Neurology Department, Shanghai No.1 People's Hospital (in the process of affiliation), School of Medicine, Tongji University, Shanghai, 200081, China.
Neuroscience Research Australia, Randwick, NSW, 2031, Australia.
Mol Brain. 2018 Sep 12;11(1):50. doi: 10.1186/s13041-018-0393-4.
Postsynaptic density - 95 kDa protein (PSD95) is an important molecule on the postsynaptic membrane. It interacts with many other proteins and plays a pivotal role in learning and memory formation. Its distribution in the brain has been studied previously using in situ hybridization as well as immunohistochemistry. However, these studies are based on 2 dimensional (2D) sections and results are presented with a few sections. The present study aims to show PSD-95 distribution in 3 dimensions (3D) without slicing the brain tissue of C57BL/6 mice into sections using the advanced CUBIC technique.
Immunofluorescent staining using a PSD-95 antibody was performed on a half of the mouse brain after clarifying it using the advanced CUBIC protocol. The brain tissue was imaged using a Zeiss Z1 light sheet microscope and 3D reconstruction was completed using the Arivis Vision 4 dimensional (4D) software.
The majority of brain nuclei have similar distribution pattern to what has been reported from in situ hybridization and immunohistochemical studies in the mouse. The signal can be easily followed in the 3D and their spatial relationship with adjacent structures clearly demarcated. In the present study, some fiber bundles also showed strong PSD-95 signal, which is different from what was shown in previous studies and need to be confirmed in future studies.
突触后密度-95kDa 蛋白（PSD95）是突触后膜上的重要分子。它与许多其他蛋白质相互作用，在学习和记忆形成中起着关键作用。其在大脑中的分布以前曾使用原位杂交和免疫组织化学进行研究。然而，这些研究是基于二维（2D）切片，结果仅呈现少数切片。本研究旨在使用先进的 CUBIC 技术，在不将 C57BL/6 小鼠的脑组织切成切片的情况下，以三维（3D）方式显示 PSD-95 的分布。
使用 PSD-95 抗体进行免疫荧光染色，在使用先进的 CUBIC 方案澄清脑组织的一半后进行。使用蔡司 Z1 光片显微镜对脑组织进行成像，并使用 Arivis Vision 4 维（4D）软件完成 3D 重建。
大多数脑核具有与在小鼠中进行的原位杂交和免疫组织化学研究中报道的相似的分布模式。可以轻松地在 3D 中跟踪信号，并清楚地标定其与相邻结构的空间关系。在本研究中，一些纤维束也显示出强烈的 PSD-95 信号，这与以前的研究不同，需要在未来的研究中进一步证实。
