Nissinen M J, Panula P
Department of Anatomy, University of Helsinki, Finland.
J Histochem Cytochem. 1993 Sep;41(9):1405-12. doi: 10.1177/41.9.8354880.
We studied the distribution of histamine (HA) immunoreactivity in endocrine cells of the acid-producing mucosa in rat stomach with pre-embedding immunoelectron microscopy (IEM) using an antiserum against HA. Four fixation modifications were compared to optimize the ultrastructural morphology and staining pattern with the antisera produced against carbodiimide-conjugated HA. Fixation with 4% 1-ethyl-3(3-dimethyl-aminopropyl) carbodiimide (EDCDI) combined with both 4% paraformaldehyde and 0.1% glutaraldehyde gave superior results compared with EDCDI alone. Enterochromaffin-like (ECL) cells were easily distinguished from other endocrine cells in optimally fixed samples. The peroxidase end-product was distributed within the cytoplasm surrounding the vesicles of the ECL cells. ECL cells comprised about 75% of all endocrine cells, and about 90% of them were HA immunoreactive (HA-IR). No other HA-IR cell types were identified by EM in the basal half of the oxyntic region of rat gastric mucosa. The results suggest that a combination of EDCDI and aldehydes is suitable for IM demonstration of HA in cells. ECL cells from a predominant portion of endocrine cells in the oxyntic glands and may constitute the only significant non-mast cell store of HA in rat gastric mucosa.
我们使用抗组胺(HA)抗血清,通过包埋前免疫电子显微镜(IEM)研究了大鼠胃产酸黏膜内分泌细胞中HA免疫反应性的分布。比较了四种固定方法的改进,以优化用针对碳二亚胺偶联HA产生的抗血清的超微结构形态和染色模式。与单独使用EDCDI相比,用4% 1-乙基-3(3-二甲基氨基丙基)碳二亚胺(EDCDI)与4%多聚甲醛和0.1%戊二醛联合固定产生了更好的结果。在最佳固定的样本中,肠嗜铬样(ECL)细胞很容易与其他内分泌细胞区分开来。过氧化物酶终产物分布在ECL细胞囊泡周围的细胞质内。ECL细胞约占所有内分泌细胞的75%,其中约90%具有HA免疫反应性(HA-IR)。在大鼠胃黏膜胃底区的下半部分,通过电子显微镜未发现其他HA-IR细胞类型。结果表明,EDCDI和醛类的组合适用于细胞中HA的免疫组织化学显示。ECL细胞是胃底腺内分泌细胞的主要部分,可能构成大鼠胃黏膜中HA唯一重要的非肥大细胞储存部位。