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儿茶酚胺能RCSN - 3细胞系:一种研究多巴胺代谢的模型。

The catecholaminergic RCSN-3 cell line: a model to study dopamine metabolism.

作者信息

Paris Irmgard, Lozano Jorge, Cardenas Sergio, Perez-Pastene Carolina, Saud Katherine, Fuentes Patricio, Caviedes Pablo, Dagnino-Subiabre Alexies, Raisman-Vozari Rita, Shimahara Takeshi, Kostrzewa John P, Chi David, Kostrzewa Richard M, Caviedes Raul, Segura-Aguilar Juan

机构信息

Molecular and Clinical Pharmacology, ICBM, Faculty of Medicine, Casilla 70000, Santiago-7, Chile.

出版信息

Neurotox Res. 2008 May-Jun;13(3-4):221-30. doi: 10.1007/BF03033505.

Abstract

RCSN-3 cells are a cloned cell line derived from the substantia nigra of an adult rat. The cell line grows in monolayer and does not require differentiation to express catecholaminergic traits, such as (i) tyrosine hydroxylase; (ii) dopamine release; (iii) dopamine transport; (iv) norepinephrine transport; (v) monoamine oxidase (MAO)-A expression, but not MAO-B; (vi) formation of neuromelanin; (vii) VMAT-2 expression. In addition, this cell line expresses serotonin transporters, divalent metal transporter, DMT1, dopamine receptor 1 mRNA under proliferating conditions, and dopamine receptor 5 mRNA after incubation with dopamine or dicoumarol. Expression of dopamine receptors D(2), D(3) and D(4) mRNA were not detected in proliferating cells or when the cells were treated with dopamine, CuSO(4), dicoumarol or dopamine-copper complex. Angiotensin II receptor mRNA was also found to be expressed, but it underwent down regulation in the presence of aminochrome. Total quinone reductase activity corresponded 94% to DT-diaphorase. The cells also express antioxidant enzymes such as superoxide dismutase, catalase and glutathione peroxidase. This cell line is a suitable in vitro model for studies of dopamine metabolism, since under proliferating conditions the cells express all the pertinent markers.

摘要

RCSN - 3细胞是一种从成年大鼠黑质中克隆得到的细胞系。该细胞系以单层形式生长,无需分化即可表达儿茶酚胺能特性,如:(i)酪氨酸羟化酶;(ii)多巴胺释放;(iii)多巴胺转运;(iv)去甲肾上腺素转运;(v)单胺氧化酶(MAO)-A表达,但不表达MAO - B;(vi)神经黑色素形成;(vii)囊泡单胺转运体2(VMAT - 2)表达。此外,该细胞系在增殖条件下表达5-羟色胺转运体、二价金属转运体、二价金属离子转运体1(DMT1)、多巴胺受体1信使核糖核酸(mRNA),在与多巴胺或双香豆素孵育后表达多巴胺受体5 mRNA。在增殖细胞中或用多巴胺、硫酸铜、双香豆素或多巴胺 - 铜复合物处理细胞时,未检测到多巴胺受体D(2)、D(3)和D(4) mRNA的表达。还发现血管紧张素II受体mRNA有表达,但在氨基色素存在下其表达下调。总醌还原酶活性与DT - 黄递酶的活性相当,为94%。这些细胞还表达抗氧化酶,如超氧化物歧化酶、过氧化氢酶和谷胱甘肽过氧化物酶。由于在增殖条件下这些细胞表达所有相关标志物,因此该细胞系是研究多巴胺代谢的合适体外模型。

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