Lampe R A, Davis L G, Gutnick M J
Medical Products Department, E. I. du Pont de Nemours and Company, Wilmington, DE 19898.
Adv Exp Med Biol. 1987;221:201-10. doi: 10.1007/978-1-4684-7618-7_15.
Xenopus laevis oocytes when injected with rat brain mRNA synthesize neuronal receptors that can be analyzed electrophysiologically. After a post-injection incubation period of 24-72 hours, L-glutamic acid, kainic acid and quisqualic acid caused a dose dependent (10-100 microM) depolarization of the oocyte membrane. The voltage and conductance changes associated with kainate activation were distinguishable from those seen for L-glutamate or quisqualate. There was no response to L-aspartate application and an inconsistent response to N-methyl-D-aspartate. Upon fractionation of the mRNA on sucrose gradients, transcripts greater than 2 Kb in length were obligatory for the synthesis of excitatory amino acid receptors. The electrophysiological response of injected oocytes exposed to L-glutamate was similar to that of native oocytes when exposed to muscarinic agents. This similarity may reflect the activation of the same ionophore and suggests that the active mRNA fraction for glutamate responsiveness either encodes for a binding protein that can be assembled along with native ion channels into the oocyte membrane or encodes for a glutamate binding site with a similar channel.
非洲爪蟾卵母细胞注射大鼠脑信使核糖核酸(mRNA)后,会合成可通过电生理学方法进行分析的神经元受体。在注射后24至72小时的孵育期后,L-谷氨酸、海人酸和quisqualic酸会引起卵母细胞膜剂量依赖性(10 - 100微摩尔)的去极化。与海人酸激活相关的电压和电导变化与L-谷氨酸或quisqualate引起的变化不同。对L-天冬氨酸的应用无反应,对N-甲基-D-天冬氨酸的反应不一致。当在蔗糖梯度上对mRNA进行分级分离时,长度大于2千碱基对(Kb)的转录本对于兴奋性氨基酸受体的合成是必需的。注射了mRNA的卵母细胞暴露于L-谷氨酸时的电生理反应与天然卵母细胞暴露于毒蕈碱剂时的反应相似。这种相似性可能反映了相同离子载体的激活,并表明对谷氨酸反应性的活性mRNA部分要么编码一种可以与天然离子通道一起组装到卵母细胞膜中的结合蛋白,要么编码一个具有相似通道的谷氨酸结合位点。