Organizing co mmittee

SEARCH FOR STIMULAS AFFECTING THE LEVEL OF 
INORGANIC POLYPHOSPHATES IN THYMOCYTES 
 
Ergasheva Diyora 
 
Сenter of advanced technologies under the Ministry of Innovative Development of 
the Republic of Uzbekistan, 
e-mail: [e-mail]
 
Phosphorus is a vital element of all living organisms, being part of the most 
important organic compounds, including nucleic acids, ATP and other nucleoside 
phosphates, phospholipids, phosphorylated proteins, and carbohydrates. An insufficient 
amount of this element in the environment is an unfavorable factor limiting growth and 
development, and its absence leads to the death of organisms. Inorganic polyphosphates 
(Poly-P) is the oldest molecule found in all living organisms from bacteria to mammals, 
they are multifunctional molecules depending on the subcellular location in different 
types of organism. 
In this regard, an important strategy for survival in changing environmental 
conditions for organisms is the accumulation of reserves of phosphorus compounds. 
These compounds are varied. These can be organic phosphorus compounds, such as 
phytin (Ca-Mg-salt of inositol phosphate) of higher plants, phosphomannan of some 
yeast species (
Hansenula capsulata
), teichoic acids of the bacterial cell wall. 
To date, it is known that Poly-P in mammalian cells play a predominantly regulatory 
role. Although the specific enzymes involved in the synthesis of Poly-P are still 
unknown. However, from the literature and the results of our studies, it is known that 
the synthesis of Poly-P is associated with the energy metabolism of the cell, as well as 
the integrity of the membrane. To date, the issue of studying the pathways of Poly-P 
metabolism in mammalian cells remains relevant. 
The aim of this work was to assess the phosphate-accumulating potential of rat 
thymocytes, as well as to study the features of the metabolism of inorganic polyP in 
these cells, which accumulate Pi most efficiently. 
To achieve this goal, thymocytes of outbred rats weighing 100-120 grams were 
isolated using differential centrifugation. The isolated thymocyte suspension was 
incubated with a 20 µM DAPI probe at room temperature for 40 minutes. During 
incubation, thymocytes were pipetted every 5-7 minutes. 
Fluorescence was performed using an Agilent Technologies fluorometer (Cary 
Eclipse Fluorescence Spectrophotometer). All recordings were made in 3 ml glass 
cuvettes with Ringer's solution without glucose (medium composition: 5 mM KCl, 135 
mM NaCl, 11 mM HEPES, 2 mM CaCl, 1 mM MgCl, pH 7.3). 
To study the features of Poly-P metabolism in thymocytes, a modified measurement 
of Poly-P fluorescence using DAPI was used. Changes in the level of Poly-P on the 
main molecule of energy metabolism, glucose, were taken as a control. Application of 
10 mM glucose to incubated thymocytes in a glucose-free medium caused an increase in 
DAPI-polyP fluorescence in thymocytes. In the incubation medium without the addition 
of glucose, a decrease in DAPI-polyP fluorescence was observed, which indicates that 
the metabolism of Poly-P is inextricably linked with the energy metabolism of the cell. 
The results obtained can be used to further study the effect of activators and 
inhibitors of polyP metabolism in thymus cells. 

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Poster presentation 
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