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The Journal of Neuroscience, November 4, 2009, 29(44):13751-13760; doi:10.1523/JNEUROSCI.2985-09.2009

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Behavioral/Systems/Cognitive
Functional Clustering of Neurons in Motor Cortex Determined by Cellular Resolution Imaging in Awake Behaving Mice

Daniel A. Dombeck,^1,3 Michael S. Graziano,^2,3 and David W. Tank^1,3

Departments of ¹Molecular Biology and ²Psychology, and ³Princeton Neuroscience Institute, Princeton University, Princeton, New Jersey 08544

Correspondence should be addressed to either Daniel A. Dombeck or David W. Tank, 270C Carl Icahn Laboratory, Washington Road, Princeton University, Princeton, NJ 08544. Email: ddombeck{at}princeton.edu or Email: dwtank{at}princeton.edu

Macroscopic (millimeter scale) functional clustering is a hallmark characteristic of motor cortex spatial organization in awake behaving mammals; however, almost no information is known about the functional micro-organization (100 µm scale). Here, we optically recorded intracellular calcium transients of layer 2/3 neurons with cellular resolution over 200-µm-diameter fields in the forelimb motor cortex of mobile, head-restrained mice during two distinct movements (running and grooming). We showed that the temporal correlation between neurons was statistically larger the closer the neurons were to each other. We further explored this correlation by using two separate methods to spatially segment the neurons within each imaging field: K-means clustering and correlations between single neuron activity and mouse movements. The two methods segmented the neurons similarly and led to the conclusion that the origin of the inverse relationship between correlation and distance seen statistically was twofold: clusters of highly temporally correlated neurons were often spatially distinct from one another, and (even when the clusters were spatially intermingled) within the clusters, the more correlated the neurons were to each other, the shorter the distance between them. Our results represent a direct observation of functional clustering within the microcircuitry of the awake mouse motor cortex.

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Received June 22, 2009; revised Aug. 24, 2009; accepted Sept. 15, 2009.

Correspondence should be addressed to either Daniel A. Dombeck or David W. Tank, 270C Carl Icahn Laboratory, Washington Road, Princeton University, Princeton, NJ 08544. Email: ddombeck{at}princeton.edu or Email: dwtank{at}princeton.edu

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