Grants and Contracts Details
The University of Kentucky group led by the co-investigator, Dr. Bret N. Smith, will conduct experiments pertaining to the NIH R01DK082625. In these experiments, visceral organs of rats (e.g., stomach, kidney, heart) will be inoculated with psudorabies virus (PRV-152) to label neurons of the brainstem nucleus of the solitary tract (NTS) that project to those visceral organs. The labeled neurons will be identified in brainstem slices 5 days after the injection under a fluorescent microscope and electrophysiological recordings will be conducted to determine differences in the expression of nicotinic acetylcholine (nAChR) and the neuronal properties of labeled and unlabeled (control) NTS neurons. Therefore, the neuronal properties and the expression of nAChRs in labeled NTS neurons will be correlates with the neuronal projection targets. This method of injections has been developed and extensively used in the laboratory of Dr. Bret N. Smith (Glatzer et al., 2003) and will allow determining responsiveness to nicotine of identified gastric-related NTS neurons (the primary hypothesis of the proposal). Similarly, NTS neurons that are involved in neuronal regulation of sympathetic pathways can be identified by inoculation of kidney. The latter pathway involves intermediate infection of the caudal ventrolateral medulla (CVLM), but not dorsal motor nucleus of the vagus (DMV), which allows separating of gastric- from sympathetic-related NTS neurons via double labeling. Labeled NTS neurons will be selected using epifluorescence and their responsiveness to 0.2 mM nicotine will be tested using patch-clamp recordings. Dr. Smith will help Dr. Uteshev to establish this in vivo injection procedure.
|Effective start/end date||11/1/11 → 6/30/14|
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