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Genetic ablation of VIAAT in glycinergic neurons causes a severe respiratory phenotype and perinatal death

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Abstract

Both glycinergic and GABAergic neurons require the vesicular inhibitory amino acid transporter (VIAAT) for synaptic vesicle filling. Presynaptic GABA concentrations are determined by the GABA-synthesizing enzymes glutamate decarboxylase (GAD)65 and GAD67, whereas the presynaptic glycine content depends on the plasma membrane glycine transporter 2 (GlyT2). Although severely impaired, glycinergic transmission is not completely absent in GlyT2-knockout mice, suggesting that other routes of glycine uptake or de novo synthesis of glycine exist in presynaptic terminals. To investigate the consequences of a complete loss of glycinergic transmission, we generated a mouse line with a conditional ablation of VIAAT in glycinergic neurons by crossing mice with loxP-flanked VIAAT alleles with a GlyT2-Cre transgenic mouse line. Interestingly, conditional VIAAT knockout (VIAAT cKO) mice were not viable at birth. In addition to the dominant respiratory failure, VIAAT cKO showed an umbilical hernia and a cleft palate. Immunohistochemistry revealed an almost complete depletion of VIAAT in the brainstem. Electrophysiology revealed the absence of both spontaneous glycinergic and GABAergic inhibitory postsynaptic currents from hypoglossal motoneurons. Our results demonstrate that the deletion of VIAAT in GlyT2-Cre expressing neurons also strongly affects GABAergic transmission and suggest a large overlap of the glycinergic and the GABAergic neuron population during early development in the caudal parts of the brain.

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Acknowledgments

This work was funded by the Cluster of Excellence and DFG Research Center Nanoscale Microscopy and Molecular Physiology of the Brain (CNMPB). SH and JH received additional support from the DFG (Hu797/7-1, 8-1; Hi1414/2-1). The authors are grateful to Anja-Annett Grützner, Astrid Zeuch, Astrid Ohle and Annette Fahrenholz for technical assistance.

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Author notes

  1. Swen Hülsmann

    Present address: Clinic for Anesthesiology, Laboratory for Experimental Neuroanesthesiology, University Hospital Göttingen, Humboldtallee 23, 37073, Göttingen, Germany

  2. Christian Schnell

    Present address: Divisions of Pathophysiology and Repair and Neuroscience, School of Biosciences, Cardiff University, Cardiff, UK

Authors and Affiliations

  1. Center for Nanoscale Microscopy and Molecular Physiology of the Brain (CNMPB), Göttingen, Germany

    Christian Schnell & Swen Hülsmann

  2. Institute for Neurophysiology and Cellular Biophysics, Georg-August-University Göttingen, Humboldtallee 23, 37073, Göttingen, Germany

    Jamilur Rahman, Christian Schnell & Swen Hülsmann

  3. Carl-Ludwig-Institute for Physiology, University of Leipzig, Liebigstrasse 27, 04103, Leipzig, Germany

    Stefanie Besser & Johannes Hirrlinger

  4. Institute for Biochemistry and Molecular Medicine, University of Erlangen, Fahrstrasse 17, 91054, Erlangen, Germany

    Volker Eulenburg

  5. Department of Molecular Neurobiology, Max Planck Institute of Experimental Medicine, Hermann-Rein-Straße 3, 37075, Göttingen, Germany

    Sonja M. Wojcik

  6. Department of Neurogenetics, Max Planck Institute of Experimental Medicine, Hermann-Rein-Str. 3, 37075, Göttingen, Germany

    Johannes Hirrlinger

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  1. Jamilur Rahman
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  2. Stefanie Besser
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  6. Sonja M. Wojcik
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Correspondence to Sonja M. Wojcik or Swen Hülsmann.

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S. Hülsmann and S. M. Wojcik contributed equally.

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Rahman, J., Besser, S., Schnell, C. et al. Genetic ablation of VIAAT in glycinergic neurons causes a severe respiratory phenotype and perinatal death. Brain Struct Funct 220, 2835–2849 (2015). https://doi.org/10.1007/s00429-014-0829-2

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