Whitaker K J, Manchester E L, Jacobson W, Wilkinson M
Brain Res Bull. 1984 Mar;12(3):215-9. doi: 10.1016/0361-9230(84)90048-0.
We have characterized and quantified the specific binding of [3H]-flunitrazepam ( FNZ ) to thick (230 micron) slices of mouse brain. The binding site has the characteristics of a benzodiazepine receptor, i.e., binding of FNZ is reversible, stereospecific, saturable and of high affinity. Clonazepam, but not R05 -4864, readily displaces the label. In contrast to results from homogenate assays, neither GABA nor bicuculline has any effect on [3H]- FNZ binding. However, as previously reported, the slice assay confirms the lower number of benzodiazepine receptors in "emotional" mouse brain. In addition, we have confirmed that the neurotoxin DSP4 can modify [3H]- FNZ binding though in our hands this compound elevates rather than reduces binding. The speed, simplicity and minimal tissue preparation involved suggests that this slice assay could be a valuable addition to neurochemical studies of neurotransmitter receptors.
我们已对[3H] -氟硝西泮(FNZ)与小鼠大脑厚(230微米)切片的特异性结合进行了表征和定量。该结合位点具有苯二氮䓬受体的特征,即FNZ的结合是可逆的、立体特异性的、可饱和的且具有高亲和力。氯硝西泮能轻易取代标记物,而R05 - 4864则不能。与匀浆测定结果不同,γ-氨基丁酸(GABA)和荷包牡丹碱对[3H] - FNZ结合均无任何影响。然而,正如先前报道的那样,切片测定证实了“情绪化”小鼠大脑中苯二氮䓬受体数量较少。此外,我们已证实神经毒素DSP4可改变[3H] - FNZ结合,不过在我们的实验中,该化合物提高而非降低了结合。该方法的速度、简便性以及所需的最少组织制备表明,这种切片测定可能会为神经递质受体的神经化学研究增添有价值的内容。