Gundersen C B, Miledi R, Parker I
Proc R Soc Lond B Biol Sci. 1984 Apr 24;221(1223):127-43. doi: 10.1098/rspb.1984.0027.
Xenopus laevis oocytes injected with poly(A)+ mRNA extracted from rat brain became sensitive to serotonin, glutamate, kainate, acetylcholine and gamma-aminobutyrate. Application of these substances to mRNA-injected oocytes elicited membrane currents. The glutamate- and acetylcholine-induced currents usually showed oscillations, while the kainate current was smooth. The current oscillations during glutamate application reversed direction at about the chloride equilibrium potential (-24 mV), but the reversal potential for the kainate current was close to 0 mV. The current-voltage relation for the glutamate-induced current oscillations showed strong rectification at hyperpolarized potentials, while that for the kainate current was nearly linear. In some oocytes, glutamate elicited smooth membrane currents, with oscillations either absent, or appearing after a delay. The reversal potential of this component was close to 0 mV, and was clearly different from that of the oscillatory component. The appearance of glutamate and kainate sensitivity in the oocyte membrane is due to the translation of the foreign messenger RNA, and not to activation of the oocytes' own genome, because oocytes still become sensitive when transcription is prevented by enucleation or by treatment with actinomycin D. It appears that mRNA from rat brain contains translationally active messengers which code for various neurotransmitter receptors. When this mRNA is injected into Xenopus oocytes, the messengers are translated and receptors are inserted into the oocyte membrane, where they form functionally active receptor-channel complexes.
将从大鼠脑中提取的多聚腺苷酸(poly(A)+)mRNA注入非洲爪蟾卵母细胞后,这些卵母细胞对血清素、谷氨酸、海人酸、乙酰胆碱和γ-氨基丁酸变得敏感。将这些物质施加到注射了mRNA的卵母细胞上会引发膜电流。谷氨酸和乙酰胆碱诱导的电流通常表现出振荡,而海人酸电流是平稳的。施加谷氨酸期间的电流振荡在大约氯离子平衡电位(-24 mV)处反转方向,但海人酸电流的反转电位接近0 mV。谷氨酸诱导的电流振荡的电流-电压关系在超极化电位下表现出强烈的整流作用,而海人酸电流的电流-电压关系几乎是线性的。在一些卵母细胞中,谷氨酸引发平稳的膜电流,要么没有振荡,要么延迟后出现振荡。该成分的反转电位接近0 mV,明显不同于振荡成分的反转电位。卵母细胞膜中谷氨酸和海人酸敏感性的出现是由于外源信使RNA的翻译,而不是由于卵母细胞自身基因组的激活,因为当通过去核或放线菌素D处理阻止转录时,卵母细胞仍然会变得敏感。似乎来自大鼠脑的mRNA包含编码各种神经递质受体的具有翻译活性的信使。当这种mRNA注入非洲爪蟾卵母细胞时,信使被翻译,受体被插入卵母细胞膜中,在那里它们形成功能活跃的受体-通道复合物。