Sender S, Böttcher K, Cetin Y, Gros G
Vegetative Physiologie, Zentrum Physiologie, Medizinische Hochschule Hannover, Hannover, Germany.
J Histochem Cytochem. 1999 Jan;47(1):43-50. doi: 10.1177/002215549904700105.
Flounders Platichthys flesus were investigated with respect to isolation, purification, and cellular localization of carbonic anhydrase (CA) in the respiratory system. CA was purified from gills and erythrocytes and was shown to exclusively represent a soluble enzyme with an apparent molecular weight of 30 kD. Inhibition constants (KI) towards acetazolamide (ACTZ) were 8.4.10(-9) M for erythrocyte CA and 7.6.10(-9) M for gill CA, indicating a high sensitivity to sulfonamides, as exhibited by human CA II. Specific CA activity did not differ significantly in seawater- and freshwater-acclimated fish. Antibodies were raised against purified gill and erythrocyte CA. Both antisera crossreacted and were used to localize CA in the gills of seawater and freshwater flounders at the light microscopic level. Independent of the salinity, a positive reaction of variable intensity was found in the following cell types: pavement cells (PVCs), forming the gill epithelial surface layer; mucous cells (MCs); pillar cells (PCs), bordering the vascular channels of the secondary lamellae; and chloride cells (CCs), mitochondria-rich cells located in the primary epithelium, the interlamellar regions, and at the bases of the secondary lamellae.(J Histochem Cytochem 47:43-50, 1999)
针对呼吸系统中碳酸酐酶（CA）的分离、纯化及细胞定位，对欧鲽（Platichthys flesus）展开了研究。CA从鳃和红细胞中纯化得到，结果表明其完全是一种表观分子量为30 kD的可溶性酶。红细胞CA对乙酰唑胺（ACTZ）的抑制常数（KI）为8.4×10⁻⁹ M，鳃CA的抑制常数为7.6×10⁻⁹ M，这表明其对磺胺类药物具有高敏感性，如同人CA II所表现出的那样。海水适应和淡水适应的鱼的特异性CA活性并无显著差异。制备了针对纯化的鳃和红细胞CA的抗体。两种抗血清发生交叉反应，并用于在光学显微镜水平上定位海水和淡水欧鲽鳃中的CA。不论盐度如何，在以下细胞类型中均发现了强度各异的阳性反应：构成鳃上皮表层的扁平细胞（PVCs）；黏液细胞（MCs）；与次生鳃小片血管通道相邻的柱状细胞（PCs）；以及位于初级上皮、鳃小片间区域和次生鳃小片基部的富含线粒体的氯细胞（CCs）。（《组织化学与细胞化学杂志》47:43 - 50, 1999）
