Deller T, Naumann T, Frotscher M
Institute of Anatomy, University of Freiburg, PO Box 111, D-79001, Freiburg, Germany.
J Neurosci Methods. 2000 Nov 15;103(1):117-26. doi: 10.1016/s0165-0270(00)00301-0.
Fiber tracts in the brain are formed by neurochemically heterogeneous neuron populations. To distinguish between the different neurons that contribute to a fiber tract it is necessary to combine anterograde and retrograde tracing techniques with immunocytochemistry. In this article, we describe two techniques which allow for the neurochemical identification of retrogradely labeled neurons and anterogradely labeled axons on the ultrastructural level. The identification of the neurotransmitter identity of retrogradely labeled neurons is achieved by combining retrograde Fluoro-Gold tracing with preembedding immunocytochemistry, while the neurotransmitter identity of anterogradely labeled axons can be revealed by combining anterograde Phaseolus vulgaris-leucoagglutinin (PHAL) tracing and postembedding immunostaining.
大脑中的纤维束由神经化学性质各异的神经元群体构成。为了区分对纤维束有贡献的不同神经元，有必要将顺行和逆行追踪技术与免疫细胞化学相结合。在本文中，我们描述了两种技术，它们能够在超微结构水平上对逆行标记的神经元和顺行标记的轴突进行神经化学鉴定。通过将逆行荧光金追踪与包埋前免疫细胞化学相结合来实现对逆行标记神经元神经递质身份的鉴定，而顺行标记轴突的神经递质身份可通过将顺行菜豆白细胞凝集素（PHA-L）追踪与包埋后免疫染色相结合来揭示。
