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A tctex1-Ca2+ channel complex for selective surface expression of Ca2+ channels in neurons

Nature Neuroscience volume 8, pages 435–442 (2005)Cite this article

Abstract

Voltage-gated Ca2+ channels (VGCCs) are important in regulating a variety of cellular functions in neurons. It remains poorly understood how VGCCs with different functions are sorted within neurons. Here we show that the t-complex testis-expressed 1 (tctex1) protein, a light-chain subunit of the dynein motor complex, interacts directly and selectively with N- and P/Q-type Ca2+ channels, but not L-type Ca2+ channels. The interaction is insensitive to Ca2+. Overexpression in hippocampal neurons of a channel fragment containing the binding domain for tctex1 significantly decreases the surface expression of endogenous N- and P/Q-type Ca2+ channels but not L-type Ca2+ channels, as determined by immunostaining. Furthermore, disruption of the tctex1–Ca2+ channel interaction significantly reduces the Ca2+ current density in hippocampal neurons. These results underscore the importance of the specific tctex1-channel interaction in determining sorting and trafficking of neuronal Ca2+ channels with different functionalities.

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Figure 1: Selective interaction between tctex1 and N- and P/Q-type Ca2+ channels.
Figure 2: Effects of Ca2+ on the tctex1-channel interaction, and identification of the tctex1 binding domain on the channel C terminus.
Figure 3: Sensitized FRET and acceptor photobleaching measurements point to interaction between tctex1 and full-length N-type Ca2+ channels.
Figure 4: Formation of the tctex1–Ca2+ channel complex in vivo.
Figure 5: Overexpression of a dominant-negative construct, NCD4-GFP, reduces surface expression of endogenous N-type Ca2+ channels.
Figure 6: Quantitative summary of the effects of overexpression of a dominant-negative construct on surface expression of three different types of endogenous Ca2+ channels.
Figure 7: Disruption of the tctex1–Ca2+ channel interaction reduces the Ca2+ current density in hippocampal neurons.
Figure 8: Quantitative analysis of the effects of NCD4-GFP on the Ca2+ current density in hippocampal neurons.

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Acknowledgements

We thank R. Pittman for the use of a Nikon Eclipse TE2000 inverted microscope; K. Campbell and C.H. Sung for anti-tctex1 antibody; M. Maronski and M. Dichter for help with hippocampal neuron cultures; P. Baas, K. Pfister and J. Meinkoth for helpful discussions and comments; and J. Field for initial help with yeast two-hybrid screening. This work was supported by grants from the US National Institutes of Health (J.F.Z. & W.A.S.), the American Heart Association (J.F.Z.) and the National Alliance for Research on Schizophrenia and Depression (Y.M.H.).

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Author notes

  1. Meizan Lai, Fushun Wang, Yuan Chen & Ji-fang Zhang

    Present address: Department of Physiology, Jefferson Medical College, 1020 Locust Street, Philadelphia, Pennsylvania, 19107, USA

  2. Yuka Maeno-Hikichi

    Present address: Department of Physiology and Biophysics, Case Western Reserve University School of Medicine, Cleveland, Ohio, 44106, USA

Authors and Affiliations

  1. Department of Pharmacology, University of Pennsylvania School of Medicine, 3620 Hamilton Walk, Philadelphia, 19104, Pennsylvania, USA

    Meizan Lai, Yuka Maeno-Hikichi, Yuan Chen & Ji-fang Zhang

  2. Department of Pharmacology, University of Colorado Health Sciences Center, P.O. Box 6511, Aurora, 80045, Colorado, USA

    Joyce G Rohan & William A Sather

  3. Department of Neurobiology, University of Alabama at Birmingham, 1719 6th Avenue South, Birmingham, 35294, Alabama, USA

    Yi Zhou

  4. Department of Molecular & Cellular Engineering, University of Pennsylvania School of Medicine, 3620 Hamilton Walk, Philadelphia, 19104, Pennsylvania, USA

    Guangping Gao

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Lai, M., Wang, F., Rohan, J. et al. A tctex1-Ca2+ channel complex for selective surface expression of Ca2+ channels in neurons. Nat Neurosci 8, 435–442 (2005). https://doi.org/10.1038/nn1418

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