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The Fruitless Gene Influences Female Mate Preference in Drosophila

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The Fruitless Gene Influences Female Mate Preference in Drosophila Western University Scholarship@Western Electronic Thesis and Dissertation Repository 7-2-2019 2:30 PM The fruitless gene influences emalef mate preference in Drosophila Tabashir A. Chowdhury The University of Western Ontario Supervisor Moehring, Amanda J. The University of Western Ontario Graduate Program in Biology A thesis submitted in partial fulfillment of the equirr ements for the degree in Doctor of Philosophy © Tabashir A. Chowdhury 2019 Follow this and additional works at: https://ir.lib.uwo.ca/etd Part of the Behavior and Ethology Commons, Evolution Commons, Genetics Commons, and the Molecular Genetics Commons Recommended Citation Chowdhury, Tabashir A., "The fruitless gene influences emalef mate preference in Drosophila" (2019). Electronic Thesis and Dissertation Repository. 6440. https://ir.lib.uwo.ca/etd/6440 This Dissertation/Thesis is brought to you for free and open access by Scholarship@Western. It has been accepted for inclusion in Electronic Thesis and Dissertation Repository by an authorized administrator of Scholarship@Western. For more information, please contact [email protected]. Abstract Species can arise as a result of reproductive barriers that prevent gene flow between diverging populations that force them to remain isolated from one another. Behavioural isolation is one of the earliest acting reproductive barriers determined by the evolution of mating preferences that prevent inter-specific matings. Several traits have been identified that contribute to behavioural isolation, but the genetic basis of interspecific female preference is yet to be determined. I used genetic mapping techniques to identify and confirm that the fruitless gene is affecting species-specific female rejection of interspecies males, contributing to the behavioural isolation between Drosophila melanogaster and D. simulans. I also determined that this species-specific female preference is caused by a non-sex specifically spliced transcript of fruitless. Transgenic rescue of fruitless expression using the GAL4/UAS system identified fruitless protein isoforms with a specific 3’ C-terminal end are likely involved in species-specific female preference. Additionally, I discovered that female rejection of interspecies males is not determined by an individual sensory modality such as male courtship song or female perception of auditory and olfactory signals, and is likely controlled by the integration of multiple modalities. Finally, I constructed transgenic RNA interference lines to silence expression of specific D. melanogaster or D. simulans fruitless transcripts. These RNAi lines can be used to knockdown fruitless expression at specific developmental stages and in specific tissues using the GAL4/UAS system, and thus can be a useful tool for characterizing the genetic and neural mechanisms that govern species-specific female rejection. i Summary Species are maintained as distinct groups by reproductive barriers that prevent interbreeding. Behavioural Isolation is one such reproductive barrier that acts via the incompatibilities in mating signals and preferences between diverged population groups. Our current understanding of the genetic mechanisms for behavioural isolation is limited. My research identifies the fruitless gene in Drosophila as a candidate gene for behavioural isolation. Through a series of genetic and molecular biological experiments I established that the fruitless gene is influencing female preference for mates within its own species, which is the first reported behavioural role in females for this gene. I also identified potential molecular and genetic mechanisms by which this gene is affecting female mating behaviour and contributing to the behavioural isolation observed between different species of Drosophila. ii Keywords Speciation, Behavioural isolation, female preference, Drosophila, fruitless. iii Co-Authorship Statement I completed the work of this dissertation under the supervision of Dr. Amanda Moehring. I received help from an undergraduate honours thesis student (A. Dhillon) in the collection of RT-PCR data (Chapter 2). Several of the deficiency and P-element lines used for deficiency mapping in Chapter 2 were completed by honours thesis students (Katrina Bruch, Co-Author, Chapter 2) and a former graduate student (Dr. Ryan Calhoun, Co-Author, Chapter 2). Transgenic deletions of fruitless exons were created by Dr. Megan Neville (Co-Author, Chapter 2). Finally, I received help from another graduate student (Joshua Issacson) in the creation of transgenic fruitless RNAi lines. iv Acknowledgments I would like to acknowledge my supervisor Dr. Amanda Moehring, former and current graduate students Dr. Rachelle Kanipayoor, Dr. Ryan Calhoun, Jalina Bielaska Da Silva, Joshua Issacson and Brendan Charles. I would also like to acknowledge all the honours thesis students and volunteers who worked on this project, including: Akashdeep Dhillon, Nick Werry, Ellen Xu, Geyi Wang, Danyal Younus, Hannah Guiang, Darshit Patel, Ramin Javaheri, Sonia Sadr and Stephen Andebirhan. Finally, I would not be able to complete this dissertation without the support and encouragement of my wife Saeba Ruslana, and my brothers Dr. Enam Chowdhury and Tanzir Chowdhury. v Table of Contents Abstract ................................................................................................................................ i Summary ............................................................................................................................. ii Keywords ........................................................................................................................... iii Co-Authorship Statement ................................................................................................... iv Acknowledgments ............................................................................................................... v Table of Contents ............................................................................................................... vi List of Tables ..................................................................................................................... ix List of Figures .................................................................................................................... xi List of Appendices ........................................................................................................... xiii List of Abbreviations ....................................................................................................... xiv Chapter 1 ............................................................................................................................. 1 1 General Introduction ...................................................................................................... 1 1.1 Speciation and Biological Species Concept ............................................................ 1 1.2 Mechanisms of reproductive isolation .................................................................... 2 1.3 Behavioural isolation .............................................................................................. 5 1.4 Genetic basis of behavioural isolation .................................................................... 6 1.5 Drosophila as a model organism ............................................................................ 8 1.6 Courtship behaviour in Drosophila ...................................................................... 12 1.7 Behavioural isolation in Drosophila ..................................................................... 14 1.8 The fruitless gene .................................................................................................. 17 1.9 Overview of dissertation ....................................................................................... 22 1.10 Literature cited ...................................................................................................... 23 Chapter 2 ........................................................................................................................... 44 vi 2 The fruitless gene affects female receptivity and behavioural isolation between D. melanogaster and D. simulans ..................................................................................... 44 2.1 Introduction ........................................................................................................... 44 2.2 Methods................................................................................................................. 48 2.3 Results ................................................................................................................... 54 2.4 Discussion ............................................................................................................. 65 2.5 Conclusion ............................................................................................................ 68 2.6 Literature Cited ..................................................................................................... 68 Chapter 3 ........................................................................................................................... 77 3 Isoforms of the fruitless gene influence species-specific female rejection .................. 77 3.1 Introduction ........................................................................................................... 77 3.2 Methods................................................................................................................
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