RESUMEN

Growth arrest-specific 1 (Gas1) is a pleiotropic protein that induces apoptosis of tumor cells and has important roles during development. Recently, the presence of two forms of Gas1 was reported: one attached to the cell membrane by a GPI anchor; and a soluble extracellular form shed by cells. Previously, we showed that Gas1 is expressed in different areas of the adult mouse CNS. Here, we report the levels of Gas1 mRNA protein in different regions and analyzed its expressions in glutamatergic, GABAergic, and dopaminergic neurons. We found that Gas1 is expressed in GABAergic and glutamatergic neurons in the Purkinje-molecular layer of the cerebellum, hippocampus, thalamus, and fastigial nucleus, as well as in dopaminergic neurons of the substantia nigra. In all cases, Gas1 was found in the cell bodies, but not in the neuropil. The Purkinje and the molecular layers show the highest levels of Gas1, whereas the granule cell layer has low levels. Moreover, we detected the expression and release of Gas1 from primary cultures of Purkinje cells and from hippocampal neurons as well as from neuronal cell lines, but not from cerebellar granular cells. In addition, using SH-SY5Y cells differentiated with retinoic acid as a neuronal model, we found that extracellular Gas1 promotes neurite outgrowth, increases the levels of tyrosine hydroxylase, and stimulates the inhibition of GSK3ß. These findings demonstrate that Gas1 is expressed and released by neurons and promotes differentiation, suggesting an important role for Gas1 in cellular signaling in the CNS.

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RESUMEN

Retinoic acid (RA), a metabolite of retinol, is one of the most biologically active forms of retinoid and plays vital roles in embryonic development and in the regulation of cell proliferation and differentiation. Knowing that liposomes simulate cell membranes and that hydrogel is an ideal delivery vehicle for topical medicine, liposome-hydrogel is a novel preparation that has synergistic advantages over each component separately. Our objective was to investigate the characteristics of RA liposome-hydrogel. For quality control of the RA-loaded liposomes, we measured their morphology, particle size, Zeta-potential, and entrapment efficiency. Then we determined the viscosity of RA liposome-hydrogel. Next, the diffusion through mouse skin was explored, followed by investigation of the mRNA expression levels of Ker18, REX1, and α-FP using Q-PCR. The results showed that RA liposome-hydrogel penetrates the mouse skin effectively. The permeation rates were: Qn (%) of RA liposome-hydrogel < Qn(%) of RA-loaded liposome < Qn (%) of RA. The mRNA expression levels were dose-dependent and the effective dose decreased between vehicles due to their different release rates. F9 mouse teratocarcinoma stem cells were an ideal model to explore the mechanism of RA liposome-hydrogel in stem cell differentiation.

O ácido retinóico (RA) é um metabolito de retinol. Ele também é uma das formas mais biologicamente ativas de retinóide. Desempenha papel vital no desenvolvimento embrionário e na regulação da proliferação e diferenciação celular. Sabendo-se que lipossomas simulam a membrana das células e que hidrogel é um sistema ideal para o medicamento tópico, o lipossoma-hidrogel é uma nova preparação, que apresenta vantagens sinérgicas em relação a cada um dos componentes separados. Nosso objetivo foi investigar as características de RA lipossoma-hidrogel. A fim de controlar a qualidade do lipossoma carregado com RA, medimos morfologia, tamanho das partículas, potencial zeta e eficiência de retenção. Em seguida, determinou-se a viscosidade de RA lipossoma-hidrogel. Em seguida, avaliou-se a sua difusão através da pele de camundongos, seguida da investigação dos níveis da expressão de mRNA de Ker18, REX e de α-FP, utilizando-se Q-PCR. Os resultados mostraram que RA lipossoma-hidrogel pode penetrar na pele do camundongo de forma eficaz. As taxas de permeação foram: Qn (%) de RA lipossoma-hidrogel<Qn(%) de lipossoma RA- carregado <Qn (%) de RA. Os níveis de expressão de mRNA foram dependentes de dose e a dose efetiva diminuiu entre os veículos devido às diferentes taxas de liberação, As células estaminais de teratocarcinoma F9 de camundongo mostraram-se como modelo ideal para explorar o mecanismo de diferenciaçãode células tronco pelo RA lipossoma-hidrogel.

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