Fluorescent labeling of GABAergic neurons in VGAT-Venus transgenic mouse forebrain
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1
University of Tokyo , Japan
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2
NIPS, Japan
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3
Gunma University, Japan
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4
Kitasato University, Japan
GABAergic neurons play an important role in the regulation and stabilization of network activities and are essential for a number of functions including cognition and memory. GABA is synthesized by glutamate decarboxylase (GAD) and is accumulated into synaptic vesicles by vesicular GABA transporter (VGAT). Two forms of GAD, namely GAD65 and GAD67 and VGAT are specifically expressed in GABAergic neurons. Because the GABAergic neurons are scattered around in the CNS including forebrain, it is difficult to identify these cells in living brain preparations. The GAD67-GFP knock-in mouse has been widely used for identifying GABAergic neurons, but their GAD67 expression as well as their GABA content was reduced compared to the wild-type mouse. To overcome such a problem and to highlight the function and morphology of GABAergic neurons, we generated VGAT-Venus transgenic mice expressing Venus fluorescent protein under the control of the mouse VGAT promoter. Western blots and spectrofluorometric measurements of forebrain showed that the VGAT-Venus transgenic mouse had stronger GFP immunoreactivity and brighter fluorescent intensity than the GAD67-GFP knock-in mouse. Double immunostaining analysis in the VGAT-Venus transgenic mouse revealed almost complete overlapping of Venus-expressing cells and GABA-positive cells in the hippocampus and cerebral cortex. These results indicate that the VGAT-Venus transgenic mouse is useful for studying functional and morphological properties of GABAergic neurons in the forebrain.
Conference:
EMBO workshop: Gaba Signalling and Brain Networks , Amsterdam, Netherlands, 30 Jun - 2 Jul, 2010.
Presentation Type:
Poster Presentation
Topic:
Posters
Citation:
Ebihara
S,
Hirabayashi
M,
Kakizaki
T,
Kato
M,
Miwa
H,
Saito
K,
Sakagami
H,
Wang
Y,
Yanagawa
Y and
Zhang
Y
(2010). Fluorescent labeling of GABAergic neurons in VGAT-Venus transgenic mouse forebrain.
Conference Abstract:
EMBO workshop: Gaba Signalling and Brain Networks .
doi: 10.3389/conf.fnins.2010.15.00026
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Received:
25 Jun 2010;
Published Online:
25 Jun 2010.
*
Correspondence:
Y. Yanagawa, Gunma University, Maebashi, Japan, yuchio@gunma-u.ac.jp