Ikejima T, Ito S
J Histochem Cytochem. 1984 Jun;32(6):625-35. doi: 10.1177/32.6.6427328.
Mouse parotid gland and saliva were studied by histochemical, immunohistochemical, and activity measurements for carbonic anhydrase. Hansson 's histochemical reaction for carbonic anhydrase revealed positive enzyme activity in the parotid acinar cell cytoplasm and little or no reaction in the secretory granules. The luminal contents in all of the glandular duct systems also reacted positively, but the duct cells themselves were only weakly positive. Ultrastructural observations confirmed the light microscope histochemical localization and, in addition, revealed luminal content activity in intercellular ducts. Purified carbonic anhydrase isolated from mouse salivary glands was used to raise antibodies in rabbits. Localization of carbonic anhydrase by direct immunolabeling with fluorescein-coupled antibody and indirect immunoperoxidase labeling revealed enzyme localization on or in the acinar cell secretory granule membrane and not in the surrounding cytoplasm. The luminal contents of the intercalated and striated ducts were also strongly positive. Stimulation of salivary secretion with phenylephrine or pilocarpine increased the amount of carbonic anhydrase in saliva. Acetatazolamide and potassium cyanate inhibited carbonic anhydrase activity. Reasons underlying the discrepancy between the histochemical and immunolabeling localization of carbonic anhydrase are discussed. It is concluded that the parotid acinar cells of mice appear to be a significant source of carbonic anhydrase in saliva but its role is enigmatic.
通过组织化学、免疫组织化学以及碳酸酐酶活性测定等方法对小鼠腮腺和唾液进行了研究。汉森的碳酸酐酶组织化学反应显示，腮腺腺泡细胞胞质中有阳性酶活性，而分泌颗粒中反应微弱或无反应。所有腺管系统的管腔内容物也呈阳性反应，但管细胞本身仅呈弱阳性。超微结构观察证实了光学显微镜下的组织化学定位，此外，还揭示了细胞间小管管腔内容物的活性。从小鼠唾液腺中分离出的纯化碳酸酐酶被用于在兔体内产生抗体。用荧光素偶联抗体直接免疫标记和间接免疫过氧化物酶标记对碳酸酐酶进行定位，结果显示酶定位于腺泡细胞分泌颗粒膜上或膜内，而非周围细胞质中。闰管和纹状管的管腔内容物也呈强阳性。用去氧肾上腺素或毛果芸香碱刺激唾液分泌可增加唾液中碳酸酐酶的含量。乙酰唑胺和氰酸钾可抑制碳酸酐酶活性。文中讨论了碳酸酐酶组织化学定位与免疫标记定位之间差异的潜在原因。得出的结论是，小鼠腮腺腺泡细胞似乎是唾液中碳酸酐酶的重要来源，但其作用尚不清楚。
