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Investigating the Co-Regulatory Role of Extramacrochaetae and Midline Transcription Factors on Neuronal Cell Fate Specification in Developing Drosophila Eye Tissues

Undergraduate #7
Discipline: Biological Sciences
Subcategory: Cell and Molecular Biology

Francisca Seiwah Donkor - Harris-Stowe State University
Co-Author(s): Dr. Sandra Leal, Harris-Stowe State University, Saint Louis, MO



Although transcription factor proteins (ID1-4) have been extensively studied as regulators of cell proliferation in mammalian tissue, the ID4 isoform remains relatively uncharacterized. Mutations of distinct ID proteins result in a variety of cancers including breast, colon and prostate cancer. The Drosophila homolog of ID4 is Extramacrochaetae (Emc). Previously, the Leal lab showed that Emc and Midline (Mid) function within the NOtch-Delta pathway to specify the cell fates of neuronal sensory organ precursor (SOP) cells in developing eye tissue (Das et al., 2013; Chen et al., 2015). The potential co-regulatory relationship between Emc and Mid within the Notch-Delta pathway has also not yet been elucidated. By using basic genetics and the UAS-Gal4 system that was developed by Brand and Perriman (1993), we are applying loss of function (LOF) and gain of function (GOF) studies to determine whether Mid and Emc interact to specify neuronal cell fates in developing eye tissues. The importance of this research is to contribute further to the validation of a transcriptional factor combinationl code implicit in regulating specific gene expression during development of tissues. In addition, Emc activity may be evolutionarily conserved with that of its homolg ID4 which will result in a comparative functional analysis to further characterize ID4 function.

Funder Acknowledgement(s): NSF HBCU UP Awarded to Dr. Smith NSF RIG Awarded to Dr. Leal

Faculty Advisor: Dr. Sandra Leal, Leals@hssu.edu

Role: I maintained Drosophila melanogaster strains on standard cornmeal-yeast-agar media on a 12 hour light-dark cycle. Oregon-R (OR) flies were used as wild-type (WT) controls. The UAS-Gal4 system (Brand and Perrimon, 1993) was applied by crossing transgenic flies carrying the eye-specific driver GMR-Gal4 balanced on chromosome III with either UAS-mid-RNAi, UAS-emc-RNAi, UAS-mid or UAS-emc transgenic lines to produce Loss-of-Function (LOF) or Gain-of-Function (GOF) mutant conditions within third-instar larval eye imaginal discs (Qian et al., 2005). I dissected OR and mutant eye imaginal discs from third-instar larva followed by fixing and immunolabeling these tissues.

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This material is based upon work supported by the National Science Foundation (NSF) under Grant No. DUE-1930047. Any opinions, findings, interpretations, conclusions or recommendations expressed in this material are those of its authors and do not represent the views of the AAAS Board of Directors, the Council of AAAS, AAAS’ membership or the National Science Foundation.

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