Bring CLARITY to Temporal Lobe Epilepsy: 3D Visualization of p-Tau(Ser262) and 14-3-3 Zeta
Qingqin Wu,
Honghong Song,
Juan Feng,
Yang Xia,
Dezhong Yao
Issue:
Volume 5, Issue 6, December 2017
Pages:
63-67
Received:
5 December 2017
Published:
6 December 2017
Abstract: CLARITY is one new technology which allows the brain tissue become transparent. It has successfully been combined with immunofluorescence staining to achieve the 3D visualization of some molecules or neuronal cells in some disease brains. The temporal lobe epilepsy (TLE) is one neuronal disease which is characterized by the sprouting of the massy fibers (MSF). Previous study has showed that MSF could be affected by the phosphorylation at site-262 of microtubule association protein Tau (p-tau (Ser262)). However, in TLE little useful information was reported concerning the 3D architecture of p-tau (Ser262) and its relationship with 14-3-3 zeta that regulated the phosphorylation of Tau in AD disease. In this paper, pilocarpine-induced epilepsy model was established and identified by Timm-staining. Double immunofluorescent staining results showed that the development of TLE gave rise to the colocalization of p-tau (Ser262) and 14-3-3 zeta protein in CA1 and CA3 zone in hippocampi. The mm-thick brain sections were passively clarified, and 3D reconstruction imaging of the immunofluorescent staining showed that p-tau (Ser262) was diverse cluster-like shape. These results proved that CLARITY could be used to study TLE, in which the 3D morphologic changes of p-tau (Ser262) and the role of 14-3-3 zeta in the regulation of Tau needed to be further investigated.
Abstract: CLARITY is one new technology which allows the brain tissue become transparent. It has successfully been combined with immunofluorescence staining to achieve the 3D visualization of some molecules or neuronal cells in some disease brains. The temporal lobe epilepsy (TLE) is one neuronal disease which is characterized by the sprouting of the massy f...
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Application of CLARITY to Investigate the 3D Architecture of 14-3-3 Zeta Protein in AD
Honghong Song,
Juan Feng,
Jian Li,
Yang Xia,
Dezhong Yao
Issue:
Volume 5, Issue 6, December 2017
Pages:
68-71
Received:
5 December 2017
Published:
6 December 2017
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.
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 tissu...
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