Babington E J, Vatanparast J, Verrall J, Blackshaw S E
Department of Human Anatomy & Genetics, University of Oxford, South Parks Road, Oxford, OX1 3QX, UK.
Invert Neurosci. 2005 Nov;5(3-4):173-82. doi: 10.1007/s10158-005-0006-7. Epub 2005 Oct 24.
Three-dimensional (3D) collagen gels provide a stable matrix in which isolated regenerating ganglia from leech and snail can be maintained for studies of the molecular and cellular mechanisms underlying the regenerative process. Segmental ganglia from leech, or supraoesophageal, suboesophageal or buccal ganglia from snail were maintained for up to 3 weeks in 3D matrices of mammalian Type I collagen. The collagen matrix supports the regenerative outgrowth of axon tracts as well as the migration of microglial cells, important elements in the repair process. Proteins or soluble factors or target tissue may be added to the basic collagen matrix to manipulate the environment of the regenerating tissue. We describe techniques for immunostaining of regenerating axons and microglial cells within the gel matrix in combination with staining of cell nuclei, and the use of intracellular labelling to distinguish axons of identified neurons within the regenerative outgrowth.
三维（3D）胶原凝胶提供了一个稳定的基质，在其中可以维持从水蛭和蜗牛分离出的再生神经节，用于研究再生过程背后的分子和细胞机制。来自水蛭的节段性神经节，或来自蜗牛的食管上神经节、食管下神经节或颊神经节，在哺乳动物I型胶原的3D基质中维持长达3周。胶原基质支持轴突束的再生生长以及小胶质细胞的迁移，这些都是修复过程中的重要元素。可以将蛋白质、可溶性因子或靶组织添加到基本的胶原基质中，以操纵再生组织的环境。我们描述了在凝胶基质中对再生轴突和小胶质细胞进行免疫染色并结合细胞核染色的技术，以及使用细胞内标记来区分再生生长中已识别神经元的轴突的方法。
