High Contrast and Resolution Labeling of Amyloid Plaques in Tissue Sections from APP-PS1 Mice and Humans with Alzheimer’s Disease with the Zinc Chelator HQ-O: Practical and Theoretical Considerations

Author(s): Larry Schmued*, James Raymick, Sumit Sarkar.

Journal Name: Current Alzheimer Research

Volume 16 , Issue 7 , 2019

  Journal Home
Translate in Chinese
Become EABM
Become Reviewer

Abstract:

Background: Various methodologies have been employed for the localization of amyloid plaques in numerous studies on Alzheimer’s disease. The majority of these stains are thought to label the plaques by virtue of their affinity for aggregated Aβ. However, plaques are known to contain numerous other components, including multivalent metals such as zinc.

Objective: This investigates whether it is possible to localize the presence of zinc in parenchymal and vascular amyloid plaques in afflicted brains. To accomplish this, a novel fluorescent zinc chelator, HQO, was investigated to determine its mechanism of binding and to optimize a stain for the high contrast and resolution histological localization of amyloid plaques.

Methods: A novel zinc chelator, HQ-O, was developed for localizing zinc within amyloid plaques. The histology involves incubating tissue sections in a dilute aqueous solution of HQ-O. Its compatibility with a variety of other fluorescent methodologies is described.

Results: All amyloid plaques are stained in fine detail and appear bright green under blue light excitation. The staining of parenchymal plaques correlates closely with that seen following staining with antibodies to Aβ, however, the HQ-O sometimes also label additional globular structures within blood vessels. In situ mechanistic studies revealed that fluorescent plaque-like structures are only observed with HQ-O when synthetic Aβx-42 is aggregated in the presence of zinc.

Conclusion: Zinc is intimately bound to all amyloid plaques, which was demonstrated by its histological localization using a novel fluorescent zinc chelator, HQ-O. Additionally, the tracer is also capable of labeling intravascular leucocytes due to their high zinc content.

Keywords: Amyloid plaques, Alzheimer's disease, zinc chelator, brain pathology, Aβ, Amylo-Glo.

[1]
Puchtler H, Sweat F. Congo red as a stain for fluorescence microscopy of amyloid. J Histochem Cytochem 13(8): 693-4. (1965)
[http://dx.doi.org/10.1177/13.8.693] [PMID: 4160077]
[2]
Kelényi G. Thioflavin S fluorescent and Congo red anisotropic stainings in the histologic demonstration of amyloid. Acta Neuropathol 7(4): 336-48. (1967)
[http://dx.doi.org/10.1007/BF00688089] [PMID: 4166287]
[3]
Schmued L, Raymick J, Tolleson W, Sarkar S, Bell-Cohn A. Introducing Amylo-Glo, a novel histochemical trracer especially suited for multiple labeling and large scale quantification studies. J Neurosci Methods 30(209): 120-6. (2012)
[http://dx.doi.org/10.1016/j.jneumeth.2012.05.019] [PMID: 22705750]
[4]
Sato K, Higuchi M, Iwata N, Saido TC, Sasamoto K. Fluoro-substituted and 13C-labeled styrylbenzene derivatives for detecting brain amyloid plaques. Eur J Med Chem 39(7): 573-8. (2004)
[http://dx.doi.org/10.1016/j.ejmech.2004.02.013] [PMID: 15236837]
[5]
Lee JY, Mook-Jung I, Koh JY. Histochemically reactive zinc in plaques of the Swedish mutant beta-amyloid precursor protein transgenic mice. J Neurosci 19(11): RC10. (1999)
[http://dx.doi.org/10.1523/JNEUROSCI.19-11-j0002.1999] [PMID: 10341271]
[6]
Falangola MF, Lee SP, Nixon RA, Duff K, Helpern JA. Histological co-localization of iron in Abeta plaques of PS/APP transgenic mice. Neurochem Res 30(2): 201-5. (2005)
[http://dx.doi.org/10.1007/s11064-004-2442-x] [PMID: 15895823]
[7]
Bourassa MW, Leskovjan AC, Tappero RV, Farquhar ER, Colton CA, Van Nostrand WE, et al. Elevated copper in the amyloid plaques and iron in the cortex are observed in mouse models of Alzheimer’s disease that exhibit neurodegeneration. Biomed Spectrosc Imaging 2(2): 129-39. (2013)
[PMID: 24926425]
[8]
Prachayasittikul V, Prachayasittikul S, Ruchirawat S, Prachayasittikul V. 8-Hydroxyquinolines: a review of their metal chelating properties and medicinal applications. Drug Des Devel Ther 7: 1157-78. (2013)
[http://dx.doi.org/10.2147/DDDT.S49763] [PMID: 24115839]
[9]
Smith GL, Jenkins RA, Gough JF. A fluorescent method for the detection and localization of zinc in human granulocytes. J Histochem Cytochem 17(11): 749-50. (1969)
[http://dx.doi.org/10.1177/17.11.749] [PMID: 5386301]
[10]
Sternberg SS, Cronin A, Philips FS. Histochemical demonstration of zinc in the dorsolateral prostate of the rat: studies with oxine and dithizone. Am J Pathol 47: 325-37. (1965)
[PMID: 14335773]
[11]
Iwatsubo T, Odaka A, Suzuki N, Mizusawa H, Nukina N, Ihara Y. Visualization of A beta 42(43) and A beta 40 in senile plaques with end-specific A beta monoclonals: evidence that an initially deposited species is A beta 42(43). Neuron 13(1): 45-53. (1994)
[http://dx.doi.org/10.1016/0896-6273(94)90458-8] [PMID: 8043280]
[12]
Cherny RA, Barnham KJ, Lynch T, Volitakis I, Li QX, McLean CA, et al. Chelation and intercalation: complementary properties in a compound for the treatment of Alzheimer’s disease. J Struct Biol 130(2-3): 209-16. (2000)
[http://dx.doi.org/10.1006/jsbi.2000.4285] [PMID: 10940226]
[13]
James SA, Churches QI, de Jonge MD, Birchall IE, Streltsov V, McColl G, et al. Iron, Copper, and Zinc Concentration in Aβ plaques in the app/ps1 mouse model of Alzheimer’s disease correlates with metal levels in the surrounding neuropil. ACS Chem Neurosci 8(3): 629-37. (2017)
[http://dx.doi.org/10.1021/acschemneuro.6b00362] [PMID: 27958708]
[14]
Bush AI, Pettingell WH Jr, de Paradis M, Tanzi RE, Wasco W. The amyloid beta-protein precursor and its mammalian homologues. Evidence for a zinc-modulated heparin-binding superfamily. J Biol Chem 269(43): 26618-21. (1994)
[PMID: 7929392]
[15]
Cherny RA, Atwood CS, Xilinas ME, Gray DN, Jones WD, McLean CA, et al. Treatment with a copper-zinc chelator markedly and rapidly inhibits beta-amyloid accumulation in Alzheimer’s disease transgenic mice. Neuron 30(3): 665-76. (2001)
[http://dx.doi.org/10.1016/S0896-6273(01)00317-8] [PMID: 11430801]
[16]
Zhang YH, Raymick J, Sarkar S, Lahiri DK, Ray B, Holtzman D, et al. Efficacy and toxicity of clioquinol treatment and A-beta42 inoculation in the APP/PSI mouse model of Alzheimer’s disease. Curr Alzheimer Res 10(5): 494-506. (2013)
[http://dx.doi.org/10.2174/1567205011310050005] [PMID: 23627708]
[17]
Tsubaki T, Honma Y, Hoshi M. Neurological syndrome associated with clioquinol. Lancet 1(7701): 696-7. (1971)
[http://dx.doi.org/10.1016/S0140-6736(71)92699-7] [PMID: 4101631]
[18]
Tyszka-Czochara M, Grzywacz A, Gdula-Argasińska J, Librowski T, Wiliński B, Opoka W. The role of zinc in the pathogenesis and treatment of central nervous system (CNS) diseases. Implications of zinc homeostasis for proper CNS function. Acta Pol Pharm 71(3): 369-77. (2014)
[PMID: 25265815]


Rights & PermissionsPrintExport Cite as

Article Details

VOLUME: 16
ISSUE: 7
Year: 2019
Page: [577 - 586]
Pages: 10
DOI: 10.2174/1567205016666190725155038

Article Metrics

PDF: 28
HTML: 4