Molecular Mapping of the Binding Sites of a Nav1.1 Activator Héctor Botella Master’s Thesis by Héctor Botella, MSc in MedIS Biomedicine, Aalborg Universitet / H. Lundbeck A/S (2018) Thesis submitted 07 - 09 - 2018 Internal Thesis supervisor Prof. Dr. Meg Duroux, Aalborg University External Thesis supervisor Dr. Tau Benned-Jensen, Lundbeck A/S Thesis committee Prof. Dr. Sebastian Frische Prof. Dr. Meg Duroux, Aalborg University Dr. Tau Benned-Jensen, Lundbeck A/S Thesis Series Aalborg University 9220 Aalborg Ø, Aalborg, Denmark Faculty of Medicine, Aalborg University Niels Jernes Vej 10 | DK - 9220 Aalborg Ø Phone: +45 9940 9940 | [email protected] School of Medicine and Health Niels Jernes Vej 12, A5 | 9220 Aalborg Ø | Aalborg, Denmark Department of Health, Science and Technology Aalborg University | Fredrik Bajers Vej 7 D2 | DK-9220, Aalborg Ø Published in Aalborg University Publication Repository (vbn.aau.dk) © Héctor J. Botella Valle Printed in Aalborg, Denmark, 2018 Nº of pages 95 2 Molecular Mapping of the Binding Sites of a Nav1.1 Activator Master’s Thesis by Héctor Botella, MSc in MedIS Biomedicine, Aalborg Universitet / H. Lundbeck A/S (2018) I. INDEX OF CONTENTS I. Index of contents ................................................................................................................................................................. 3 II. Preface ................................................................................................................................................................................. 5 III. Abstract ............................................................................................................................................................................... 6 IV. List of abbreviations .......................................................................................................................................................... 7 1. INTRODUCTION .................................................................................................................................................... 1.1. - The nervous system ....................................................................................................................................................... 8 1.2. - The CNS ........................................................................................................................................................................... 8 1.3. - The brain ......................................................................................................................................................................... 9 1.4. - The neurons ..................................................................................................................................................................10 1.4.1. - Interneurons ..............................................................................................................................................................11 1.5. - Diseases of the CNS ......................................................................................................................................................12 1.5.1. - Schizophrenia ............................................................................................................................................................12 1.5.2. - Dravet Syndrome (SMEI)...........................................................................................................................................13 1.6. - Other types of neural cells: the glia ............................................................................................................................14 1.7. - Neuronal synapses .......................................................................................................................................................15 1.8. - Action potentials ..........................................................................................................................................................16 1.9. - Ion channels ..................................................................................................................................................................18 1.9.1. - Architecture of ion channels ....................................................................................................................................18 1.9.2. - The sodium ion channel family .................................................................................................................................19 1.9.2.bis. - Activators and inactivators .................................................................................................................................22 1.9.2.1. - Nav1.1 .....................................................................................................................................................................24 1.9.2.2. - Nav1.4 .....................................................................................................................................................................25 1.9.3. - The potassium ion channel family ............................................................................................................................26 1.9.3.1. - Kv2.1 ........................................................................................................................................................................27 1.10. -Electrophysiology ........................................................................................................................................................28 1.10.1. - Voltage-Clamp .........................................................................................................................................................29 2. AIM OF THIS STUDY .......................................................................................................................................... 30 3. RESULTS ........................................................................................................................................................... 32 3.1. – Prelude to results ........................................................................................................................................................32 3.2. - Analysis of native channels ..........................................................................................................................................33 3.2.1. - Nav1.1 – Published literature ....................................................................................................................................33 3.2.1.1. - Nav1.1 – Manual patch-clamp ...............................................................................................................................34 3.2.1.1.1. - Nav1.1 – MPI ........................................................................................................................................................35 3.2.1.1.2. - Nav1.1 – AUC .......................................................................................................................................................36 3.2.1.1.3. - Nav1.1 – tau .........................................................................................................................................................37 3.2.1.1.4. - Nav1.1 – I/V relation ...........................................................................................................................................38 3.2.1.1.5. - Nav1.1 – Activation .............................................................................................................................................38 3.2.1.1.6. - Nav1.1 – Inactivation ...........................................................................................................................................39 3.2.2. - Nav1.4 – Published literature ....................................................................................................................................40 3.2.2.1. - Nav1.4 – Manual patch-clamp ...............................................................................................................................40 3.2.2.1.1. - Nav1.4 – MPI ........................................................................................................................................................41 3.2.2.1.2. - Nav1.4 – AUC .......................................................................................................................................................41 3.2.2.1.3. - Nav1.4 – tau .........................................................................................................................................................43 3.2.2.1.4. - Nav1.4 – I/V relation ...........................................................................................................................................43 3.2.2.1.5. - Nav1.4 – Activation .............................................................................................................................................44 3.2.2.1.6. - Nav1.4 – Inactivation ...........................................................................................................................................45 3.2.3. - Endogenous currents in control cells .......................................................................................................................45 3.2.3.1. - Endogenous