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vglut2 and gad expression reveal distinct patterns of dual GABAergic versus glutamatergic cotransmitter phenotypes of dopaminergic and noradrenergic neurons in the zebrafish brain.

Filippi A, Mueller T, Driever W - J. Comp. Neurol. (2014)

Bottom Line: Our results show that most dopaminergic neurons also express GABAergic markers, including the dopaminergic groups of the olfactory bulb (homologous to mammalian A16) and the subpallium, the hypothalamic groups (A12, A14), the prethalamic zona incerta group (A13), the preoptic groups (A15), and the pretectal group.All together, our results demonstrate that all catecholaminergic groups in zebrafish are either GABAergic or glutamatergic.We compare our results with those that have been described for mammalian systems, discuss the phenomenon of transmitter dualism in the context of developmental specification of GABAergic and glutamatergic regions in the brain, and put this phenomenon in an evolutionary perspective.

View Article: PubMed Central - PubMed

Affiliation: Developmental Biology, Institute of Biology I, Faculty of Biology, Albert-Ludwigs-University Freiburg, 79104, Freiburg, Germany.

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Localization of gad1b/2- or vglut2-expressing cells and TH-ir neurons in the telencephalon of the juvenile brain. A: Lateral overview of a juvenile brain stained for th by in situ hybridization (image taken from Filippi et al., 2010), which serves as anatomical reference to indicate the levels of the sections shown in B–G. B–G: Fluorescent in situ hybridization to detect gad1b/2 (C,F; green) or vglut2 (D,G; green) expression combined with anti-TH immunofluorescence (red). In each row, the image at the left shows an overview of the considered section and displays only TH immunostaining. Higher magnifications of the approximate area framed by the white square are shown to the right, together with either gad1b/2 (C,F) or vglut2 (D,G) staining. All images are 6–10-μm confocal z-projections of areas comprising TH-ir cells in the olfactory bulb (OB; B–D) and in the subpallium (SP; E-G). All telencephalic THir neurons express gad1b/2 (C,F) but not vglut2 (D,G), indicating that they are GABAergic. To better display the colocalization of the markers, C1–C3 and F1–F3 show the individual channels of the areas framed in C and F, respectively. For abbreviations, see list. A magenta–green copy of this figure is available as Supporting Information Figure S3. Scale bar = 50 μm in B–G.
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fig03: Localization of gad1b/2- or vglut2-expressing cells and TH-ir neurons in the telencephalon of the juvenile brain. A: Lateral overview of a juvenile brain stained for th by in situ hybridization (image taken from Filippi et al., 2010), which serves as anatomical reference to indicate the levels of the sections shown in B–G. B–G: Fluorescent in situ hybridization to detect gad1b/2 (C,F; green) or vglut2 (D,G; green) expression combined with anti-TH immunofluorescence (red). In each row, the image at the left shows an overview of the considered section and displays only TH immunostaining. Higher magnifications of the approximate area framed by the white square are shown to the right, together with either gad1b/2 (C,F) or vglut2 (D,G) staining. All images are 6–10-μm confocal z-projections of areas comprising TH-ir cells in the olfactory bulb (OB; B–D) and in the subpallium (SP; E-G). All telencephalic THir neurons express gad1b/2 (C,F) but not vglut2 (D,G), indicating that they are GABAergic. To better display the colocalization of the markers, C1–C3 and F1–F3 show the individual channels of the areas framed in C and F, respectively. For abbreviations, see list. A magenta–green copy of this figure is available as Supporting Information Figure S3. Scale bar = 50 μm in B–G.

Mentions: For microscopic observation, larvae were mounted in 80% glycerol/PBS containing 1% agarose. Confocal z-stacks of whole-mount larvae and sectioned brains were recorded by using a Zeiss LSM 5 DUO laser scanning confocal microscope. Then z-projections from subsets of focal planes were generated with the Zeiss LSM software and exported as image format files. The images were assembled into figures and processed with the Adobe (San Jose, CA) Photoshop CS2 9.0 software. Adjustments to contrast and brightness were made by using the Photoshop Levels functions. All adjustments were made to whole images including the enlarged sections, except for Figures 1I1–I3, 3C1–C3, 3F1–F3, and 4F1–F3, for which in the whole enlarged section red and green levels were adjusted to better visualize red and green channel colocalization.


vglut2 and gad expression reveal distinct patterns of dual GABAergic versus glutamatergic cotransmitter phenotypes of dopaminergic and noradrenergic neurons in the zebrafish brain.

Filippi A, Mueller T, Driever W - J. Comp. Neurol. (2014)

Localization of gad1b/2- or vglut2-expressing cells and TH-ir neurons in the telencephalon of the juvenile brain. A: Lateral overview of a juvenile brain stained for th by in situ hybridization (image taken from Filippi et al., 2010), which serves as anatomical reference to indicate the levels of the sections shown in B–G. B–G: Fluorescent in situ hybridization to detect gad1b/2 (C,F; green) or vglut2 (D,G; green) expression combined with anti-TH immunofluorescence (red). In each row, the image at the left shows an overview of the considered section and displays only TH immunostaining. Higher magnifications of the approximate area framed by the white square are shown to the right, together with either gad1b/2 (C,F) or vglut2 (D,G) staining. All images are 6–10-μm confocal z-projections of areas comprising TH-ir cells in the olfactory bulb (OB; B–D) and in the subpallium (SP; E-G). All telencephalic THir neurons express gad1b/2 (C,F) but not vglut2 (D,G), indicating that they are GABAergic. To better display the colocalization of the markers, C1–C3 and F1–F3 show the individual channels of the areas framed in C and F, respectively. For abbreviations, see list. A magenta–green copy of this figure is available as Supporting Information Figure S3. Scale bar = 50 μm in B–G.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig03: Localization of gad1b/2- or vglut2-expressing cells and TH-ir neurons in the telencephalon of the juvenile brain. A: Lateral overview of a juvenile brain stained for th by in situ hybridization (image taken from Filippi et al., 2010), which serves as anatomical reference to indicate the levels of the sections shown in B–G. B–G: Fluorescent in situ hybridization to detect gad1b/2 (C,F; green) or vglut2 (D,G; green) expression combined with anti-TH immunofluorescence (red). In each row, the image at the left shows an overview of the considered section and displays only TH immunostaining. Higher magnifications of the approximate area framed by the white square are shown to the right, together with either gad1b/2 (C,F) or vglut2 (D,G) staining. All images are 6–10-μm confocal z-projections of areas comprising TH-ir cells in the olfactory bulb (OB; B–D) and in the subpallium (SP; E-G). All telencephalic THir neurons express gad1b/2 (C,F) but not vglut2 (D,G), indicating that they are GABAergic. To better display the colocalization of the markers, C1–C3 and F1–F3 show the individual channels of the areas framed in C and F, respectively. For abbreviations, see list. A magenta–green copy of this figure is available as Supporting Information Figure S3. Scale bar = 50 μm in B–G.
Mentions: For microscopic observation, larvae were mounted in 80% glycerol/PBS containing 1% agarose. Confocal z-stacks of whole-mount larvae and sectioned brains were recorded by using a Zeiss LSM 5 DUO laser scanning confocal microscope. Then z-projections from subsets of focal planes were generated with the Zeiss LSM software and exported as image format files. The images were assembled into figures and processed with the Adobe (San Jose, CA) Photoshop CS2 9.0 software. Adjustments to contrast and brightness were made by using the Photoshop Levels functions. All adjustments were made to whole images including the enlarged sections, except for Figures 1I1–I3, 3C1–C3, 3F1–F3, and 4F1–F3, for which in the whole enlarged section red and green levels were adjusted to better visualize red and green channel colocalization.

Bottom Line: Our results show that most dopaminergic neurons also express GABAergic markers, including the dopaminergic groups of the olfactory bulb (homologous to mammalian A16) and the subpallium, the hypothalamic groups (A12, A14), the prethalamic zona incerta group (A13), the preoptic groups (A15), and the pretectal group.All together, our results demonstrate that all catecholaminergic groups in zebrafish are either GABAergic or glutamatergic.We compare our results with those that have been described for mammalian systems, discuss the phenomenon of transmitter dualism in the context of developmental specification of GABAergic and glutamatergic regions in the brain, and put this phenomenon in an evolutionary perspective.

View Article: PubMed Central - PubMed

Affiliation: Developmental Biology, Institute of Biology I, Faculty of Biology, Albert-Ludwigs-University Freiburg, 79104, Freiburg, Germany.

Show MeSH