Glycine transporter isoforms show differential subcellular localization in PC12 cells

Authors

  • Arjan Geerlings,

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      The present address of Arjan Geerlings is Puleva Biotech, Camino de Purchil 66, 18004 Granada, Spain.

  • Enrique Núñez,

    1. Centro de Biología Molecular ‘Severo Ochoa’, Facultad de Ciencias, Universidad Autónoma de Madrid, Consejo Superior de Investigaciones Científicas, Madrid, Spain
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  • Lara Rodenstein,

    1. Centro de Biología Molecular ‘Severo Ochoa’, Facultad de Ciencias, Universidad Autónoma de Madrid, Consejo Superior de Investigaciones Científicas, Madrid, Spain
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  • Beatriz López-Corcuera,

    1. Centro de Biología Molecular ‘Severo Ochoa’, Facultad de Ciencias, Universidad Autónoma de Madrid, Consejo Superior de Investigaciones Científicas, Madrid, Spain
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  • Carmen Aragón

    1. Centro de Biología Molecular ‘Severo Ochoa’, Facultad de Ciencias, Universidad Autónoma de Madrid, Consejo Superior de Investigaciones Científicas, Madrid, Spain
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Address correspondence and reprint requests to Carmen Aragón, Centro de Biología Molecular ‘Severo Ochoa’, Facultad de Ciencias, Universidad Autónoma de Madrid, Consejo Superior de Investigaciones Científicas, 28049 Madrid, Spain. E-mail: caragon@cbm.uam.es

Abstract

The subcellular localization of glycine transporters one (GLYT1) and two (GLYT2) stably expressed in PC12 cells has been studied. To facilitate visualization, enhanced green fluorescent protein (GFP) was fused to the amino terminus of both glycine transporters. Functional analysis of the GFP–GLYT1 and GFP–GLYT2 stable cell lines demonstrated that they exhibited high affinity for glycine and the characteristic properties of both glycine transporter subtypes. The GFP-coupled transporters were differently distributed throughout the cell. GFP–GLYT1 was mainly localized on the plasma membrane, whereas most of GFP–GLYT2 was present on large dense-core vesicles and endosomes. Both transporters were absent from the synaptic vesicle population in PC12 cells.

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