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Imaging synaptic inhibition in transgenic mice expressing the chloride indicator, Clomeleon

  • Brain Cell Technology
  • Published:
Brain Cell Biology

Abstract

We describe here a molecular genetic approach for imaging synaptic inhibition. The thy-1 promoter was used to express high levels of Clomeleon, a ratiometric fluorescent indicator for chloride ions, in discrete populations of neurons in the brains of transgenic mice. Clomeleon was functional after chronic expression and provided non-invasive readouts of intracellular chloride concentration ([Cl−]i) in brain slices, allowing us to quantify age-dependent declines in resting [Cl−]i during neuronal development. Activation of hippocampal interneurons caused [Cl−]i to rise transiently in individual postsynaptic pyramidal neurons. [Cl−]i increased in direct proportion to the amount of inhibitory transmission, with peak changes as large as 4 mM. Integrating responses over populations of pyramidal neurons allowed sensitive detection of synaptic inhibition. Thus, Clomeleon imaging permits non-invasive, spatiotemporally resolved recordings of [Cl−]i in a large variety of neurons, opening up new opportunities for imaging synaptic inhibition and other forms of chloride signaling.

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Acknowledgments

We thank R. L. Dunbar for participating in initial experiments; W. Drake, E. Ellison, and T. Schweizer for technical assistance; W. Denk for providing access to a spinning-disk confocal microscope; G. Giese for help with confocal microscopy; B. Sakmann and P. Seeburg for generous support; F. Galeffi, K. Tanaka, and R. Yasuda for their comments on this manuscript; and A. Frick for advice on preparation of hippocampal slices. K.B. was a Research Fellow of the Japan Society for the Promotion of Science. This work was partially supported by DFG grant KU1983/1-2 (FOR 577) awarded to T.K., NIH Grants awarded to G.J.A. and G.F., as well as an Alfred P. Sloan Research Fellowship, a Klingenstein Fellowship Award in the Neuroscience, and a Beckman Young Investigator Award to G.F.

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Authors and Affiliations

  1. Department of Neurobiology, Duke Univ. Medical Center, Box 3209, Durham, NC, 27710, USA

    Ken Berglund, Li Shen Loo, Dongqing Wang, Nell B. Cant, Guoping Feng & George J. Augustine

  2. Department of Cell Physiology, Max Planck Institute for Medical Research, Jahnstrasse 29, 69120, Heidelberg, Germany

    Wolfram Schleich, Patrik Krieger & Thomas Kuner

  3. Institute of Anatomy and Cell Biology, University of Heidelberg, Im Neuenheimer Feld 307, 69120, Heidelberg, Germany

    Thomas Kuner

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  1. Ken Berglund
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  2. Wolfram Schleich
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  4. Li Shen Loo
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Correspondence to George J. Augustine or Thomas Kuner.

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Berglund, K., Schleich, W., Krieger, P. et al. Imaging synaptic inhibition in transgenic mice expressing the chloride indicator, Clomeleon . Brain Cell Bio 35, 207–228 (2006). https://doi.org/10.1007/s11068-008-9019-6

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  • DOI: https://doi.org/10.1007/s11068-008-9019-6

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