Genetic requirement of p47phox for superoxide production by murine microglia.
Genetic requirement of p47phox for superoxide production by murine microglia.
复制标题
p47phox 小鼠小胶质细胞产生超氧化物的遗传要求。
DOI：
10.1096/fj.00-0608fje
复制
发表时间：
2001
期刊：
影响因子：
--
通讯作者：
Leto,TL
中科院分区：
文献类型：
--
作者：
Lavigne,MC;Malech,HL;Holland,SM;Leto,TL
NADPH氧化酶被认为在中枢神经系统的小胶质细胞中起作用。这些结论基于药理学和免疫化学证据，尽管这些方法是间接的，并提出了特异性问题。例如，二亚苯基碘鎓抑制多种黄素酶，包括黄嘌呤氧化酶、NADH脱氢酶和NADPH氧化酶。在这里，我们提供的遗传证据表明，p47 phox，吞噬细胞NADPH氧化酶的一个重要组成部分，需要从小胶质细胞释放超氧阴离子。来自新生野生型小鼠的小胶质细胞，而不是来自新生p47 phox缺陷（“敲除”;-/-）小鼠的小胶质细胞，在调理酵母聚糖或佛波醇肉豆蔻酸酯刺激后产生超氧化物。内源性p47 phox仅在野生型小胶质细胞中检测到，与这些细胞中选择性超氧化物的产生一致。在用人p47 phoxcDNA逆转录病毒转导的p47 phox缺陷型小胶质细胞中恢复了超氧化物释放。观察到类似的超氧化物生成动力学，与野生型和恢复的小胶质细胞中相同的酶功能一致。p47 phoxin转导细胞的免疫检测证实，超氧化物释放的恢复与重组蛋白的产生相关。这些数据提供了遗传学证据，p47 phoxis所必需的超氧化物释放的小胶质细胞，并表明，系统相关的吞噬细胞氧化酶是活跃的，在这些细胞。
An NADPH oxidase is thought to function in microglial cells of the central nervous system. These conclusions are based on pharmacological and immunochemical evidence, although these approaches are indirect and raise issues of specificity. For example, diphenyleneiodonium inhibits a variety of flavoenzymes, including xanthine oxidase, NADH dehydrogenase, and NADPH oxidase. Here, we provide genetic evidence that p47phox, an essential component of the phagocyte NADPH oxidase, is required for superoxide anion release from microglia. Microglia derived from newborn wild‐type mice, but not from newborn p47phox‐deficient (“knockout”; ‐/‐) mice, produced superoxide after stimulation by opsonized zymosan or phorbol myristate acetate. Endogenous p47phoxwas detected only in wild‐type microglia, consistent with selective superoxide production in these cells. Superoxide release was restored in p47phox‐deficient microglia that were retrovirally transduced with human p47phoxcDNA. Similar kinetics of superoxide generation were observed, consistent with the same enzyme functioning in wild‐type and restored microglia. Immuno‐detection of p47phoxin transduced cells confirmed that restoration of superoxide release correlated with production of recombinant protein. These data provide genetic proof that p47phoxis necessary for superoxide release by microglial cells and indicate that a system related to the phagocyte oxidase is active in these cells.