Identification of common biological markers underlying establishment of drug dependence and an attempt to develop pharmacogenomic therapy

鉴定药物依赖性建立的常见生物标志物并尝试开发药物基因组疗法

基本信息

批准号：
18500302
负责人：
KATSURA Masashi
金额：
$ 2.55万
依托单位：
Kawasaki Medical School
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

项目摘要

In this research project, we have been carried out to identify the common biological markers underlying establishment of drug dependence and an attempt to develop pharmacogenomic therapy.Functional alterations in L-type high voltage-gated calcium channels (Cav1) after short-term (24h) exposure of mouse cerebrocortical neurons to drugs of abuse were examined as compared with those in psychological dependent mouse brains. KC1 (30 mM)-stimulated [^<45>Ca^<2+>] influx into the neurons increased with increasing the duration of the drugs exposure and its concentrations. This enhancement was completely abolished by Cav1 inhibitors, and antagonists for ryanodine and IP_3 receptors. Cav1.2 and Cav α2-δ proteins were increased in abused drugs-treated neurons. Increased binding of [^<3>H]PN200-110 after 24 h exposure to drugs of abuse was due to decreased Kd value. Similar changes in the binding parameters and protein expressions were obtained in cerebral cortices as well as nucleus accumbens fro … More m psychological dependent mice. These results indicate that stimulation of calcium-induced calcium release by ryanodine receptors and subsequent activations of IP_3 receptors induces Cav1 up-regulation.Furthermore, we examined the functional involvement of accessory proteins of Cav 3 subunits in ethanol (EtOH)-induced Cav1 up-regulation. Short-term exposure of the neurons to EtOH significantly increased in Cav β3 subunit levels, whereas Cav β1, β2 and β4 subunits showed no changes. High potassium-stimulated [^<45>Ca^<2+>] influx into the neurons significantly increased by EtOH exposure and this increase was not observed in the neurons pretreated with morpholino oligomers specific for Cav β3 subunits. Decreased Kd value of [^3H] P: N200-110 after EtOH exposure was also abolished by Cav β3 subunits knockdown. Similar changes in protein expressions were obtained in cerebral cortices from psychological dependent mice. These results indicate that short-term exposure of the neurons to EtOH up-regulates Cav1 functions via increased expression of Cav β3 subunit proteins in the neuronal membrane. Less

在这个研究项目中，我们已经确定了药物依赖建立的常见生物标志物，并尝试开发药物基因组疗法。短期（24小时）后L型高压门控钙通道（Cav1）的功能改变) 与心理依赖性 KC1 (30 mM) 刺激的 [^<45>Ca^<2+>] 流入相比，检查了小鼠大脑皮质神经元暴露于滥用药物的情况。随着药物暴露时间和浓度的增加，这种增强作用被 Cav1 抑制剂完全消除，而兰尼定和 IP_3 受体拮抗剂在滥用药物处理的神经元中增加。暴露于滥用药物24小时后[^ 3 H]PN200-110的结合增加是由于Kd值降低，在大脑皮质中获得了类似的变化。这些结果表明，刺激钙-钙诱导兰尼碱受体的释放以及随后 IP_3 受体的激活诱导 Cav1 上调。此外，我们检查了 Cav 辅助蛋白的功能参与。乙醇 (EtOH) 中的 3 亚基诱导的 Cav1 上调神经元短期暴露于 EtOH，Cav β3 亚基水平显着增加，而 Cav β1、β2 和 β2 亚基水平显着增加。 β4亚基没有表现出变化，由于EtOH暴露，高钾刺激的[^ 45 Ca ^ 2+ ]流入神经元显着增加，并且在用Cav β3亚基特异性吗啉寡聚物预处理的神经元中没有观察到这种增加。 EtOH 暴露后 [^3H] P: N200-110 的 Kd 值降低也被 Cav β3 亚基敲低所消除，蛋白质表达也发生了类似的变化。这些结果表明，神经元短期暴露于 EtOH 通过增加神经元膜中 Cav β3 亚基蛋白的表达来上调 Cav1 功能。