Human embryonic teratocarcinoma-derived Ntera2/cl.D1 (NT2) cells recapitulate many features of embryonic neuronal progenitor cells. Upon retinoic acid (RA) treatment they terminally differentiate into post-mitotic neuron-like cells (NT2-N), akin to human fetal neurons, thus representing an in vitro model of human neuron terminal differentiation. Experimental evidence also indicate NT2-N cultures as a potential source for cell transplantation therapy. The neurosteroids progesterone and its metabolite 3 alpha-hydroxy-5 alpha-pregnan-20-one (3 alpha,5 alpha-THP) promote neurogenesis and show anti-neurodegenerative properties. This study's aim was to assess the neurosteroidogenic competence of NT2 cells during RA-induced neuronal differentiation. Radioimmunoassay measurements revealed progesterone only in NT2-N Cultures (4 week RA). Accordingly, progesterone synthesis from H-3-pregnenolone was absent in NT2 cells and increased during RA exposure, being highest in NT2-N+ [H-3]-pregnenolone metabolism, yielding [H-3]-progesterone and [H-3]-5 alpha-dihydroprogesterone ([H-3]-5 alpha-DHP), was time-dependent and inhibited by trilostane, a 3 beta-hydroxysteroid-dehydrogenase (3 beta-HSD) inhibitor. Conversely, H-3-progesterone metabolism, which yielded [H-3]-5 alpha-DHP > [H-3]-3 beta,5 alpha-THP > [H-3]-3 alpha,5 alpha-THP, occurred at all time points examined, though showing a nadir in cultures treated with RA for 1 and 2 weeks. The differentiation-dependent increase of progesterone accumulation matched 3 beta-HSD type I mRNA expression and 3 beta-HSD immunoreactivity, that co-localized with Map2a/b- and GAD67 in NT2-N. Hence, in vitro differentiated human neurons, while retaining progesterone metabolic activity, also become competent in progesterone synthesis. These findings suggest an autocrine/paracrine role of neuronal progesterone, either on its own or through its 5 alpha-reduced metabolites, in fetal brain development and allow speculation that NT2-N-produced neurosteroids may contribute to the encouraging results of NT2-N transplants in animal models of neurodegenerative diseases. (C) 2009 Elsevier Inc. All rights reserved.

Pistritto, G., Papacleovoulou, G., Ragone, G., Di Cesare, S., Papaleo, V., Mason, J.I., et al. (2009). Differentiation-dependent progesterone synthesis and metabolism in NT2-N human neurons. EXPERIMENTAL NEUROLOGY, 217(2), 302-311 [10.1016/j.expneurol.2009.03.008].

Differentiation-dependent progesterone synthesis and metabolism in NT2-N human neurons

PISTRITTO, GIUSEPPA;BARBACCIA, MARIA LUISA
2009

Abstract

Human embryonic teratocarcinoma-derived Ntera2/cl.D1 (NT2) cells recapitulate many features of embryonic neuronal progenitor cells. Upon retinoic acid (RA) treatment they terminally differentiate into post-mitotic neuron-like cells (NT2-N), akin to human fetal neurons, thus representing an in vitro model of human neuron terminal differentiation. Experimental evidence also indicate NT2-N cultures as a potential source for cell transplantation therapy. The neurosteroids progesterone and its metabolite 3 alpha-hydroxy-5 alpha-pregnan-20-one (3 alpha,5 alpha-THP) promote neurogenesis and show anti-neurodegenerative properties. This study's aim was to assess the neurosteroidogenic competence of NT2 cells during RA-induced neuronal differentiation. Radioimmunoassay measurements revealed progesterone only in NT2-N Cultures (4 week RA). Accordingly, progesterone synthesis from H-3-pregnenolone was absent in NT2 cells and increased during RA exposure, being highest in NT2-N+ [H-3]-pregnenolone metabolism, yielding [H-3]-progesterone and [H-3]-5 alpha-dihydroprogesterone ([H-3]-5 alpha-DHP), was time-dependent and inhibited by trilostane, a 3 beta-hydroxysteroid-dehydrogenase (3 beta-HSD) inhibitor. Conversely, H-3-progesterone metabolism, which yielded [H-3]-5 alpha-DHP > [H-3]-3 beta,5 alpha-THP > [H-3]-3 alpha,5 alpha-THP, occurred at all time points examined, though showing a nadir in cultures treated with RA for 1 and 2 weeks. The differentiation-dependent increase of progesterone accumulation matched 3 beta-HSD type I mRNA expression and 3 beta-HSD immunoreactivity, that co-localized with Map2a/b- and GAD67 in NT2-N. Hence, in vitro differentiated human neurons, while retaining progesterone metabolic activity, also become competent in progesterone synthesis. These findings suggest an autocrine/paracrine role of neuronal progesterone, either on its own or through its 5 alpha-reduced metabolites, in fetal brain development and allow speculation that NT2-N-produced neurosteroids may contribute to the encouraging results of NT2-N transplants in animal models of neurodegenerative diseases. (C) 2009 Elsevier Inc. All rights reserved.
Pubblicato
Rilevanza internazionale
Articolo
Sì, ma tipo non specificato
Settore BIO/14
English
Con Impact Factor ISI
Ntera2/cl.D1(+) NT2-N; Retinoic acid; Neuronal differentiation; Neurosteroids; Progesterone; 3 alpha-hydroxy-5 alpha-pregnan-20-one (3 alpha-hydroxy-5 alpha-tetrahydroprogesterone/3 alpha-5 alpha-THP); 3 beta-hydroxysteroid-dehydrogenase (3 beta-HSD)
Pistritto, G., Papacleovoulou, G., Ragone, G., Di Cesare, S., Papaleo, V., Mason, J.I., et al. (2009). Differentiation-dependent progesterone synthesis and metabolism in NT2-N human neurons. EXPERIMENTAL NEUROLOGY, 217(2), 302-311 [10.1016/j.expneurol.2009.03.008].
Pistritto, G; Papacleovoulou, G; Ragone, G; Di Cesare, S; Papaleo, V; Mason, J; Barbaccia, Ml
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/2108/39197
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