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The Hippocampus Participates in a Pharmacological Rat Model of Absence Seizures

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The Hippocampus Participates in a Pharmacological Rat Model of Absence Seizures Western University Scholarship@Western Electronic Thesis and Dissertation Repository 12-11-2013 12:00 AM The Hippocampus Participates in a Pharmacological Rat Model of Absence Seizures Justin Andrew Arcaro The University of Western Ontario Supervisor Dr. Stan Leung The University of Western Ontario Joint Supervisor Dr. Seyed Mirsattari The University of Western Ontario Graduate Program in Neuroscience A thesis submitted in partial fulfillment of the equirr ements for the degree in Master of Science © Justin Andrew Arcaro 2013 Follow this and additional works at: https://ir.lib.uwo.ca/etd Part of the Molecular and Cellular Neuroscience Commons Recommended Citation Arcaro, Justin Andrew, "The Hippocampus Participates in a Pharmacological Rat Model of Absence Seizures" (2013). Electronic Thesis and Dissertation Repository. 1824. https://ir.lib.uwo.ca/etd/1824 This Dissertation/Thesis is brought to you for free and open access by Scholarship@Western. It has been accepted for inclusion in Electronic Thesis and Dissertation Repository by an authorized administrator of Scholarship@Western. For more information, please contact [email protected]. THE HIPPOCAMPUS PARTICIPATES IN A PHARMACOLOGICAL RAT MODEL OF ABSENCE SEIZURES (Thesis format: Monograph) by Justin Arcaro Graduate Program in Neuroscience A thesis submitted in partial fulfillment of the requirements for the degree of Master of Science The School of Graduate and Postdoctoral Studies The University of Western Ontario London, Ontario, Canada © Justin Arcaro 2013 ABSTRACT The thalamocortical network is responsible for the generation of spike-and-wave discharges (SWDs) in absence epilepsy. Recent studies suggest a potential involvement of the hippocampus, which may explain the variability in the extent of cognitive deficits among patients with absence epilepsy. I hypothesize that the hippocampus may become entrained in spike-and-wave discharges following thalamocortical activation. The gamma-butyrolactone (GBL) rat model of absence seizures was used in this thesis. Following GBL injection, SWDs of 4 to 6 Hz developed in the spontaneous local field potentials (LFPs) recorded by depth electrodes in the thalamus, neocortex and hippocampus. Synchronization of hippocampal, thalamic and neocortical SWDs was revealed by coherence analysis of the LFPs, and multiple unit activity of hippocampal neurons occurred within 250 msec prior to the negative peak of thalamic SWDs. Functional magnetic resonance imaging (fMRI) demonstrated functional connectivity between the hippocampus and the thalamocortical network. Thus, electrophysiological and fMRI activity of the hippocampus were shown to be time-locked to the thalamocortical SWDs, suggesting functional connectivity of the hippocampus and thalamocortical network during GBL-induced absence seizures. Keywords: spike-and-wave discharges, absence epilepsy, gamma-butyrolactone, local field potentials, functional magnetic resonance imaging, functional connectivity, thalamocortical network ii ACKNOWLEDGEMENTS I would like to thank everyone who was supportive throughout my graduate career. First, I offer my gratitude and appreciation to my supervisors, Dr. Stan Leung and Dr. Seyed Mirsattari for their guidance and the opportunity to develop my knowledge on the fundamental principles of neuroscience and applications to the study of epilepsy. Thanks also to Dr. Jingyi Ma for her support with animal care and assistance in the lab. I would also like to thank the members of my advisory committee, Dr. Michael Poulter and Dr. Terry Peters. I would like to thank all members of my lab for their support and assistance. In particular, I would like thank Min-Ching Kuo for her patience in teaching me the necessary skills to conduct surgery and electrophysiological experiments. I would like to extend my appreciation specifically to Matt Hutchison for his expertise in functional magnetic resonance imaging and data analysis. I am greatly indebted to both Min-Ching and Matt for their advice, guidance and support. iii TABLE OF CONTENTS Abstract ............................................................................................................................. ii. Acknowledgements ......................................................................................................... iii. Table of Contents ............................................................................................................ iv. List of Figures and Tables .............................................................................................. vi. 1.0 INTRODUCTION........................................................................................................1 1.1 Epilepsy & Seizures ..................................................................................................1 1.1.1 Classification of Seizures ............................................................................... 2 1.1.2 Absence Seizures ............................................................................................2 1.2 Neurobiology of the Thalamocortical Network .....................................................4 1.2.1 Neurotransmitters ............................................................................................4 1.2.2 Principal Neurons of the Thalamocortical Network .......................................7 1.2.3 Reticular Thalamic Nucleus ............................................................................8 1.2.4 T-type Calcium Ion Channels .......................................................................10 1.3 The Role of the Thalamocortical Network in Absence Seizures ........................12 1.3.1 Competing Theories on the Pathophysiology of Absence Seizures .............16 1.4 Rodent Model of Typical Absence Seizures .........................................................18 1.5 Current Study .........................................................................................................21 2.0 MATERIALS & METHODS....................................................................................24 2.1 Animals ....................................................................................................................24 2.2 Local Field Potentials & Neuronal Unit Recordings ...........................................24 2.2.1 Surgery & Electrode Implantation: Local Field Potentials & Behaviour .....24 2.2.2 Surgery & Electrode Implantation: Neuronal Unit Recordings in relation to Spike-and-Wave Discharges .........................................................................25 2.2.3 Local Field Potentials & Behaviour ..............................................................26 2.2.4 Neuronal Unit Recordings in relation to Spike-and-Wave Discharges ........26 2.2.5 Perfusion, Histology & Staining ...................................................................27 2.2.6 Data Analysis: Local Field Potentials & Behaviour ....................................29 iv 2.2.7 Data Analysis: Neuronal Unit Recordings in relation to Spike-and-Wave Discharges .....................................................................................................30 2.3 Functional Magnetic Resonance Imaging (fMRI) ...............................................32 2.3.1 Acclimation Procedure..................................................................................32 2.3.2 MRI Acquisition ...........................................................................................33 2.3.3 Data Analysis: fMRI .....................................................................................34 3.0 RESULTS ...................................................................................................................36 3.1 Local Field Potentials and Behaviour ......................................................................36 3.2 Single Unit Recordings ............................................................................................42 3.3 Functional Connectivity ...........................................................................................51 4.0 DISCUSSION .............................................................................................................54 4.1 Coherence of Hippocampal, Thalamic and Neocortical Local Field Potentials .....54 4.2 Time-locked Hippocampal Neuronal Firing ............................................................58 4.3 Functional Connectivity between the Hippocampus and Thalamocortical Network ....................................................................................................................59 4.4 Anatomical Connections between the Hippocampus, Thalamus and Neocortex ....62 4.5 Conclusion ...............................................................................................................65 5.0 REFERENCES ...........................................................................................................68 APPENDIX A ...................................................................................................................78 Animal Use Protocol (AUS) Approval Form .................................................................79 Curriculum Vitae .............................................................................................................80 v LIST OF FIGURES & TABLES FIGURES Figure 1: Traces of electroencephalographic spike-and-wave discharge in the human feline ........................................................................................................................................
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