University of New Mexico UNM Digital Repository Biology ETDs Electronic Theses and Dissertations Fall 12-12-2018 PHYSELLA ACUTA, COMPARATIVE IMMUNOLOGY AND EVOLUTIONARY ASPECTS OF GASTROPOD IMMUNE FUNCTION Jonathan H. Schultz University of New Mexico - Main Campus Follow this and additional works at: https://digitalrepository.unm.edu/biol_etds Part of the Bioinformatics Commons, Biology Commons, Computational Biology Commons, Genomics Commons, Immunity Commons, and the Parasitology Commons Recommended Citation Schultz, Jonathan H.. "PHYSELLA ACUTA, COMPARATIVE IMMUNOLOGY AND EVOLUTIONARY ASPECTS OF GASTROPOD IMMUNE FUNCTION." (2018). https://digitalrepository.unm.edu/biol_etds/311 This Dissertation is brought to you for free and open access by the Electronic Theses and Dissertations at UNM Digital Repository. It has been accepted for inclusion in Biology ETDs by an authorized administrator of UNM Digital Repository. For more information, please contact [email protected]. Jonathan H. Schultz_______________________ candidate Biology_________________________________ Department This dissertation is approved, and is acceptable in quality and form for publication: Approved by the Dissertation Committee: Dr. Coenraad M. Adema, Chairperson_____________________________________________ Dr. Eric S. Loker_____________________________________________________ Dr. Irene Salinas_____________________________________________________ Dr. Patrick Hanington_________________________________________________ i PHYSELLA ACUTA, COMPARATIVE IMMUNOLOGY AND EVOLUTIONARY ASPECTS OF GASTROPOD IMMUNE FUNCTION By Jonathan H. Schultz B.A., Biology, State University of New York at Geneseo, 2012 DISSERTATION Submitted in Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy Ph.D. in Biology Center for Evolutionary and Theoretical Immunology (CETI) The University of New Mexico Albuquerque, New Mexico May, 2019 ii DEDICATION For my nephews Jackson Jonathan and Ian James. The next generation of critical thinkers. iii ACKNOWLEDGMENTS I’d like to especially thank my advisor, colleague, and friend Dr. Coen M. Adema. I knew very little about the scientific endeavor entering graduate school and I thank you for accepting me into your program and elevating my career in science. I look forward to continuing our impassioned discussions of comparative immunobiology. Thank you to my committee members Dr. Irene Salinas, Dr. Eric S. Loker, and Dr. Patrick Hanington. Irene, thank you for your continued support and guidance, especially the push to attend NACI in North Carolina, which set the course of direction for my postdoctoral studies. Sam, thank you for opening my eyes to the world of evolutionary immunology during my very first semester at UNM and thank you for the support from CETI throughout the years. Patrick, thank you for the enlightening discussions on snail immunity and for supporting me through the end of my PhD. Thank you to Dr. Susan Bandoni-Muench, my first mentor in science. The drive to pursue a career as a scientist started because of taking your parasitology course at SUNY Geneseo and led me on my path to UNM. I am forever grateful. Thank you to my lab and office mates near and far: Dr. Janeth Peña, Lijun Lu, Journey Nolan, and Erin Watson. We shared some of the highs and lows of graduate school together and there isn’t a better group of people to spend the time with. I wish you all continued success and happiness. The utmost gratitude to my closest friends Arthur Tackman, Ben Wheaton, Bishoy Kamel, Danielle Griegos, Erica and Dan Ebbs, and Nate Holscher. Thank you for all of the support and friendship that will last beyond my years in Albuquerque. A very special shout out to my roommate and confidant Joshua Lynn. I couldn’t ask for a better friend. iv Love and thanks to my family. My brother Justin, Nikki, and JJ, thank you for everything, especially the nightly vid chats. There just aren’t enough words. To my Mom, Dad, and Aunt Ronni, for your continued support and for travelling all those miles to see me defend. And lastly, thanks to my ride or die PhD pup and travel buddy, Bo (a.k.a. THE BEANZ), seeing me through it all. v PHYSELLA ACUTA, COMPARATIVE IMMUNOLOGY AND EVOLUTIONARY ASPECTS OF GASTROPOD IMMUNE FUNCTION By Jonathan H. Schultz B.A., Biology, State University of New York at Geneseo, 2012 Ph.D., Biology, University of New Mexico, 2019 ABSTRACT Gastropod immunobiology has benefitted from investigations focused on the planorbid snail Biomphalaria glabrata, intermediate host for the human parasite Schistosoma mansoni. Though such concentrated efforts have elucidated fascinating aspects of invertebrate immunity, they have not provided full knowledge regarding the evolution of immune function among other gastropod species. This dissertation presents the importance of making strategic choices regarding which organisms to select for comparative immunology. Herein, the choice was made to investigate the immunobiology of Physella acuta, a freshwater snail species of the Physidae, a sister family to Planorbidae to which B. glabrata belongs. Benefiting greatly from the use of next-generation sequencing (NGS), the immunobiology of P. acuta was studied using 454 pyrosequencing, Illumina RNA-seq, experimental infections with Echinostoma paraensei (trematode parasite), and other molecular techniques. These analyses revealed that many components of gastropod immunity have been conserved among physid and planorbid snails. Also, P. acuta displays differences in immune function, such as the use of fibrinogen-related proteins in response to trematode parasite exposure. Remarkably, P. acuta differentially expressed relatively large immune-relevant gene families (CD109/TEP, dermatopin, GTPase IMAP, among others) after exposure to E. paraenesi. Inspection of the individual members that represent these gene families vi demonstrated complex transcriptional profiles that suggest parasite influence on host immune function and the capacity of a host to maintain homeostasis while supporting parasite development, an extended phenotype of E. paraenesi. These lab-based studies represent the first large scale characterizations of P. acuta immune function. The immune factors described through NGS approaches enable investigations of the ecoimmunology of P. acuta snails collected from the field. This approach uncovered many sequences that are differentially expressed by P. acuta naturally in the field relative to the lab environment. There is variation in the expression of certain antimicrobial factors and genes governing biological processes. Overall, this dissertation has expanded the scope of gastropod immunity and provides resources and insights that are accessible for continued development and understanding of evolutionary and comparative immunology concepts. vii TABLE OF CONTENTS CHAPTER 1: ……………………….…………………………………………………………………1 INTRODUCTION……………………………………………………………………….1 COMPARATIVE IMMUNOLOGY………………………………………………………………1 GASTROPOD IMMUNOLOGY………………………………………………………………….3 PHYSELLA ACUTA………………………………………………………………………………..5 SIGNIFICANCE…………………………………………………………………………………...6 REFERENCES…………………………………………………………………………………...10 CHAPTER 2: …………………………………………………………………….………………15 COMPARATIVE IMMUNOGENOMICS OF MOLLUSCS………………………..15 ABSTRACT……………………………………………………………………………………..16 INTRODUCTION………………………………………………………………………………17 MOLLUSCAN IMMUNITY…………………………………………………………………..18 BEYOND “FACTOROLOGY”, IMMUNOGENOMICS PHASE 1……………………...22 GENOMES AND NEXT-GENERATION SEQUENCING, IMMUNOGENOMICS PHASE 2…………………………………………………………….28 LIMITATIONS AND POTENTIAL, FUTURE IMMUNOGENOMICS………………...34 ACKNOWLEDGEMENTS……………………………………………………………………38 FIGURES AND TABLES……………………………………………………………………..39 REFERENCES………………………………………………………………………………….42 CHAPTER 3 ………………………………………………………………………………………….63 COMPARATIVE IMMUNOLOGICAL STUDY OF THE SNAIL Physella acuta (HYGROPHILA,PULMONATA) REVEALS SHARED AND UNIQUE ASPECTS OF GASTROPOD IMMUNOBIOLOGY……..…………………………………………………..........................63 viii ABSTRACT……………………………………………………………………………………..64 INTRODUCTION………………………………………………………………………………65 MATERIALS AND METHODS……………………………………………………………...69 RESULTS………………………………………………………………………………………..77 DISCUSSION…………………………………………………………………………………...82 ACKNOWLEDGEMENTS……………………………………………………………………90 APPENDIX A…………………………………………………………………………………...90 FIGURES AND TABLES……………………………………………………………………..91 REFERENCES………………………………………………………………………………….97 CHAPTER 4…………………………………………………………………………………………...107 CHARACTERIZATION OF THE EARLY HOST RESPONSE OF Physella acuta TO THE DIGENETIC TREMATODE Echinostoma paraensei……………………………………………………………………………………….107 ABSTRACT……………………………………………………………………………………108 INTRODUCTION…………………………………………………………………………..…109 MATERIALS AND METHODS…………………………………………………………….112 RESULTS………………………………………………………………………………………115 DISCUSSION………………………………………………………………………………….119 ACKNOWLEDGEMENTS………………………………………………………………..…127 FIGURES AND TABLES……………………………………………………………………128 REFERENCES………………………………………………………………………………...134 ix CHAPTER 5: ………………………………………………………………………………………….145 FROM LAB TO FIELD: INITIAL INVESTIGATION OF THE ECOIMMUNOLOGY OF Physella acuta SNAILS………………………………………………………………………………………...145 ABSTRACT……………………………………………………………………………………146 INTRODUCTION…………………………………………………………………………..…147 MATERIALS AND METHODS ……………………………………………………………149 RESULTS…………………………………………………………………………………..…..152 DISCUSSION………………………………………………………………………………….155 ACKNOWLEDGEMENTS………………………………………………………………..…159 FIGURES AND TABLES.…………………………………………………………………...160 REFERENCES………………………………………………………………………………...171 CHAPTER 6:…………………………………………………………………………………………175