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大鼠气管上皮浆液细胞分泌颗粒中的降钙素基因相关肽

Calcitonin gene-related peptide in secretory granules of serous cells in the rat tracheal epithelium.

作者信息

Baluk P, Nadel J A, McDonald D M

机构信息

Cardiovascular Research Institute, University of California, San Francisco 94143-0130.

出版信息

Am J Respir Cell Mol Biol. 1993 Apr;8(4):446-53. doi: 10.1165/ajrcmb/8.4.446.

DOI:10.1165/ajrcmb/8.4.446
PMID:7682823
Abstract

The tracheal epithelium of pathogen-free rats consists mainly of serous-type secretory cells, ciliated cells, basal cells, and a few neuroendocrine cells. Mucus-containing goblet cells are rare. Calcitonin gene-related peptide (CGRP) is known to exist in the neuroendocrine cells and in sensory nerves of the tracheal mucosa and is released into the airway lumen by sensory nerve stimulation. In this study, we determined whether epithelial serous cells are another source of CGRP. Tracheas of adult male specific pathogen-free F344 rats were immunostained by an avidin-biotin technique either as whole mounts or as cryostat sections using two different polyclonal primary antibodies to rat CGRP. Some specimens were stained for CGRP-like immunofluorescence and examined with a confocal microscope. CGRP immunoreactivity was present in granules of serous cells throughout the trachea. In whole mounts, the stained cells were most abundant between the cartilaginous rings, especially in the rostral trachea, where they constituted 56% of the epithelial cells in contact with the tracheal lumen. Serous cells were easily distinguished from neuroendocrine cells and nerve fibers with CGRP immunoreactivity. In evidence that the CGRP immunoreactivity was specific, the staining of serous cells was abolished by omitting the primary antibody and by absorption with 10 micrograms/ml CGRP. Antibodies to substance P, vasoactive intestinal polypeptide, and tyrosine hydroxylase did not stain epithelial serous cells. An antibody to protein gene product 9.5 labeled neuroendocrine cells, but not serous cells. Injection of capsaicin (150 micrograms/kg intravenously), a substance known to degranulate epithelial serous cells, reduced the staining of the serous cells for CGRP.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

无特定病原体大鼠的气管上皮主要由浆液型分泌细胞、纤毛细胞、基底细胞和少数神经内分泌细胞组成。含黏液的杯状细胞很少见。降钙素基因相关肽(CGRP)已知存在于气管黏膜的神经内分泌细胞和感觉神经中,并通过感觉神经刺激释放到气道腔中。在本研究中,我们确定上皮浆液细胞是否是CGRP的另一个来源。成年雄性无特定病原体F344大鼠的气管,采用抗生物素蛋白-生物素技术进行免疫染色,制成整装片或冰冻切片,使用两种不同的抗大鼠CGRP多克隆一抗。一些标本进行CGRP样免疫荧光染色,并用共聚焦显微镜检查。整个气管的浆液细胞颗粒中均存在CGRP免疫反应性。在整装片中,染色细胞在软骨环之间最为丰富,尤其是在气管前部,它们占与气管腔接触的上皮细胞的56%。浆液细胞很容易与具有CGRP免疫反应性的神经内分泌细胞和神经纤维区分开来。为证明CGRP免疫反应性具有特异性,通过省略一抗和用10微克/毫升CGRP吸收,浆液细胞的染色被消除。P物质、血管活性肠肽和酪氨酸羟化酶的抗体未对上皮浆液细胞染色。蛋白基因产物9.5的抗体标记神经内分泌细胞,但不标记浆液细胞。静脉注射辣椒素(150微克/千克),一种已知可使上皮浆液细胞脱颗粒的物质,可减少浆液细胞对CGRP的染色。(摘要截短于250字)

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