B6.FVB(129)-Tg(Slc6a5-EGFP)1Uze/J

Stock number: 038516
Product name: GlyT2-EGFP
Research areas: Neurobiology Research, Research Tools

Description

GlyT2-EGFP transgenic mice express EGFP under the control of the murine Slc6a5 promoter. They may be useful for identifying glycinergic neurons.

Detailed description

GlyT2-EGFP transgenic mice express EGFP under the control of the murine solute carrier family 6 (neurotransmitter transporter, glycine), member 5 (Slc6a5) promoter. GlyT2 encodes a membrane glycoprotein that is an established marker of glycinergic neurons. GlyT2 is expressed in the central nervous system (CNS) in two splice variants (GlyT2a and GlyT2b), which differ in their N-termini as a result of the use of alternative first exons (exon 1a and exon 1b).

In this strain, EGFP cDNA was inserted in frame into the beginning of exon 2 of the Slc6a5 locus of the BAC (and exon 2 was subsequently removed). In these GlyT2-EGFP transgenic mice, EGFP fluorescence is strong and specific to neurons in the brain and spinal cord that are shown by immunohistochemical analysis and direct immunofluorescence to express markers of glycinergic neurons. Specifically, EGFP is evident in dendrites and axons in the spinal cord, brainstem, and cerebellum. The hypothalamus, intralaminar nuclei of the thalamus, and basal forebrain also display 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.

Hemizygous mice are viable and fertile, it is unknown if homozygous mice are viable and fertile.

Development

A bacterial artificial chromosome (BAC) clone, 365E4, from C57BL/6 BAC library RPCI-23 was used to obtain mouse genomic DNA containing the entire mouse solute carrier family 6 (neurotransmitter transporter, glycine), member 5 (Slc6a5) gene. This BAC was modified to insert EGFP cDNA and a bovine growth hormone polyadenylation (polyA) into the middle of exon 2, and exon 2 was then removed. The linearized BAC was microinjected into fertilized FVB oocytes. GlyT2-EGFP founder mice were bred to 129/SVJ mice and the colony was subsequently maintained by breeding transgenic mice with C57BL/6J mice (Stock No. 000664) for at least 5 generations by the donating laboratory. Upon arrival at The Jackson Laboratory, sperm was cryopreserved. To establish a live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes.

Allele

Symbol: Tg(Slc6a5-EGFP)1Uze
Name: transgene insertion 1, University of Zurich
MGI accession ID: MGI:3835459
Generation method: Transgenic
Attributes: Reporter
Synonyms: GlyT2-EGFP; Slc6a5^tg(EGFP)Uze; TgN(GlyT2-EGFP)
Marker symbol: Tg(Slc6a5-EGFP)1Uze
Marker name: transgene insertion 1, University of Zurich
Marker synonyms: GlyT2-EGFP; Slc6a5^tg(EGFP)Uze
Chromosome: UN
Strain of origin: FVB
Molecular note: The EGFP coding sequence, with the bovine growth hormone polyadenylation signal, was inserted into the middle of exon 2 of the Sl6a5 gene in the BAC clone 365E4 from C57BL/6 BAC library RPCI-23. This BAC contains the entire coding sequence for both isoforms of the protein (also known as glycine transporter 2, GlyT2) and 105 kb of 5' and 21 kb of 3' flanking genomic sequence. In transgenic mice, EGFP fluorescence is strong and specific to neurons in the brain and spinal cord that are shown by immunohistochemical analysis to express markers of glycinergic neurons.
General note: This record represents seven founder lines: lines 13 and 16 have integrated more than four copies of the transgene; lines 14 and 15 a single copy; and the rest between two and four copies. EGFP expression patterns were examined in detail in one high copy number and one single copy number line, lines 13 and 14, respectively. While these differed somewhat in fluorescence intensity, both exhibited significant signal in neurons identified by immunohistochemical analysis as glycinergic neurons. (J:145521)