle.
- starved
- Dissected females show a strong resorption of fatbody predominately in the abdomen and the oogenesis is blocked. These are considered as eventually lethal phenotypes and the blocked oogenesis probably a secondary defect due to starvation.
- eclosion not fulfilled
- The emergence of the adult from the pupa stage is interrupted. This phenotype shows pupal as well as adult features.
number of eggs/embryos/larvae on slide: < 50
larval musculature pattern irregular -
Penetrance (percentage of animals/eggs on slide): more than 80%
larval musculature fluorescence decreased - Penetrance (percentage of animals/eggs on slide): 30% - 50%
affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): more than 80%
remarks: muscle pattern potentially obscured by segmentation defect,
larval musculature fluorescence decreased - Penetrance (percentage of animals/eggs on slide): 30% - 50%
affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): more than 80%
remarks: muscle pattern potentially obscured by segmentation defect,
number of eggs/embryos/larvae on slide: > 50
larva segment central constricted - Penetrance (percentage of animals/eggs on slide): 30% - 50%
head shape irregular -
Penetrance (percentage of animals/eggs on slide): 30% - 50%
thorax shape irregular - Penetrance (percentage of animals/eggs on slide): 30% - 50%
affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): more than 80%
larva segment central constricted - Penetrance (percentage of animals/eggs on slide): 30% - 50%
| Comment: Constriction of abdomen segment is major phenotype but with this phenotype there is also head & thorax appendages shape irregular & larvae segements are randomly partially not present in all three tagmata. |
thorax shape irregular - Penetrance (percentage of animals/eggs on slide): 30% - 50%
affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): more than 80%
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