University of Massachusetts Amherst ScholarWorks@UMass Amherst Doctoral Dissertations Dissertations and Theses July 2016 Spider Brain Morphology & Behavior Skye M. Long University of Massachusetts Amherst Follow this and additional works at: https://scholarworks.umass.edu/dissertations_2 Part of the Biology Commons, Entomology Commons, and the Other Neuroscience and Neurobiology Commons Recommended Citation Long, Skye M., "Spider Brain Morphology & Behavior" (2016). Doctoral Dissertations. 707. https://doi.org/10.7275/8128500.0 https://scholarworks.umass.edu/dissertations_2/707 This Open Access Dissertation is brought to you for free and open access by the Dissertations and Theses at ScholarWorks@UMass Amherst. It has been accepted for inclusion in Doctoral Dissertations by an authorized administrator of ScholarWorks@UMass Amherst. For more information, please contact [email protected]. SPIDER BRAIN MORPHOLOGY & BEHAVIOR A Dissertation Presented by SKYE MICHELLE LONG Submitted to the Graduate School of the University of Massachusetts Amherst in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY May 2016 The Program in Organismic and Evolutionary Biology © Copyright by Skye Long 2016 All Rights Reserved SPIDER BRAIN MORPHOLOGY AND BEHAVIOR A Dissertation Presented By SKYE MICHELLE LONG Approved as to style and content by: Elizabeth M. Jakob, Chair Jeffrey Podos, Member Elizabeth R. Dumont, Member Susan Fahrbach, Member Elizabeth R. Dumont, Director Interdepartmental Graduate Programs DEDICATION To my beloved husband Robert. You make me want to be the person you believe I am. EPITAPH “It's still magic even if you know how it's done.” ― Terry Pratchett, A Hat Full of Sky ACKNOWLEDGEMENTS First of all, I would never thank all the thousands of people who have helped me in large and small ways get to this point in my career, but please accept this partial list. I thank my advisor, Beth Jakob, for her incredible support for this dissertation. She provided an environment where I could explore risky ideas and follow my passions. I could not have done this without her help. She provided an unlimited number of revisions not only this dissertation, but numerous grants, letters and emails. The exception being the acknowledgements, all mistakes within are mine and mine alone. In addition, her enthusiastic support allowed me to visit numerous labs across the country to learn techniques and attempt to morph from an animal behaviorist into a larval neurobiologist. I would like to thank Chris Maher and Ken Weber for giving me the skills and confidence to even apply to graduate school. Jeff Podos, Betsy Dumont and Susan Fahrbach for all of their advice and direction as part of my committee. I know the path was not always clear. I would like to thank the many people who provided advice and direction. Marc Seid for helping me with dissection techniques and introducing me to Reconstruct. Susan Fahrbach for helping me with embedding advice, as well as intellectual support as a member of my committee. Ron Hoy, Gil Menda and Paul Shamble for evaluating my early work and pushing me further. Glen Marrs for his work on developing the tissue processing method. Christine Divine for introducing me to DiI. Tom Eiting for sharing the secrets of paraffin sectioning with me. The resources of many labs were shared in the completion of this dissertation. Ethan Graf and Dominic Poccia for allowing me to access to the confocal, without which Chapters 3 and 4 would not have been possible. Tobias Baskin for the use of his vibratome. Kate Dorfman for allowing me lab and hood space. Robbie Cairl for donating invaluable supplies. vi I received support for this work from numerous agencies: The University of Massachusetts Amherst, The Graduate Program in Organismic & Evolutionary Biology, Sigma Xi and The American Arachnological Society. In addition, this work was supported by NSF IOS 0952822 to my advisor Elizabeth Jakob. My graduate school experience would not have been the same without the many amazing people I have met both here at OEB and at the many wonderful meetings I have attended. In particular, Lexi Brown, Rachel & Matt Bolus, Sarah Goodwin, Dina Navon, Moira Concannon, Tom Eiting, Chi-Yun Kuo, Michael Rosario, Kit Harcourt, Casey Gilman, Annie Leonard, Anne Stengle, Jamie Church, Zane Barlow, Dana Moseley, Yi-Fen Lin, Jamie Tanner, Israel Del Toro and Andy Smith. In addition, I have had the opportunity to work with some truly incredible undergraduates, both as students and lab assistants. Finally, Penny Jaques for the thousands of times she helped me find my way out of the woods. All of you helped me more than you could ever know. vii ABSTRACT SPIDER BRAIN MORPHOLOGY & BEHAVIOR MAY 2016 SKYE MICHELLE LONG, B.A., UNIVERSITY OF SOUTHERN MAINE Ph.D., UNIVERSITY OF MASSACHUSETTS AMHERST Directed by: Professor Elizabeth M. Jakob Spiders are ideal model animals for experimental and comparative studies of behavior, learning and perception. They display many complex behaviors, such as the multimodal mating dances of lycosid spiders, the stealthy hunting strategies of the jumping spider Portia sp., to the labile sociality of theridiids. Spiders also demonstrate a wide range of cognitive capabilities. Spiders perceive their environment using multiple sensory modalities including: chemosensory organs; lyriform and slit-sense organs and specialized hairs that detect vibration and air movement; and up to eight eyes that vary in function, some able to detect polarization and a broad spectrum of light, including ultraviolet. While much is known about the behavior and external morphology of spiders, little is known about the spider’s nervous system. Early in the 20th century researchers, such as Saint- Remy, Hanström and Legendre, began the process of cataloging the variety of form and function within the arachnid brain. Unfortunately, these studies were limited by techniques and sample quality and much of the information is difficult to access and place into a modern context. In modern research the focus on comparative studies of spider brain morphology disappeared and was replaced with more focused research on a single species, the wandering spider Cupiennius salei. While much has been learned from these studies, C. salei represents only a small fraction of the spectrum of behaviors and sensory system morphologies that may be reflected in brain viii morphology. Current advances in techniques and collecting methods, combined with the framework of knowledge gained from C. salei allow for meaningful comparative work on spider neurobiology. The four chapters of my dissertation explore spider behavior, learning and neuromorphology and present two novel protocols for their study. In Chapter 1, I present a behavioral study in which I explore the effect of firefly flashing on the predatory behavior of spiders. In Chapter 2, I present a novel protocol for aversive learning trials in spiders. In Chapter 3, I present a novel method for producing whole-head DiI stained spider cephalothorax sections. In Chapter 4, I describe the variation in the visual processing pathways in spiders representing 19 different families. ix TABLE OF CONTENTS Page ACKNOWLEDGEMENTS ................................................................................................................... vi ABSTRACT ....................................................................................................................................... viii LIST OF TABLES ................................................................................................................................ xv LIST OF FIGURES ............................................................................................................................. xvi CHAPTER 1. FIREFLY FLASHING AND JUMPING SPIDER PREDATION ............................................................... 1 1.1 Introduction .................................................................................................................. 1 1.2 Methods ........................................................................................................................ 4 1.2.1 Study organisms ............................................................................................ 4 1.2.2 Firefly Disturbance and Flash Behavior ........................................................ 5 1.2.3 Dusk Activity Trials ........................................................................................ 6 1.2.4 Tests of the Startle Hypothesis for Flashing ................................................. 6 1.2.5 Effect of Flashing on Spiders’ Prey Choice .................................................... 8 1.2.6 Effect of Flashing on Avoidance of Unpalatable Prey Over Multiple Encounters ................................................................................................ 8 1.3 Results ......................................................................................................................... 10 1.3.1 Firefly Disturbance and Flash Behavior ...................................................... 10 1.3.2 Dusk Activity Trials ...................................................................................... 10 1.3.3 Tests of the Startle Hypothesis for Flashing ............................................... 10 1.3.4 Effect of Flashing on Spiders’ Prey Choice .................................................. 11 1.3.5 Effect of Flashing on Avoidance of Unpalatable Prey Over Multiple Encounters