Arterial Chemoreceptors: Mal(adaptive) Responses: O2 Dependent and Independent Mechanisms

Preface
Contents
Contributors
1: Transcriptomics of the Carotid Body
	1.1	 Introduction
		1.1.1	 Notes of Caution
	1.2	 Overview of Carotid Body Transcriptomic Studies
	1.3	 The Known Unknowns of the Carotid Body’s Transcriptome
	1.4	 Summary: Future Directions
	References
2: The Adult Carotid Body: A Germinal Niche at the Service of Physiology
	2.1	 Introduction
	2.2	 The CB Contains Intermediate Restricted Progenitors from Both Vascular and Neuronal Lineages, to Accelerate Adaptation to Chronic Hypoxia
	2.3	 Mature Glomus Cells as Master Regulators of the Adult Carotid Body Germinal Niche
	2.4	 Clinical Implications and Concluding Remarks
	References
3: Evidences That Sympathetic Overactivity and Neurogenic Hypertension Correlate with Changes in the Respiratory Pattern in Rodent Models of Experimental Hypoxia
	3.1	 Introduction
	3.2	 Wistar Ribeirão Preto Rats Submitted to Chronic Intermittent Hypoxia
		3.2.1	 Male Rats Submitted to Chronic Intermittent Hypoxia
		3.2.2	 Female Rats Submitted to Chronic Intermittent Hypoxia
	3.3	 Wistar Ribeirão Preto Rats Submitted to Sustained Hypoxia
	3.4	 Wistar Hannover Rats Submitted to Sustained Hypoxia
	3.5	 Sprague-Dawley Rats Submitted to Sustained Hypoxia
	3.6	 Mice Submitted to Sustained Hypoxia
	3.7	 Summary
	References
4: Control of Arterial Hypertension by the AhR Blocker CH-223191: A Chronopharmacological Study in Chronic Intermittent Hypoxia Conditions
	4.1	 Introduction
	4.2	 Material and Methods
		4.2.1	 In Vivo Experiments
			4.2.1.1	 Ethics
			4.2.1.2	 Animals
			4.2.1.3	 Chronic Intermittent Hypoxia Paradigm
			4.2.1.4	 Study Design
				4.2.1.4.1 Evaluation of the Chronopharmacology of the Antihypertensive Efficacy of the AhR Blocker CH-223191 in CIH Conditions
				4.2.1.4.2 Circadian Variation of AhR Activation in the Kidney Cortex Under Normoxic Conditions
			4.2.1.5	 Terminal Surgeries
		4.2.2	 Assessment of AhR Activation Through Western Blot Analysis of CYP1A1 Levels
		4.2.3	 Statistical Analysis
	4.3	 Results
	4.4	 Discussion
	References
5: Three Days of Chronic Intermittent Hypoxia Induce β1-Adrenoceptor Dependent Increases in Left Ventricular Contractility
	5.1	 Introduction
	5.2	 Methods
		5.2.1	 Ethical Approval
		5.2.2	 Chronic Intermittent Hypoxia Protocol
		5.2.3	 Anesthetized In Vivo Preparation
		5.2.4	 Gene Expression
		5.2.5	 Data and Statistical Analysis
	5.3	 Results
		5.3.1	 Baseline Cardiovascular Parameters
		5.3.2	 Left Ventricular Contractility in Response to Chemostimulation
		5.3.3	 Cardiovascular Response to β-Adrenoceptor Blockade
		5.3.4	 Sympathetic Nervous System Inhibition
		5.3.5	 Gene Expression of the β1-Adrenoceptor Pathway
		5.3.6	 Catecholamine Concentrations
	5.4	 Discussion
		5.4.1	 Hypoxia and the Cardiovascular System
	5.5	 Conclusion
	References
6: The Beneficial Effect of the Blockade of Stim-Activated TRPC-ORAI Channels on Vascular Remodeling and Pulmonary Hypertension Induced by Intermittent Hypoxia Is Independent of Oxidative Stress
	6.1	 Introduction
	6.2	 Methods
		6.2.1	 Animals and Intermittent Hypoxia Protocol
			6.2.1.1	 2-APB Treatment
		6.2.2	 Right Ventricular Systolic Pressure Measurement
		6.2.3	 STOC Pulmonary Gene Expression
		6.2.4	 Systemic and Pulmonary Oxidative Stress Measurement
		6.2.5	 Vascular Remodeling and Immunohistochemistry
		6.2.6	 Data Analysis, Pearson Correlation, and Statistical Analyses
	6.3	 Results
		6.3.1	 Pearson’s Correlation of Physiological Variables Related to Right Ventricle Systolic Pressure
		6.3.2	 Pearson’s Correlation of Physiological Variables Related to MDA Concentrations at the Pulmonary Level
	6.4	 Discussion
	References
7: Intermittent Hypoxia and Weight Loss: Insights into the Etiology of the Sleep Apnea Phenotype
	7.1	 Introduction
	7.2	 Methods
		7.2.1	 Animals and Experimental Groups
			7.2.1.1	 Intermittent Hypoxia Protocol
		7.2.2	 Animal Monitoring and Experimental Measurements
			7.2.2.1	 Ventilatory Measurements
			7.2.2.2	 Assessment of Respiratory Reflexes
			7.2.2.3	 Data Analysis
			7.2.2.4	 Blood Sampling and Biochemical Analyses
		7.2.3	 Statistical Analysis
	7.3	 Results
		7.3.1	 Moderate IH Augments Respiratory Instability During Sleep and “Basal” Arterial Blood Pressure
		7.3.2	 IH Increases the Chemoreflex Response
		7.3.3	 IH Induces Weight and Fat Loss
		7.3.4	 IH Reduces ACTH and Testosterone Levels and Promotes Inflammation
	7.4	 Discussion
		7.4.1	 Efficiency of the Intermittent Hypoxia Protocol
		7.4.2	 Intermittent Hypoxia Reduces ACTH and Favors Weight Loss
		7.4.3	 Intermittent Hypoxia and Leptin
	7.5	 Conclusions
	References
8: Effects of Gestational Intermittent Hypoxia on Placental Morphology and Fetal Development in a Murine Model of Sleep Apnea
	8.1	 Introduction
	8.2	 Methods
		8.2.1	 Animal Models and Anesthesia
		8.2.2	 Macroscopic and Microscopic Study of Placentas
		8.2.3	 Statistical Analysis
	8.3	 Results
		8.3.1	 Maternal, Placenta and Fetus Body Weight
		8.3.2	 Macroscopic and Microscopic Study of Placentas
	8.4	 Discussion
	References
9: Ventilatory Effects of Acute Intermittent Hypoxia in Conscious Dystrophic Mice
	9.1	 Introduction
	9.2	 Materials and Methods
		9.2.1	 Ethical Approval
		9.2.2	 Experimental Animals
		9.2.3	 Whole-Body Plethysmography
		9.2.4	 Data and Statistical Analysis
	9.3	 Results
		9.3.1	 Effect of AIH on Ventilation
		9.3.2	 Effect of AIH on Metabolism
		9.3.3	 Effect of AIH on the Ventilatory Equivalent
	9.4	 Discussion
	References
10: Intermittent Hypoxia and Diet-Induced Obesity on the Intestinal Wall Morphology in a Murine Model of Sleep Apnea
	10.1	 Introduction
	10.2	 Methods
		10.2.1	 Animal Protocols
		10.2.2	 Tissue Collection
		10.2.3	 Statistical Analysis
	10.3	 Results
		10.3.1	 Body Weight Gain and Visceral Fat Deposits
		10.3.2	 Basal Glycemia and Markers of Sympathetic and Inflammatory Activity
		10.3.3	 Morphology of Jejunum Wall
	10.4	 Discussion
	References
11: Enhanced Peripheral Chemoreflex Drive Is Associated with Cardiorespiratory Disorders in Mice with Coronary Heart Disease
	11.1	 Introduction
	11.2	 Methodology
		11.2.1	 Animal Model
		11.2.2	 Resting Breathing and Chemoreflex Function
		11.2.3	 Electrocardiogram and Autonomic Balance
		11.2.4	 Data Analysis
	11.3	 Results
		11.3.1	 SR-B1−/−/HypoApoE Mice Display Increased Peripheral Chemoreflex Drive
		11.3.2	 SR-B1−/−/HypoApoE Mice Show Breathing Pattern Irregularity
		11.3.3	 Cardiac Sympathetic Tone Is Enhanced in SR-B1−/−/HypoApoE Mice
	11.4	 Discussion
	References
12: Role of Peripheral Chemoreceptors on Enhanced Central Chemoreflex Drive in Nonischemic Heart Failure
	12.1	 Introduction
	12.2	 Methodology
		12.2.1	 Animals
		12.2.2	 Heart Failure Model
		12.2.3	 Echocardiography
		12.2.4	 Plethysmography
		12.2.5	 Carotid Body Denervation
		12.2.6	 Statistical Analysis
	12.3	 Results
		12.3.1	 Cardiac Morphology and Carotid Body Denervation in Heart Failure
		12.3.2	 Carotid Body Resection Restores Normal Hypercapnic Ventilatory Responses and Breathing Disorders in CHF Rats
	12.4	 Discussion
	References
13: Effect of Carotid Body Denervation on Systemic Endothelial Function in a Diabetic Animal Model
	13.1	 Introduction
	13.2	 Methods
		13.2.1	 Animals
		13.2.2	 Evaluation of Endothelial Function
		13.2.3	 Nitric Oxide Quantification in Plasma and Aorta
		13.2.4	 Western Blot Analyses of eNOS, Inos, and PGF2αR Protein Levels in Aorta Artery
		13.2.5	 Statistical Analysis
	13.3	 Results
		13.3.1	 Effect of HFHSu Diet and CSN Resection on In Vivo Metabolic Parameters
		13.3.2	 Effect of HFHSu Diet and of CSN Resection on Vasoconstrictor Responses and Endothelial Function in Aorta Artery
		13.3.3	 Effect of HFHSu Diet and of CSN Resection on NO Levels in Plasma and Aorta Artery
		13.3.4	 Effect of HFHSu Diet and of CSN Resection on eNOS, iNOS, and PGF2αR Levels in Aorta Artery
	13.4	 Discussion
	References
14: Contribution of Carotid Bodies on Pulmonary Function During Normoxia and Acute Hypoxia
	14.1	 Introduction
	14.2	 Methods
		14.2.1	 Ethical Considerations and Animals
		14.2.2	 Carotid Body Denervation
		14.2.3	 Noninvasive Measurement of Pulmonary Function
		14.2.4	 Invasive Measurement of Pulmonary Function
		14.2.5	 Lung Histology
		14.2.6	 Statistical Analysis
	14.3	 Results
		14.3.1	 Carotid Body Ablation and Resting Ventilatory Parameters
		14.3.2	 Carotid Body Ablation Blunted the Hypoxic Ventilatory Response in Mice
		14.3.3	 Lung Mechanics in Normoxia and the Effect of Carotid Body Denervation
		14.3.4	 Hypoxia and Lung Mechanics Following Carotid Body Denervation in Mice
		14.3.5	 Alveolar Morphology and Carotid Body Denervation
	14.4	 Discussion
	References
15: Increased Abdominal Perimeter Differently Affects Respiratory Function in Men and Women
	15.1	 Introduction
	15.2	 Methods
		15.2.1	 Ethical Approval
		15.2.2	 Subjects and Study Design
		15.2.3	 Statistical Analysis
	15.3	 Results
		15.3.1	 Demographic and Clinical Information of the Participants
		15.3.2	 Effect of Overweight and Obesity on Basal Ventilation
		15.3.3	 Effect of Increased Abdominal Circumference on Basal Ventilation
	15.4	 Discussion
	References
16: Carotid Body Resection Prevents Short-Term Spatial Memory Decline in Prediabetic Rats Without Changing Insulin Signaling in the Hippocampus and Prefrontal Cortex
	16.1	 Introduction
	16.2	 Methods
		16.2.1	 Animals
		16.2.2	 Insulin Tolerance Test (ITT) and Glucose Tolerance Test (OGTT)
		16.2.3	 Whole-Body Plethysmography Recordings of Ventilation
		16.2.4	 Y- Maze Test
		16.2.5	 Protein Analysis (Western Blot)
		16.2.6	 Statistical Analysis
	16.3	 Results
		16.3.1	 Impact of HFHSu Diet and CSN Resection on Glycaemia, Insulin Sensitivity, and Glucose Tolerance
		16.3.2	 Effect of HFHSu Diet and CSN Resection on the Responses to Hypoxia and Hypercapnia
		16.3.3	 Effect of HFHSu Diet Consumption and CSN Resection on Short-Term Spatial Memory
		16.3.4	 Impact of HFHSu Diet and CSN Resection on Insulin Signaling-Related Proteins in the Hippocampus and Prefrontal Cortex
	16.4	 Discussion
	References
17: Constitutive Expression of Hif2α Confers Acute O2 Sensitivity to Carotid Body Glomus Cells
	17.1	 Introduction
	17.2	 HIF2α-Dependent Gene Expression Profile in Carotid Body Cells
	17.3	 Selective Inhibition of Acute Responsiveness to Hypoxia in Hif2α-Deficient Glomus Cells
	17.4	 Conclusions and Perspectives
	References
18: Of Mice and Men and Plethysmography Systems: Does LKB1 Determine the Set Point of Carotid Body Chemosensitivity and the Hypoxic Ventilatory Response?
	18.1	 Introduction
	18.2	 Results and Discussion
		18.2.1	 LKB1 Determines a Set Point for Carotid Body Chemosensitivity
		18.2.2	 LKB1 and the AMPK-Related Kinases: From Synaptic Transmission to Gene Expression Regulation
		18.2.3	 LKB1, AMPK, and the Hypoxic Ventilatory Response
	18.3	 Conclusion
	References
19: Analyzing Angiotensin II Receptor Type 1 Clustering in PC12 Cells in Response to Hypoxia Using Direct Stochastic Optical Reconstruction Microscopy (dSTORM)
	19.1	 Introduction
	19.2	 Methods
		19.2.1	 PC12 Cell Culture, Hypoxic Protocol, and Immunocytochemistry
		19.2.2	 Direct Stochastic Optical Reconstruction Microscopy (dSTORM) Imaging and Cluster Analysis
		19.2.3	 Statistical Analysis
	19.3	 Results
		19.3.1	 AT1Rs Are Clustered on the Cell Membrane of PC12 Cells with Measurable Characteristics
		19.3.2	 Maximum AT1R Cluster Area Is Increased by Hypoxia
		19.3.3	 Maximum AT1R Cluster Area Is Increased by Hypoxia
	19.4	 Discussion
	References
20: The Carotid Body “Tripartite Synapse”: Role of Gliotransmission
	20.1	 Introduction
	20.2	 Methods
		20.2.1	 Cell Culture
		20.2.2	 Fura-2 Ratiometric Ca2+ Imaging
		20.2.3	 Solutions and Drugs
	20.3	 Results
		20.3.1	 Selective Chemoexcitants for Carotid Body Type I Versus Type II Cells
		20.3.2	 Crosstalk from Type I to Type II Cells During Chemotransduction: Paracrine Roles for ATP and Angiotensin II
		20.3.3	 Inhibitory Roles of Dopamine and Histamine in Type I to Type II Cross Talk
		20.3.4	 Evidence for ATP as a Type II Cell “Gliotransmitter”
	20.4	 Discussion
	References
21: Carotid Body-Mediated Chemoreflex Function in Aging and the Role of Receptor-Interacting Protein Kinase
	21.1	 Introduction
	21.2	 Methodology
		21.2.1	 Animals
		21.2.2	 Breathing and Chemoreflex Function
		21.2.3	 Statistical Analysis
	21.3	 Results
		21.3.1	 Resting Ventilatory Physiological Parameters
		21.3.2	 Loss of RIPK3 Signaling Improves Peripheral and Central Chemoreflex Function in Aged Animals
		21.3.3	 Absence of RIPK3 Decreased the Incidence of Breathing Disorders in Aged Mice
	21.4	 Discussion
	References
22: Chronic Metformin Administration Does Not Alter Carotid Sinus Nerve Activity in Control Rats
	22.1	 Introduction
	22.2	 Methods
		22.2.1	 Ethical Approval
		22.2.2	 Animal Procedures
		22.2.3	 CSN Electrophysiological Recordings
		22.2.4	 Statistical Analysis
	22.3	 Results
	22.4	 Discussion
	References
Concluding Remarks