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Hamostaseologie
DOI: 10.1055/a-2773-1622
Original Article

Heterogeneity of Single-platelet Calcium Responses to Activation

Autor*innen

  • Fedor A. Balabin‡

    1   Center for Theoretical Problems of Physico-Chemical Pharmacology, Russian Academy of Sciences, Moscow, Russian Federation (Ringgold ID: RIN307756)

  • Sofia V. Galkina‡

    1   Center for Theoretical Problems of Physico-Chemical Pharmacology, Russian Academy of Sciences, Moscow, Russian Federation (Ringgold ID: RIN307756)
    2   Dmitry Rogachev National Medical Research Center of Pediatric Hematology, Oncology and Immunology, Moscow, Russian Federation

  • Ravilya Dzhamaliddinova

    3   Moscow Medical Research Center named after A.S. Loginov, Moscow, Russian Federation (Ringgold ID: RIN307756)

  • Leonid Vedernikov

    1   Center for Theoretical Problems of Physico-Chemical Pharmacology, Russian Academy of Sciences, Moscow, Russian Federation (Ringgold ID: RIN307756)

  • Irina E. Zhizhaikina

    4   Sechenov First Moscow State Medical University, Moscow, Russian Federation

  • Ludmila G. Zhukova

    3   Moscow Medical Research Center named after A.S. Loginov, Moscow, Russian Federation (Ringgold ID: RIN307756)

  • Ekaterina V. Shamova 

    5   National Academy of Sciences of Belarus, Institute of Biophysics and Cell Engineering, Minsk, Belarus (Ringgold ID: RIN428469)

  • Mikhail Panteleev

    1   Center for Theoretical Problems of Physico-Chemical Pharmacology, Russian Academy of Sciences, Moscow, Russian Federation (Ringgold ID: RIN307756)
    2   Dmitry Rogachev National Medical Research Center of Pediatric Hematology, Oncology and Immunology, Moscow, Russian Federation
    6   M.V. Lomonosov Moscow State University, Moscow, Russian Federation

  • Anastasia N. Sveshnikova

    1   Center for Theoretical Problems of Physico-Chemical Pharmacology, Russian Academy of Sciences, Moscow, Russian Federation (Ringgold ID: RIN307756)
    2   Dmitry Rogachev National Medical Research Center of Pediatric Hematology, Oncology and Immunology, Moscow, Russian Federation
    6   M.V. Lomonosov Moscow State University, Moscow, Russian Federation

The study was supported by the Russian Science Foundation, grant 23-45-10039, and Belarusian Republican Foundation for Fundamental Research, grant B23RSF-162.
Gefördert durch: Russian Science Foundation 23-45-10039
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Abstract

Cytosolic calcium oscillations play a central role in platelet activation. However, signaling heterogeneity between platelets of the same individual, or between individuals, is poorly characterized. We utilized total internal reflection fluorescence microscopy of calcium fluorophore-loaded, surface-attached human platelets to monitor single-platelet calcium responses to collagen, ADP, and thrombin. For all activation types in healthy adult donors, four types of platelet calcium dynamics (“activation groups) were distinguished: (I) isolated spikes; (II) oscillations with a period of 3–10 s; (III) clusters of spikes following each other with calcium levels never returning to baseline; and (IV) a sustained high calcium level. The activation Groups I and II were predominant in the immobilized platelets of healthy adults (46 ± 22% and 33 ± 10%, respectively), with 18 ± 13% of platelets in Group III. Stimulation with ADP shifted the activation pattern, with Group I fraction falling to 15 ± 9% and Group III fraction rising to 43 ± 13% instead. For stimulation with ADP plus thrombin or collagen, Group III was predominant (71 ± 11% for thrombin, 46 ± 18% for collagen). A combination of all three agonists mainly produced Level III (69 ± 15%) and Level IV (18 ± 12%) platelets. Confocal microscopy revealed a gradual increase in all activation markers (including integrin activation, granule secretion, and spreading type) when one goes from Group I to Group IV. In the triple-negative breast cancer patients samples before therapy, the responses were shifted toward Group I indicating that their platelets were refractory. These results indicate the importance of platelet heterogeneity analysis and suggest a novel technique to investigate minor populations of refractory or hyperactive platelets.

Keywords

Platelet activation - Calcium signaling - TIRF microscopy - Flow chambers - Breast cancer

Data Availability Statement

All data generated or analyzed during this study are included in this article. Raw microscopy data could be found in FigShare.com (https://figshare.com/projects/Single_platelet_calcium_test_Healthy_adults_and_children_/192752). Further enquiries can be directed to the corresponding author.




Publikationsverlauf

Eingereicht: 28. Juni 2025

Angenommen nach Revision: 15. Dezember 2025

Artikel online veröffentlicht:
30. Januar 2026

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