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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

GnRH receptors are expressed in undifferentiated neurospheres. RT-PCR analyses of the four zfGnRHRs and zfAR. All zfGnRHRs are expressed in the brain of adult zebrafish. zfGnRHR1, zfGnRHR2 and zfGnRHR4 are expressed in undifferentiated neurospheres (7 days in vitro). UNS, neurospheres; RT+, positive reverse-transcription; RT−, negative reverse transcription. β-actin and Gapdh: house-keeping genes.
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BIO010447F7: GnRH receptors are expressed in undifferentiated neurospheres. RT-PCR analyses of the four zfGnRHRs and zfAR. All zfGnRHRs are expressed in the brain of adult zebrafish. zfGnRHR1, zfGnRHR2 and zfGnRHR4 are expressed in undifferentiated neurospheres (7 days in vitro). UNS, neurospheres; RT+, positive reverse-transcription; RT−, negative reverse transcription. β-actin and Gapdh: house-keeping genes.

Mentions: Because the neurospheres responded to testosterone and GnRH we examined whether the undifferentiated neurospheres (Fig. 2A4) express one or more of the androgen and/or GnRH receptors. We amplified zfGnRHR1–zfGnRHR4 from undifferentiated neurospheres and whole brain of adult fish (Fig. 7) using previously published primers (Tello et al., 2008). The products resulting from the amplifications were sequenced and their identity confirmed. In agreement with the literature, all four zfGnRHRs were expressed in the whole brain tissue with zfGnRHR3 being expressed at very low levels (Fig. 7, Brain). In contrast, only zfGnRHR1, -2, and -4 were expressed in undifferentiated neurospheres (Fig. 7, UNS). The absence of zfGnRHR3 expression in vitro is consistent with previous reports showing zfGnRHR3 is highly expressed in the eye but is almost absent in the brain (Tello et al., 2008).Fig. 7.


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

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

GnRH receptors are expressed in undifferentiated neurospheres. RT-PCR analyses of the four zfGnRHRs and zfAR. All zfGnRHRs are expressed in the brain of adult zebrafish. zfGnRHR1, zfGnRHR2 and zfGnRHR4 are expressed in undifferentiated neurospheres (7 days in vitro). UNS, neurospheres; RT+, positive reverse-transcription; RT−, negative reverse transcription. β-actin and Gapdh: house-keeping genes.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

BIO010447F7: GnRH receptors are expressed in undifferentiated neurospheres. RT-PCR analyses of the four zfGnRHRs and zfAR. All zfGnRHRs are expressed in the brain of adult zebrafish. zfGnRHR1, zfGnRHR2 and zfGnRHR4 are expressed in undifferentiated neurospheres (7 days in vitro). UNS, neurospheres; RT+, positive reverse-transcription; RT−, negative reverse transcription. β-actin and Gapdh: house-keeping genes.
Mentions: Because the neurospheres responded to testosterone and GnRH we examined whether the undifferentiated neurospheres (Fig. 2A4) express one or more of the androgen and/or GnRH receptors. We amplified zfGnRHR1–zfGnRHR4 from undifferentiated neurospheres and whole brain of adult fish (Fig. 7) using previously published primers (Tello et al., 2008). The products resulting from the amplifications were sequenced and their identity confirmed. In agreement with the literature, all four zfGnRHRs were expressed in the whole brain tissue with zfGnRHR3 being expressed at very low levels (Fig. 7, Brain). In contrast, only zfGnRHR1, -2, and -4 were expressed in undifferentiated neurospheres (Fig. 7, UNS). The absence of zfGnRHR3 expression in vitro is consistent with previous reports showing zfGnRHR3 is highly expressed in the eye but is almost absent in the brain (Tello et al., 2008).Fig. 7.

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