NSF Award
1451350
This project is directed at developing a novel technology, robotic microscopy (RM), to "bridge multiple spatial, temporal, and organizational scales to provide fundamental insights into the emergent properties of neural circuitry that ultimately lead to behavior and cognition." There are two primary benefits of the proposed studies. First, the experiments will result in the development of an RM instrument that can monitor the molecular dynamics of individual neurons in slices of living brain tissue over weeks at a time. Second, the experiments will use RM to study for the first time how learning and memory change the fundamental properties of specific neurons in situ to enhance synaptic plasticity, providing new insights into the mechanisms involved in learning and memory. In the past, RM was used to study neurons in culture. The primary activity in the proposed studies will be to validate the utility of RM to monitor neuronal dynamics in brain slices that maintain their natural physiological connectivity and architecture. Other benefits will be access of the scientific community to a technology that can examine the biochemistry of neurons and other cell types longitudinally and the development of new instrumentation that can be used in academic courses that teach students, postdoctoral fellows and research scientists novel imaging approaches to study brain circuits.<br/><br/>The problem to be addressed is whether RM can be used to study neurons expressing the Arc gene in situ in hippocampal brain slices. Arc is important for long-term memory consolidation and synaptic plasticity, and its activity is greatly enhanced in specific neuron populations by stimuli that affect learning. The methods to be employed will involve the use of novel Arc genetic probes and transgenic mice to determine if RM can identify selective Arc-expressing neurons activated in brain slices by long-term potentiation (LTP) and long-term depression (LTD) or in brain in vivo during learning and memory consolidation. The goals of the studies will be to determine if different Arc neuronal circuits mediate LTP and LTD, and if the same Arc expressing neurons activated in vivo in the brain during learning can be monitored by RM in situ in brain slices. The scope of the studies will determine if RM can identify and study different Arc neuronal circuits activated in vitro and in vivo by different forms of learning and memory consolidation.
