Targeted neutralization of calcineurin, by expression of an inhibitor peptide under the control of a cholinergic specific promoter in PC12 cells, promotes neurite outgrowth in the presence of NGF.
Targeted neutralization of calcineurin, by expression of an inhibitor peptide under the control of a cholinergic specific promoter in PC12 cells, promotes neurite outgrowth in the presence of NGF.
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通过在 PC12 细胞中胆碱能特异性启动子的控制下表达抑制肽，靶向中和钙调神经磷酸酶，在 NGF 存在的情况下促进神经突生长。
DOI：
10.1007/bf02680014
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发表时间：
2000
期刊：
影响因子：
--
通讯作者：
Dedman,JR
中科院分区：
文献类型：
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作者：
Naciff,JM;King,KL;Dedman,JR
我们已经确定了小鼠囊泡性乙酰胆碱转运体(Vacht)/胆碱乙酰转移酶(ChAT)基因的一个区域，它是Vacht和ChAT基因表达的胆碱能特异性启动子，以及体内的报告基因(LacZ)。我们已经使用这个启动子来指导来自钙调神经磷酸酶(Caln)自身调节域的抑制肽的表达，以直接中和Caln的功能，以确定这种钙/钙调蛋白调节的磷酸酶在轴突生长中的作用。在NGF存在的情况下，靶向抑制Caln可促进PC12细胞突起生长，最早在转染后24小时。抑制CaN介导的促进PC12细胞突起生长的作用在转染后的前4~6天达到最大，当高表达长达12天时不会引起不良反应。非靶向Caln抑制剂环孢素A可使模型细胞的轴突数目增加1.5倍，而在PC12细胞中，表达Caln抑制肽可使轴突数目增加1.8倍。这些数据表明，Caln是胆碱能细胞神经营养反应的重要调节因子，在开发促进神经损伤恢复的治疗策略方面可能被证明有价值。
We have characterized a region of the mouse vesicular acetylcholine transporter(VAChT)/choline acetyltransferase (ChAT) gene locus that serves as a cholinergic-specific promoter for the expression of both VAChT and ChAT genes, as well as a reporter gene (LacZ)in vivo. We have used this promoter to direct the expression of an inhibitor peptide, derived from the calcineurin (CalN) autoregulatory domain, to directly neutralize the function of CalN to define the role of this Ca2+/Calmodulin regulated phosphatase in neurite outgrowth. Targeted inhibition of CalN promotes neurite outgrowth in PC12 cells in the presence of NGF, as early as 24 h after transfection. Inhibition of CalN-mediated enhancement of neurite outgrowth in PC12 cells reaches a maximum effect within the first 4 to 6 days after transfection, and does not cause adverse effects when highly expressed for up to 12 days. Cyclosporin A, a nontargeted CalN inhibitor, increases the number of neurites in mock transfected cells by 1.5 fold, while in transfected PC12 cells, the expression of the CalN inhibitor peptide increases the neurite number by 1.8 fold. These data demonstrate that CalN is an important regulator of the neurotrophic response in cholinergic cells and may prove valuable in developing treatment strategies to promote recovery from neurological Injury.