Noronha J G, Bell J M, Rapoport S I
Laboratory of Neurosciences, NIA, NIH, Bethesda, MD 20892.
J Neurosci Res. 1990 Jun;26(2):196-208. doi: 10.1002/jnr.490260209.
The distribution of radioactivity within brain metabolic compartments was examined following the intravenous injection of [9,10-3H]palmitate into awake rats. Brain radioactivity reached a maximum value by 15 min after [9,10-3H]palmitate injection and remained unchanged for at least 4 hr. Regional differences in radioactivity could be determined with high resolution by quantitative autoradiography, at the level of cell layers within the hippocampus and cerebral cortex, and between striosomes of the caudate nucleus. Regional brain radioactivities were converted to normalized regional radioactivities (k) by dividing them by the integrated plasma fatty acid radioactivity (integrated over the time course of the experiment). These values reflected incorporation mainly into brain phospholipids; radioactivity due to nonlipid components was minimal. Indeed, about 85% of brain radioactivity was within lipids between 5 min and 4 hr postinjection, the remainder being equally divided between protein-associated pellet and aqueous-soluble metabolites. The major lipids labeled were phospholipids, particularly phosphatidylcholine, which contained about 75% of phospholipid radioactivity. The results show that [9,10-3H]palmitate can be used to examine incorporation of plasma palmitate into individual brain regions via quantitative autoradiography. Furthermore, the tracer is a rather selective marker for phosphatidylcholine and can be used to examine turnover and synthesis of this phospholipid. [9,10-3H]palmitate has advantages over [U-14C]palmitate for autoradiographic studies of incorporation; following the 14C-tracer, significant label even at 4 hr after injection is in nonlipid compartments (glutamate and aspartate), and the long path length of 14C limits resolution at the cell layer level.
将[9,10 - 3H]棕榈酸静脉注射到清醒大鼠体内后,检测了脑代谢区室中的放射性分布。注射[9,10 - 3H]棕榈酸后15分钟,脑放射性达到最大值,并至少保持4小时不变。通过定量放射自显影术可在高分辨率下确定放射性的区域差异,在海马体和大脑皮层的细胞层水平以及尾状核的纹状体之间。将区域脑放射性除以实验时间过程中积分的血浆脂肪酸放射性,从而将其转换为归一化区域放射性(k)。这些值主要反映了掺入脑磷脂中的情况;非脂质成分产生的放射性极小。实际上,注射后5分钟至4小时内,约85%的脑放射性存在于脂质中,其余部分在与蛋白质相关的沉淀和水溶性代谢物之间平均分配。标记的主要脂质是磷脂,尤其是磷脂酰胆碱,其含有约75%的磷脂放射性。结果表明,[9,10 - 3H]棕榈酸可用于通过定量放射自显影术检测血浆棕榈酸掺入各个脑区的情况。此外,该示踪剂是磷脂酰胆碱的一种相当有选择性的标记物,可用于检测这种磷脂的周转和合成。对于掺入的放射自显影研究,[9,10 - 3H]棕榈酸比[U - 14C]棕榈酸具有优势;使用14C示踪剂时,即使在注射后4小时,非脂质区室(谷氨酸和天冬氨酸)中仍有显著标记,并且14C的长路径长度限制了细胞层水平的分辨率。