Limits...
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

Hypothalamic progenitors generate glial and neuronal cells. (A-F) Neurospheres differentiated after 7 days in vitro. (A-C) Differentiated cells surround the core of cells positive for nestin (arrow head) and extend cellular projections that are reactive to GFAP (arrows). (D-F) The reduced number of Sox2 positive cells (arrows) and the high number of cell processes reactive to Vimentin (arrow heads) suggests that progenitors have differentiated into glial cells. (G-L) Neurospheres reactive to GFAP show (G-I) a reduced number of cells positive for PCNA (arrows), (J-L) high number of neurons (arrows) and differentiated glial cells (arrow heads). Scale bar 25 μm, n=3 plates derived from different cell cultures.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4582115&req=5

BIO010447F4: Hypothalamic progenitors generate glial and neuronal cells. (A-F) Neurospheres differentiated after 7 days in vitro. (A-C) Differentiated cells surround the core of cells positive for nestin (arrow head) and extend cellular projections that are reactive to GFAP (arrows). (D-F) The reduced number of Sox2 positive cells (arrows) and the high number of cell processes reactive to Vimentin (arrow heads) suggests that progenitors have differentiated into glial cells. (G-L) Neurospheres reactive to GFAP show (G-I) a reduced number of cells positive for PCNA (arrows), (J-L) high number of neurons (arrows) and differentiated glial cells (arrow heads). Scale bar 25 μm, n=3 plates derived from different cell cultures.

Mentions: In order to determine whether the neurospheres could generate different cell types, they were cultured in differentiation media (see Materials and Methods) on a L-poly-l-lysine/laminin coated substrate (Fig. 2A4-6, upper panel). Cells were then fixed after 7 days and processed for different markers. The expression pattern of the IF proteins nestin (Fig. 4A,C, green), GFAP (Fig. 4B,C, red) and vimentin (Fig. 4E,F red, arrowhead) changed as glia differentiated and began to elongate their process (Fig. 4A-C arrows, D-F, arrowheads). Concurrently, the number of cells expressing Sox2 (Fig. 4D,F, green, arrow) and PCNA (Fig. 4G,I, green, arrow) decreased from 0.87±0.09 to 0.06±0.05 (Sox2+/total cells) and from 0.32±0.09 to 0.07±0.04 (PCNA+/total cells), respectively. In agreement with the differentiation of newborn neurons, the number of cells expressing HuC (Fig. 4J,L, green, arrows) increased (0.61±0.04 HuC+/total cells). The change in morphology and in expression of markers correlated with the differentiation of neurons and glia was observed in all differentiated cultures (n=3), and is consistent with media-induced differentiation observed in other studies (Louis and Reynolds, 2005).Fig. 4.


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

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

Hypothalamic progenitors generate glial and neuronal cells. (A-F) Neurospheres differentiated after 7 days in vitro. (A-C) Differentiated cells surround the core of cells positive for nestin (arrow head) and extend cellular projections that are reactive to GFAP (arrows). (D-F) The reduced number of Sox2 positive cells (arrows) and the high number of cell processes reactive to Vimentin (arrow heads) suggests that progenitors have differentiated into glial cells. (G-L) Neurospheres reactive to GFAP show (G-I) a reduced number of cells positive for PCNA (arrows), (J-L) high number of neurons (arrows) and differentiated glial cells (arrow heads). Scale bar 25 μm, n=3 plates derived from different cell cultures.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

BIO010447F4: Hypothalamic progenitors generate glial and neuronal cells. (A-F) Neurospheres differentiated after 7 days in vitro. (A-C) Differentiated cells surround the core of cells positive for nestin (arrow head) and extend cellular projections that are reactive to GFAP (arrows). (D-F) The reduced number of Sox2 positive cells (arrows) and the high number of cell processes reactive to Vimentin (arrow heads) suggests that progenitors have differentiated into glial cells. (G-L) Neurospheres reactive to GFAP show (G-I) a reduced number of cells positive for PCNA (arrows), (J-L) high number of neurons (arrows) and differentiated glial cells (arrow heads). Scale bar 25 μm, n=3 plates derived from different cell cultures.
Mentions: In order to determine whether the neurospheres could generate different cell types, they were cultured in differentiation media (see Materials and Methods) on a L-poly-l-lysine/laminin coated substrate (Fig. 2A4-6, upper panel). Cells were then fixed after 7 days and processed for different markers. The expression pattern of the IF proteins nestin (Fig. 4A,C, green), GFAP (Fig. 4B,C, red) and vimentin (Fig. 4E,F red, arrowhead) changed as glia differentiated and began to elongate their process (Fig. 4A-C arrows, D-F, arrowheads). Concurrently, the number of cells expressing Sox2 (Fig. 4D,F, green, arrow) and PCNA (Fig. 4G,I, green, arrow) decreased from 0.87±0.09 to 0.06±0.05 (Sox2+/total cells) and from 0.32±0.09 to 0.07±0.04 (PCNA+/total cells), respectively. In agreement with the differentiation of newborn neurons, the number of cells expressing HuC (Fig. 4J,L, green, arrows) increased (0.61±0.04 HuC+/total cells). The change in morphology and in expression of markers correlated with the differentiation of neurons and glia was observed in all differentiated cultures (n=3), and is consistent with media-induced differentiation observed in other studies (Louis and Reynolds, 2005).Fig. 4.

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