Park Yang Hae, Kantor Lana, Guptaroy Bipasha, Zhang Minjia, Wang Kevin K W, Gnegy Margaret E
Department of Pharmacology, University of Michigan School of Medicine, Ann Arbor, Michigan 48109, USA.
J Neurochem. 2003 Dec;87(6):1546-57. doi: 10.1046/j.1471-4159.2003.02127.x.
Repeated intermittent treatment with amphetamine (AMPH) induces both neurite outgrowth and enhanced AMPH-stimulated dopamine (DA) release in PC12 cells. We investigated the role of protein kinases in the induction of these AMPH-mediated events by using inhibitors of protein kinase C (PKC), mitogen activated protein kinase (MAP kinase) or protein kinase A (PKA). PKC inhibitors chelerythrine (100 nm and 300 nm), Ro31-8220 (300 nm) and the MAP kinase kinase inhibitor, PD98059 (30 micro m) inhibited the ability of AMPH to elicit both neurite outgrowth and the enhanced AMPH-stimulated DA release. The direct-acting PKC activator, 12-O-tetradecanoyl phorbol 13-acetate (TPA, 250 nm) mimicked the ability of AMPH to elicit neurite outgrowth and enhanced DA release. On the contrary, a selective PKA inhibitor, 100 micro m Rp-8-Br-cAMPS, blocked only the development of AMPH-stimulated DA release but not the neurite outgrowth. Treatment of the cells with acute AMPH elicited an increase in the activity of PKC and MAP kinase but not PKA. These results demonstrated that AMPH-induced increases in MAP kinase and PKC are important for induction of both the enhancement in transporter-mediated DA release and neurite outgrowth but PKA was only required for the enhancement in AMPH-stimulated DA release. Therefore the mechanisms by which AMPH induces neurite outgrowth and the enhancement in AMPH-stimulated DA release can be differentiated.
用苯丙胺(AMPH)进行反复间歇性处理可诱导PC12细胞中的神经突生长,并增强AMPH刺激的多巴胺(DA)释放。我们通过使用蛋白激酶C(PKC)、丝裂原活化蛋白激酶(MAP激酶)或蛋白激酶A(PKA)的抑制剂,研究了蛋白激酶在这些AMPH介导事件诱导中的作用。PKC抑制剂白屈菜红碱(100 nM和300 nM)、Ro31 - 8220(300 nM)以及MAP激酶激酶抑制剂PD98059(30 μM)均抑制了AMPH引发神经突生长和增强AMPH刺激的DA释放的能力。直接作用的PKC激活剂12 - O - 十四烷酰佛波醇13 - 乙酸酯(TPA,250 nM)模拟了AMPH引发神经突生长和增强DA释放的能力。相反,选择性PKA抑制剂100 μM Rp - 8 - Br - cAMPS仅阻断了AMPH刺激的DA释放的发展,而不影响神经突生长。用急性AMPH处理细胞会导致PKC和MAP激酶活性增加,但不会导致PKA活性增加。这些结果表明,AMPH诱导的MAP激酶和PKC增加对于诱导转运体介导的DA释放增强和神经突生长均很重要,但PKA仅对AMPH刺激的DA释放增强是必需的。因此,AMPH诱导神经突生长和AMPH刺激的DA释放增强的机制可以区分。