检测淀粉样沉淀蛋白的三种染色方法比较：抗体染色、Gallyas银染和硫磺素S染色

施新泽; 魏轩; 沙龙泽; 许琪

doi:10.24920/03476

Your Location：

Home >

Browse articles >

检测淀粉样沉淀蛋白的三种染色方法比较：抗体染色、Gallyas银染和硫磺素S染色

Chinese Medical Sciences Journal 2018年33卷第3期页码：167-173

研究论文 | Updated：2024-04-10

- 检测淀粉样沉淀蛋白的三种染色方法比较：抗体染色、Gallyas银染和硫磺素S染色
- Affiliations：
  
  1 State Key Laboratory of Medical Molecular Biology and Neuroscience Center, Institute of Basic Medical Sciences Chinese Academy of Medical Sciences & School of Basic Medicine Peking Union Medical College, Beijing 100005, China;
  2 High School Attached to Beijing University of Technology,Beijing 100022, China
- Author bio：
  
  * Tel: 86-10-69156432, E-mail: xuqi@pumc.edu.cn
- Funds：
  
  Supported by the 2016 Major Collaborative Innovation Program of the Chinese Academy of Medical Sciences(2016-I2M-1004)
- DOI：10.24920/03476
  中图分类号：
- 网络出版日期：2018-09-20，
  
  纸质出版日期：2018-09-20
- Accepted：
Scan QR Code
施新泽, 魏轩, 沙龙泽, 等. 检测淀粉样沉淀蛋白的三种染色方法比较：抗体染色、Gallyas银染和硫磺素S染色[J]. Chinese Medical Sciences Journal, 2018,33(3):167-173.

Shi Xinze, Wei Xuan, Sha Longze, et al. Comparison of β-Amyloid Plaque Labeling Methods: Antibody Staining, Gallyas Silver Staining, and Thioflavin-S Staining[J]. Chinese medical sciences journal, 2018, 33(3): 167-173.
施新泽, 魏轩, 沙龙泽, 等. 检测淀粉样沉淀蛋白的三种染色方法比较：抗体染色、Gallyas银染和硫磺素S染色[J]. Chinese Medical Sciences Journal, 2018,33(3):167-173. DOI： 10.24920/03476.

Shi Xinze, Wei Xuan, Sha Longze, et al. Comparison of β-Amyloid Plaque Labeling Methods: Antibody Staining, Gallyas Silver Staining, and Thioflavin-S Staining[J]. Chinese medical sciences journal, 2018, 33(3): 167-173. DOI： 10.24920/03476.

摘要

目的

探究阿尔茨海默病小鼠模型的淀粉样沉淀的形成过程。比较抗体染色

Gallyas银染和硫磺素S染色3种染色技术检测淀粉样沉淀的效果。

方法

检测不同月龄的APPswe/PS1dE9转基因小鼠（APP/PS1）大脑淀粉样沉积的时空变化。对小鼠同一脑区的临近切片行抗体染色

Gallyas银染和硫磺素S染色

并对比其染色效果。

结果

随着模型鼠年龄的增大

淀粉样沉积首先出现在的大脑皮层

随后扩散至整个大脑。与Gallyas银染和硫磺素S染色比较

6E10抗体染色能检测到更多的淀粉样斑块沉积（P

0.05）。Gallyas银染和硫磺素S染色的检测结果之间没有显著的统计学意义（P = 0.0033）。

结论

APP/PS1转基因小鼠模型能够模拟阿尔茨海默病患者脑内的淀粉样沉积过程。抗体染色得到的结果相较于其它两种检测方法更为可靠。

Abstract

Objective

To evaluate senile plaque formation and compare the sensitivity of three different β-amyloid (Aβ) labeling methods (antibody staining

Gallyas silver staining

and thioflavin-S staining) to detect Aβ deposition.

Methods

APPswe/PSEN1dE9 transgenic mice (APP/PS1) of different ages were used to examine spatiotemporal changes in Aβ plaque deposition. Antibody staining

Gallyas silver staining

and thioflavin-S staining were used to detect Aβ plaque deposition in the same brain region of adjacent slices from model mice

and the results were compared.

Results

With aging

Aβ plaques first appeared in the cortex and then the deposition increased throughout the whole brain. Significantly greater plaque deposition was detected by 6E10 antibody than that analyzed with Gallyas silver staining or thioflavin-S staining (

0.05). Plaque deposition did not show significant difference between the APP/PS1 mice brains assayed with Gallyas silver staining and ones with thioflavin-S staining (

=0.0033).

Conclusions

The APP/PS1 mouse model of Alzheimer’s disease could mimick the progress of Aβ plaques occurred in patients with Alzheimer’s disease. Antibody detection of Aβ deposition may be more sensitive than chemical staining methods.

关键词

Keywords

references

Scheltens P , Blennow K , BretelerM M , et al . Alzheimer’s disease . Lancet 2016 ; 388 ( 10043 ): 505 - 17 . doi: 10.1016/S0140-6736(15)01124-1 10.1016/S0140-6736(15)01124-1 . DOI: 10.1016/S0140-6736(15)01124-1 http://doi.org/10.1016/S0140-6736(15)01124-1 https://linkinghub.elsevier.com/retrieve/pii/S0140673615011241 https://linkinghub.elsevier.com/retrieve/pii/S0140673615011241

Strooper BD , Karran E . The cellular phase of Alzheimer’s disease . Cell 2016 ; 164 ( 4 ): 603 - 15 . doi: 10.1016/j.cell.2015.12.056 10.1016/j.cell.2015.12.056 . DOI: 10.1016/j.cell.2015.12.056 http://doi.org/10.1016/j.cell.2015.12.056 https://linkinghub.elsevier.com/retrieve/pii/S0092867415017201 https://linkinghub.elsevier.com/retrieve/pii/S0092867415017201

Goedert M . Neurodegeneration. Alzheimer’s and Parkinson’s diseases: the prion concept in relation to assembled Aβ, tau, and α-synuclein . Science 2015 ; 349 ( 6248 ): 1255555 . doi: 10.1126/science.1255555 10.1126/science.1255555 . DOI: 10.1126/science.1255555 http://doi.org/10.1126/science.1255555 http://www.sciencemag.org/cgi/doi/10.1126/science.1255555 http://www.sciencemag.org/cgi/doi/10.1126/science.1255555

Lindberg DJ , Wenger A , Sundin E , et al . Binding of thioflavin-T to amyloid fibrils leads to fluorescence self-quenching and fibril compaction . Biochemistry 2017 ; 56 ( 16 ): 2170 . doi: 10.1021/acs.biochem.7b00035 10.1021/acs.biochem.7b00035 . DOI: 10.1021/acs.biochem.7b00035 http://doi.org/10.1021/acs.biochem.7b00035 http://pubs.acs.org/doi/10.1021/acs.biochem.7b00035 http://pubs.acs.org/doi/10.1021/acs.biochem.7b00035

Luna-Muñoz J , Peralta-Ramirez J , Chávez-Macías L , et al . Thiazin red as a neuropathological tool for the rapid diagnosis of Alzheimer’s disease in tissue imprints . Acta Neuropathol 2008 ; 116 ( 5 ): 507 - 15 . doi: 10.1007/s00401-008-0431-x 10.1007/s00401-008-0431-x . DOI: 10.1007/s00401-008-0431-x http://doi.org/10.1007/s00401-008-0431-x http://link.springer.com/10.1007/s00401-008-0431-x http://link.springer.com/10.1007/s00401-008-0431-x

Menter T , Bachmann M , Grieshaber S , et al . A more accurate approach to amyloid detection and subtyping: combining in situ Congo Red staining and immunohistochemistry . Pathobiology 2008 ; 84 ( 1 ): 49 - 55 . doi: 10.1159/000447304 10.1159/000447304 .

Uchihara T , Tsuchiya K , Nakamura A , et al . Silver staining profiles distinguish Pick bodies from neurofibrillary tangles of Alzheimer type: comparison between Gallyas and Campbell-Switzer methods . Acta Neuropathol 2005 ; 109 ( 5 ): 483 - 9 . doi: 10.1007/s00401-005-0988-6 10.1007/s00401-005-0988-6 . DOI: 10.1007/s00401-005-0988-6 http://doi.org/10.1007/s00401-005-0988-6 http://link.springer.com/10.1007/s00401-005-0988-6 http://link.springer.com/10.1007/s00401-005-0988-6

Mavrogiorgou P , Gertz HJ , Ferszt R , et al . Are routine methods good enough to stain senile plaques and neurofibrillary tangles in different brain regions of demented patients? Psychiatr Danub 2011 ; 23 ( 4 ): 334 - 9 .

Harding AJ , Kril JJ , Halliday GM . Practical measures to simplify the Braak tangle staging method, for routine pathological screening . Acta Neuropathol 2000 ; 99 ( 2 ): 199 - 208 . DOI: 10.1007/PL00007425 http://doi.org/10.1007/PL00007425 http://link.springer.com/10.1007/PL00007425 http://link.springer.com/10.1007/PL00007425

Kuninaka N , Kawaguchi M , Ogawa M , et al . Simplification of the modified Gallyas method . Neuropathology 2015 ; 35 ( 1 ): 10 - 5 . doi: 10.1111/neup.12144 10.1111/neup.12144 . DOI: 10.1111/neup.12144 http://doi.org/10.1111/neup.12144 http://doi.wiley.com/10.1111/neup.2015.35.issue-1 http://doi.wiley.com/10.1111/neup.2015.35.issue-1

Bhattacharya S , Haertel C , Maelicke A , et al . Galantamine slows down plaque formation and behavioral decline in the 5XFAD mouse model of Alzheimer’s disease . PLoS One 2014 ; 9 ( 2 ): e89454 . doi: 10.1371/journal.pone.0089454.eCollection 2014 10.1371/journal.pone.0089454.eCollection 2014 . DOI: 10.1371/journal.pone.0089454 http://doi.org/10.1371/journal.pone.0089454 http://dx.plos.org/10.1371/journal.pone.0089454 http://dx.plos.org/10.1371/journal.pone.0089454

Grienberger C , Rochefort NL , Adelsberger H , et al . Staged decline of neuronal function in vivo in an animal model of Alzheimer’s disease . Nature Communications 2012 ; 3 : 774 . doi: 10.1038/ncomms1783 10.1038/ncomms1783 . DOI: 10.1038/ncomms1783 http://doi.org/10.1038/ncomms1783 http://www.nature.com/articles/ncomms1783 http://www.nature.com/articles/ncomms1783

Kawai R , Araki M , Yoshimura M , et al . Core binding site of a thioflavin-T-derived imaging probe on amyloid β fibrils predicted by computational methods . ACS Chem Neurosci 2018 ; 9 ( 5 ): 957 - 66 . doi: 10.1021/acschemneuro.7b00389 10.1021/acschemneuro.7b00389 . DOI: 10.1021/acschemneuro.7b00389 http://doi.org/10.1021/acschemneuro.7b00389 http://pubs.acs.org/doi/10.1021/acschemneuro.7b00389 http://pubs.acs.org/doi/10.1021/acschemneuro.7b00389

Bacskai BJ , Hickey GA , Skoch J , et al . Four-dimensional multiphoton imaging of brain entry, amyloid binding, and clearance of an amyloid-β ligand in transgenic mice . Proc Natl Acad Sci U S A 2003 ; 100 ( 21 ): 12462 - 7 . doi: 10.1073/pnas.2034101100 10.1073/pnas.2034101100 . DOI: 10.1073/pnas.2034101100 http://doi.org/10.1073/pnas.2034101100 http://www.pnas.org/cgi/doi/10.1073/pnas.2034101100 http://www.pnas.org/cgi/doi/10.1073/pnas.2034101100

Villemagne VL , Burnham S , Bourgeat P , et al . Amyloid β deposition, neurodegeneration, and cognitive decline in sporadic Alzheimer’s disease: a prospective cohort study . Lancet Neurol 2013 ; 12 ( 4 ): 357 . doi: 10.1016/S1474-4422(13)70044-9 10.1016/S1474-4422(13)70044-9 . DOI: 10.1016/S1474-4422(13)70044-9 http://doi.org/10.1016/S1474-4422(13)70044-9 http://linkinghub.elsevier.com/retrieve/pii/S1474442213700449 http://linkinghub.elsevier.com/retrieve/pii/S1474442213700449

Sun A , Xuan VN , Bing G . Comparative analysis of an improved Thioflavin-S Stain, Gallyas Silver Stain, and immunohistochemistry for neurofibrillary tangle demonstration on the same sections . J Histochem Cytochem 2002 ; 50 ( 4 ): 463 - 72 . doi: 10.1177/002215540205000403 10.1177/002215540205000403 . DOI: 10.1177/002215540205000403 http://doi.org/10.1177/002215540205000403 http://journals.sagepub.com/doi/10.1177/002215540205000403 http://journals.sagepub.com/doi/10.1177/002215540205000403

Czasch S , Paul S , Baumgärtner W . A comparison of immunohistochemical and silver staining methods for the detection of diffuse plaques in the aged canine brain . Neurobiol Aging 2006 ; 27 ( 2 ): 293 - 305 . doi: 10.1016/j.neurobiolaging.2005.02.017 10.1016/j.neurobiolaging.2005.02.017 . DOI: 10.1016/j.neurobiolaging.2005.02.017 http://doi.org/10.1016/j.neurobiolaging.2005.02.017 http://linkinghub.elsevier.com/retrieve/pii/S0197458005001661 http://linkinghub.elsevier.com/retrieve/pii/S0197458005001661

Wilcock GK , Matthews SM , Moss T . Comparison of three silver stains for demonstrating neurofibrillary tangles and neuritic plaques in brain tissue stored for long periods . Acta Neuropathol 1990 ; 79 ( 5 ): 566 - 8 . DOI: 10.1007/BF00296118 http://doi.org/10.1007/BF00296118 http://link.springer.com/10.1007/BF00296118 http://link.springer.com/10.1007/BF00296118

Radde R , Bolmont T , Kaeser SA , et al . Abeta42-driven cerebral amyloidosis in transgenic mice reveals early and robust pathology . EMBO Rep 2006 ; 7 ( 9 ): 940 - 6 . doi: 10.1038/sj.embor.7400784 10.1038/sj.embor.7400784 . DOI: 10.1038/sj.embor.7400784 http://doi.org/10.1038/sj.embor.7400784 http://embor.embopress.org/cgi/doi/10.1038/sj.embor.7400784 http://embor.embopress.org/cgi/doi/10.1038/sj.embor.7400784

浏览量

2462

Downloads

CSCD

文章被引用时，请邮件提醒。

Submit

关联资源

暂无数据