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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.

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Related in: MedlinePlus

Hypothalamic progenitors can differentiate GnRH neurons. (A-C) Neurospheres differentiated for 7 days in vitro without hormonal stimulus. Neurospheres generate GnRH neurons (arrows). (D-F) Differentiated 7 days in vitro neurospheres, stimulated for 5 days with 10 μM testosterone. Testosterone increases the number of GnRH neurons in NSC cultures (arrows). (G-I) Differentiated 7 days in vitro neurospheres, stimulated for 5 days with 10 nM GnRH increases the number of GnRH neurons in NSC culture (arrows). Scale bar 25 μm.
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BIO010447F5: Hypothalamic progenitors can differentiate GnRH neurons. (A-C) Neurospheres differentiated for 7 days in vitro without hormonal stimulus. Neurospheres generate GnRH neurons (arrows). (D-F) Differentiated 7 days in vitro neurospheres, stimulated for 5 days with 10 μM testosterone. Testosterone increases the number of GnRH neurons in NSC cultures (arrows). (G-I) Differentiated 7 days in vitro neurospheres, stimulated for 5 days with 10 nM GnRH increases the number of GnRH neurons in NSC culture (arrows). Scale bar 25 μm.

Mentions: Once the differentiation of neurons and glia was confirmed, subsequent experiments were carried out to determine whether the differentiated neurospheres were immunoreactive for GnRH. Cells from disaggregated neurospheres differentiated 7 days in vitro without hormonal stimulus (Fig. 2A3-6, bottom panel), were fixed and processed for GnRH immunoreactivity using the BB8 antibody (Kah et al., 1986). Strikingly, differentiated progenitor cultures contained GnRH positive cells (Fig. 5B,C, red, arrow) that were also positive for neurofilament (Fig. 5A,C, green, arrow) confirming their identity as GnRH positive neurons.Fig. 5.


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 can differentiate GnRH neurons. (A-C) Neurospheres differentiated for 7 days in vitro without hormonal stimulus. Neurospheres generate GnRH neurons (arrows). (D-F) Differentiated 7 days in vitro neurospheres, stimulated for 5 days with 10 μM testosterone. Testosterone increases the number of GnRH neurons in NSC cultures (arrows). (G-I) Differentiated 7 days in vitro neurospheres, stimulated for 5 days with 10 nM GnRH increases the number of GnRH neurons in NSC culture (arrows). Scale bar 25 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

BIO010447F5: Hypothalamic progenitors can differentiate GnRH neurons. (A-C) Neurospheres differentiated for 7 days in vitro without hormonal stimulus. Neurospheres generate GnRH neurons (arrows). (D-F) Differentiated 7 days in vitro neurospheres, stimulated for 5 days with 10 μM testosterone. Testosterone increases the number of GnRH neurons in NSC cultures (arrows). (G-I) Differentiated 7 days in vitro neurospheres, stimulated for 5 days with 10 nM GnRH increases the number of GnRH neurons in NSC culture (arrows). Scale bar 25 μm.
Mentions: Once the differentiation of neurons and glia was confirmed, subsequent experiments were carried out to determine whether the differentiated neurospheres were immunoreactive for GnRH. Cells from disaggregated neurospheres differentiated 7 days in vitro without hormonal stimulus (Fig. 2A3-6, bottom panel), were fixed and processed for GnRH immunoreactivity using the BB8 antibody (Kah et al., 1986). Strikingly, differentiated progenitor cultures contained GnRH positive cells (Fig. 5B,C, red, arrow) that were also positive for neurofilament (Fig. 5A,C, green, arrow) confirming their identity as GnRH positive neurons.Fig. 5.

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