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Kinetic analysis of recombinant mammalian α1 and α1β glycine receptor channels

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Abstract

To analyze the influence of the β-subunit on the kinetic properties of GlyR channel currents, α1-subunits and α1β-subunits were transiently expressed in HEK 293 cells. A piezo dimorph was used for fast application of glycine to outside-out patches. The rise time of activation was dose dependent for both receptors and decreased with increasing glycine concentrations. Subunit composition had no effect on the time course of activation. Coexpression of α1- and β-subunits resulted in a significantly lower EC50 and a reduced slope of the dose–response curve of glycine compared with expression of α1-subunits alone. For both receptor subtypes, the time course of desensitization was concentration dependent. Desensitization was best fitted with a single time constant at 10–30 µM, with two at 0.1 mM, and at saturating concentrations (0.3–3 mM) with three time constants. Desensitization of homomeric α1-receptor channels was significantly slower than that of α1β-receptor channels. The time course of current decay after the end of glycine pulses was tested at different pulse durations of 1 mM glycine. It was best fitted with two time constants for both α1 and α1β GlyR channels, and increased significantly with increasing pulse duration.

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Acknowledgements

We thank Prof. H. Betz, Frankfurt, for the kind gift of cDNAs of glycine receptors, U. Jensen for expert technical assistance, and A. Niesel and J. Kilian for technical support. This study was supported by grants from the Deutsche Forschungsgemeinschaft and the Medizinische Hochschule Hannover.

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Authors and Affiliations

  1. Department of Neurology, Medical SchoolUniversity Hannover, Carl-Neuberg-Str. 1, 30625, Hannover, Germany

    B. Mohammadi, K. Krampfl, C. Cetinkaya, H. Moschref, J. Grosskreutz, R. Dengler & J. Bufler

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  1. B. Mohammadi
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  2. K. Krampfl
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  3. C. Cetinkaya
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  4. H. Moschref
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  5. J. Grosskreutz
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  6. R. Dengler
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  7. J. Bufler
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Correspondence to J. Bufler.

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Mohammadi, B., Krampfl, K., Cetinkaya, C. et al. Kinetic analysis of recombinant mammalian α1 and α1β glycine receptor channels. Eur Biophys J 32, 529–536 (2003). https://doi.org/10.1007/s00249-003-0286-y

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  • DOI: https://doi.org/10.1007/s00249-003-0286-y

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