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雏鸡十二指肠中维生素D依赖性钙结合蛋白的免疫细胞化学定位

Immunocytochemical localization of the vitamin D-dependent calcium binding protein in chick duodenum.

作者信息

Thorens B, Roth J, Norman A W, Perrelet A, Orci L

出版信息

J Cell Biol. 1982 Jul;94(1):115-22. doi: 10.1083/jcb.94.1.115.

Abstract

The vitamin D-dependent calcium binding protein (CaBP) of chick duodenum has been localized by immunocytochemistry and by radioimmunoassay. Light microscopically, CaBP was seen to be present in the absorptive cells of the villi while in other cell types of the villi and the crypts, including goblet cells and endocrine cells, no CaBP was seen. At the electron microscopic level, CaBP was shown to be localized in the cytosol and the euchromatin of the nucleus but not in membrane-bounded cytoplasmic compartments. Quantitative evaluation of the immunocytochemical protein A-gold label showed that the terminal web and the cytosol of basal cellular regions were most highly labeled while the brush border was weakly labeled. The radioimmunoassay evaluation of intestinal subcellular fractions indicated that 96% of the homogenate CaBP is in the cytosol high-speed supernatant fraction. Collectively, these results support the hypothesis that the vitamin D-dependent intestinal CaBP may play a role in either regulation of intracellular calcium concentration or movement of calcium across the brush border membrane from the gut lumen.

摘要

通过免疫细胞化学和放射免疫测定法对雏鸡十二指肠中维生素D依赖性钙结合蛋白(CaBP)进行了定位。在光学显微镜下,可见CaBP存在于绒毛的吸收细胞中,而在绒毛和隐窝的其他细胞类型中,包括杯状细胞和内分泌细胞,则未观察到CaBP。在电子显微镜水平上,CaBP显示定位于细胞质溶胶和细胞核的常染色质中,而不在膜结合的细胞质区室中。对免疫细胞化学蛋白A-金标记的定量评估表明,终末网和基底细胞区域的细胞质溶胶标记程度最高,而刷状缘标记较弱。对肠亚细胞组分的放射免疫测定评估表明,匀浆中96%的CaBP存在于细胞质溶胶高速上清液组分中。总体而言,这些结果支持以下假说:维生素D依赖性肠CaBP可能在调节细胞内钙浓度或钙从肠腔穿过刷状缘膜的转运过程中发挥作用。

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