Effects of Domino Mutations on Cell Proliferation in the Drosophila Eye Pubblico
Callaway, Heather Merrick (2013)
Abstract
Domino is a critical component of the Notch signaling pathway, which regulates cell proliferation. Domino has also been implicated as a Notch-independent regulator of cell proliferation in the Drosophila eye. Recently, it was reported that domino may be pleiotropic for cell proliferation in the wing, such that both domino loss-of-function and gain-of-function mutations enhance nicking in the wing margin. In the present study, domino mutant strains, over-expression strains, and RNAi strains were crossed to an eye hyperproliferation strain (GMR-YkiS168A) in order to determine whether domino behaves the same way in the eye as it has previously been reported to behave in the wing. Notch loss-of-function and gain-of-function mutants were also crossed to the eye hyperproliferation strain. Offspring of the crosses were analyzed for visually-scoreable phenotypic differences, differences in penetrance, and for statistically significant differences in eye size compared to controls. The offspring from both sets of crosses were compared to each other in order to determine whether phenotypes created by the different domino strains might be due to Domino's interactions with Notch.
We found that two domino RNAi strains and one domino over-expression strain showed suppression of the hyperproliferation eye phenotype, but that the remaining strains did not show consistent, statistically significant differences either in eye size or in the penetrance of the hyperproliferation phenotype compared to control crosses. Notch loss-of-function strains also showed suppression of proliferation when crossed to w1118 (wild-type) or GMR-GAL4 (over-expression of the enhancer GAL4 in the eye) flies. Notch gain-of-function mutations were lethal when crossed to GMR-YkiS168A. When Notch loss-of-function mutants were crossed to GMR-YkiS168A, the resulting offspring had different eye phenotypes than those of the GMR-YkiS168A x domino experimental strains, suggesting that Domino's effects on cell proliferation may be independent of Notch.
It is not possible to definitively conclude that domino behaves the same way in the eye tissue as it does in the wing tissue. Our results that domino loss-of-function and domino gain-of-function suppress hyperproliferation in the eye, however, are congruent to previous observations that domino RNAi and domino over-expression strains both enhance a hypo-proliferation phenotype in the wing.
Table of Contents
Introduction...........................................................................................1
Methods.................................................................................................7
i) Genetic
crosses...................................................................................7
ii)Drosophila
strains...............................................................................8
a) The Domino Modifier Genetic
Analysis......................................................8
b) Test for Environmentally Caused Variations in Eye
Size..............................10
c) Genetic Crosses with GMR-YkiS168A and Domino Mutant
Strains.................10
d) Genetic Crosses with GMR-YkiS168A and Notch Mutant
Strains...................12
iii) Data
collection..................................................................................12
a) Scoring for Wing
Nicks...........................................................................13
b) Scoring for the Yorkie Eye
Phenotype.......................................................14
iv) Data
analysis.....................................................................................14
Results...................................................................................................16
i) The Domino Modifier Genetic
Analysis.................................................16
ii) Test for Environmentally Caused Variations in Eye
Size......................19
iii) Genetic Crosses with GMR-YkiS168A and Domino
Mutant Strains......21
iv) Genetic Crosses with GMR-YkiS168A and Notch
Mutant Strains.........28
Discussion..............................................................................................31
Conclusion..............................................................................................38
References..............................................................................................40
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