Lysz T W, Needleman P, Ferrendelli J A
J Neurosci Res. 1983;10(3):317-26. doi: 10.1002/jnr.490100310.
Assay conditions sensitive enough to measure prostaglandin (PG) biosynthetic capacity in discrete brain areas have been developed. Serial sections of the rabbit hippocampal formation were dissected from the brain, frozen, and 16 micron sections were cut and freeze dried. The assay was performed with 40-200 micrograms dry weight of tissue in a 30 microliter incubation volume containing high specific activity arachidonic acid. The whole hippocampal formation slice or its subdivisions (subiculum, dentate gyrus, and hippocampus) when assayed individually demonstrated the ability to produce approximately 30-50 pmol/mg dry weight PGF2 alpha, PGE2, and lesser amounts of PGD2 (in a ratio of 1:0.8:0.3). A modest yet significant increase in PGE2 production was measured in the dentate gyrus when compared to PGE2 formation in the subiculum or hippocampus. An intact PGE2 isomerase was confirmed in the hippocampal formation slice by incubations with PGH2 endoperoxide. These results demonstrate that quantitative histochemical techniques can be used to measure PG metabolism in microscopic regions of brain.
已经开发出了足够灵敏的检测条件,用于测量离散脑区中前列腺素(PG)的生物合成能力。从兔脑中切下海马结构的连续切片,冷冻后切成16微米的切片并冻干。检测在30微升孵育体积中进行,使用40 - 200微克干重的组织,其中含有高比活性的花生四烯酸。单独检测整个海马结构切片或其各个亚区(下托、齿状回和海马体)时,显示出能够产生约30 - 50皮摩尔/毫克干重的前列腺素F2α、前列腺素E2,以及少量的前列腺素D2(比例为1:0.8:0.3)。与下托或海马体中的前列腺素E2生成相比,在齿状回中检测到前列腺素E2生成有适度但显著的增加。通过与前列腺素H2内过氧化物孵育,在海马结构切片中证实了完整的前列腺素E2异构酶。这些结果表明,定量组织化学技术可用于测量脑微观区域中的PG代谢。