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Central Neural Mechanisms of Salt-Sensitive Hypertension Michigan Technological University Digital Commons @ Michigan Tech Dissertations, Master's Theses and Master's Reports 2016 CENTRAL NEURAL MECHANISMS OF SALT-SENSITIVE HYPERTENSION Robert Larson Michigan Technological University, [email protected] Copyright 2016 Robert Larson Recommended Citation Larson, Robert, "CENTRAL NEURAL MECHANISMS OF SALT-SENSITIVE HYPERTENSION", Open Access Dissertation, Michigan Technological University, 2016. https://doi.org/10.37099/mtu.dc.etdr/188 Follow this and additional works at: https://digitalcommons.mtu.edu/etdr Part of the Cardiovascular Diseases Commons CENTRAL NEURAL MECHANISMS OF SALT-SENSITIVE HYPERTENSION By Robert A. Larson A DISSERTATION Submitted in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY In Biological Sciences MICHIGAN TECHNOLOGICAL UNIVERSITY 2016 © 2016 Robert A. Larson This dissertation has been approved in partial fulfillment of the requirements for the Degree of DOCTOR OF PHILOSOPHY in Biological Sciences Department of Biological Sciences Dissertation Advisor: Dr. Qing-Hui Chen Committee Member: Dr. Jason R. Carter Committee Member: Dr. Tarun K. Dam Committee Member: Dr. Zhiying Shan Department Chair: Dr. Chandrashekhar P. Joshi Table of Contents List of Figures ................................................................................................ vi List of Tables ................................................................................................ viii Preface .......................................................................................................... ix Acknowledgments .......................................................................................... x List of Abbreviations ..................................................................................... xii Abstract ....................................................................................................... xiv Chapter 1. Literature Review .......................................................................... 1 1.1 Introduction ........................................................................................... 1 1.1.1 Blood Pressure ............................................................................... 1 1.1.2 Blood Pressure Regulation ............................................................. 1 1.1.3 Sodium Homeostasis ..................................................................... 2 1.2 Salt-Sensitive Hypertension .................................................................. 4 1.2.1 Renal Mechanism ........................................................................... 4 1.2.2 Vascular Mechanism ...................................................................... 6 1.2.3 Central Neural Mechanism ............................................................. 6 1.2.4 Angiotensin II-High Salt Model ....................................................... 8 1.3 Central Autonomic Regulation ............................................................ 12 1.3.1 Circumventricular Organs ............................................................. 12 1.3.2 Paraventricular Nucleus ............................................................... 14 1.3.3 Mechanisms Contributing to Augmented PVN Activity ................. 14 1.3.4 Synaptic Mechanisms .................................................................. 15 1.3.5 Intrinsic Mechanisms .................................................................... 15 1.4 Endoplasmic Reticulum ...................................................................... 19 1.4.1 ER Stress ..................................................................................... 19 1.4.2 Intracellular and ER Ca2+ Regulation ........................................... 20 1.5 Summary and Hypothesis ................................................................... 22 iii Chapter 2. ..................................................................................................... 24 2.1 Introduction ......................................................................................... 24 2.2 Materials and Methods ........................................................................ 25 2.2.1 Animals......................................................................................... 25 2.2.2 Protocols of Animal Experimental Model ...................................... 26 2.2.3 Experiment Preparation ................................................................ 26 2.2.4 Recording of Sympathetic Nerve Activity ..................................... 27 2.2.5 PVN Microinjection ....................................................................... 27 2.2.6 Punched Brain Tissues from Rats ................................................ 28 2.2.7 Western Blot Analysis of SK Channel Protein .............................. 28 2.2.8 Data Analysis ............................................................................... 29 2.3 Results ................................................................................................ 29 2.3.1 AngII-salt HTN .............................................................................. 29 2.3.2 Effects of PVN-Injected SK Channel Blocker on SSNA, RSNA, MAP and HR ......................................................................................... 30 2.3.3 Effects of PVN-Injected SK Channel Activator on SSNA, RSNA, MAP and HR ......................................................................................... 34 2.3.4 Histological Analysis ..................................................................... 35 2.3.5 Comparison of SK Channel Protein Expression ........................... 35 2.4 Discussion........................................................................................... 38 2.5 Perspectives ....................................................................................... 43 2.6 Acknowledgments ............................................................................... 43 Chapter 3. ..................................................................................................... 44 3.1 Introduction ......................................................................................... 44 3.2 Methods .............................................................................................. 45 3.2.1 Animals......................................................................................... 45 3.2.2 Microinjection Experiment Preparation ......................................... 46 3.2.3 Recording of Sympathetic Nerve Activity (SNA) ........................... 46 3.2.4 Paraventricular Nucleus (PVN) Microinjection .............................. 46 3.2.5 Retrograde Labeling ..................................................................... 47 3.2.6 Electrophysiology ......................................................................... 47 3.2.7 Testing Neuronal Excitability ........................................................ 48 3.2.8 Western Blot measurements of SERCA Protein ........................... 49 iv 3.2.9 Chemicals ..................................................................................... 49 3.2.10 Data Analysis ............................................................................. 50 3.3 Results ................................................................................................ 50 3.3.1 Depletion of PVN ER Ca2+ stores Augments SNA and ABP ........ 50 3.3.2 HS Diet Disrupts PVN ER Ca2+ Store Function Contributing to Sympathoexcitation ............................................................................... 53 3.3.3 HS Diet Disrupts ER Ca2+ Store Function Contributing to Augmented Excitability of PVN-RVLM Neurons .................................... 55 3.3.4 HS Disruption of ER Ca2+ Store Reduces Spike-Frequency Adaptation in PVN-RVLM Neurons ....................................................... 56 3.3.5 Comparison of PVN SERCA Protein Expression ......................... 59 3.3.6 Histology....................................................................................... 60 3.4 Discussion........................................................................................... 61 3.5 Perspectives ....................................................................................... 66 3.6 Acknowledgments ............................................................................... 66 Chapter 4. ..................................................................................................... 67 4.1 Summary ............................................................................................ 67 4.2 Limitations ........................................................................................... 68 4.3 Future Studies ..................................................................................... 70 4.4 Conclusions ........................................................................................ 71 References ................................................................................................... 73 Appendix A. Raw Data for Study 1. .............................................................. 96 Appendix B. Summary Statistics for Study 1. ............................................. 101 Appendix C. Raw Data for Study 2. ............................................................ 107 Appendix D. Summary Statistics for Study 2. ............................................. 121 Appendix E. Chemical Structures ..............................................................
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