Belloni-Olivi L, Bressler J P, Goldstein G W
Kennedy Research Institute, Johns Hopkins Medical Institutions, Baltimore, MD.
Curr Eye Res. 1992 Mar;11(3):203-11. doi: 10.3109/02713689209001771.
In this investigation we localized and compared the level of gamma-glutamyl transpeptidase (GGTP) activity in retinal and brain preparations using histochemical, enzymatic and in situ hybridization assays. We compared GGTP distribution to another microvessel specific enzyme, alkaline phosphatase (AP). In the rat brain, GGTP activity was observed in microvessels and choroid plexus by a histochemical method. Similar studies in the rat retina revealed activity in the pigment epithelium but only a very weak reaction in microvessels. Histochemical staining for alkaline phosphatase was observed in both retinal and brain microvessels choroid plexus and pigment epithelium. Biochemical analysis verified that GGTP activity was significantly lower in retinal than brain microvessels, while alkaline phosphatase activity was similar in both types of microvessels. GGTP specific activity of bovine brain and retinal microvessels was 185 +/- 39 mUnits and 8.5 +/- 1.5 mUnits (p less than 0.001), respectively. By contrast, alkaline phosphatase specific activity in brain and retinal microvessels was 732 +/- 139 and 471 +/- 114 (p greater than 0.1), respectively. Choroid plexus and retinal pigment epithelium exhibited similar levels of GGTP and alkaline phosphatase. Differences in GGTP expression between retinal and brain microvessels were also observed on the mRNA level. In situ hybridization studies revealed that brain microvessels expressed four times more GGTP specific mRNA than retinal microvessels. We conclude that retinal microvessels do not express high levels of GGTP which may make them more vulnerable than brain microvessels to injuries mediated by leukotrienes and oxidative stress.
在本研究中,我们使用组织化学、酶学和原位杂交分析方法,对视网膜和脑组织样本中γ-谷氨酰转肽酶(GGTP)的活性水平进行了定位和比较。我们将GGTP的分布与另一种微血管特异性酶——碱性磷酸酶(AP)进行了比较。通过组织化学方法,在大鼠脑的微血管和脉络丛中观察到了GGTP活性。在大鼠视网膜中进行的类似研究显示,色素上皮中有活性,但微血管中的反应非常微弱。在视网膜和脑的微血管、脉络丛以及色素上皮中均观察到了碱性磷酸酶的组织化学染色。生化分析证实,视网膜微血管中的GGTP活性显著低于脑微血管,而两种类型微血管中的碱性磷酸酶活性相似。牛脑和视网膜微血管的GGTP比活性分别为185±39 mUnits和8.5±1.5 mUnits(p<0.001)。相比之下,脑和视网膜微血管中的碱性磷酸酶比活性分别为732±139和471±114(p>0.1)。脉络丛和视网膜色素上皮表现出相似水平的GGTP和碱性磷酸酶。在mRNA水平上也观察到了视网膜和脑微血管之间GGTP表达的差异。原位杂交研究显示,脑微血管表达的GGTP特异性mRNA比视网膜微血管多四倍。我们得出结论,视网膜微血管不表达高水平的GGTP,这可能使它们比脑微血管更容易受到白三烯和氧化应激介导的损伤。