Tags

Type your tag names separated by a space and hit enter

Fluorescent labeling of both GABAergic and glycinergic neurons in vesicular GABA transporter (VGAT)-venus transgenic mouse.
Neuroscience 2009; 164(3):1031-43N

Abstract

Inhibitory neurons play important roles in a number of brain functions. They are composed of GABAergic neurons and glycinergic neurons, and vesicular GABA transporter (VGAT) is specifically expressed in these neurons. Since the inhibitory neurons are scattered around in the CNS, it is difficult to identify these cells in living brain preparations. The glutamate decarboxylase (GAD) 67-GFP knock-in mouse has been widely used for the identification of GABAergic neurons, but their GAD67 expression was decreased compared to the wild-type mice. To overcome such a problem and to highlight the function and morphology of inhibitory neurons, we generated four lines of VGAT-Venus transgenic mice (lines #04, #29, #39 and #49) expressing Venus fluorescent protein under the control of mouse VGAT promoter. We found higher expression level of Venus transcripts and proteins as well as brighter fluorescent signal in line #39 mouse brains, compared to brains of other lines examined. By Western blots and spectrofluorometric measurements of forebrain, the line #39 mouse showed stronger GFP immunoreactivity and brighter fluorescent intensity than the GAD67-GFP knock-in mouse. In addition, Venus was present not only in somata, but also in neurites in the line #39 mouse by histological studies. In situ hybridization analysis showed that the expression pattern of Venus in the line #39 mouse was similar to that of endogenous VGAT. Double immunostaining analysis in line #39 mouse showed that Venus-expressing cells are primarily immunoreactive for GABA in cerebral cortex, hippocampus and cerebellar cortex and for GABA or glycine in dorsal cochlear nucleus. These results demonstrate that the VGAT-Venus line #39 mouse should be useful for studies on function and morphology of inhibitory neurons in the CNS.

Authors+Show Affiliations

Department of Genetic and Behavioral Neuroscience, Gunma University Graduate School of Medicine, Maebashi 371-8511, Japan.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

19766173

Citation

Wang, Y, et al. "Fluorescent Labeling of Both GABAergic and Glycinergic Neurons in Vesicular GABA Transporter (VGAT)-venus Transgenic Mouse." Neuroscience, vol. 164, no. 3, 2009, pp. 1031-43.
Wang Y, Kakizaki T, Sakagami H, et al. Fluorescent labeling of both GABAergic and glycinergic neurons in vesicular GABA transporter (VGAT)-venus transgenic mouse. Neuroscience. 2009;164(3):1031-43.
Wang, Y., Kakizaki, T., Sakagami, H., Saito, K., Ebihara, S., Kato, M., ... Yanagawa, Y. (2009). Fluorescent labeling of both GABAergic and glycinergic neurons in vesicular GABA transporter (VGAT)-venus transgenic mouse. Neuroscience, 164(3), pp. 1031-43. doi:10.1016/j.neuroscience.2009.09.010.
Wang Y, et al. Fluorescent Labeling of Both GABAergic and Glycinergic Neurons in Vesicular GABA Transporter (VGAT)-venus Transgenic Mouse. Neuroscience. 2009 Dec 15;164(3):1031-43. PubMed PMID: 19766173.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Fluorescent labeling of both GABAergic and glycinergic neurons in vesicular GABA transporter (VGAT)-venus transgenic mouse. AU - Wang,Y, AU - Kakizaki,T, AU - Sakagami,H, AU - Saito,K, AU - Ebihara,S, AU - Kato,M, AU - Hirabayashi,M, AU - Saito,Y, AU - Furuya,N, AU - Yanagawa,Y, Y1 - 2009/09/17/ PY - 2009/01/30/received PY - 2009/08/19/revised PY - 2009/09/04/accepted PY - 2009/9/22/entrez PY - 2009/9/22/pubmed PY - 2010/2/9/medline SP - 1031 EP - 43 JF - Neuroscience JO - Neuroscience VL - 164 IS - 3 N2 - Inhibitory neurons play important roles in a number of brain functions. They are composed of GABAergic neurons and glycinergic neurons, and vesicular GABA transporter (VGAT) is specifically expressed in these neurons. Since the inhibitory neurons are scattered around in the CNS, it is difficult to identify these cells in living brain preparations. The glutamate decarboxylase (GAD) 67-GFP knock-in mouse has been widely used for the identification of GABAergic neurons, but their GAD67 expression was decreased compared to the wild-type mice. To overcome such a problem and to highlight the function and morphology of inhibitory neurons, we generated four lines of VGAT-Venus transgenic mice (lines #04, #29, #39 and #49) expressing Venus fluorescent protein under the control of mouse VGAT promoter. We found higher expression level of Venus transcripts and proteins as well as brighter fluorescent signal in line #39 mouse brains, compared to brains of other lines examined. By Western blots and spectrofluorometric measurements of forebrain, the line #39 mouse showed stronger GFP immunoreactivity and brighter fluorescent intensity than the GAD67-GFP knock-in mouse. In addition, Venus was present not only in somata, but also in neurites in the line #39 mouse by histological studies. In situ hybridization analysis showed that the expression pattern of Venus in the line #39 mouse was similar to that of endogenous VGAT. Double immunostaining analysis in line #39 mouse showed that Venus-expressing cells are primarily immunoreactive for GABA in cerebral cortex, hippocampus and cerebellar cortex and for GABA or glycine in dorsal cochlear nucleus. These results demonstrate that the VGAT-Venus line #39 mouse should be useful for studies on function and morphology of inhibitory neurons in the CNS. SN - 1873-7544 UR - https://www.unboundmedicine.com/medline/citation/19766173/Fluorescent_labeling_of_both_GABAergic_and_glycinergic_neurons_in_vesicular_GABA_transporter__VGAT__venus_transgenic_mouse_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0306-4522(09)01485-7 DB - PRIME DP - Unbound Medicine ER -