Application of CLARITY to Investigate the 3D Architecture of 14-3-3 Zeta Protein in AD

Abstract

CLARITY (Clear Lipid–exchanged Acrylamide–hybridized Rigid Imaging/ Immunostaining/ in situ–hybridization–compatible Tissue hydrogel) is a powerful, innovative, whole brain-clearing technology, and it has been successfully combined with the immunofluorescence staining to achieve the 3D visualization of some proteins or cells in mm-thick brain tissue or even the intact brains. These 3D information help to gain deeper understanding on the pathologic mechanism of some neuronal diseases (for example Parkinson and Alzheimer). 14-3-3 zeta is a highly-expressed protein in Alzheimer’s disease (AD) brain, which was closely related with the formation of Tau aggregation and neurofibrillary tangles. However, little useful information has been available concerning the 3D architecture of 14-3-3 zeta in AD disease. In this paper, the transgenic AD mice were used and the 1mm-thick brain slices were passively clarified. Immunofluorescence staining results showed that different from the control group, 14-3-3 zeta was mainly present around the hippocampus in AD mice. Additionally, the morphology of 14-3-3 zeta protein was filamentous with different lengths. This result will be helpful for exploring the in vivo role of 14-3-3 zeta protein during the progression of AD.