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Glycinergic neurons expressing enhanced green fluorescent protein in bacterial artificial chromosome transgenic mice.

Abstract

Although glycine is a major inhibitory transmitter in the mammalian CNS, the role of glycinergic neurons in defined neuronal circuits remains ill defined. This is due in part to difficulties in identifying these cells in living slice preparations for electrophysiological recordings and visualizing their axonal projections. To facilitate the morphological and functional analysis of glycinergic neurons, we generated bacterial artificial chromosome (BAC) transgenic mice, which specifically express enhanced green fluorescent protein (EGFP) under the control of the promotor of the glycine transporter (GlyT) 2 gene, which is a reliable marker for glycinergic neurons. Neurons expressing GlyT2-EGFP were intensely fluorescent, and their dendrites and axons could be visualized in great detail. Numerous positive neurons were detected in the spinal cord, brainstem, and cerebellum. The hypothalamus, intralaminar nuclei of the thalamus, and basal forebrain also received a dense GlyT2-EGFP innervation, whereas in the olfactory bulb, striatum, neocortex, hippocampus, and amygdala positive fibers were much less abundant. No GlyT2-EGFP-positive cell bodies were seen in the forebrain. On the subcellular level, GlyT2-EGFP fluorescence was colocalized extensively with glycine immunoreactivity in somata and dendrites and with both glycine and GlyT2 immunoreactivity in axon terminals, as shown by triple staining at all levels of the neuraxis, confirming the selective expression of the transgene in glycinergic neurons. In slice preparations of the spinal cord, no difference between the functional properties of EGFP-positive and negative neurons could be detected, confirming the utility of visually identifying glycinergic neurons to investigate their functional role in electrophysiological studies.

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  • Authors+Show Affiliations

    ,

    Department of Experimental and Clinical Pharmacology and Toxicology, University of Erlangen-Nürnberg, 91054 Erlangen, Germany.

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    Source

    The Journal of comparative neurology 482:2 2005 Feb 07 pg 123-41

    MeSH

    Amino Acid Transport Systems, Neutral
    Animals
    Biomarkers
    Brain
    Chromosomes, Artificial, Bacterial
    Female
    Gene Expression Regulation
    Genetic Engineering
    Glycine
    Glycine Plasma Membrane Transport Proteins
    Green Fluorescent Proteins
    Immunohistochemistry
    Interneurons
    Luminescent Agents
    Male
    Mice
    Mice, Inbred C57BL
    Mice, Transgenic
    Neural Pathways
    Neurons
    Promoter Regions, Genetic
    Spinal Cord

    Pub Type(s)

    Journal Article
    Research Support, Non-U.S. Gov't

    Language

    eng

    PubMed ID

    15611994

    Citation

    Zeilhofer, Hanns Ulrich, et al. "Glycinergic Neurons Expressing Enhanced Green Fluorescent Protein in Bacterial Artificial Chromosome Transgenic Mice." The Journal of Comparative Neurology, vol. 482, no. 2, 2005, pp. 123-41.
    Zeilhofer HU, Studler B, Arabadzisz D, et al. Glycinergic neurons expressing enhanced green fluorescent protein in bacterial artificial chromosome transgenic mice. J Comp Neurol. 2005;482(2):123-41.
    Zeilhofer, H. U., Studler, B., Arabadzisz, D., Schweizer, C., Ahmadi, S., Layh, B., ... Fritschy, J. M. (2005). Glycinergic neurons expressing enhanced green fluorescent protein in bacterial artificial chromosome transgenic mice. The Journal of Comparative Neurology, 482(2), pp. 123-41.
    Zeilhofer HU, et al. Glycinergic Neurons Expressing Enhanced Green Fluorescent Protein in Bacterial Artificial Chromosome Transgenic Mice. J Comp Neurol. 2005 Feb 7;482(2):123-41. PubMed PMID: 15611994.
    * Article titles in AMA citation format should be in sentence-case
    TY - JOUR T1 - Glycinergic neurons expressing enhanced green fluorescent protein in bacterial artificial chromosome transgenic mice. AU - Zeilhofer,Hanns Ulrich, AU - Studler,Barbara, AU - Arabadzisz,Dimitrula, AU - Schweizer,Claude, AU - Ahmadi,Seifollah, AU - Layh,Beate, AU - Bösl,Michael R, AU - Fritschy,Jean-Marc, PY - 2004/12/22/pubmed PY - 2005/4/19/medline PY - 2004/12/22/entrez SP - 123 EP - 41 JF - The Journal of comparative neurology JO - J. Comp. Neurol. VL - 482 IS - 2 N2 - Although glycine is a major inhibitory transmitter in the mammalian CNS, the role of glycinergic neurons in defined neuronal circuits remains ill defined. This is due in part to difficulties in identifying these cells in living slice preparations for electrophysiological recordings and visualizing their axonal projections. To facilitate the morphological and functional analysis of glycinergic neurons, we generated bacterial artificial chromosome (BAC) transgenic mice, which specifically express enhanced green fluorescent protein (EGFP) under the control of the promotor of the glycine transporter (GlyT) 2 gene, which is a reliable marker for glycinergic neurons. Neurons expressing GlyT2-EGFP were intensely fluorescent, and their dendrites and axons could be visualized in great detail. Numerous positive neurons were detected in the spinal cord, brainstem, and cerebellum. The hypothalamus, intralaminar nuclei of the thalamus, and basal forebrain also received a dense GlyT2-EGFP innervation, whereas in the olfactory bulb, striatum, neocortex, hippocampus, and amygdala positive fibers were much less abundant. No GlyT2-EGFP-positive cell bodies were seen in the forebrain. On the subcellular level, GlyT2-EGFP fluorescence was colocalized extensively with glycine immunoreactivity in somata and dendrites and with both glycine and GlyT2 immunoreactivity in axon terminals, as shown by triple staining at all levels of the neuraxis, confirming the selective expression of the transgene in glycinergic neurons. In slice preparations of the spinal cord, no difference between the functional properties of EGFP-positive and negative neurons could be detected, confirming the utility of visually identifying glycinergic neurons to investigate their functional role in electrophysiological studies. SN - 0021-9967 UR - https://www.unboundmedicine.com/medline/citation/15611994/Glycinergic_neurons_expressing_enhanced_green_fluorescent_protein_in_bacterial_artificial_chromosome_transgenic_mice_ L2 - https://doi.org/10.1002/cne.20349 DB - PRIME DP - Unbound Medicine ER -