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通过激活培养的CHO-K1细胞中的内源性P2嘌呤受体增加细胞内钙含量。

Increases in intracellular calcium via activation of an endogenous P2-purinoceptor in cultured CHO-K1 cells.

作者信息

Iredale P A, Hill S J

机构信息

Department of Physiology and Pharmacology, Medical School, Queen's Medical Centre, Nottingham.

出版信息

Br J Pharmacol. 1993 Dec;110(4):1305-10. doi: 10.1111/j.1476-5381.1993.tb13960.x.

Abstract
  1. Increases in intracellular calcium ([Ca2+]i) were measured in chinese hamster cultured ovary cells (clone, CHO-K1), by use of the fluorescent, calcium-sensitive dye, fura-2. 2. Addition of both ATP and UTP elicited rapid increases in [Ca2+]i due to mobilization from intracellular stores and calcium entry across the plasma membrane. 3. Omission of calcium from the extracellular medium and pre-incubation with the inorganic calcium channel blocker, nickel (Ni2+) prevented the calcium entry components of the responses. 4. Investigation of the concentration-response relationships of various analogues of ATP suggests the presence of a purinoceptor which cannot be characterized as P2X or P2Y. In addition, there appears to be a sub-population of P2Y-purinoceptors which do not cross-react with the 'nucleotide' receptor population. 5. Cross-desensitization and additivity experiments suggest that both ATP and UTP activate the same receptor. 6. Pre-incubation with the tumour-promoting agent, beta-phorbol-12,13 dibutyrate (PDBu), caused a reduction in the increases in [Ca2+]i, suggesting a role for protein kinase C in feedback inhibition of purinoceptor responses in this cell line. 7. In summary, we present evidence for the existence of an endogenous P2U-purinoceptor (or 'nucleotide receptor') which is linked to increases in [Ca2+]i in CHO-K1 cells.
摘要
  1. 利用荧光钙敏染料fura - 2,在中国仓鼠卵巢培养细胞(克隆株,CHO - K1)中测量细胞内钙浓度([Ca2+]i)的增加情况。

  2. ATP和UTP的添加均引起[Ca2+]i的快速增加,这是由于细胞内钙库释放以及钙通过质膜内流所致。

  3. 去除细胞外培养基中的钙并预先用无机钙通道阻滞剂镍(Ni2+)孵育,可阻止反应中的钙内流成分。

  4. 对ATP各种类似物浓度 - 反应关系的研究表明存在一种嘌呤受体,其不能被归类为P2X或P2Y。此外,似乎存在一群不与“核苷酸”受体群体发生交叉反应的P2Y嘌呤受体。

  5. 交叉脱敏和相加实验表明ATP和UTP激活相同的受体。

  6. 预先用肿瘤促进剂β - 佛波醇 - 12,13 - 二丁酸酯(PDBu)孵育,导致[Ca2+]i增加减少,表明蛋白激酶C在该细胞系中对嘌呤受体反应的反馈抑制中起作用。

  7. 总之,我们提供了证据表明存在一种内源性P2U嘌呤受体(或“核苷酸受体”),其与CHO - K1细胞中[Ca2+]i的增加相关。

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