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J Neurophysiol (February 15, 2007). doi:10.1152/jn.01207.2006
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Submitted on November 15, 2006
Accepted on February 6, 2007

Two-photon calcium imaging of network activity in XFP expressing neurons in the mouse

Jennifer M Wilson1, Daniel A Dombeck2, Manuel Diaz Rios3, Ronald M. Harris-Warrick4, and Robert M. Brownstone5*

1 Anatomy & Neurobiology, Dalhousie University, Halifax, Canada
2 Dept of Neurobiology & Behavior, Cornell University, Ithaca, New York, United States; Molecular Biology Department, Princeton University, princeton, New Jersey, United States
3 Dept of Neurobiology & Behavior, Cornell University, Ithaca, New York, United States; Anatomy/Institute of Neurobiology, University of Puerto Rico, San Juan, Puerto Rico
4 Dept of Neurobiology & Behavior, Cornell University, Ithaca, New York, United States
5 Surgery (Neurosurgery), Dalhousie University, Halifax, Canada; Anatomy & Neurobiology, Dalhousie University, Halifax, Canada

* To whom correspondence should be addressed. E-mail: rob.brownstone{at}dal.ca.

Fluorescent protein (XFP) expression in postnatal neurons allows the anatomical and physiological investigation of identified subpopulations of interneurons with established techniques. However, the spatiotemporal pattern of activity of these XFP neurons within a network and their role in the functional output of the network are more challenging issues to investigate. Here we apply two-photon excitation laser scanning microscopy to mouse spinal cord locomotor networks, and present the methodology by which calcium activity can be recorded in XFP expressing neurons. Such activity can be studied both in relation to neighbouring non-XFP neurons in a spinal cord slice preparation, and in relation to functional locomotor output monitored by ventral root activity in the intact in vitro spinal cord. Thus, the network properties and functional correlates with locomotion of identified populations of interneurons can be studied simultaneously.




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