Fluorescent characterization of amyloid deposits in the kidneys of mdx mice

  • Valeriia Gusel'nikova Institute of Experimental Medicine, St. Petersburg, Russian Federation. http://orcid.org/0000-0002-9499-8275
  • Olga Antimonova Institute of Experimental Medicine, St. Petersburg, Russian Federation.
  • Elena Fedorova Institute of Experimental Medicine, St. Petersburg, Russian Federation. http://orcid.org/0000-0002-0190-885X
  • Mikhail Shavlovsky Institute of Experimental Medicine, St. Petersburg State Medical University academician I.P. Pavlov, Russian Academy of Sciences, Northwestern State Medical University I.I. Mechnikov, Russian Federation.
  • Aleksandr Krutikov Northwestern Federal Medical Research Center V.A. Almazov, Russian Federation.
  • Ekaterina Mikhailova | 4evamkh@gmail.com Russian Academy of Sciences, Institute of Cytology, Department of Protein Stability, Dynamics and Folding, Russian Federation. http://orcid.org/0000-0002-6982-0360
  • Aleksandra Gudkova Northwestern Federal Medical Research Center V.A. Almazov, Russian Academy of Sciences, Russian Federation. http://orcid.org/0000-0003-0156-8821
  • Vyacheslav Mikhailov Russian Academy of Sciences, Institute of Cytology, Department of Protein Stability, Dynamics and Folding, Russian Federation. http://orcid.org/0000-0002-3937-8655
  • Dmitry Korzhevskii Institute of Experimental Medicine, St. Petersburg State Medical University academician I.P. Pavlov, Russian Federation. http://orcid.org/0000-0002-2456-8165

Abstract

Amyloidosis is a group of diseases that occurs when amyloid proteins are deposited in tissues and organs. The traditional way of identifying amyloid in tissue sections is staining with Congo red. However, this method has a number of limitations including background staining (background fluorescence), low fluorescence intensity and false-positive staining. Therefore, a complex of fluorescence-based methods should be applied to characterize tissue localization of amyloid deposits. The aim of this study was to identify amyloid deposits in the kidneys of dystrophin-deficient mdx mice using different fluorescent dyes. We examined 8 cases of renal amyloidosis in aged mdx mice. In all cases, we used traditional methods for amyloid detection (Congo red and Thioflavin T), as well as a new fluorescent dye, disodium salt of 2,7- (1-amino-4-sulfo-2-naphthylazo) fluorene (DSNAF). In our study, we confirmed the amyloid structure of protein deposits in kidneys of aging mdx mice by several fluorescence-based staining methods. We found that fixation method has profound effects on downstream staining procedures, and demonstrated that the application of specific fixative, zinc-ethanol-formaldehyde (ZEF), instead of traditional NBF allow to reduce the background fluorescence. We also illustrated the usefulness of novel fluorescent dye DSNAF for detection of amyloid deposits in mouse tissues. Our results confirmed the strong affinity and high specificity of this dye for amyloid fibrils. The verification of DSNAF for detecting amyloid in human tissues will provide a conclusion on the applicability of the developed staining method in clinical research practice.

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Author Biographies

Valeriia Gusel'nikova, Institute of Experimental Medicine, St. Petersburg
Laboratory of Functional Morphology of the Central and Peripheral Nervous System
Olga Antimonova, Institute of Experimental Medicine, St. Petersburg

Laboratory of Human Molecular Genetics

Elena Fedorova, Institute of Experimental Medicine, St. Petersburg

Laboratory of Functional Morphology of the Central and Peripheral Nervous System

Mikhail Shavlovsky, Institute of Experimental Medicine, St. Petersburg State Medical University academician I.P. Pavlov, Russian Academy of Sciences, Northwestern State Medical University I.I. Mechnikov

Laboratory of Human Molecular Genetics

Aleksandr Krutikov, Northwestern Federal Medical Research Center V.A. Almazov

Research Department of the Infiltrative Heart Diseases

Aleksandra Gudkova, Northwestern Federal Medical Research Center V.A. Almazov, Russian Academy of Sciences

Cardiomyopathy Laboratory

Vyacheslav Mikhailov, Russian Academy of Sciences, Institute of Cytology, Department of Protein Stability, Dynamics and Folding

Group of Cell populations genetics

Dmitry Korzhevskii, Institute of Experimental Medicine, St. Petersburg State Medical University academician I.P. Pavlov

Laboratory of Functional Morphology of the Central and Peripheral Nervous System

Published
2018-04-03
Section
Original Papers
Keywords:
amyloidosis, mdx mice, Congo red, Thioflavin T, fluorene
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How to Cite
Gusel’nikova, V., Antimonova, O., Fedorova, E., Shavlovsky, M., Krutikov, A., Mikhailova, E., Gudkova, A., Mikhailov, V., & Korzhevskii, D. (2018). Fluorescent characterization of amyloid deposits in the kidneys of mdx mice. European Journal of Histochemistry, 62(2). https://doi.org/10.4081/ejh.2018.2870