大鼠肝线粒体在原位和体外固定后谷草转氨酶活性的生化与组织化学研究。
A biochemical and histochemical study of glutamic oxalacetic transaminase activity of rat hepatic mitochondria fixed in situ and in vitro.
作者信息
Lee S H, Torack R M
出版信息
J Cell Biol. 1968 Dec;39(3):725-32. doi: 10.1083/jcb.39.3.725.
Rat liver perfused in situ briefly with a glutaraldehyde-formaldehyde mixture was homogenized in isotonic sucrose. The mitochondria, isolated from a homogenate of the perfused liver by differential centrifugation, assumed a slender and compact appearance similar to those often seen in an intact cell. The glutamic oxalacetic transaminase (GOT) activity of this mitochondrial fraction survived an additional formaldehyde fixation and was studied by biochemical and histochemical methods. The biochemical assay of the enzyme activity revealed that the activity was only slightly less than that of an unfixed mitochondrial fraction. The reaction product due to mitochondrial GOT activity was found to be localized to the cristae, as had been demonstrated in an intact liver cell. GOT activity of the mitochondrial fraction isolated from fresh liver tissue homogenate in 0.25 M sucrose was inactivated readily by either glutaraldehyde or formaldehyde and was no longer demonstrable by biochemical and histochemical methods after fixation.
用戊二醛 - 甲醛混合物对大鼠肝脏进行原位短暂灌注后，在等渗蔗糖中匀浆。通过差速离心从灌注肝脏的匀浆中分离出的线粒体呈现出细长且紧密的外观，类似于在完整细胞中常见的线粒体。该线粒体部分的谷氨酸草酰乙酸转氨酶（GOT）活性在额外的甲醛固定后仍能保留，并通过生化和组织化学方法进行研究。酶活性的生化测定表明，该活性仅略低于未固定的线粒体部分。正如在完整肝细胞中所证明的那样，线粒体GOT活性产生的反应产物定位于嵴。从新鲜肝组织在0.25 M蔗糖中匀浆分离出的线粒体部分的GOT活性很容易被戊二醛或甲醛灭活，固定后通过生化和组织化学方法不再能检测到。
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