Seeley P J, Field P M
Laboratory of Neurobiology and Development, National Institute for Medical Research, London, U.K.
Brain Res. 1988 May 24;449(1-2):177-91. doi: 10.1016/0006-8993(88)91036-0.
We have made stable complexes between wheat germ agglutinin and either 5 or 10 nm particles of colloidal gold. These complexes were phagocytosed by neuronal and glial cells in embryonic rat hippocampal cultures and the incorporated gold gave intense, low-background staining in the light microscope either directly, for the most heavily labelled cells, or after intensification by physical development of silver. Cells were labelled in a punctate fashion over perikarya and processes. In the electron microscope, particles of gold were observed in lysosomal vesicles, frequently in an aggregated form. Gold complex incorporated into cells in culture was retained by those cells over periods up to 20 days. Embryonic hippocampal cells were labelled in suspension culture by incorporation of wheat germ agglutinin-gold complexes and transplanted into the brains of syngeneic adult host rats. Grafted neurons and glia were observed in the electron microscope to retain high levels of gold label over periods up to 30 days. Receipt of synaptic connections by transplanted neurones was observed. Complexes of wheat germ agglutinin with 10 nm gold particles were injected unilaterally into field CA3 of the hippocampus of adult rats. Specific retrograde transport of gold was observed in the light and electron microscopes to pyramidal and hilar neurones of the contralateral hippocampus and to neurones of the medial septal nucleus. Colloidal gold-wheat germ agglutinin complexes appear to be useful cellular markers that can be visualized at both light and electron microscope levels.
我们已制备出小麦胚凝集素与5纳米或10纳米胶体金颗粒的稳定复合物。这些复合物在胚胎大鼠海马体培养物中被神经元和神经胶质细胞吞噬，所摄入的金在光学显微镜下，对于标记最密集的细胞可直接产生强烈的低背景染色，或者在通过银的物理显影增强后产生这种效果。细胞在胞体和突起上呈点状标记。在电子显微镜下，金颗粒见于溶酶体囊泡中，常呈聚集形式。培养物中摄入细胞的金复合物在长达20天的时间内被这些细胞保留。胚胎海马体细胞在悬浮培养中通过摄入小麦胚凝集素 - 金复合物进行标记，然后移植到同基因成年宿主大鼠的大脑中。在电子显微镜下观察到移植的神经元和神经胶质细胞在长达30天的时间内保留高水平的金标记。观察到移植神经元接受突触连接。将小麦胚凝集素与10纳米金颗粒的复合物单侧注射到成年大鼠海马体的CA3区。在光学显微镜和电子显微镜下均观察到金的特异性逆行运输至对侧海马体的锥体细胞和门区神经元以及内侧隔核的神经元。胶体金 - 小麦胚凝集素复合物似乎是有用的细胞标记物，可在光学显微镜和电子显微镜水平上观察到。
