Atrial fibrillation (AF) is the most common supraventricular tachyarrhythmia that can present without cardiovascular disease (lone AF). Frequent high-intensity endurance exercise is a risk factor for lone AF, and the pathophysiology of AF induced by intense endurance exercise is unknown. We found that after 6 weeks of intense swimming and running, mice were far more susceptible to AF, but not ventricular arrhythmias. Exercise induced atrial fibrosis, inflammation and slowed conduction without detectible changes in ventricles. Since AF is associated with stretch and since a tumor necrosis factor-α (TNFα) is a mechanosensitive inflammatory factor, mice were treated with the TNFα inhibitor etanercept. Etanercept treatment blocked inflammation, fibrosis, and AF vulnerability in the exercised mice. Consistent with these findings, we found that exercise caused large elevations in atrial pressures. Our findings support the conclusion that mechanical loading of atria during exercise induces TNFα release, leading to structural remodeling and enhanced AF vulnerability.

In the mature dentate gyrus (DG) new neurons are continuously produced in a process known as adult neurogenesis. These neurons are thought to contribute to learning and memory processes, but their precise function is still not fully understood. The rate of neuronal production and the development of young neurons are affected by stimuli such as physical exercise and learning events. The neurotransmitter γ-aminobutyric acid (GABA) has recently emerged as a possible key mediator of this activity-dependent regulation. My study examined the role of a subtype of GABAA receptors that contains the δ subunit (δGABA(A)R) in the regulation of adult hippocampal neurogenesis. Mice lacking the δ subunit (Gabrd−/− or δ-null mice) displayed decreased maturation, migration and dendritic complexity of adult-born neurons. Conversely, following treatment with a selective δGABA(A)R agonist, neuronal maturation was promoted in wild-type, but not δ-null mice. These results indicate a key role for δGABA(A)R in activity-dependent regulation of adult neurogenesis. When administered to rats, δGABA(A)R agonists promoted neuronal survival as well as maturation. The effect on maturation was blocked by the N-Methyl-D-aspartate (NMDA) blocker AP-5, suggesting that some aspects of δGABA(A)R-mediated regulation require the activation of NMDA receptors. To further understand the contribution of adult born neurons to memory function, neurogenesis in rats was alternatively suppressed using ionizing radiation, or enhanced by allowing the rats to engage in running. The rats were then trained and tested in a behavioral paradigm designed to assess their susceptibility to memory interference, a phenomenon which occurs when similar memories are not sufficiently distinguished from one another. Irradiated rats exhibited increased susceptibility to memory interference, indicating an important role for adult-born neurons in the encoding and/or retrieval of distinct memories. Remarkably, irradiated rats that engaged in running exhibited increased neuronal growth and a complete reversal of the memory impairment,emphasizing the beneficial effects of physical exercise on cognitive functions. Taken together, the findings reported in this dissertation offer novel information about the process of adult neurogenesis and its physiological significance.

Glucagon-like peptide-1 (GLP-1) is an incretin hormone produced by the enteroendocrine L-cell that potently stimulates insulin secretion. Although signaling pathways promoting GLP-1 secretion are well characterized, the mechanism by which GLP-1 containing granules fuse to the L-cell membrane remain elusive. RT-PCR and protein analysis indicate that vesicle-associated membrane protein 2 (VAMP2) is expressed and localized to secretory granules in the murine GLUTag L-cell model. VAMP2, but not VAMP1, interacted with the core SNARE complex protein, Syntaxin 1a, in GLUTag cells. Tetanus toxin (TetX) cleavage of VAMP2 in GLUTag cells prevented glucose-dependent insulinotropic peptide (GIP)- and oleic acid (OA)-stimulated GLP-1 secretion, as well as K+-stimulated exocytosis from GLUTag cells. Although components of membrane rafts were detected in GLUTag cells, their role in GLP-1 secretion remains to be determined. Together, these findings indicate an essential role for VAMP2 in GLP-1 exocytosis from the GLUTag cell.

Glucagon-like peptide-1 (GLP-1) is an incretin hormone produced by the enteroendocrine L-cell that potently stimulates insulin secretion. Although signaling pathways promoting GLP-1 secretion are well characterized, the mechanism by which GLP-1 containing granules fuse to the L-cell membrane remain elusive. RT-PCR and protein analysis indicate that vesicle-associated membrane protein 2 (VAMP2) is expressed and localized to secretory granules in the murine GLUTag L-cell model. VAMP2, but not VAMP1, interacted with the core SNARE complex protein, Syntaxin 1a, in GLUTag cells. Tetanus toxin (TetX) cleavage of VAMP2 in GLUTag cells prevented glucose-dependent insulinotropic peptide (GIP)- and oleic acid (OA)-stimulated GLP-1 secretion, as well as K+-stimulated exocytosis from GLUTag cells. Although components of membrane rafts were detected in GLUTag cells, their role in GLP-1 secretion remains to be determined. Together, these findings indicate an essential role for VAMP2 in GLP-1 exocytosis from the GLUTag cell.

In the mature dentate gyrus (DG) new neurons are continuously produced in a process known as adult neurogenesis. These neurons are thought to contribute to learning and memory processes, but their precise function is still not fully understood. The rate of neuronal production and the development of young neurons are affected by stimuli such as physical exercise and learning events. The neurotransmitter γ-aminobutyric acid (GABA) has recently emerged as a possible key mediator of this activity-dependent regulation. My study examined the role of a subtype of GABAA receptors that contains the δ subunit (δGABA(A)R) in the regulation of adult hippocampal neurogenesis. Mice lacking the δ subunit (Gabrd−/− or δ-null mice) displayed decreased maturation, migration and dendritic complexity of adult-born neurons. Conversely, following treatment with a selective δGABA(A)R agonist, neuronal maturation was promoted in wild-type, but not δ-null mice. These results indicate a key role for δGABA(A)R in activity-dependent regulation of adult neurogenesis. When administered to rats, δGABA(A)R agonists promoted neuronal survival as well as maturation. The effect on maturation was blocked by the N-Methyl-D-aspartate (NMDA) blocker AP-5, suggesting that some aspects of δGABA(A)R-mediated regulation require the activation of NMDA receptors. To further understand the contribution of adult born neurons to memory function, neurogenesis in rats was alternatively suppressed using ionizing radiation, or enhanced by allowing the rats to engage in running. The rats were then trained and tested in a behavioral paradigm designed to assess their susceptibility to memory interference, a phenomenon which occurs when similar memories are not sufficiently distinguished from one another. Irradiated rats exhibited increased susceptibility to memory interference, indicating an important role for adult-born neurons in the encoding and/or retrieval of distinct memories. Remarkably, irradiated rats that engaged in running exhibited increased neuronal growth and a complete reversal of the memory impairment,emphasizing the beneficial effects of physical exercise on cognitive functions. Taken together, the findings reported in this dissertation offer novel information about the process of adult neurogenesis and its physiological significance.

Severe hypoglycemia poses the greatest challenge to glycemic control in diabetic patients, especially children with type 1 diabetes mellitus. Although in vivo animal models exist for investigating the effects of hypoglycemia, few studies examine repeated hypoglycemia and none investigate within the context of a juvenile animal model. The main objective of this thesis was to examine electroencephalographic (EEG) and behavioral abnormalities manifesting as a result of repeated hypoglycemia in juvenile diabetic and non-diabetic mouse models. Using a novel implantation technique, the hippocampal and cortical EEG were recorded during repeated insulin-induced hypoglycemia. It was discovered that repeated hypoglycemia exacerbated behavioral convulsion severity and promoted epileptiform EEG activity within the hippocampus and cortex of both diabetic and non-diabetic animals. Furthermore, sustained hypoglycemia caused a significant decrease in hippocampal EEG activity in diabetic animals compared with non-diabetics. These results suggest recurrent hypoglycemia may promote and worsen seizures associated with hypoglycemia in diabetic children.

The simultaneous application of interlaced innocuous warm and cool stimuli (a thermal grill stimulus, TGS) can elicit sensations of burning heat (the Thermal Grill Illusion, TGI). The TGS is thought to alter the central interactions between somatosensory sub-modalities (i.e. cold-inhibition of pain). Previous psychophysical findings point to body site differences in perceptual thermal thresholds. The primary objective of this study was to evaluate whether, using the same TGS, a TGI can be elicited at body sites other than the upper extremity. The present findings indicate that the TGI can be induced at the palm, back, calf, and foot. Pain and unpleasantness in response to the TGS were more frequent and intense following stimulation of the palm and back than the calf and foot. Lower cold pain thresholds were associated with lower pain intensity ratings in response to the TGS. These two findings may reflect differences in central integrative processes.

The mechanisms underlying the transition to seizure are still unresolved. Proposed mechanisms include excitatory GABAergic drive, loss of interneuron-mediated inhibition, and glutamatergic input potentiation. The objective of this thesis was to investigate the relative contributions of synchronized glutamatergic and GABAergic inputs and their functional roles during ictogenesis in the epileptic neonatal (postnatal days 6-12) mouse hippocampus, induced with 0.25mM Mg2+/5mM K+ perfusion. Simultaneous field and whole-cell patch-clamp recordings were obtained from CA3 stratum-oriens interneurons and pyramidal cells. The antagonists for GABAA and glutamate receptors abolished the preictal and ictal discharges, respectively, suggesting that the preictal state is mediated by the coherent discharges of GABAergic inhibitory interneurons, whereas the recurrent excitatory inputs are required for ictogenesis. Synaptic charge transfers underlying the synchronized discharges showed a dynamic change in the balance between the inputs: GABAergic currents markedly diminished by ictal onset whereas glutamatergic currents dominated at ictal onset and throughout the ictus.

Ventilation may limit exercise. Wearing a gas mask may further compromise ventilation. Continuous positive airway pressure (CPAP) improves ventilation by reducing airway resistance and thus the work of breathing. We investigated the effects of wearing a gas mask with and without CPAP on the work of breathing (WOB) during resting and exercise conditions to determine (a) whether wearing a gas mask increases the WOB and (b) whether the application of CPAP to a gas mask will mitigate (reduce) said increase to the WOB. Ten healthy males completed two test protocols with three stages each, and in three mask conditions. Physiological and dyspnea parameters were measured. Wearing a gas mask increased the metabolic cost and work of breathing. When the portable CPAP device was applied, there was no change in the calculated work of breathing, but metabolic cost of breathing was significantly reduced. CPAP also significantly reduced the sensation of dyspnea.

Infection with the gut-pathogen Helicobacter pylori is the single, most important risk factor in the development of gastric cancer. Although there is a rising incidence in mortality resulting from this malignancy, the exact mechanism underlying the initiation and progression of bacterial-induced gastric tumorigenesis is still not completely understood. Several studies implicate the activation of the Signal Transducer and Activator of Transcription 3 (STAT3) signaling pathway as a cellular trigger for promoting carcinogenes. In this thesis, I studied the role of the STAT3 signaling pathway in H. pylori mediated tumorigenesis, and attempted to delineate mechanisms involved. I have found that H. pylori activates the STAT3 signaling pathway both in vitro and in vivo, to promote carcinogenesis. Pivotal for H. pylori mediated STAT3 activation are the bacterial effector protein CagA and host receptor components, the gp130 and the IL-6αR subunits. Further investigation into the mechanism of STAT3 induction identified a key role for cholesterol-enriched membrane lipid rafts. Bacterial invasion and CagA injection into host cells was also dependent on lipid raft integrity. Co-fractionation via the use of sucrose gradients, which permits the isolation of lipid rafts, identified H. pylori CagA to be associated with these membrane microdomains. CagA, once injected into the cell, appears to interact with the inner leaflet of the host plasma membrane via a charge association that either directly or indirectly anchors it to the negatively charged anionic lipids in the cytoplasmic membrane. In addition, janus kinases were recruited to rafts upon H. pylori infection. In this thesis, I present a dynamic model of STAT3 activation, which requires the interaction of lipid raft associated proteins, H. pylori CagA and recruited JAKs with non-lipid raft receptor components to support STAT3 signaling. This study is significant since it provides insight into the possible mechanisms by which H. pylori induces gastric cancer and furthermore, it facilitates the development of novel therapeutic targets directed against bacterial induced carcinogenesis.