Clusterization of Inositol Trisphosphate Receptors Determines the Shape of the Calcium Oscillation Peak in Platelet Cytosol

Balabin, F. A.; Морозова, Д. С.; Mayorov, A. S.; Martyanov, A. A.; Panteleev, Mikhail A.; Sveshnikova, Anastasia N.

doi:10.3103/s0027134918050041

Cited by 4 publications

(2 citation statements)

References 21 publications

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“…Parallel-plate flow chambers, used here, were described earlier [21]. Monoclonal antibody to CD31 covered glass coverslips were used for platelet immobilization.…”

Section: Methodsmentioning

confidence: 99%

On calcium fluorophore’s impact in platelet signaling studies

Galkina

Balabin

2021

SBPR

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Observationof calcium signaling in platelets -blood cells designed to be involved in stopping bleeding and forming blood clots -is an important part of fundamental research in hemostasis. However, such a study is possible only with the use of calcium fluorophores -small molecules that penetrate the platelet membrane due to their hydrophobic -AM part, which is then hydrolyzed by cytosol esterases. In this work, we consider the phenomenon of inhomogeneous loading of calcium fluorophores into platelets.We used platelets from healthy adult donors loaded with various fluorescent probes (CalBryte590, DiOC6 (3), Fura Red, Fluo-4 and CellTracker Violet BMQC) and immobilized on antibodies to CD31 in parallel plane flow chambers. Total internal reflection fluorescence (TIRF) microscopy was used for observations.We demonstrated that all studied probes are loaded heterogeneously, with 30% platelets being loaded with a probe 2-6 times higher than the population median value. Using the CalBryte590 probe as an example, we have shown that a decrease in the incubation temperature, the addition of Pluronic 127 to the incubation medium, or membrane cholesterol depletion significantly reduces the heterogeneity of the probe distribution in the population. By looking at platelet activation from the surface, we have shown that the probability of experiencing strong activation, as measured by the intensity of calcium oscillations, correlates with the amount of probe in the platelet.Thus, we conclude that the type of fluorophore used and the conditions of its loading into platelets can significantly affect the results of experiments on the observation of calcium signaling in platelets.

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“…Parallel-plate flow chambers, used here, were described earlier [21]. Monoclonal antibody to CD31 covered glass coverslips were used for platelet immobilization.…”

Section: Methodsmentioning

confidence: 99%

On calcium fluorophore’s impact in platelet signaling studies

Galkina

Balabin

2021

SBPR

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“…Cells were resuspended in AllGrad Wash buffer to an approximate density of 10 8 cells/mL. For epifluorescent microscopy, spermatozoa were immobilized on poly- l -lysine-covered glass slides and investigated in flow chambers [ 58 ]. An inverted Nikon Eclipse Ti-E microscope equipped with 100×/1.49 NA TIRF oil objective was used.…”

Section: Methodsmentioning

confidence: 99%

Occurrence of Calcium Oscillations in Human Spermatozoa Is Based on Spatial Signaling Enzymes Distribution

Korobkin

Balabin

Yakovenko

et al. 2021

IJMS

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In human spermatozoa, calcium dynamics control most of fertilization events. Progesterone, present in the female reproductive system, can trigger several types of calcium responses, such as low-frequency oscillations. Here we aimed to identify the mechanisms of progesterone-induced calcium signaling in human spermatozoa. Progesterone-induced activation of fluorophore-loaded spermatozoa was studied by fluorescent microscopy. Two computational models were developed to describe the spermatozoa calcium responses: a homogeneous one based on a system of ordinary differential equations and a three-dimensional one with added space dimensions and diffusion for the cytosolic species. In response to progesterone, three types of calcium responses were observed in human spermatozoa: a single transient rise of calcium concentration in cytosol, a steady elevation, or low-frequency oscillations. The homogenous model provided qualitative description of the oscillatory and the single spike responses, while the three-dimensional model captured the calcium peak shape and the frequency of calcium oscillations. The model analysis demonstrated that an increase in the calcium diffusion coefficient resulted in the disappearance of the calcium oscillations. Additionally, in silico analysis suggested that the spatial distribution of calcium signaling enzymes governs the appearance of calcium oscillations in progesterone-activated human spermatozoa.

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Personalization of a computational systems biology model of blood platelet calcium signaling

Balabin,

Korobkina,

Galkina

et al. 2024

BIOMED KHIM

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Anuclear blood cells, platelets, are the basis for the formation of blood clots in human vessels. While antiplatelet therapy is most often used after ischemic events, there is a need for its personalization due to the limited effectiveness and risks of bleeding. Previously, we developed a series of computational models to describe intracellular platelet signaling and a set of experimental methods to characterize the platelets of a given patient. To build a personalized model of platelet signaling, we also conducted research on platelet proteomics. The aim of this study was to personalize the central module of intracellular platelet signaling responsible for the formation of calcium oscillations in response to activation. The model consists of 26 ordinary differential equations. To personalize the model, proteomics data were used and unknown model parameters were selected based on experimental data on the shape and frequency of calcium oscillations in single platelets. As a result of the study, it has been shown that the key personalized parameters of the platelet oscillatory response are the degree of asymmetry of a single calcium spike and the maximum frequency of oscillations. Based on the listed experimentally determined parameters and proteomics data, an algorithm for personalization of the model has been proposed. Here we considered three healthy pediatric donors of different ages. Based on the models, personal curves of platelet calcium response to activation were obtained. The analysis of the models has shown that while there is a large heterogeneity of individual indicators of intracellular signaling, such as the activity of calcium pumps (SERCA) and inositoltriphosphate (IP₃) receptors (IP₃R), these indicators compensate each other in each donors. This observation is confirmed by the analysis of proteomics data from 15 healthy patients: this analysis demonstrates a correlation between the total amount of SERCA and IP₃R. Thus, several new features of human platelet calcium signaling are shown and an algorithm for personalizing its model is proposed.

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Clusterization of Inositol Trisphosphate Receptors Determines the Shape of the Calcium Oscillation Peak in Platelet Cytosol

Cited by 4 publications

References 21 publications

On calcium fluorophore’s impact in platelet signaling studies

On calcium fluorophore’s impact in platelet signaling studies

Occurrence of Calcium Oscillations in Human Spermatozoa Is Based on Spatial Signaling Enzymes Distribution

Personalization of a computational systems biology model of blood platelet calcium signaling

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