Overexpression of the myristoylated alanine‐rich C kinase substrate decreases uptake and K+‐evoked release of noradrenaline in the human neuroblastoma SH‐SY5Y
Overexpression of the myristoylated alanine‐rich C kinase substrate decreases uptake and K+‐evoked release of noradrenaline in the human neuroblastoma SH‐SY5Y
复制标题
人神经母细胞瘤 SH-SY5Y 中肉豆蔻酰化富含丙氨酸的 C 激酶底物的过度表达降低了去甲肾上腺素的摄取和 K+ 诱发的释放
DOI：
10.1046/j.0953-816x.2001.01466.x
复制
发表时间：
2001
影响因子：
3.4
通讯作者：
P. Vaughan
中科院分区：
文献类型：
--
作者：
M. Hartness;J. Wade;J. Walker;P. Vaughan
本研究的目的是研究肉豆蔻酰化富含丙氨酸的C激酶底物（MARCKS）在人神经母细胞瘤细胞系SH‐ SY 5 Y中去甲肾上腺素摄取和释放机制中的可能作用。 通过用含有正义方向的MARCKS cDNA的pCEP 4转染SH-SY 5 Y制备显示MARCKS两倍过表达的稳定细胞系。与未转染的SH-SY 5 Y和仅用pCEP 4转染的SH-SY 5 Y（模拟转染）相比，该细胞系显示神经丝或去甲肾上腺素能大致密核心囊泡标志物的表达没有变化。类似地，在这三种细胞系之间未检测到细胞生长速率的差异。相比之下，与模拟转染的SH-SY 5 Y相比，过表达MARCKS的细胞系对去甲肾上腺素的特异性摄取和去极化诱发（100 mm K+）释放的抑制约为50%。 用12-O-十四酰基佛波醇13-乙酸酯（100 nm）预处理增强的K+诱发的去甲肾上腺素释放也被抑制50%。 相反，卡巴胆碱诱发的去甲肾上腺素释放不受影响。因此，在SH-SY 5 Y细胞中，MARCKS的过表达导致K+诱发的去甲肾上腺素释放减少，可能是通过增加肌动蛋白交联阻止含有去甲肾上腺素的大致密核心囊泡响应去极化而移动到质膜。
The aim of this study was to investigate a possible role of the myristoylated alanine‐rich C kinase substrate (MARCKS) in the mechanism of noradrenaline uptake and release in the human neuroblastoma cell line SH‐SY5Y. A stable cell line showing a twofold overexpression of MARCKS was prepared by transfecting SH‐SY5Y with pCEP4 containing MARCKS cDNA in the sense orientation. This cell line showed no changes in the expression of neurofilaments or markers of noradrenergic large dense‐cored vesicles compared with both untransfected SH‐SY5Y and SH‐SY5Y transfected with pCEP4 only (mock transfected). Similarly, no differences in the rate of cell growth could be detected between these three cell lines. In contrast, specific uptake and depolarization‐evoked (100 mm K+) release of noradrenaline from the cell line overexpressing MARCKS was inhibited by approximately 50% compared with mock‐transfected SH‐SY5Y. K+‐evoked noradrenaline release enhanced by pretreatment with 12‐O‐tetradecanoylphorbol 13‐acetate (100 nm) was also inhibited by 50%. In contrast, carbachol‐evoked noradrenaline release was unaffected. Thus, in SH‐SY5Y cells, overexpression of MARCKS leads to a decrease in the K+‐evoked noradrenaline release possibly by increased actin cross‐linking preventing the movement of noradrenaline containing large dense‐cored vesicles to the plasma membrane in response to depolarization.
登录
查看更多内容
DOI：
10.1016/s0021-9258(19)67742-2
发表时间：
1991-03
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
B. McIlroy;J. Walters;P. Blackshear;J. Johnson
通讯作者：
B. McIlroy;J. Walters;P. Blackshear;J. Johnson
DOI：
10.1016/s0021-9258(18)98698-9
发表时间：
1991-08
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
D. Harlan;J. Graff;D. Stumpo;D. Stumpo;R. Eddy;T. Shows;J. Boyle;P. Blackshear
通讯作者：
D. Harlan;J. Graff;D. Stumpo;D. Stumpo;R. Eddy;T. Shows;J. Boyle;P. Blackshear
影响因子：
11.4
作者：
ALLEN, LAH;ADEREM, A
通讯作者：
ADEREM, A
影响因子：
3.9
作者：
ADEREM, A
通讯作者：
ADEREM, A