Saarnio J, Parkkila S, Parkkila A K, Waheed A, Karttunen T, Sly W S
Department of Surgery, University of Oulu, Oulu, Finland.
J Histochem Cytochem. 1999 Apr;47(4):517-24. doi: 10.1177/002215549904700410.
Mitochondrial carbonic anhydrase V (CA V) in liver provides HCO3- to pyruvate carboxylase for the first step in gluconeogenesis and HCO3- to carbamyl phosphate synthetase I for the first step in ureagenesis. Because carbamyl phosphate synthetase I and ornithine transcarbamylase are also expressed in enterocytes, we tested the hypothesis that CA V is expressed in the gastrointestinal tract in addition to liver. Polyclonal rabbit antisera were raised against a polypeptide of 17 C-terminal amino acids of human CA V and against purified recombinant mouse isozyme and were used in Western blotting and immunoperoxidase staining of human and rat tissues. Immunohistochemistry showed that CA V is expressed cell-specifically in the alimentary canal mucosa from stomach to rectum. Immunoreactions for CA V were detected in the parietal cells and gastrin-producing G-cells of the stomach and in intestinal enterocytes. Western blotting of human and rat gastrointestinal tissues with isozyme-specific antibodies showed positive signals for CA V with the expected molecular mass. The findings in human tissues paralleled those in rat. The cell-specific pattern of CA V expression suggests a role for CA V in alimentary canal physiology. We propose that mitochondrial CA V participates in the detoxification of ammonia produced in the gastrointestinal tract by providing bicarbonate to carbamyl phosphate synthetase I. (J Histochem Cytochem 47:517-524, 1999)
肝脏中的线粒体碳酸酐酶V(CA V)为糖异生的第一步反应中的丙酮酸羧化酶提供HCO₃⁻,并为尿素生成第一步反应中的氨甲酰磷酸合成酶I提供HCO₃⁻。由于氨甲酰磷酸合成酶I和鸟氨酸转氨甲酰酶也在肠上皮细胞中表达,因此我们检验了以下假说:CA V除了在肝脏中表达外,在胃肠道中也有表达。我们制备了针对人CA V C末端17个氨基酸多肽以及纯化的重组小鼠同工酶的兔多克隆抗血清,并将其用于人和大鼠组织的蛋白质免疫印迹及免疫过氧化物酶染色。免疫组织化学显示,CA V在从胃到直肠的消化道黏膜中呈细胞特异性表达。在胃的壁细胞和产生胃泌素的G细胞以及肠上皮细胞中检测到了CA V的免疫反应。用同工酶特异性抗体对人和大鼠胃肠道组织进行蛋白质免疫印迹分析,显示CA V呈现出预期分子量的阳性信号。人体组织中的结果与大鼠的结果相似。CA V表达的细胞特异性模式提示其在消化道生理学中发挥作用。我们提出,线粒体CA V通过为氨甲酰磷酸合成酶I提供碳酸氢盐,参与胃肠道中产生的氨的解毒过程。(《组织化学与细胞化学杂志》47:517 - 524, 1999)