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Zebrafish adult-derived hypothalamic neurospheres generate gonadotropin-releasing hormone (GnRH) neurons.

Cortés-Campos C, Letelier J, Ceriani R, Whitlock KE - Biol Open (2015)

Bottom Line: Using this system, we show that neurospheres derived from the adult hypothalamus can be maintained in culture and subsequently differentiate glia and neurons.Finally, we show in vivo that a neurogenic niche in the hypothalamus contains GnRH positive neurons.Thus, we demonstrated for the first time that neurospheres can be derived from the hypothalamus of the adult zebrafish and that these neural progenitors are capable of producing GnRH containing neurons.

View Article: PubMed Central - PubMed

Affiliation: Centro Interdisciplinario de Neurociencia de Valparaíso (CINV), Facultad de Ciencias, Universidad de Valparaíso, Pasaje Harrington 269, Valparaíso 2340000, Chile Whitehead Institute for Biomedical Research (WIBR), 9 Cambridge Center, Cambridge, MA 02142, USA.

No MeSH data available.


Related in: MedlinePlus

The hypothalamus of the adult zebrafish contains newly differentiated GnRH cells. Horizontal sections showing the anterior region of the parvocellular preoptic nucleus (PPa), with DAPI labeling (blue). (A-C) HuC labeling in a HuC:GFP background (green, arrows) and vimentin labeling (red, arrowhead) do not co-localize. The vimentin positive cells line the edges of the ventricles. (D-F) HuC (green) and Sox2 labeling (red, arrows) co-localizes in some cells (arrows) located closer to the border of the ventricle (arrows). (G-I) Section showing GnRH cell body labeling (green, box, asterisk) and labeling of processes (green, arrows), Sox2 labeling (red, box, asterisk) in cells located several cell diameters from the border of the ventricle (arrows) co-localizes with the GnRH labeling (merge). All Images: anterior is to the left. Scale bar=30 μm.
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BIO010447F8: The hypothalamus of the adult zebrafish contains newly differentiated GnRH cells. Horizontal sections showing the anterior region of the parvocellular preoptic nucleus (PPa), with DAPI labeling (blue). (A-C) HuC labeling in a HuC:GFP background (green, arrows) and vimentin labeling (red, arrowhead) do not co-localize. The vimentin positive cells line the edges of the ventricles. (D-F) HuC (green) and Sox2 labeling (red, arrows) co-localizes in some cells (arrows) located closer to the border of the ventricle (arrows). (G-I) Section showing GnRH cell body labeling (green, box, asterisk) and labeling of processes (green, arrows), Sox2 labeling (red, box, asterisk) in cells located several cell diameters from the border of the ventricle (arrows) co-localizes with the GnRH labeling (merge). All Images: anterior is to the left. Scale bar=30 μm.

Mentions: To investigate whether the hypothalamus of the intact adult zebrafish contains potential GnRH progenitors we used adult male zebrafish that had been selected via the mating training protocol and sacrificed them to obtain brains for sectioning. Cryostat sections were analyzed using antibodies against GnRH, Sox2, and HuC. Cells positive for HuC labeling (Fig. 8A, arrows) were localized in the anterior region of the parvocellular preoptic nucleus (PPa) and these cells were distinct from the vimentin positive glia (tanycytes) lining the ventricle (Fig. 8B,C, red, arrowheads). Immuno-localization of the progenitor marker Sox2 showed nuclear label in the cells lining the ventricle (Fig. 8E, boxed area for example) and cytoplasmic localization in cells located several cell diameters from the ventricle (Fig. 8E,F, arrows). A subset of the Sox2 positive cells co-localized with HuC positive cells (Fig. 8F) a marker for newly generated neurons in agreement the role of Sox2 as a neural progenitor. While cells lying adjacent to the ventricle had nuclear localization Sox2, cells lying more distant, such as HuC positive cells show a cytoplasmic expression of Sox2. Immuno-labeling for GnRH localized cell bodies (Fig. 8G, green, box, asterisk) and processes (Fig. 8G, green, arrows). Strikingly, a small population of cells positive for GnRH also immuno-labeled with Sox2 (Fig. 8H,I, boxed area, asterisk). Like HuC, GnRH co-localizes with cytoplasmic expression of Sox2 in cells located several cell diameters from the ventricular surface (Fig. 8A,D,G). Thus GnRH positive neurons are located in a neurogenic niche in the PPa of the hypothalamus in the brain of the adult zebrafish.Fig. 8.


Zebrafish adult-derived hypothalamic neurospheres generate gonadotropin-releasing hormone (GnRH) neurons.

Cortés-Campos C, Letelier J, Ceriani R, Whitlock KE - Biol Open (2015)

The hypothalamus of the adult zebrafish contains newly differentiated GnRH cells. Horizontal sections showing the anterior region of the parvocellular preoptic nucleus (PPa), with DAPI labeling (blue). (A-C) HuC labeling in a HuC:GFP background (green, arrows) and vimentin labeling (red, arrowhead) do not co-localize. The vimentin positive cells line the edges of the ventricles. (D-F) HuC (green) and Sox2 labeling (red, arrows) co-localizes in some cells (arrows) located closer to the border of the ventricle (arrows). (G-I) Section showing GnRH cell body labeling (green, box, asterisk) and labeling of processes (green, arrows), Sox2 labeling (red, box, asterisk) in cells located several cell diameters from the border of the ventricle (arrows) co-localizes with the GnRH labeling (merge). All Images: anterior is to the left. Scale bar=30 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4582115&req=5

BIO010447F8: The hypothalamus of the adult zebrafish contains newly differentiated GnRH cells. Horizontal sections showing the anterior region of the parvocellular preoptic nucleus (PPa), with DAPI labeling (blue). (A-C) HuC labeling in a HuC:GFP background (green, arrows) and vimentin labeling (red, arrowhead) do not co-localize. The vimentin positive cells line the edges of the ventricles. (D-F) HuC (green) and Sox2 labeling (red, arrows) co-localizes in some cells (arrows) located closer to the border of the ventricle (arrows). (G-I) Section showing GnRH cell body labeling (green, box, asterisk) and labeling of processes (green, arrows), Sox2 labeling (red, box, asterisk) in cells located several cell diameters from the border of the ventricle (arrows) co-localizes with the GnRH labeling (merge). All Images: anterior is to the left. Scale bar=30 μm.
Mentions: To investigate whether the hypothalamus of the intact adult zebrafish contains potential GnRH progenitors we used adult male zebrafish that had been selected via the mating training protocol and sacrificed them to obtain brains for sectioning. Cryostat sections were analyzed using antibodies against GnRH, Sox2, and HuC. Cells positive for HuC labeling (Fig. 8A, arrows) were localized in the anterior region of the parvocellular preoptic nucleus (PPa) and these cells were distinct from the vimentin positive glia (tanycytes) lining the ventricle (Fig. 8B,C, red, arrowheads). Immuno-localization of the progenitor marker Sox2 showed nuclear label in the cells lining the ventricle (Fig. 8E, boxed area for example) and cytoplasmic localization in cells located several cell diameters from the ventricle (Fig. 8E,F, arrows). A subset of the Sox2 positive cells co-localized with HuC positive cells (Fig. 8F) a marker for newly generated neurons in agreement the role of Sox2 as a neural progenitor. While cells lying adjacent to the ventricle had nuclear localization Sox2, cells lying more distant, such as HuC positive cells show a cytoplasmic expression of Sox2. Immuno-labeling for GnRH localized cell bodies (Fig. 8G, green, box, asterisk) and processes (Fig. 8G, green, arrows). Strikingly, a small population of cells positive for GnRH also immuno-labeled with Sox2 (Fig. 8H,I, boxed area, asterisk). Like HuC, GnRH co-localizes with cytoplasmic expression of Sox2 in cells located several cell diameters from the ventricular surface (Fig. 8A,D,G). Thus GnRH positive neurons are located in a neurogenic niche in the PPa of the hypothalamus in the brain of the adult zebrafish.Fig. 8.

Bottom Line: Using this system, we show that neurospheres derived from the adult hypothalamus can be maintained in culture and subsequently differentiate glia and neurons.Finally, we show in vivo that a neurogenic niche in the hypothalamus contains GnRH positive neurons.Thus, we demonstrated for the first time that neurospheres can be derived from the hypothalamus of the adult zebrafish and that these neural progenitors are capable of producing GnRH containing neurons.

View Article: PubMed Central - PubMed

Affiliation: Centro Interdisciplinario de Neurociencia de Valparaíso (CINV), Facultad de Ciencias, Universidad de Valparaíso, Pasaje Harrington 269, Valparaíso 2340000, Chile Whitehead Institute for Biomedical Research (WIBR), 9 Cambridge Center, Cambridge, MA 02142, USA.

No MeSH data available.


Related in: MedlinePlus