Joost Sarah, Kobayashi Kazuto, Wree Andreas, Haas Stefan Jean-Pierre
Department of Anatomy, Rostock University Medical Center, Gertrudenstraße 9, 18057 Rostock, Germany.
Department of Molecular Genetics, Institute of Biomedical Sciences, Fukushima Medical University School of Medicine, Fukushima 960-1295, Japan.
J Neurosci Methods. 2017 Jun 15;285:49-57. doi: 10.1016/j.jneumeth.2017.05.003. Epub 2017 May 4.
The nigrostriatal pathway is of great importance for the execution of movements, especially in the context of Parkinson's disease. In research, analysis of this pathway often requires the application of severe animal experiments. Organotypic nigrostriatal slice cultures offer a resource-saving alternative to animal experiments for research on the nigrostriatal system.
We have established a time-saving protocol for the preparation of murine sagittal nigrostriatal slice cultures by using a tissue chopper and agarose embedding instead of a vibratome. Furthermore, we developed the first murine co-culture model and the first co-culture utilising sagittal slices for modelling the nigrostriatal pathway.
Sagittal nigrostriatal slice cultures show good overall tissue preservation and a high number of morphologically unimpaired dopaminergic neurons in the substantia nigra. Sagittal-frontal co-culture demonstrates massive outgrowth of dopaminergic fibres from the substantia nigra into co-cultured tissue.
The use of a tissue chopper instead of a vibratome allows notable time-saving during culture preparation, therefore allowing optimisation of the preparation time. Sagittal co-cultures offer the opportunity to study dopaminergic fibres in their physiological environment and in co-cultured tissue from a different animal in the same culture system.
We here present a possibility to optimise the slice culture preparation process with the simple means of using a tissue chopper and fast agarose embedding. Furthermore, our sagittal-frontal co-culture system is suitable for the observation of dopaminergic outgrowth in both co-cultured tissues.
黑质纹状体通路对于运动的执行非常重要，尤其是在帕金森病的背景下。在研究中，对该通路的分析通常需要进行严苛的动物实验。器官型黑质纹状体切片培养为黑质纹状体系统的研究提供了一种节省资源的动物实验替代方法。
我们建立了一种省时的方案，通过使用组织切片机和琼脂糖包埋而非振动切片机来制备小鼠矢状黑质纹状体切片培养物。此外，我们开发了首个小鼠共培养模型以及首个利用矢状切片对黑质纹状体通路进行建模的共培养方法。
矢状黑质纹状体切片培养物显示出良好的整体组织保存情况，并且黑质中存在大量形态未受损的多巴胺能神经元。矢状-额状共培养显示多巴胺能纤维从黑质大量生长到共培养组织中。
使用组织切片机而非振动切片机可在培养物制备过程中显著节省时间，从而优化制备时间。矢状共培养提供了在同一培养系统中研究多巴胺能纤维在其生理环境以及来自不同动物的共培养组织中的情况的机会。
我们在此展示了一种通过使用组织切片机和快速琼脂糖包埋这种简单方法来优化切片培养制备过程的可能性。此外，我们的矢状-额状共培养系统适用于观察两种共培养组织中的多巴胺能生长情况。
