Matute C, Nguyen Q T, Miledi R
Department of Psychobiology, University of California, Irvine 92717.
J Neurosci Res. 1993 Aug 15;35(6):652-63. doi: 10.1002/jnr.490350608.
Levels of mRNAs encoding neurotransmitter receptors in the visual cortex, lateral geniculate nucleus, and superior colliculus of the rabbit and rat, and properties of the receptors expressed, were studied using Xenopus laevis oocytes. mRNA extracted from these areas was injected into the oocytes, which then acquired functional receptors. Electrical recordings of neurotransmitter-induced membrane currents reflect the relative amounts of mRNAs encoding the corresponding receptors. Receptors to gamma aminobutyric acid (GABA), kainate, glutamate, and serotonin exhibited uniformly high levels of expression, whereas expression of receptors to glycine and N-methyl-D-aspartate was uniformly low. In contrast, the expression of receptors to acetylcholine and substance P was highly non-uniform. Expression of acetylcholine receptors was high in oocytes injected with mRNA from the visual cortex, low for the lateral geniculate nucleus, and very low or absent for the superior colliculus. Conversely, the currents elicited by substance P were large in oocytes injected with superior colliculus mRNA, but were small or absent in oocytes injected with mRNAs from the other regions. Immunohistochemical analysis, at the light and electron microscopic levels, was used to localize choline acetyltransferase, the acetylcholine-synthesizing enzyme, and substance P-containing synaptic boutons in the three visual areas. Their presence closely paralleled the potency of mRNAs coding for acetylcholine and substance P receptors. The ability of rat mRNA, from each visual area, to induce neurotransmitter receptors was similar to that observed in the corresponding rabbit mRNAs. In addition to the marked differential distribution of mRNA encoding neurotransmitter receptors in the visual system, our findings reveal the probable existence of as yet uncharacterized receptors, whose new molecular forms may be revealed by further study. Our results also provide the basic information required for subsequent studies on the effect of monocular deprivation on the expression of neurotransmitter receptors in the visual system.
利用非洲爪蟾卵母细胞研究了兔和大鼠视觉皮层、外侧膝状体核及上丘中编码神经递质受体的mRNA水平及其所表达受体的特性。从这些区域提取的mRNA被注入卵母细胞,卵母细胞随后获得功能性受体。神经递质诱导的膜电流的电记录反映了编码相应受体的mRNA的相对含量。γ-氨基丁酸(GABA)、海人藻酸、谷氨酸和5-羟色胺的受体表现出一致的高表达水平,而甘氨酸和N-甲基-D-天冬氨酸受体的表达则一致较低。相反,乙酰胆碱和P物质受体的表达高度不均一。注射来自视觉皮层mRNA的卵母细胞中乙酰胆碱受体的表达较高,外侧膝状体核中的表达较低,而上丘中的表达则非常低或不存在。相反,注射上丘mRNA的卵母细胞中由P物质引发的电流较大,而注射来自其他区域mRNA的卵母细胞中的电流较小或不存在。利用光镜和电镜水平的免疫组织化学分析来定位胆碱乙酰转移酶(乙酰胆碱合成酶)和三个视觉区域中含P物质的突触小体。它们的存在与编码乙酰胆碱和P物质受体的mRNA的效力密切平行。来自每个视觉区域的大鼠mRNA诱导神经递质受体的能力与在相应兔mRNA中观察到的相似。除了视觉系统中编码神经递质受体的mRNA存在明显的差异分布外,我们的研究结果还揭示了可能存在尚未鉴定的受体,其新的分子形式可能通过进一步研究得以揭示。我们的结果还为后续关于单眼剥夺对视觉系统中神经递质受体表达影响的研究提供了所需的基础信息。