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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 anterior diencephalon of the juvenile brain. A: Overview of section planes shown in B–M (for details, see Fig. 3A legend). B–M: Fluorescent in situ hybridization to detect gad1b/2 (C,F,I,L; green) or vglut2 (D,G,J,M; green) expression combined with anti-TH immunofluorescence (red). The DA clusters in the magnocellular preoptic nucleus (PM; B–D), in the prethalamus and suprachiasmatic nucleus (VL, SC; E–G), in the pretectum (Pr; H–J), and in the posterior tuberculum (PT; K–M) are shown. In each row, the image at the left shows an overview of the considered section and displays only TH immunostaining. The pretectal, prethalamic, and suprachiasmatic clusters are included in the same overview, shown twice in E and H. Higher magnifications of the approximate area framed by the white square in the overviews are shown at their right, together with either gad1b/2 (C,F,I,L) or vglut2 (D,G,J,M) staining. The majority of the images are 4–15-μm confocal z-projections, except for C1–C3, G, and I1–I3, which are single planes. All DA clusters considered in this figure are also GABAergic, as demonstrated by coexpression of TH with gad1b/2, but not with vglut2. The only exception is represented by the suprachiasmatic nucleus, in which some DA neurons express gad1b/2 (arrowhead in F1–F3) and some do not (arrow in F1–F3). For abbreviations, see list. A magenta–green copy of this figure is available as Supporting Information Figure S4. Scale bar = 50 μm in B–M.
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fig04: Localization of gad1b/2- or vglut2-expressing cells and TH-ir neurons in the anterior diencephalon of the juvenile brain. A: Overview of section planes shown in B–M (for details, see Fig. 3A legend). B–M: Fluorescent in situ hybridization to detect gad1b/2 (C,F,I,L; green) or vglut2 (D,G,J,M; green) expression combined with anti-TH immunofluorescence (red). The DA clusters in the magnocellular preoptic nucleus (PM; B–D), in the prethalamus and suprachiasmatic nucleus (VL, SC; E–G), in the pretectum (Pr; H–J), and in the posterior tuberculum (PT; K–M) are shown. In each row, the image at the left shows an overview of the considered section and displays only TH immunostaining. The pretectal, prethalamic, and suprachiasmatic clusters are included in the same overview, shown twice in E and H. Higher magnifications of the approximate area framed by the white square in the overviews are shown at their right, together with either gad1b/2 (C,F,I,L) or vglut2 (D,G,J,M) staining. The majority of the images are 4–15-μm confocal z-projections, except for C1–C3, G, and I1–I3, which are single planes. All DA clusters considered in this figure are also GABAergic, as demonstrated by coexpression of TH with gad1b/2, but not with vglut2. The only exception is represented by the suprachiasmatic nucleus, in which some DA neurons express gad1b/2 (arrowhead in F1–F3) and some do not (arrow in F1–F3). For abbreviations, see list. A magenta–green copy of this figure is available as Supporting Information Figure S4. Scale bar = 50 μm in B–M.

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 anterior diencephalon of the juvenile brain. A: Overview of section planes shown in B–M (for details, see Fig. 3A legend). B–M: Fluorescent in situ hybridization to detect gad1b/2 (C,F,I,L; green) or vglut2 (D,G,J,M; green) expression combined with anti-TH immunofluorescence (red). The DA clusters in the magnocellular preoptic nucleus (PM; B–D), in the prethalamus and suprachiasmatic nucleus (VL, SC; E–G), in the pretectum (Pr; H–J), and in the posterior tuberculum (PT; K–M) are shown. In each row, the image at the left shows an overview of the considered section and displays only TH immunostaining. The pretectal, prethalamic, and suprachiasmatic clusters are included in the same overview, shown twice in E and H. Higher magnifications of the approximate area framed by the white square in the overviews are shown at their right, together with either gad1b/2 (C,F,I,L) or vglut2 (D,G,J,M) staining. The majority of the images are 4–15-μm confocal z-projections, except for C1–C3, G, and I1–I3, which are single planes. All DA clusters considered in this figure are also GABAergic, as demonstrated by coexpression of TH with gad1b/2, but not with vglut2. The only exception is represented by the suprachiasmatic nucleus, in which some DA neurons express gad1b/2 (arrowhead in F1–F3) and some do not (arrow in F1–F3). For abbreviations, see list. A magenta–green copy of this figure is available as Supporting Information Figure S4. Scale bar = 50 μm in B–M.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4288968&req=5

fig04: Localization of gad1b/2- or vglut2-expressing cells and TH-ir neurons in the anterior diencephalon of the juvenile brain. A: Overview of section planes shown in B–M (for details, see Fig. 3A legend). B–M: Fluorescent in situ hybridization to detect gad1b/2 (C,F,I,L; green) or vglut2 (D,G,J,M; green) expression combined with anti-TH immunofluorescence (red). The DA clusters in the magnocellular preoptic nucleus (PM; B–D), in the prethalamus and suprachiasmatic nucleus (VL, SC; E–G), in the pretectum (Pr; H–J), and in the posterior tuberculum (PT; K–M) are shown. In each row, the image at the left shows an overview of the considered section and displays only TH immunostaining. The pretectal, prethalamic, and suprachiasmatic clusters are included in the same overview, shown twice in E and H. Higher magnifications of the approximate area framed by the white square in the overviews are shown at their right, together with either gad1b/2 (C,F,I,L) or vglut2 (D,G,J,M) staining. The majority of the images are 4–15-μm confocal z-projections, except for C1–C3, G, and I1–I3, which are single planes. All DA clusters considered in this figure are also GABAergic, as demonstrated by coexpression of TH with gad1b/2, but not with vglut2. The only exception is represented by the suprachiasmatic nucleus, in which some DA neurons express gad1b/2 (arrowhead in F1–F3) and some do not (arrow in F1–F3). For abbreviations, see list. A magenta–green copy of this figure is available as Supporting Information Figure S4. Scale bar = 50 μm in B–M.
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