完整牛肾组织和体外培养肾皮质细胞中甲状旁腺素结合的免疫反应性。

A Niendorf, H Arps, M Sieck, M Dietel
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引用次数: 6

摘要

pth结合位点的定位已经在完整的肾脏切片和牛肾皮质的培养细胞中进行了检查。组织切片与10(-7)M牛甲状旁腺激素(1-84)在37℃下孵育2小时,细胞孵育15分钟。使用羧基末端特异性抗甲状旁腺激素抗血清(s478)通过免疫细胞化学观察甲状旁腺激素结合情况。作为对照,使用竞争性的10(-7)牛甲状旁腺激素(1-84)和10倍过量的合成1-34甲状旁腺激素片段进行细胞培养孵育,该片段不具有s478的抗原。这导致没有染色。近端和远端小管细胞均可见pth结合,Henle’s袢厚升肢细胞中pth结合程度较低。在收集管中,PTH特异性染色也存在,这仅限于定位在其他没有PTH结合位点的单个细胞之间。肾小球、亨利氏袢细肢、血管、结缔组织未见染色。数据表明肾元的大部分含有甲状旁腺素结合位点,尽管数量不同。这与甲状旁腺激素在肾脏中的许多作用是一致的。在收集片段中,揭示了不同功能状态或细胞异质性的明显细胞间差异。
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Immunoreactivity of PTH-binding in intact bovine kidney tissue and cultured cortical kidney cells indicative for specific receptors.

Localization of PTH-binding sites has been examined in intact kidney sections and cultured cells derived from bovine kidney cortex. Tissue sections were incubated with 10(-7) M bovine PTH (1-84) for 2 h, cells for 15 min, at 37 degrees C. Visualization of PTH-binding was achieved by immunocytochemistry using a carboxy-terminal specific anti-PTH antiserum (S 478). For control, cell culture incubations were performed applying competitively 10(-7) bovine PTH (1-84) and a 10-fold excess of synthetic 1-34 PTH fragment, not antigenic for S 478. This resulted in a lack of staining. PTH-binding was found in all cells of the proximal and the distal tubule, and with less intensity in the thick ascending limb of Henle's loop. In collecting ducts a PTH specific staining was also present, which was confined to single cells localized between others without PTH binding sites. No staining was seen in glomerula, the thin limb of Henle's loop, in blood vessels, and in connective tissue. The data suggest that large parts of the nephron contain PTH-binding sites, although in different amounts. This is in agreement with the numerous actions of PTH in the kidney. In the collecting segment a distinct cell-to-cell difference was disclosed indicative for different functional states or cellular heterogeneity.

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