Institute of Cell Biophysics of the Russian Academy of Sciences

Laboratory of Molecular Cell Physiology

Biophysics Cell Biology Physiology

C-8468-2015

7101992625

Head of Laboratory

Kolesnikov, Stasnislav S

DSc in Biological/biomedical sciences, Professor, Associate member of the Russian Academy of Sciences

Publications

Citations

2 116

h-index

C-8468-2015

7101992625

Authorization required.

Lab website

In the course of life, cells perceive external signals and generate responses to them. These processes are interconnected by an endless network of ordered and interconnected transformations of intracellular molecules, which are generically called intracellular signaling. Our laboratory is studying the mechanisms of signal transmission from the receptor deep into the cell using physiological experiments on single cells.

The most interesting object for the study of intracellular signaling systems are sensory cells. Taste cells, in response to stimulation by taste molecules, are excited and transmit information about its qualities to the taste nerve. The main stages of the mechanism of forming a taste stimulus signal have been established, but there are many questions that have not yet been answered. Our work is designed to fill in the gaps in understanding the intracellular processes underlying the formation of taste at the earliest stages of perception

Non-excitable cells also perceive external signals, and the mechanisms of their transmission into the cell have not been fully studied. Unlimited possibilities for the study of such processes are provided by cells of culture lines, into which new molecules can be "embedded" or existing ones can be "turned off".

It should be noted that the methods of studying intracellular signaling are universal and can be applied to cells of any type. Our laboratory has developed unique techniques for recording cell ion currents (patch clamp), monitoring intracellular calcium (Ca2+-imaging) and other signaling molecules, photolysis of chemical groups (uncaging), as well as their simultaneous use. Using the methods of molecular and cellular biology, we "embed" or "turn off" the receptors or other molecules of interest in cells, and we also get new cell lines carrying fluorescent sensors of key signaling molecules. Using these methods and tools in a complex, we obtain unique data on the mechanisms of intracellular signal transmission, which expand fundamental concepts and form the basis of mathematical models of cell functioning.

Mathematical modeling
The method of local potential fixation (Patch clamp)
Monitoring of intracellular molecules using fluorescent probes and sensors (Ca2+, cAMP, etc.)
Work with eukaryotic cells: transfection, obtaining stable cell lines
CRISPR/Cas9 Genome Editing Technology

Stasnislav Kolesnikov

Head of Laboratory

Polina Kotova 🥼

Leading researcher

Klim Sladkov 🥼

Junior researcher

Research directions

Testing of medicinal compounds using sensor cells

Ideally, medicinal substances should act on the target molecule and related signaling processes, but not affect other molecules. However, in most cases, medicinal compounds do not have such specificity, and their use leads to the development of side effects. Therefore, it is important to have as complete an understanding as possible of the effect of the compounds used on intracellular processes, and the development of methods and tools for conducting appropriate research is a very urgent task. We are creating cell lines that carry genetically encoded fluorescent sensors of the most important signaling molecules. Such cells allow monitoring of one or more intracellular molecules in real time. For example, the sensor cells we obtained allowed us to establish that inhibitors of the intracellular enzyme PI3 kinase, used for the treatment of oncological diseases, affect not only the target molecule, but also have additional targets. These compounds block cell receptors for acetylcholine, histamine and serotonin, which leads to the suppression of important signaling processes throughout the body. The studied compounds cause a wide range of side effects, some of which may be related precisely to their inappropriate effect that we have discovered.

Obtaining model cellular systems for the study of signaling processes

Traditionally, the mechanisms of signal transmission inside the cell are investigated in physiological experiments using inhibitory analysis, using receptor agonists and antagonists, ion channel blockers and modulators, enzyme inhibitors and activators. However, one stimulus can activate several signaling processes in a cell at the same time, which in turn can influence each other. In this regard, the results of physiological experiments are often difficult to interpret unambiguously, which makes it much more difficult to study the mechanisms of intracellular signaling. Difficulties of this kind can sometimes be overcome with the help of artificially created cellular systems. For example, we are able to "embed" molecules of interest into an uncharacteristic cellular system, where they will not be subjected to specific regulation. We can also "turn off" the molecule to confirm its participation in the process under study or to stop its effect on this process. To create such model cell systems, we use advanced methods of molecular and cellular biology, including the creation of plasmid vectors containing genes of proteins of interest, CRISPR/Cas9 genome editing technology and techniques for obtaining monoclonal eukaryotic cell lines.

Signaling processes in the taste bud

Taste sensations are formed in the organ of taste - the taste bud. A couple of cells in the kidney register sour, a couple dozen more register bitter, sweet and umami, other cells register salty and help others perform their functions. No one really knows how they do it. Our laboratory has established a technique for extracting individual taste cells from the epithelium of the mouse tongue and their further study. The electrical activity of cells (by the patch-clamp method) and signaling processes (microphotometry, photolysis of chemical groups) are recorded. The results are compared with the developed computer model of the taste bud. If you are wondering what taste is, we are the only ones in Russia who answer this question in the language of molecules and cellular communications.