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Dye Coupling and Immunostaining of Astrocyte-Like Glia Following Intracellular Injection of Fluorochromes in Brain Slices of the Grasshopper, Schistocerca gregaria

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Brain Development

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1082))

Abstract

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.

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Acknowledgements

This work was supported by DFG grant BO 1434/3–5 and the Graduate School of Systemic Neuroscience, University of Munich. We thank S. Götz for assistance with dye injection and immunolabeling of glia (Fig. 2d–f).

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Authors and Affiliations

  1. Developmental Neurobiology Group, Biocenter, Ludwig-Maximilians-Universität, Martinsried, Germany

    George Boyan & Yu Liu

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  1. George Boyan
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  2. Yu Liu
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Editors and Affiliations

  1. Department of Biology, University of Fribourg, Fribourg, Switzerland

    Simon G Sprecher

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Boyan, G., Liu, Y. (2014). Dye Coupling and Immunostaining of Astrocyte-Like Glia Following Intracellular Injection of Fluorochromes in Brain Slices of the Grasshopper, Schistocerca gregaria . In: Sprecher, S. (eds) Brain Development. Methods in Molecular Biology, vol 1082. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-655-9_7

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  • DOI: https://doi.org/10.1007/978-1-62703-655-9_7

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-654-2

  • Online ISBN: 978-1-62703-655-9

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