The Basolateral Amygdala and Superior Colliculus Regulate Physiological and Behavioral Measures of Avoidance and Approach
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THE BASOLATERAL AMYGDALA AND SUPERIOR COLLICULUS REGULATE PHYSIOLOGICAL AND BEHAVIORAL MEASURES OF AVOIDANCE AND APPROACH A Dissertation submitted to the Faculty of the Graduate School of Arts and Sciences of Georgetown University in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Neuroscience By Catherine Elorette, B.S. Washington, DC June 13, 2019 Copyright 2019 by Catherine Elorette All Rights Reserved ii THE BASOLATERAL AMYGDALA AND SUPERIOR COLLICULUS REGULATE PHYSIOLOGICAL AND BEHAVIORAL MEASURES OF AVOIDANCE AND APPROACH Catherine Elorette, B.S. Thesis Advisors: Ludise Malkova, Ph.D; Patrick Forcelli, Ph.D ABSTRACT The superior colliculus (SC) and amygdala are central components of the defensive system within the brain. The SC processes conscious and non-conscious visual information and directs orienting and defensive behavioral responses. The amygdala exerts “top down” control over SC and may interact with the SC to regulate emotional response to visual stimuli. To determine the anatomical connectivity between the SC and the amygdala, I injected retrograde tracers into the amygdala and anterograde tracers into the SC and examined regions of colocalization within the pulvinar, a vision associated region. Anterograde tracers labeled axonal projections that colocalized with cell bodies labeled by the retrograde tracer, concentrated in the medial portions of the medial (PM), oral (PO) and inferior pulvinar (PI). These data support the existence of a "bottom up” pathway mediating fast processing of emotionally salient information projecting from the SC to the amygdala through the pulvinar. To determine the causal association of activity in the amygdala on (1) autonomic activity, (2) emotional reactivity to social and threatening stimuli, and (3) social dominance, I used a pharmacological microinjection approach. I inhibited the amygdala via GABA A agonist muscimol (MUS) and activated it with GABA A antagonist bicuculline methiodide (BMI). Saline was injected as a control. Following drug infusion, animals iii were tested in the above behavioral domains. Activity in the amygdala was associated with decreased heart rate, nonspecific avoidance behavior, and decreased dominance. Inhibition of the amygdala was associated with decreased heart rate, reduced responsiveness to a threatening stimulus, increased responsiveness to a social stimulus, and decreased dominance. To determine if activity within the SC regulated autonomic responses, I microinjected BMI into the SC and measured heart rate and blood pressure. Consistent with its laminar structure and segregated zones of anatomical inputs and outputs, subregions of the SC showed a diversity of responses, but primarily decreased heart rate and blood pressure, localized in intermediate and deep layers. I have shown that BLA and SC separably regulate autonomic outputs. BLA additionally regulates behavioral response to threats and challenges to established behavioral patterns, reflecting alterations in larger networks involved in recognizing and responding appropriately to environmental cues. iv Acknowledgments First and foremost, I want to thank my family for supporting me, both during my graduate education and in the years leading up to it. My mother, Debbie, my father, Greg, and my brother Gregory – thank you for helping me, listening to me, and giving me space to figure it all out. You have been my tireless champions. To my mentors, Ludise and Patrick, who guided me with grace, patience, occasional bribery, and minimal exasperation, and provided me with advice, support, opportunities, and experience that helped me to grow both personally and academically. I could not have asked for a better pair of people to guide me on this journey. I owe a great deal to all the teachers, mentors, and administrators that got me to where I am now, including the McNulty Foundation, which awarded me a scholarship for my undergraduate education. My greatest thanks must go to my friends, who are the family we choose for ourselves. I have many people I must acknowledge here – I am lucky to have many, many wonderful party members in this adventure of life. To my childhood crew, Morgan, Megan, Catherine, Sarah, Melissa – you have all shaped me and helped me to grow, both early on and to the present day. I know you will have my back whatever may come next. To university friends, Addie, Brit, Tori, Bernadette, Shannon, and many others –I will always appreciate your love and emotional support that kept me centered. To friends I met along the way, Violet, Mary, Lauren, Jane, Kayla, Julie, Zeda, and many, MANY more – thank you for enriching my life. You’re all close to my heart. To Vanessa, thank you for helping me to grow. To IPN and associates, my classmates, program mates, and others (Joey, Katie, Kelly, Sri, Emily, Laya, Cam, Pat, Lorenzo, Sikoya, Joscelin, Shawn, Paige, Erin, Aileen, Allison, this section will take up half the page if I let it) thank you for your jokes, serious late night talks, board game nights and movie marathons. I’m going to miss you all. To George, your care and support have kept me grounded, and our discussions have helped me to reach higher. Thank you – and here’s to many more adventures. To my monkey ladies, Brittany, Elyssa, and Hannah – thanks for taking this journey together with me. You’ve kept me going through early morning procedures and late night check-ins. I couldn’t have done it without you all by my side. To the smol nugs, Laura, Adam, Kevin, and Jayson, I love you all dearly. Thank you for always being there for me, helping me work through my struggles and celebrating my successes. I hope we can continue to grow together for the rest of our lives. With love, Catherine Elorette v TABLE OF CONTENTS CHAPTER 1: INTRODUCTION ...................................................................................................................................... 1 1.1 STRUCTURE OF THE AMYGDALA ................................................................................................................................. 3 1.2 EMOTIONAL REGULATION .............................................................................................................................................. 6 1.3 SOCIAL COGNITION .......................................................................................................................................................... 9 1.4 STRUCTURE OF THE SUPERIOR COLLICULUS ...................................................................................................... 12 1.5 BLINDSIGHT ....................................................................................................................................................................... 15 1.6 BEHAVIORAL RESPONSES OF SUPERIOR COLLICULUS .................................................................................... 16 1.7 ACUTE MICROINJECTION ............................................................................................................................................. 18 1.8 GOALS OF THE DISSERTATION .................................................................................................................................. 20 CHAPTER 2: ANATOMY OF THE PROJECTIONS FROM SUPERIOR COLLICULUS TO AMYGDALA ............................................................................................................................................................................ 23 2.1 INTRODUCTION ................................................................................................................................................................. 23 2.2 METHODS ........................................................................................................................................................................... 26 2.3 RESULTS ............................................................................................................................................................................ 32 2.4 DISCUSSION ...................................................................................................................................................................... 36 CHAPTER 3: THE ROLE OF THE AMYGDALA IN PHYSIOLOGICAL AND AFFECTIVE AROUSAL ................................................................................................................................................................................ 52 3.1 INTRODUCTION ................................................................................................................................................................. 52 3.2 METHODS ........................................................................................................................................................................... 54 3.3 RESULTS ............................................................................................................................................................................ 64 3.4 DISCUSSION ...................................................................................................................................................................... 73 CHAPTER 4: THE ROLE OF SUPERIOR COLLICULUS IN AUTONOMIC RESPONSE .............. 90 4.1 INTRODUCTION ................................................................................................................................................................. 90 4.2 METHODS ..........................................................................................................................................................................