Boland C R, Kraus E R, Scheiman J M, Black C, Deshmukh G D, Dobbins W O
Research Service, Veterans Administration Medical Center, Ann Arbor 48105.
Am J Physiol. 1990 May;258(5 Pt 1):G774-87. doi: 10.1152/ajpgi.1990.258.5.G774.
Mucin is a critical component of the protective layer secreted by gastrointestinal mucous cells. A detailed understanding of the molecular processing of gastric mucin and the physiology of its secretion has been limited by the lack of an adequate model for their study. We have developed a primary culture system of canine gastric mucous cells that has permitted us to study their synthetic and secretory functions. It was found that [3H]glucosamine used for metabolic labeling studies was incorporated into both mucin and lipid components of gastric mucus. To measure mucin with this model, a new immunoassay was developed to quantitate canine gastric mucin. Mucin was purified from the canine stomach, a polyclonal antibody was generated, and an enzyme-linked immunosorbent assay for gastric mucin was established. Mast cells were frequent contaminants of the gastric mucous cell preparation, and two methods were developed to limit their contamination. A new culture system has been developed for the study of gastric mucous cells. These cells synthesize and secrete both mucin and phospholipids. This system will permit us to study the molecular processing of mucin and the physiology of its production and release.
粘蛋白是胃肠道粘液细胞分泌的保护层的关键组成部分。由于缺乏合适的研究模型，对胃粘蛋白的分子加工及其分泌生理学的详细了解一直受到限制。我们开发了一种犬胃粘液细胞原代培养系统，这使我们能够研究它们的合成和分泌功能。研究发现，用于代谢标记研究的[3H]葡萄糖胺被整合到胃粘液的粘蛋白和脂质成分中。为了用该模型测量粘蛋白，开发了一种新的免疫测定法来定量犬胃粘蛋白。从犬胃中纯化粘蛋白，制备多克隆抗体，并建立了胃粘蛋白的酶联免疫吸附测定法。肥大细胞是胃粘液细胞制备过程中常见的污染物，为此开发了两种方法来限制其污染。已开发出一种用于研究胃粘液细胞的新培养系统。这些细胞合成并分泌粘蛋白和磷脂。该系统将使我们能够研究粘蛋白的分子加工及其产生和释放的生理学。
