This proposal is an innovative effort to develop a novel tool that could revolutionize the execution and application of in vivo sharp electrode intracellular electrophysiology, the fundamental approach for examining cellular and synaptic mechanisms of systems-level functions. This tool is a reporter gene that will be detectible by electrophysiological methods in real-time by sharp electrodes in vivo, without the need for secondary factors (i.e., light stimulus or exogenous ligands). This could provide a necessary link to enable a more thorough incorporation of classic-type systems physiology with the reduced preparations and lower species that make up the majority of current experimental approaches used to understand genetic manipulations. An ideal electrophysiological reporter is one that is genetically-encoded, is "visible" to electrophysiological methods, has a signal that is easily recognized and distinguished from native processes in real time, and does not interfere with normal physiological function. We have identified a lab-generated mutant ion channel that fits these criteria, and appears to be an excellent candidate. We will make use of new viral vector technology to deliver candidate gene to adult rats.
Aim 1 will use both in vivo and in vitro single-cell electrophysiological approaches to determine if the signal from the reporter gene can be detected against the well characterized background of the model systems (dorsal root ganglion sensory neurons and spinal motoneurons).
Aim 2 is hypothesis driven and will determine if constitutive signaling by trkC, the receptor for neurotrophin-3, plays a necessary role in maintenance of the cellular and synaptic properties of adult motoneurons. The issue of maintenance of cellular properties by target-derived factors is in a question of fundamental importance in terms of the principles of neuroscience/neurobiology, and also in terms of biomedical issues in the adult nervous system such as aging, neurodegeneration, traumatic injury, and learning and memory.
This project is aimed at developing a new tool for research - a gene that will report the status of genetic manipulations to electrophysiological probes. If successful, it will greatly facilitate the execution of, and interpretation of data from, systems-level in vivo neurophysiology, an experimental approach that is particularly relevant to functional studies of the role of specific genes/cell-types in systems-level processes such as aging, locomotion, sensori-motor integration, models of psychiatric disorders and addiction/substance abuse, and nervous system disease/injury/insult and the responses to treatments. This project also Aims to determine the effect of altering the expression of trkC (the receptor for neurotrophin-3) on the cellular and synaptic electrophysiological properties of a subpopulation of neurons, which is relevant to conditions or disease states that may include reduced neurotrophin-receptor signaling in the pathophysiology, such as aging, diabetic neuropathy, traumatic nervous system injury, and neurodegeneration.
|Harrison, Benjamin J; Venkat, Gayathri; Lamb, James L et al. (2016) The Adaptor Protein CD2AP Is a Coordinator of Neurotrophin Signaling-Mediated Axon Arbor Plasticity. J Neurosci 36:4259-75|
|Harrison, Benjamin J; Venkat, Gayathri; Hutson, Thomas et al. (2015) Transcriptional changes in sensory ganglia associated with primary afferent axon collateral sprouting in spared dermatome model. Genom Data 6:249-52|
|Hougland, M Tyler; Harrison, Benjamin J; Magnuson, David S K et al. (2012) The Transcriptional Response of Neurotrophins and Their Tyrosine Kinase Receptors in Lumbar Sensorimotor Circuits to Spinal Cord Contusion is Affected by Injury Severity and Survival Time. Front Physiol 3:478|