Oliver D L, Beckius G E, Ostapoff E M
Department of Anatomy, University of Connecticut Health Center, Farmington 06030-3405.
J Neurosci Methods. 1994 Jul;53(1):23-7. doi: 10.1016/0165-0270(94)90140-6.
The components of a neural circuit are usually distinguished in separate experiments to identify long connections, presynaptic, and postsynaptic components. We describe a procedure to visualize these components in the same experiment. Neurons in the inferior colliculus the axons of which project to the medial geniculate body were identified by retrograde transport of latex microspheres, while their innervation from the cochlear nucleus was simultaneously visualized by anterograde transport of dextrans. In aldehyde-fixed slices, the microsphere-labeled neurons near dextran-labeled axons were injected with biotinylated Lucifer Yellow. Subsequent avidin-biotin histochemistry allowed permanent visualization. The specific neurons involved in this circuit and the axonal contacts they received were easily visualized with the light microscope. This method allows the study of complex innervation patterns in the mammalian central nervous system.
神经回路的组成部分通常在单独的实验中加以区分，以识别长连接、突触前和突触后成分。我们描述了一种在同一实验中可视化这些成分的方法。通过乳胶微球的逆行运输来识别下丘中轴突投射到内侧膝状体的神经元，同时通过葡聚糖的顺行运输来同时可视化它们来自耳蜗核的神经支配。在醛固定切片中，对靠近葡聚糖标记轴突的微球标记神经元注射生物素化的路西法黄。随后的抗生物素蛋白-生物素组织化学实现了永久可视化。参与该回路的特定神经元及其所接受的轴突接触很容易在光学显微镜下观察到。这种方法有助于研究哺乳动物中枢神经系统中的复杂神经支配模式。
