The experiments in this research grant proposal will identify the cellular and synaptic effects of endogenous cannabinoid (eCB) ligands and of glucocorticoid-induced release of eCBs in the dorsal motor nucleus of the vagus (DMV). Neurons in the DMV regulate parasympathetic output to most of the subdiaphragmatic viscera and therefore critically control feeding, digestion, glucose and insulin secretion, and other metabolic functions. Their activity is largely controlled by synaptic input to the DMV, which is modulated by locally released chemicals and circulating hormones. Regulation of DMV neurons by cannabinoids, vanilloids, and glucocorticoids has been suggested;when applied centrally these compounds profoundly alter parasympathetic function. Several eCB ligands, which are thought to be released from cell membranes in a retrograde fashion, activate both cannabinoid type 1 receptors (CB1R) and transient receptor potential vanilloid type 1 (TRPV1). In the DMV, activation of TRPV1 enhances neurotransmitter release, whereas CB1R tends to inhibit synapses. Both effects occur by activation of receptors on presynaptic terminals. Preliminary evidence suggests that eCB ligands are released from DMV neurons, and that glucocorticoids or depolarization can induce this release. Thus, eCB activity in the DMV may modulate both TRPV1 and CB1R activity. Neither the type(s) of eCB ligands released, the effects of most eCB ligands on synaptic activity, nor the trigger or mechanism of eCB release in the DMV are known. We will use whole-cell patch-clamp recordings from DMV neurons in brainstem slices to identify effects of eCB ligands on cellular activity in the DMV, and will also identify the compounds released by cells in the area using pharmacological and biochemical methods. The experiments will be guided by three specific aims: 1) Differentiate effects of eCB ligands on CB1R and TRPV1 in the DMV;2) Determine the eCB involvement in mediating rapid effects of glucocorticoids on local circuitry in the DMV;and 3) Identify the cellular pathway of the glucocorticoid effect. We will test the hypotheses that eCBs alter TRPV1 and CBR1 activity in the DMV in specific and predictable spatial, temporal, functionally relevant, and activity-dependent patterns, and that glucocorticoids induce eCB release from DMV cells by acting at membrane-bound G protein-coupled receptors on DMV neurons. Drugs based on the eCB system are being investigated for therapeutic use in a variety of nervous system pathologies, including disorders related to feeding, digestion, and obesity. Glucocorticoids, which release eCBs in some systems, are widely prescribed, and are also released by stressful stimuli. Results of these studies will be critical to predicting and understanding how these compounds interact with each other and affect parasympathetic function. Possible translational benefits also exist because of the benefit in controlling eCB levels in the vagal system of patients with elevated glucocorticoids.
Endogenous cannabinoid compounds have particularly complex actions in the area of the brainstem most responsible for maintaining functions of organ systems, and a number of drugs therapies based on altering these compounds in the brain are being developed without knowledge about their effects in this critical brain area. The experiments in this research grant proposal will identify the characteristics of these drugs, how they are released by glucocorticoid hormones, and what actions they have on the neurons. Results of these studies will be critical for predicting and understanding how these compounds interact to affect digestive functions, and they will also be applicable to multiple other neuronal systems.
|Derbenev, Andrei V; Smith, Bret N (2013) Dexamethasone rapidly increases GABA release in the dorsal motor nucleus of the vagus via retrograde messenger-mediated enhancement of TRPV1 activity. PLoS One 8:e70505|
|Zsombok, Andrea; Bhaskaran, Muthu D; Gao, Hong et al. (2011) Functional plasticity of central TRPV1 receptors in brainstem dorsal vagal complex circuits of streptozotocin-treated hyperglycemic mice. J Neurosci 31:14024-31|
|Stinchcomb, Audra L (2010) The role of entrepreneurial activities in academic pharmaceutical science research. J Pharm Sci 99:2532-7|