Alkaline phosphatase vs luciferase as secreted reporter molecules in vivo
Alkaline phosphatase vs luciferase as secreted reporter molecules in vivo
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DOI：
10.1016/j.ab.2005.01.023
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发表时间：
2005-04-15
影响因子：
2.9
通讯作者：
Kitamura, M
中科院分区：
文献类型：
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作者：
Hiramatsu, N;Kasai, A;Kitamura, M
分泌型碱性磷酸酶（SEAP）和Metridia荧光素酶（MLuc）是体外有用的报告分子，但对它们在体内的有用性知之甚少。在本研究中，我们研究了重组SEAP和MLuc在血液和尿液中的体内活性。当将SEAP转染细胞或重组SEAP注射到大鼠中时，观察到血清SEAP水平的显著增加。相反，在注射SEAP转染细胞的大鼠的尿中未检测到SEAP活性，或者在注射SEAP的Nagase无白蛋白血症大鼠的尿中也未检测到重组SEAR SEAP活性，其中肾小球对大分子的渗透性增强。当MLuc转染细胞植入大鼠体内时，不仅在尿液中，而且在病血清中也检测不到MLuc的活性。即使在静脉注射重组MLuc后立即。血清中未检测到MLuc活性。随后的实验表明，与SEAP相反，MLuc被大鼠血清、胎牛血清迅速灭活。或者人类血清。白蛋白被鉴定为负责抑制MLuc活性的分子。这些数据阐明了分泌型报告分子SEAP和MLuc在体内情况下的优点和局限性。(c)2005年爱思唯尔公司All rights reserved.
Secreted alkaline phosphatase (SEAP) and Metridia luciferase (MLuc) are useful reporter molecules in vitro, but little is understood about their usefulness ill vivo. Ill this Study, we investigated in vivo activity of recombinant SEAP and MLuc in blood and Urine. When SEAP-transfected cells or recombinant SEAP were injected into rats, Substantial increase in the level of serum SEAP was observed. In contrast, activity of SEAP was not detected in urine of rats injected with either the SEAP-transfected cells or recombinant SEAR SEAP activity was also undetectable in urine of SEAP-injected Nagase analbuminemic rats in which glomerular permeability to macromolecules is enhanced. When MLuc-transfected cells were implanted into rats, activity of MLuc was undetectable not only in urine but also ill serum. Even immediately after intravenous injection of recombinant MLuc. activity of MLuc was not detected in serum. Subsequent experinients revealed that, in contrast to SEAP, MLuc was rapidly inactivated either by rat serum, fetal bovine serum. Or human serum. Albumin was identified Lis the molecule responsible for the inhibition of MLuc activity. These data elucidated advantages and limitations of secreted reporter molecules SEAP and MLuc under in vivo Situations. (c) 2005 Elsevier Inc. All rights reserved.