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The Effect of Temperature on Sea Lamprey (Petromyzon Marinus): Ecological and Cellular

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The Effect of Temperature on Sea Lamprey (Petromyzon Marinus): Ecological and Cellular The effect of temperature on sea lamprey (Petromyzon marinus): ecological and cellular implications By Joshua Sutherby A Thesis submitted to the Faculty of Graduate Studies of The University of Manitoba In partial fulfilment of the requirements of the degree of MASTER OF SCIENCE Department of Biological Science University of Manitoba Winnipeg Copyright © 2019 by Joshua Sutherby Abstract Understanding how species respond to thermal challenges is important for forecasting future species distributions, and to provide information for population management. Sea lamprey (Petromyzon marinus), a jawless representative of a basal branch of the vertebrates, successfully invaded the Laurentian Great Lakes in the 20th century to the detriment of the local fishery, resulting in an intensive population control effort since the late 1950s. Sea lamprey are therefore an excellent study species not only for the practical implications of population management, but also as an outgroup to the vertebrates and for testing predictions of invasion theory. Here, the critical thermal maximum (CTMax) of a native population of sea lamprey larvae from New Brunwick was determined in individuals acclimated to 5 °C, 13.5 °C, and 20 °C, and gene expression profiles were determined via reverse-transcription quantitative polymerase chain reaction (qPCR) for genes involved in the cellular stress response (CSR) in response to an acute thermal shock. Subsequently, similar experiments were performed on an invasive landlocked population of sea lamprey larvae acclimated to similar temperatures, and a population comparison was performed. The CTMax results demonstrated that sea lamprey possess a relatively low acclimation capacity, as their CTMax increased only 0.12 °C for each 1 °C increase in acclimation temperature in both populations; however, the landlocked population had a consistently higher CTMax temperature. Sublethal thresholds appear to exist at ~24 °C – 26 °C and ~30 °C in the New Brunswick population, where the CSR is initiated and then shifts to an extreme response, respectively. Invasion theory predicts increased plasticity in invasive populations, and evidence for this was found in the transcription profiles of heat shock proteins in the landlocked population relative to the New Brunswick population. Temperatures above 30 °C are already being recorded in historical sea lamprey spawning streams in the southern end of ii their native distribution, suggesting that a northerly range shift may occur. Despite their greater upper thermal tolerance, this suggests the landlocked population may be more susceptible for future warming, as they already exploit their entire range in the Great Lakes Basin. iii Acknowledgements First and foremost, I would like to thank my advisor, Dr. Kenneth Jeffries for taking the chance on me. You have been a great teacher and role model, and your extensive patience with me has been greatly appreciated. I’d like to thank my lab-mates, William Bugg, McKenzie Hauger, Matthew Thorstensen, and Tina Schaefer for their support, help, and friendship. My thanks go to Dr. Mike Wilkie for allowing me to come into in his lab at Wilfrid Laurier University during the past two summers. You showed me great hospitality and were an excellent mentor. I’d also like to thank Dr. Oana Birceanu, also of WLU, for her help in matters great and small. In addition, this research would not have been possible without the students in Dr. Wilkie’s lab, especially Phil Pham-Ho, Laura Tessier, Julia Sunga, and Darren Foubister. My thanks are also due to Dr. Margaret Docker, for writing the book on lamprey biology, laying much of the foundation for this study, and for her helpful feedback. I’d like to thank my family for their support through this experience, and for all that they have done for me. This research would not have been possible without the financial support provided through an NSERC Discovery Grant, a University of Manitoba Start-up Grant, a Field work Support Program grant, and a University Research Grants Project all awarded to Dr. Kenneth Jeffries. I would also like to thank the Canadian Conference for Fisheries Research, the Faculty of Graduate Studies, and the Faculty of Science for providing funding which allowed me to present my thesis research at CCFFR 2019 in London Ontario. iv Dedication This thesis is dedicated to my Mother and Father. v Table of Contents Abstract .................................................................................................................................................... ii Acknowledgements ................................................................................................................................ iv Dedication ................................................................................................................................................ v Table of Contents ................................................................................................................................... vi List of Tables ........................................................................................................................................ viii List of Figures ......................................................................................................................................... ix List of Abbreviations ............................................................................................................................. xi List of Equations ................................................................................................................................... xii Chapter 1: General Introduction .................................................................................................................. 1 1.1 Upper Thermal Limits ..................................................................................................................... 1 1.2 The Cellular Stress Response .......................................................................................................... 4 1.3 Invasion Theory and Phenotypic Plasticity..................................................................................... 7 1.4 Sea Lamprey .................................................................................................................................... 8 Chapter 2: Response in gene expression to an acute heat shock in an anadromous population of sea lamprey ....................................................................................................................................................... 15 2.1 Abstract ............................................................................................................................................ 15 2.2 Introduction ..................................................................................................................................... 16 2.3 Materials and Methods ................................................................................................................... 19 2.3.1 Animal Collection and Holding ................................................................................................. 19 2.3.2 CTMax Procedure ...................................................................................................................... 20 2.3.3 Thermal Challenge and Sampling Procedure ............................................................................ 21 2.3.4 RNA Extraction and cDNA Synthesis ......................................................................................... 23 2.3.5 Primer Design ............................................................................................................................ 25 2.3.6 qPCR Protocol ........................................................................................................................... 26 2.3.7 qPCR Data Analysis................................................................................................................... 27 2.3.8 Potomac River Temperature ...................................................................................................... 29 2.4 Results .............................................................................................................................................. 30 2.4.1 CTMax ........................................................................................................................................ 30 2.4.2 Gene Expression ........................................................................................................................ 30 2.5 Discussion......................................................................................................................................... 34 2.5.1 CTMax and Acclimation Response Ratio ................................................................................... 35 vi 2.5.2 Gene Expression Response to Acute Thermal Shock ................................................................. 38 2.5.3 Physiological Thresholds ........................................................................................................... 44 2.5.4 Summary .................................................................................................................................... 46 2.6 Figures and Tables .......................................................................................................................... 48 Chapter 3: Thermal tolerance in a New Brunswick anadromous population and a landlocked
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