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Hypothalamic neurospheres differentiate to mature neurons expressing appetite-related neuropeptides.

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posted on 2011-05-11, 01:50 authored by Lígia Sousa-Ferreira, Ana Rita Álvaro, Célia Aveleira, Magda Santana, Inês Brandão, Sebastian Kügler, Luís Pereira de Almeida, Cláudia Cavadas

Images of hypothalamic neurons cultures obtained from plated neurospheres after 18 days in differentiation conditions. NPY vesicular immunoreactivity (arrow-heads) (A) and NPY-positive neurons (arrows) (A-merge); AGRP vesicular immunoreactivity (arrow-heads) (C) and AGRP-positive neurons (arrows) (C-merge); POMC vesicular immunoreactivity (arrow-heads) (E) and POMC-positive neurons (arrows) (E-merge); CART vesicular immunoreactivity (arrow-heads) (G) and CART-positive neurons (arrows) (G-merge); Orexin-A immunoreactivity (I) and Orexin-A positive neurons (arrows) (I-merge). (B, D, F, H, J) Relative mRNA expression of feeding-related neuropeptides in hypothalamic neuronal cultures. The expression of NPY, CART and Orexin-A increases in differentiated hypothalamic cultures while the expression of AGRP and POMC is unchanged. Mean ± SEM. ns p>0.05, * p<0.05, ** p<0.01 and *** p<0.001, compared to non-differentiated hypothalamic neurospheres. n = 4/5; t-test. DAPI, nuclear staining; non-dif, non-differentiated hypothalamic neurospheres; 18d-dif, hypothalamic neurons culture after 18 days of differentiation.