突触素在注射大鼠脑mRNA的非洲爪蟾卵母细胞多巴胺胞吐释放中的作用
Role of synaptophysin in exocytotic release of dopamine from Xenopus oocytes injected with rat brain mRNA.
作者信息
Shibaguchi H, Takemura K, Kan S, Kataoka Y, Kaibara M, Saito N, Taniyama K
机构信息
Department of Pharmacology, Nagasaki University School of Medicine, Japan.
出版信息
Cell Mol Neurobiol. 2000 Jun;20(3):401-8. doi: 10.1023/a:1007022428041.
Abstract
- The role of synaptophysin in the exocytotic release of dopamine (DA) was examined in Xenopus laevis oocytes injected with rat brain mRNA. 2. The mRNA-injected oocytes showed DA uptake which depended on the incubation time and external DA concentrations. 3. Stimulation with KCl (10-50 mM) of mRNA-injected oocytes preloaded with DA evoked external Ca2+ -dependent release of DA. The noninjected and water-injected oocytes did not produce uptake of DA and stimulation-evoked release of DA. 4. The high-KCl (50 mM)-stimulated release of DA decreased in the oocytes injected with rat brain mRNA together with antibody to synaptophysin. 5. Immunoblot analysis demonstrated that synaptophysin was expressed in the brain mRNA-injected oocytes but not in the noninjected and water-injected oocytes. 6. Thus, uptake and release machinery similar to native dopaminergic nerve terminals was expressed in Xenopus oocytes by injecting mRNA-extracted from the rat brain, and synaptophysin may play a role in the exocytotic release of DA.
摘要
- 在注射了大鼠脑mRNA的非洲爪蟾卵母细胞中研究了突触素在多巴胺（DA）胞吐释放中的作用。2. 注射了mRNA的卵母细胞表现出DA摄取，这取决于孵育时间和外部DA浓度。3. 用KCl（10 - 50 mM）刺激预先加载了DA的注射了mRNA的卵母细胞，可诱发外部Ca2+依赖性的DA释放。未注射和注射水的卵母细胞不产生DA摄取和刺激诱发的DA释放。4. 在注射了大鼠脑mRNA并同时注射突触素抗体的卵母细胞中，高KCl（50 mM）刺激的DA释放减少。5. 免疫印迹分析表明，突触素在注射了脑mRNA的卵母细胞中表达，但在未注射和注射水的卵母细胞中不表达。6. 因此，通过注射从大鼠脑提取的mRNA，在非洲爪蟾卵母细胞中表达了类似于天然多巴胺能神经末梢的摄取和释放机制，并且突触素可能在DA的胞吐释放中起作用。
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