膜结合碳酸酐酶IV、IX和XIV在小鼠心脏中的表达。
Expression of membrane-bound carbonic anhydrases IV, IX, and XIV in the mouse heart.
作者信息
Scheibe Renate J, Gros Gerolf, Parkkila Seppo, Waheed Abdul, Grubb Jeffrey H, Shah Gul N, Sly William S, Wetzel Petra
机构信息
Zentrum Physiologie, Medizinische Hochschule Hannover, Carl-Neuberg-Str. 1, 30625 Hannover, Germany.
出版信息
J Histochem Cytochem. 2006 Dec;54(12):1379-91. doi: 10.1369/jhc.6A7003.2006.
Expression of membrane-bound carbonic anhydrases (CAs) of CA IV, CA IX, CA XII, and CA XIV has been investigated in the mouse heart. Western blots using microsomal membranes of wild-type hearts demonstrate a 39-, 43-, and 54-kDa band representing CA IV, CA IX, and CA XIV, respectively, but CA XII could not be detected. Expression of CA IX in the CA IV/CA XIV knockout animals was further confirmed using matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Cardiac cells were immunostained using anti-CA/FITC and anti-alpha-actinin/TRITC, as well as anti-CA/FITC and anti-SERCA2/TRITC. Subcellular CA localization was investigated by confocal laser scanning microscopy. CA localization in the sarcolemmal (SL) membrane was examined by double immunostaining using anti-CA/FITC and anti-MCT-1/TRITC. CAs showed a distinct distribution pattern in the sarcoplasmic reticulum (SR) membrane. CA XIV is predominantly localized in the longitudinal SR, whereas CA IX is mainly expressed in the terminal SR/t-tubular region. CA IV is present in both SR regions, whereas CA XII is not found in the SR. In the SL membrane, only CA IV and CA XIV are present. We conclude that CA IV and CA XIV are associated with the SR as well as with the SL membrane, CA IX is located in the terminal SR/t-tubular region, and CA XII is not present in the mouse heart. Therefore, the unique subcellular localization of CA IX and CA XIV in cardiac myocytes suggests different functions of both enzymes in excitation-contraction coupling.
在小鼠心脏中对碳酸酐酶IV（CA IV）、碳酸酐酶IX（CA IX）、碳酸酐酶XII（CA XII）和碳酸酐酶XIV（CA XIV）这几种膜结合碳酸酐酶（CAs）的表达情况进行了研究。使用野生型心脏的微粒体膜进行的蛋白质免疫印迹分析显示，分别有代表CA IV、CA IX和CA XIV的39 kDa、43 kDa和54 kDa条带，但未检测到CA XII。使用基质辅助激光解吸电离飞行时间质谱法进一步证实了CA IX在CA IV/CA XIV基因敲除动物中的表达。使用抗CA/FITC和抗α-肌动蛋白/TRITC，以及抗CA/FITC和抗肌浆网钙ATP酶2（SERCA2）/TRITC对心脏细胞进行免疫染色。通过共聚焦激光扫描显微镜研究亚细胞CA的定位。使用抗CA/FITC和抗单羧酸转运蛋白1（MCT-1）/TRITC进行双重免疫染色，检查CA在肌膜（SL）中的定位。CAs在肌浆网（SR）膜中呈现出明显的分布模式。CA XIV主要定位于纵向SR，而CA IX主要表达于终末SR/横管区域。CA IV存在于两个SR区域，而在SR中未发现CA XII。在SL膜中，仅存在CA IV和CA XIV。我们得出结论，CA IV和CA XIV与SR以及SL膜相关，CA IX位于终末SR/横管区域，且CA XII在小鼠心脏中不存在。因此，CA IX和CA XIV在心肌细胞中独特的亚细胞定位表明这两种酶在兴奋 - 收缩偶联中具有不同功能。
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