Construction of the subtracted cDNA library of striatal neurons treated with long-term morphine / 中国医学科学杂志(英文版)
Chinese Medical Sciences Journal
; (4): 54-59, 2011.
Artículo
en Inglés
| WPRIM (Pacífico Occidental)
| ID: wpr-299413
Biblioteca responsable:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To construct a morphine tolerance model in primarily cultured striatal neurons, and screen the differentially expressed genes in this model using suppression subtractive hybridization (SSH).</p><p><b>METHODS</b>Sbtracted cDNA libraries were constructed using SSH from normal primarily cultured striatal neurons and long-term morphine treated striatal neurons (10-5 mol/L for 72 hours). To check reliability of the cell culture model, RT-PCR was performed to detect the cAMP-responsive element-binding protein (CREB) mRNA expression. The subtracted clones were prescreened by PCR. The clones containing inserted fragments from forward libraries were sequenced and submitted to GenBank for homology analysis. And the expression levels of genes of interest were confirmed by RT-PCR. Results CREB mRNA expression showed a significant increase in morphine treated striatal neurons (62.85 ± 1.98) compared with normal striatal neurons (28.43 ± 1.46, P < 0.01). Thirty-six clones containing inserted fragments were randomly chosen for sequence analysis. And the 36 clones showed homology with 19 known genes and 2 novel genes. The expression of 2 novel genes, mitochondrial carrier homolog 1 (Mtch1; 96.81 ± 2.04 vs. 44.20 ± 1.31, P < 0.01) and thymoma viral proto-oncogene 1 (Akt1; 122.10 ± 2.17 vs. 50.11 ± 2.01, P < 0.01), showed a significant increase in morphine-treated striatal neurons compared with normal striatal neurons.</p><p><b>CONCLUSIONS</b>A reliable differential cDNA library of striatal neurons treated with long-term morphine is constructed. Mtch1 and Akt1 might be the candidate genes for the development of morphine tolerance.</p>
Texto completo:
Disponible
Base de datos:
WPRIM (Pacífico Occidental)
Asunto principal:
Farmacología
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Fisiología
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Datos de Secuencia Molecular
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Biblioteca de Genes
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Células Cultivadas
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Ratas Wistar
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Cuerpo Estriado
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Reacción en Cadena de la Polimerasa de Transcriptasa Inversa
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Perfilación de la Expresión Génica
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Biología Celular
Límite:
Animales
Idioma:
Inglés
Revista:
Chinese Medical Sciences Journal
Año:
2011
Tipo del documento:
Artículo