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Inhibitory neuron-specific Cre-dependent red fluorescent labeling using VGAT BAC-based transgenic mouse lines with identified transgene integration sites
- Source :
- Journal of Comparative Neurology. 526:373-396
- Publication Year :
- 2017
- Publisher :
- Wiley, 2017.
-
Abstract
- Inhibitory neurons are crucial for shaping and regulating the dynamics of the entire network, and disturbances in these neurons contribute to brain disorders. Despite the recent progress in genetic labeling techniques, the heterogeneity of inhibitory neurons requires the development of highly characterized tools that allow accurate, convenient, and versatile visualization of inhibitory neurons in the mouse brain. Here, we report a novel genetic technique to visualize the vast majority and/or sparse subsets of inhibitory neurons in the mouse brain without using techniques that require advanced skills. We developed several lines of Cre-dependent tdTomato reporter mice based on the vesicular GABA transporter (VGAT)-BAC, named VGAT-stop-tdTomato mice. The most useful line (line #54) was selected for further analysis based on two characteristics: the inhibitory neuron-specificity of tdTomato expression and the transgene integration site, which confers efficient breeding and fewer adverse effects resulting from transgene integration-related genomic disruption. Robust and inhibitory neuron-specific expression of tdTomato was observed in a wide range of developmental and cellular contexts. By breeding the VGAT-stop-tdTomato mouse (line #54) with a novel Cre driver mouse line, Galntl4-CreER, sparse labeling of inhibitory neurons was achieved following tamoxifen administration. Furthermore, another interesting line (line #58) was generated through the unexpected integration of the transgene into the X-chromosome and will be used to map X-chromosome inactivation of inhibitory neurons. Taken together, our studies provide new, well-characterized tools with which multiple aspects of inhibitory neurons can be studied in the mouse.
- Subjects :
- 0301 basic medicine
Genetically modified mouse
Vesicular Inhibitory Amino Acid Transport Proteins
VESICULAR GABA TRANSPORTER
Transgene
Mice, Transgenic
Nerve Tissue Proteins
Biology
Genetic Technique
Inhibitory postsynaptic potential
Mice
03 medical and health sciences
0302 clinical medicine
Inhibitory neuron
Animals
Integrases
Glutamate Decarboxylase
General Neuroscience
PAX2 Transcription Factor
Estrogen Antagonists
Brain
Neural Inhibition
Molecular biology
Cell biology
Mice, Inbred C57BL
Luminescent Proteins
Tamoxifen
Fluorescent labelling
Ki-67 Antigen
030104 developmental biology
Gene Expression Regulation
Mutation
N-Acetylgalactosaminyltransferases
Somatostatin
RGS Proteins
030217 neurology & neurosurgery
Subjects
Details
- ISSN :
- 00219967
- Volume :
- 526
- Database :
- OpenAIRE
- Journal :
- Journal of Comparative Neurology
- Accession number :
- edsair.doi.dedup.....4e797e22cbefe007332c93b917d22366