Establishment of model cell lines for analyzing relation of angiotensin II-caused diseases with long term potentiation

分析血管紧张素II引起的疾病与长时程增强关系的模型细胞系的建立

• 批准号: 05660384
• 负责人: MIYAZAKI Hitoshi
• 金额: $ 1.34万
• 依托单位: University of Tsukuba
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05660384/
• 关键词: Angiotensin II Receptor; AtT20; ACTH; Phospholipase C; Calcium Channel; Adenylate Cyclase; アンジオテンシンII; 受容体; 長期増強

1.We have established an AtT20 cell line stably expressing the recombinant human angiotensin II (Ang II) receptor type 1 (AT1) to provide a model for analyzing the relation of Ang II-caused diseases with long term potentiation. AtT20 cells are derived from mouse anterior pituitary cells secreting adrenocorticotropin (ACTH). Following finding were obtained concerning characteristics of this cell line, which are essential for the purpose of this study.(1) In these transfected cells, Ang II induced activation of phospholipase C and voltage-dependent calcium channel, and inhibition of adenylate cyclase activity through the AT1 receptor. Among these the former two pathways were found to relate with Ang II-induced ACTH secretion.(2) This cell line was found to be a good model for elucidating the cross-talk between Ang II and corticotropin releasing factor (CRF) signalling pathways, because Ang II had a synergistic effect with CRF,whose functions are mediated by cAMP,on ACTH secretion.2.We tried to develop a microperfusion system to investigate whether Ang II induces a phenomenon in these endocrine cells corresponding to long term potentiation observed in neural cells. Unfortunately, reliable results were not obtained in this system since high levels of basal ACTH secretion prevented us from selectively detecting the amount of Ang II-induced ACTH secretion. We are now looking for optimal conditions in which the long term potentiation is observed using the cells cultured in dishes but not those filled in micropipette in the microperfusion system.

1.我们建立了稳定表达重组人血管紧张素II （Ang II）受体1型（AT1）的at20细胞系，为分析Ang II引起的疾病与长期增强的关系提供了模型。AtT20细胞来源于分泌促肾上腺皮质激素（ACTH）的小鼠垂体前叶细胞。关于该细胞系的特性，获得了以下发现，这对于本研究的目的至关重要。(1)在这些转染的细胞中，Ang II通过AT1受体诱导磷脂酶C和电压依赖性钙通道的激活，并抑制腺苷酸环化酶的活性。其中发现前两条通路与Ang ii诱导的ACTH分泌有关。(2)由于Ang II与促肾上腺皮质激素释放因子（CRF）具有协同作用，而CRF的功能是由cAMP介导的，因此该细胞系被发现是阐明Ang II与促肾上腺皮质激素释放因子（CRF）信号通路之间的串扰的良好模型。我们试图开发一种微灌注系统来研究Ang II是否在这些内分泌细胞中诱导了一种与在神经细胞中观察到的长期增强相对应的现象。不幸的是，在这个系统中没有得到可靠的结果，因为高水平的基底ACTH分泌阻止了我们选择性地检测Ang ii诱导的ACTH分泌量。我们现在正在寻找最佳条件，在这种条件下，用培养皿培养的细胞来观察长期增强，而不是在微灌注系统中用微管填充的细胞。

项目成果

Nishimatsu Shin-ichiro: "Isolation and characterization of two alternatively spliced complementary DNAs encoding a xenopus laevis angiotensin II receptor" Biochim.Biophys.Acta. (accepted). (1994)

Nishimatsu Shin-ichiro：“编码非洲爪蟾血管紧张素 II 受体的两个选择性剪​​接互补 DNA 的分离和表征”Biochim.Biophys.Acta。