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Alpha-Synuclein Fibrils Interact with Dopamine Reducing its Cytotoxicity on PC12 Cells

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Abstract

Aggregated alpha-synuclein (α-SYN) is the major component of Lewy bodies and Lewy neurites, two of the pathological hallmarks of Parkinson’s disease (PD). Aggregation of α-SYN leads to toxic species involved in the degeneration of dopaminergic neurons in the midbrain. Different studies suggest a strong association between the presence of dopamine (DA) and the cell specific degeneration caused by α-SYN aggregates in PD. Despite extensive studies on the effect of DA on α-SYN fibrillation, it remains unclear how the simultaneous presence of DA and α-SYN influences the degeneration of dopaminergic neurons. In this study we show that separate treatments with specific doses of DA or early stage α-SYN aggregates (ESAA) are both cytotoxic to PC12 cells. Surprisingly, simultaneous treatment of cells with DA and ESAA significantly decreased this toxicity. This cytotoxicity was further reduced by the presence of heavier particles of α-SYN aggregates with more fibrillogenic characteristics. Spectrometric analysis revealed that α-SYN fibrils interact with DA even after the sample was dialyzed for 48 h, suggesting a strong interaction. Interestingly, digestion of unprotected N- and C-α-SYN-fibril terminals by proteinase K did not affect this interaction. Our results suggest that fibrillar forms of α-SYN with localized expanded active surfaces may interact with DA and moderate its cytotoxicity. Thus, highlighting the importance of fibrillar proteins in developing clinical approaches for amyloid diseases.

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Abbreviations

AFM:

Atomic Force Microscopy

α-SYN:

α-Synuclein

CD:

Circular Dichroism

DA:

Dopamine

DAQ:

Dopamine quinone

ESAA:

Early Stages α-SYN Aggregates

FTIR:

Fourier Transform infrared

MTT:

3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide

OD:

Optical Density

PD:

Parkinson’s Disease

PK:

Proteinase K

PI:

Propidium Iodide

ROS:

Reactive Oxygen Species

SNpc:

Substantia Nigra pars compacta

ThT:

Thioflavin T

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Acknowledgments

This work was supported by the National Institute of Genetic Engineering and Biotechnology.

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Authors and Affiliations

  1. Department of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran

    Masoome Khalife, Dina Morshedi, Farhang Aliakbari, Amir Tayaranian Marvian, Hossein Mohammad Beigi & Sadegh Azimzadeh Jamalkandi

  2. Department of Medical Biotechnology, Semnan University of Medical Sciences, Semnan, Iran

    Farhang Aliakbari

  3. Department of Chemical Engineering, Tarbiat Modares University, Tehran, Iran

    Hossein Mohammad Beigi

  4. Chemical Injuries Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran

    Sadegh Azimzadeh Jamalkandi

  5. Munich Cluster for Systems Neurology, SyNergy Clinician Scientist Group, Klinikum der Universität München, Munich, Germany

    Francisco Pan-Montojo

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  1. Masoome Khalife
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Correspondence to Dina Morshedi.

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10930_2015_9625_MOESM1_ESM.pdf

Study of ASN fibrillation. Kinetic of the fibrillation monitored by (a) changes in ThT fluorescence intensity or (b) Congo red absorbance. The inset indicates maximum intensity of fluorescence. (c) Far-UV CD spectra of ASN obtained after 0-hours and 48-hours. (d) Fluorescence images of ASN fibrils dyed with ThT (PDF 214 kb)

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Khalife, M., Morshedi, D., Aliakbari, F. et al. Alpha-Synuclein Fibrils Interact with Dopamine Reducing its Cytotoxicity on PC12 Cells. Protein J 34, 291–303 (2015). https://doi.org/10.1007/s10930-015-9625-y

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