Differential expression by nerve growth factor of two types of Ca2+ channels in rat phaeochromocytoma cell lines.
Differential expression by nerve growth factor of two types of Ca2+ channels in rat phaeochromocytoma cell lines.
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神经生长因子在大鼠嗜铬细胞瘤细胞系中两种类型 Ca2+ 通道的差异表达。
DOI：
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发表时间：
1990
期刊：
影响因子：
--
通讯作者：
H. Reuter
中科院分区：
文献类型：
--
作者：
M. M. Usowicz;Hartmiut Porzig;C. Becker;H. Reuter
1. Two clones of rat phaeochromocytoma PC12 cells have been used to study the expression of Ca2+ channels and their possible involvement in neuronal differentiation. One clone differentiated morphologically when exposed to nerve growth factor (NGF) for 4 days (PC12 cells), while the other clone was insensitive to NGF, but differentiated morphologically in the presence of ouabain (0.1 mM) for 7 days (PC12‐mutant cells). 2. Whole‐cell Ba2+ currents through Ca2+ channels were measured in PC12 cells at a test potential (Et) of +10 mV, from two holding potentials (Eh) of ‐90 and ‐30 mV (I‐90 and I‐30). NGF‐induced differentiation increased I‐90 by 248% and I‐30 by 133%. The cells that differentiate in the presence of ouabain had only small, if any, Ba2+ currents that did not appear to change during morphological differentiation or after the addition of NGF. 3. Barium currents in PC12 cells could be separated into two components by selective antagonists. The component of I‐90 that could be inhibited by omega‐conotoxin GVIA (omega‐CgTX) in NGF‐differentiated cells was 458 +/‐ 84 pA (mean +/‐ S.E.M.), compared with 79 +/‐ 44 pA in native cells. I‐30 was reduced by 50 +/‐ 17 pA in NGF‐treated cells and was virtually insensitive to the toxin in native cells. By contrast, the dihydropyridine (DHP) isradipine reduced I‐30 in NGF‐treated cells by 30 +/‐ 8 pA and in native cells by 20 +/‐ 3 pA. 4. Radioligand binding studies with 125I‐omega‐CgTX in PC12 cell membrane fragments and in PC12 cells showed a 2‐ to 3‐fold increase in maximal binding capacity after NGF exposure, while mutant cells showed no such change in binding capacity after treatment with NGF or ouabain. Staurosporine inhibited the effect of NGF on 125I‐omega‐CgTX binding. [3H](+)‐isradipine binding capacity was increased 1.8‐fold by NGF in depolarized PC12 cells while no change was observed in mutant cells after NGF or ouabain. There was no interaction between omega‐CgTX and DHP binding sites. 5. Both the electrophysiological and the binding data indicate a preferential expression of omega‐CgTX‐sensitive Ca2+ channels (N type) over isradipine‐sensitive channels (L type) in PC12 cells treated with NGF. By contrast, ouabain‐induced differentiation of a mutant PC12 cell line, that lacks functional NFG receptors, was not associated with the expression of Ca2+ channels.
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钙通道拮抗剂 ( )-[3H]PN200-110 与完整培养的 PC12 细胞的去极化依赖性结合。
DOI：
--
发表时间：
1986
期刊：
The Journal of pharmacology and experimental therapeutics
影响因子：
--
作者：
Greenberg,DA;Carpenter,CL;Messing,RO
通讯作者：
Messing,RO
DOI：
10.1073/pnas.85.3.924
发表时间：
1988-02-01
影响因子：
11.1
作者：
MANDEL, G;COOPERMAN, SS;BREHM, P
通讯作者：
BREHM, P
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DOI：
--
发表时间：
1982
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Toll,L
通讯作者：
Toll,L
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DOI：
10.1016/0006-291x(86)90439-0
发表时间：
1986
影响因子：
3.1
作者：
Messing,RO;Carpenter,CL;Greenberg,DA
通讯作者：
Greenberg,DA