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Boyan, George; Liu, Yu (2014): Dye Coupling and Immunostaining of Astrocyte-Like Glia Following Intracellular Injection of Fluorochromes in Brain Slices of the Grasshopper, Schistocerca gregaria. In: Sprecher, Simon G. (ed.) : Brain development ; methods and protocols. Methods in molecular biology, Vol. 1082. New York: Humana Press. pp. 99-113
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Injection of fluorochromes such as Alexa Fluor® 568 into single cells in brain slices reveals a network of dye-coupled cells to be associated with the central complex. Subsequent immunolabeling shows these cells to be repo positive/glutamine synthetase positive/horseradish peroxidase negative, thus identifying them as astrocyte-like glia. Dye coupling fails in the presence of n-heptanol indicating that dye spreads from cell to cell via gap junctions. A cellular network of dye-coupled, astrocyte-like, glia surrounds and infiltrates developing central complex neuropils. Intracellular dye injection techniques complement current molecular approaches in analyzing the functional properties of such networks.