Chemoreception: A Consideration of Olfactory Signalling Proteins in Non-Olfactory Systems Yuliya Makeyeva A thesis in fulfillment of the requirements for the degree of Doctor of Philosophy School of Women’s and Children’s Health Faculty of Medicine November 2018 Thesis/Dissertation Sheet Australia's Global UNSWSYDNEY University Surname/Family Name Makeyeva Given Name/s Yuliya Abbreviation for degree as given in the University calendar PhD Faculty Faculty of Medicine School School of Women's and Children's Health Thesis Title Chemoreception: A Consideration of Olfactory Signalling Proteins in Non­ Olfactory Systems Abstract 350 words maximum: Chemoreception is a biological process whereby cells and/or organisms are stimulated by chemicals in their environment. The detectibn of chemicals triggers a response that can be the attraction or aversion by a single II cell or a highly integrative process that initiates a complex behaviour. Olfactory receptors (ORs) are a multigene family of molecules used to monitor extracellular chemical cues. They were originally described in the olfactory system as mediators of the sense of smell but have since been observed in a number of non-olfactory tissues. OR-mediated chemoreception in the olfactory system involves a G protein (Gait), adenylyl cyclase Ill (AC3), and OMP. ORs are difficultto study so little is known about their functions and expression profiles in these tissues. I, Olfactory marker protein (OMP) is a highly expressed protein found in mature olfactory sensory neurons of all vertebrates, co-labels with individual ORs, and is involved in intracellular signal transduction. OMP expression may indicate OR-mediated chemoreception in non-olfactory systems. Cells that are mobile are optimal candidates because they might use chemoreception to guide their movements. My thesis used OMP, Gait, and AC3 to identify chemoreception in non-olfactory cells: spermatozoa, mast cells, and Leydig cells. The distribution of these proteins was investigated using immunohistochemistry and microscopic image analysis. In rats, expression of these proteins was documented in spermatozoa, mast cells, and Leydig cells. In humans, expression was investigated in spermatozoa in three modes of activation: control, activated, and hyper-activated. The results showed that OMP, Gait, and AC3 were co-expressed in spermatozoa, mast cells, and Leydig cells. Due to the adverse reaction of tissues to a high-fat diet, the response of mast cells, the I olfactory epithelium, and olfactory bulb was tested and found to be rapidly altered. This presence of an OR-mediated signalling pathway suggests chemoreception and guidance in diverse cells of non-olfactory tissues. A high-fat diet, even in the short term, adversely affected immunity and chemoreception. My data reveal the widespread and complex interactions among diverse systems that are mediated by chemoreception. Declaration relating to disposition of project thesis/dissertation I hereby grant to the University of New South Wales or its agents the right to archive and to make available my thesis or dissertation in whole or in part in the University libraries in all forms of media, now or here after known, subject to the provisions of the Copyright Act 1968. I retain all property rights, such as patent rights. I also retain the right to use in future works (such as articles or books) all or part of this thesis or dissertation. I also authorise University Microfilms to use the 350 word abstract of my thesis in Dissertation Abstracts International (this is applicable to doctoral theses only). ....i.f ... !.{.49/.8 Witness Signature Date The University recognises that there may be exceptional circumstances requiring restrictions on copying or conditions on us�. Requests for restriction for a period of up to 2 years must be made in writing. Requests for a longer period of restriction may be considered in exceptional circumstances and require the approval of the Dean of Graduate Research. OR OFFICE USE ONLY Date of completion of requirements for Award: COPYRIGHT STATEMENT ‘I hereby grant the University of New South Wales or its agents the right to archive and to make available my thesis or dissertation in whole or part in the University libraries in all forms of media, now or here after known, subject to the provisions of the Copyright Act 1968. I retain all proprietary rights, such as patent rights. I also retain the right to use in future works (such as articles or books) all or part of this thesis or dissertation. I also authorise University Microfilms to use the 350 word abstract of my thesis in Dissertation Abstract International (this is applicable to doctoral theses only). 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Publications can be used in their thesis in lieu of a Chapter if: • The student contributed greater than 50% of the content in the publication and is the "primary author'', ie. the student was responsible primarily for the planning, execution and preparation of the work for publication • The student has approval to include the publication in their thesis in lieu of a Chapter from their supervisor and Postgraduate Coordinator. • The publication is not subject to any obligations or contractual agreements with a third party that would constrain its inclusion in the thesis Please indicate whether this thesis contains published material or not. This thesis contains no publications, either published or submitted for publication (if this box is checked, you may delete all the material on page 2) Some of the oork described in this thesis has been published and it has been documented in the relevant Chapters Vvith acknowedgement (if this box is D checked, you may delete all the material on page 2) This thesis has publications (either published or submitted for publication) incorporated into it in lieu of a chapter and the details are presented below D l, CANDIDATE 'S DE CLARATION I declare that: • I have complied with the Thesis Examination Procedure • where I have used a publication in lieu of a Chapter, the listed publication(s) below meet(s) the requirements to be included in the thesis. Name Date (dd/mmfyy) \du)1_('.j Q_, N f\� '::!E V 14- l60Y.;l0(9 Postgraduate Coordinator's Declaration (to be filled in where publications are used in lieu of Chapters) I declare that: • the information below is accurate • where listed 'publication(s) have been used in lieu of Chapter(s), their use complies with the Thesis Examination Procedure • the minimum requirements for the format of the thesis have been met. PGC'sName PGC's Signature Date (dd/mmfyy) Originality Statement 'I hereby declare that this submission is my own work and to the best of my knowledge it contains no materials previously published or written by another person, or substantial proportions of material which have been accepted for the award of any other degree or diploma at UNSW or any other educational institution, except where due acknowledgement is made in the thesis. Any contribution made to the research by others, with whom I have worked at UNSW or elsewhere, is explicitly acknowledged in the thesis. I also declare that the intellectual content of this thesis is the product of my own wofk, except to the extent that assistance from others in the project's design and conception or in style, presentation and linguistic expression is acknowledged.' Signed: Date: iii Supervisors Supervisor: Professor William Ledger School of Women’s and Children’s Health UNSW Sydney Co-Supervisor: Professor David Ryugo Hearing Research Garvan Institute of Medical Research and UNSW Sydney Co-Supervisor: Dr Steven Leach School of Women’s and Children’s Health UNSW Sydney iv Dedication For my father Valentine, my mother Elmira, and my daughter Kseniya—for their unflagging love and support v Acknowledgements I wish to thank the following people for their assistance with my PhD. My sincere thanks go: to Professor Laura Poole-Warren for her support through my PhD despite considerable difficulties; to my primary supervisor, Professor William Ledger, for inspiring my interest in reproduction and for his support and guidance during the final stages of my PhD. He gave me the strength and motivation to complete what has been an arduous journey; to my co-supervisor, Professor David Ryugo, for allowing me to complete my PhD in his laboratory, for his encouragement and feedback. I am grateful for his gentle guidance and the freedom he gave me to pursue my aims and the confidence to keep going. I also thank him for helpful comments and discussions on my thesis; to Dr Steven Leach, my co-supervisor, for his helpful assistance in interpreting