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P物质及其前体形式在豚鼠迷走神经、气管和肺组织中的表达。

Expression of substance P and its precursor forms in vagal, tracheal, and lung tissues of the guinea pig.

作者信息

Kream R M, Marchand J E, O'Connor S T, Bloomquist E I

机构信息

Department of Anesthesiology, Tufts University School of Medicine, Boston, Massachusetts.

出版信息

Am J Physiol. 1994 Dec;267(6 Pt 1):L807-14. doi: 10.1152/ajplung.1994.267.6.L807.

Abstract

Steady-state levels of the prototypic tachykinin neuropeptide substance P (SP) and its major precursor form substance P-glycine (SP-G) were detected and authenticated in guinea pig vagal and respiratory tissues by radioimmunoassay (RIA), combined high-performance liquid chromatography (HPLC)/RIA analyses, and immunohistochemistry. Four antisera were employed: anti-SP that recognizes the amidated COOH-terminal of SP and is specific for the mature peptide, anti-SP4-10 that recognizes the midportion 4-10 amino acid sequence of SP and is highly specific for both mature SP and extended precursor forms of SP, anti-SP-G that is highly specific for the unamidated COOH-terminal of SP-G, and affinity-purified anti-SP-G-K that is capable of detecting SP-G and minor forms of SP precursor in immunohistochemical analyses. In all examined areas, the content of substance P4-10-like immunoreactivity (SP4-10-LI) quantified by RIA with the use of anti-SP4-10 was greater than that quantified by RIA with the use of anti-SP serum, thereby providing biochemical evidence of steady-state expression of extended precursor forms of SP. Immunohistochemical analyses demonstrated labeled axonal profiles indicating the presence of immunoreactive SP as well as immunoreactive forms of SP precursor within lung hilum and in small fibers in the parenchyma, with no evidence of labeled neuronal cell bodies in these same areas.

摘要

通过放射免疫测定(RIA)、高效液相色谱(HPLC)/RIA联合分析以及免疫组织化学方法,在豚鼠迷走神经和呼吸组织中检测并鉴定了典型速激肽神经肽P物质(SP)及其主要前体形式P物质-甘氨酸(SP-G)的稳态水平。使用了四种抗血清:识别SP酰胺化COOH末端且对成熟肽具有特异性的抗SP;识别SP中部4-10个氨基酸序列且对成熟SP和SP的延伸前体形式均具有高度特异性的抗SP4-10;对SP-G未酰胺化COOH末端具有高度特异性的抗SP-G;以及在免疫组织化学分析中能够检测SP-G和SP前体的次要形式的亲和纯化抗SP-G-K。在所有检测区域中,使用抗SP4-10通过RIA定量的P物质4-10样免疫反应性(SP4-10-LI)含量高于使用抗SP血清通过RIA定量的含量,从而为SP延伸前体形式的稳态表达提供了生化证据。免疫组织化学分析显示,在肺门和实质中的小纤维内存在标记的轴突轮廓,表明存在免疫反应性SP以及SP前体的免疫反应形式,而在这些相同区域未发现标记的神经元细胞体。

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