Development of the technique for expression of peptide receptors in the oocyte membrane

卵母细胞膜肽受体表达技术的开发

• 批准号: 60870005
• 负责人: KUNO Motoi
• 金额: $ 6.08万
• 依托单位: Kyoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research
• 财政年份: 1985
• 资助国家: 日本
• 起止时间: 1985 至 1986
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-60870005/
• 关键词: Peptide; Receptor; mRNA; Tachykinin; Oocyte; Xenopus; Transmitter; 卯母細胞; イオン

While peptide-receptive cells are widely distributed in both the central and peripheral nervous systems, these cells are too small to analyze detailed ionic mechanisms associated with activation of the receptors. The present study has been addressed to the analysis of ionic mechanisms underlying activation of the peptide receptors expressed in Xenopus oocytes which have a diameter of about 1 mm. For this purpose, mRNA isolated from the rat brain or the bovine stomach was injected into oocytes, and the response of the injected oocytes to a variety of peptides was recorded under voltage clamp following incubation for a few days. While the receptors to five different peptides could be expressed by this technique, the study was focused on the substance P receptor. Substance P is a member of the tachykinin family. At present, in addition to substance P, two other tachykinins, substance K and neuromedin K, are known to exist in the mammalian nervous system. In accord with the presence of mul … More tiple tachykinins, the existence of three subtypes of tachykinin receptors has been suggested. The three subtypes of tachykinin receptors could be distinguished by potency ranking of different tachykinin agonists. On this basis, the tachykinin receptor induced by rat brain mRNA was found to correspond to the SP-P tachykinin receptor, whereas that directed by bovine stomach mRNA corresponded to the SP-K tachykinin receptor. Thus, two different types of tachykinin receptors could be induced in the same expression system, depending upon the source of exogenous mRNA injected. This indicates that the two receptors are made of different molecules. Therefore, the difference in nature between the two receptors cannot be attributed to the possible posttranslational modification occurring at their destination tissue alone. On the other hand, the ionic mechanisms responsible for activation of the two receptors are both based on an increase in the <Cl^-> conductance. This suggests that activation of the two receptors uses the same intracellular second messenger, which in turn, activates <Cl^-> ionic channels in the oocyte membrane. The nature of the second messenger may be different between oocytes and mammalian neurons. Therefore, the ionic mechanisms analyzed in oocytes cannot be generalized to activation of tachykinin receptors present in other species. Less

虽然肽受体细胞广泛分布在中枢和外周神经系统中，但这些细胞太小，无法分析与受体激活相关的详细离子机制。本研究已被寻址到激活的肽受体在非洲爪蟾卵母细胞，具有约1毫米的直径。为此目的，从大鼠大脑或牛胃中分离的mRNA被注射到卵母细胞中的离子机制的分析，并记录下电压钳孵育几天后的各种肽注射的卵母细胞的反应。虽然可以通过这种技术表达五种不同肽的受体，但研究重点是P物质受体。P物质是速激肽家族的一员。目前，除了P物质外，已知哺乳动物神经系统中还存在另外两种速激肽，即K物质和神经介肽K。与穆尔的存在雅阁 ...更多信息 除了速激肽外，还存在三种亚型的速激肽受体。速激肽受体的三种亚型可以通过不同速激肽激动剂的效力排序来区分。在此基础上，发现大鼠脑mRNA诱导的速激肽受体对应于SP-P速激肽受体，而牛胃mRNA诱导的速激肽受体对应于SP-K速激肽受体。因此，两种不同类型的速激肽受体可以在同一表达系统中诱导，这取决于注射的外源mRNA的来源。这表明这两种受体是由不同的分子组成的。因此，两种受体之间的性质差异不能归因于仅在其目的组织中发生的可能的翻译后修饰。另一方面，负责激活两种受体的离子机制都是基于<Cl^->电导的增加。这表明两种受体的激活使用了相同的细胞内第二信使，而第二信使又激活了卵母细胞膜上的<Cl^->离子通道。第二信使的性质可能是不同的卵母细胞和哺乳动物的神经元。因此，在卵母细胞中分析的离子机制不能推广到其他物种中存在的速激肽受体的激活。少

项目成果

HARADA, Yoshio et al.: "Expression of substance P and substance K receptors in Xenopus oocytes by exogenous mRNA" J. Neurosci.

HARADA、Yoshio 等人：“外源 mRNA 在爪蟾卵母细胞中表达 P 物质和 K 物质受体”J. Neurosci。

日本生理学雑誌. 47. (1986)

日本生理学杂志。47。（1986）

HARADA, Yoshio et al.: "Expression of substance P receptor in Xenopus oocytes by injections of poly(A) RNA extracted from the rat brain" Proc. International Union Physiol. Sci.30. 64 (1986)

HARADA、Yoshio 等人：“通过注射从大鼠脑中提取的聚 (A) RNA 来表达爪蟾卵母细胞中 P 物质受体”Proc.

原田嘉夫: J. Neurosci.

原田芳夫：神经科学杂志。

HARADA, Yoshio et al.: "Substance P receptors expressed in Xenopus oocytes from rat brain mRNA" J. Physiol. Soc. Japan. 48. 210 (1986)

HARADA、Yoshio 等人：“爪蟾卵母细胞中大鼠脑 mRNA 表达的 P 物质受体”J. Physiol。