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Harmful impact on presynaptic glutamate and GABA transport by carbon dots synthesized from sulfur-containing carbohydrate precursor

Environmental Science and Pollution Research volume 24, pages 17688–17700 (2017)Cite this article

Abstract

Carbon nanoparticles that may be potent air pollutants with adverse effects on human health often contain heteroatoms including sulfur. In order to study in detail their effects on different physiological and biochemical processes, artificially produced carbon dots (CDs) with well-controlled composition that allows fluorescence detection may be of great use. Having been prepared from different types of organic precursors, CDs expose different atoms at their surface suggesting a broad variation of functional groups. Recently, we demonstrated neurotoxic properties of CDs synthesized from the amino acid β-alanine, and it is of importance to analyze whether CDs obtained from different precursors and particularly those exposing sulfur atoms induce similar neurotoxic effects. This study focused on synthesis of CDs from the sulfur-containing precursor thiourea-CDs (TU-CDs) with a size less than 10 nm, their characterization, and neuroactivity assessment. Neuroactive properties of TU-CDs were analyzed based on their effects on the key characteristics of glutamatergic and γ-aminobutyric acid (GABA) neurotransmission in isolated rat brain nerve terminals. It was observed that TU-CDs (0.5–1.0 mg/ml) attenuated the initial velocity of Na+-dependent transporter-mediated uptake and accumulation of l-[14C]glutamate and [3H]GABA by nerve terminals in a dose-dependent manner and increased the ambient level of the neurotransmitters. Starting from the concentration of 0.2 mg/ml, TU-CDs evoked a gradual dose-dependent depolarization of the plasma membrane of nerve terminals measured with the cationic potentiometric dye rhodamine 6G. Within the concentration range of 0.1–0.5 mg/ml, TU-CDs caused an “unphysiological” step-like increase in fluorescence intensity of the рН-sensitive fluorescent dye acridine orange accumulated by synaptic vesicles. Therefore, despite different surface properties and fluorescent features of CDs prepared from different starting materials (thiourea and β-alanine), their principal neurotoxic effects are analogous but displayed at a different level of efficiency. Sulfur-containing TU-CDs exhibit lower effects (by ~30%) on glutamate and GABA transport in the nerve terminals in comparison with sulfur-free β-alanine CDs. Our results suggest considering that an uncontrolled presence of carbon-containing particulate matter in the human environment may pose a toxicity risk for the central nervous system.

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Acknowledgements

This work was supported by the Science and Technology Center in Ukraine (STCU) grant no. 6055, the Programs of National Academy of Science of Ukraine “Molecular and cellular biotechnology for medicine, industry and agriculture,” the “Sensors for medicine, ecology, industry, and technology,” and the Horizon 2020 ERA-PLANET. We thank Dr. A. Chunihin from our institute for help in liposome size assessment, Makogonenko E.M. and Bereznytsky G.K. for help with SPR experiment. The study was supported by the International Research and Innovation in Medicine Program of Cedars-Sinai Medical Center, the RECOOP HST Association (Association for Regional Cooperation in the Fields of Health, Science and Technology) and the participating CRRC (Cedars-Sinai Medical Center—RECOOP Research Centers).

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Authors and Affiliations

  1. Dept. Neurochemistry and Lab. of Nanobiotechnologies Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, 9 Leontovicha Street, Kiev, 01601, Ukraine

    Tatiana Borisova, Mariia Dekaliuk, Natalia Pozdnyakova, Artem Pastukhov, Marina Dudarenko, Arsenii Borysov & Alexander P. Demchenko

  2. International Research and Innovation in Medicine Program, Cedars-Sinai Medical Center, Los Angeles, CA, USA

    Sandor G. Vari

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  1. Tatiana Borisova
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  2. Mariia Dekaliuk
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  3. Natalia Pozdnyakova
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  4. Artem Pastukhov
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  5. Marina Dudarenko
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  6. Arsenii Borysov
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  8. Alexander P. Demchenko
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Contributions

Synthesis of TU-CDs and measurements of their spectroscopic properties were carried out by MD. Isolation of synaptosomes, l-[14C]glutamate and [3H]GABA uptake/release experiments, and spectrofluorimetry with TU-CDs using synaptosomes were performed by NP, MD, AP, and AB. Data analysis was done by ТB, AD, NP, and MD; manuscript writing by ТB and AD; and discussion of results and editing the final manuscript by SV.

Corresponding author

Correspondence to Tatiana Borisova.

Ethics declarations

The experimental procedures were conducted according to “Scientific Requirements and Research Protocols” and “Research Ethics Committees” from the Declaration of Helsinki. Experimental protocols were approved by the Animal Care and Use Committee of the Institute (the Protocol from September 19, 2011).

Competing interests

The authors declare that they have no competing interests.

Additional information

Responsible editor: Philippe Garrigues

Availability of data and materials

The datasets supporting the conclusions of this article are available in the PubChem BioAssay; caNanoLab repository (https://cananolab.nci.nih.gov/caNanoLab/#/; http://www.ncbi.nlm.nih.gov/pcassay)

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Borisova, T., Dekaliuk, M., Pozdnyakova, N. et al. Harmful impact on presynaptic glutamate and GABA transport by carbon dots synthesized from sulfur-containing carbohydrate precursor. Environ Sci Pollut Res 24, 17688–17700 (2017). https://doi.org/10.1007/s11356-017-9414-6

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  • DOI: https://doi.org/10.1007/s11356-017-9414-6

Keywords

  • Carbon dots from thiourea
  • Health risks
  • Na+-dependent uptake of GABA and glutamate
  • Extracellular level of neurotransmitters
  • Brain nerve terminals