Laboratories IKBFU

Neurobiology and Medical Physics Research Laboratory

General information

Current lab project

UNDERSTANDING FUNCTIONAL DIVERSITY OF ASTROGLIA

Neuroscience

Project goal

The goal of this research program is to reveal functional differences between astrocytes residing in different parts of the central nervous system.

Project objectives

  • To determine regional differences in gene expression between cortical, hippocampal and brainstem astrocytes

  • To determine regional differences in the profile of (glio)transmitters released in response to optogenetic or chemogenetic activation of cortical, hippocampal and brainstem astrocytes.

  • To evaluate differences in sensitivity to hypoxia and ischaemia/reperfusion injury between astrocytes residing in the cerebral cortex, hippocampus and the brainstem and determine the mechanisms underlying these differences.

Anticipated project results

The results of the proposed research are expected to contribute towards our understanding of the fundamental principles of brain organization and function and will be made available to a wider scientific community via publications in the leading scientific journals and presentations at scientific conferences around the world, therefore rising the overall competitiveness and profile of Science conducted in Russia. This project is expected to increase our understanding of the mechanisms underlying ischaemia/reperfusion injury and, therefore, may potentially lead to the development of novel treatments to protect the brain during stroke and central hypoxia.

Description of the proposed research project

Aims of the project

This project aims to advance our understanding of the general principles of brain organization and function. The key challenge of modern neuroscience is to understand the complexity of mammalian neural circuits – how brain networks process information and how they control behaviour. This task has become significantly more complex over the last several years by emerging evidence indicating that neuronal activity can be actively modulated by astrocytes – the most numerous glial cells of the brain. Our data obtained from studies on the CNS control of the respiratory system gave further impetus to these ideas by demonstrating in vivo an active control of an intact neural circuit (respiratory control network) by its astroglial neighbours (Gourine et al., 2010).

Our recent evidence obtained in collaboration between London and Kaliningrad suggested that astrocytes residing in different parts of the brain are functionally specialized. In contrast to cortical astrocytes, brainstem astrocytes possess signalling properties which are functionally relevant – they are able to sense physiological decreases in pH and respond to acidification with vesicular release of signalling molecules. This project builds upon these results and includes a comprehensive program of experimental work designed to increase our understanding of the functional differences between astrocytes residing in different parts of the CNS. Novel gene sequencing, imaging, chemo- and optogenetic tools will be used in several experimental paradigms to address experimentally the project objectives.

Other interests

  • Brain regulation of body functions

  • Functional mapping of the Brain

  • Functional diversity of astroglia

  • Neuronal vascular coupling

  • Astroglial control of vital functions

Other projects

Hardware and software experimental sample production for remote wireless diagnostics of person’s cardiovascular system function based on photoplethysmography method. Hardware device is a wireless bracelet worn on the patient's body. CVS state information transmitting to the doctor via Bluetooth module and a cellular phone thru. The information transmission of the patient's state to the doctor is realize in a real time with doctor’s feedback option.

Formation and creation brain-machine interface based on biometric control channel and multimodal feedback for human systems. This interface will be applied in the neuro-electronic systems and exoskeleton design for fills and complements of human’s motor function.

Lab summary

Nucleic acids research: DNA/RNA extraction from blood, tissues, single cell etc.

PCR, real time qPCR, digital PCR (a new approach to nucleic acid detection and quantification that uses molecular counting.

Illumina system for: Whole-genome sequencing, Targeted resequencing , Whole-transcriptome sequencing, de novo sequencing, Epigenetic regulation

PyroMark system: Pyrosequencing technology for real-time, sequence-based detection and quantification of sequence variants and epigenetic methylation. Protein research: proteins extraction from tissue, blood, cells etc.

2D electrophoresis: Mixtures of proteins are separated by two properties in two dimensions on 2D gels.

Bruker Autoflex Speed MALDI based mass spectrometer. MALDI-TOF has been used for many years as a standard in protein and peptide

Immunology. Automatic cell sorting, cell separation on magnetic columns (Meltiniy), cell sorting on magnetic cell sorter (Meltiniy), FACS analysis in flow cytometry.

Quantitative detection of different neurotransmitters and active molecules such as Ach, Lactate, ATP, Glutamate etc. in biological samples with colorimetric assay kits and high perfomance liquid chromatography.

Confocal microscopy and epifluorescent imaging.

Cell culture facilities.

Equipment

GelDoc XR+, Bio-Rad

ECX-F20,Vilber Lourmat

Boxing of microbiological safety of BMB-II- «Laminar-C» tipe A2

Nucleofector™ 2b Device

Microscope Primovert, Carl Zeiss

Axio Imager A2, Carl Zeiss

Eppendorf TransferMan 4r

Ion PGM™ System for Next-Generation Sequencing

MiSeq Personal Sequencer

Liquid handling robotic workstation Neon

PyroMark Q24 System

AmaZon SL, Bruker Daltonik

MALDI-TOF/TOF Autoflex, Bruker Daltonik

ACQUITY UPLC I-Class System

VersaDoc Model 4000, Bio-RAD

Zeiss LSM 780 confocal microscope - ZMBH

PALM MicroBeam ZEISS Microscopy

Research Team

Lab head - Philip Haytovich, PhD, visiting professor

Areas of research:

  • Molecular mechanisms of human longevity and aging

  • Molecular basis of human-specific cognition

  • General principles of regulatory evolution with a focus on humans and closely related primate species

Sergey Kasparov, PhD, visiting professor

Vitaly Kasymov, PhD

Oksana Tuchina, PhD

Publications

1. Angelova PR, Kasymov V, Christie I, Sheikhbahaei S, Turovsky E, Marina N, Korsak A,Zwicker J, Teschemacher AG, Ackland GL, Funk GD, Kasparov S, Abramov AY, Gourine AV. “Functional Oxygen Sensitivity of Astrocytes.” J Neurosci. 2015 Jul 22;35(29):10460-73.

2. Marina N, Ang R, Machhada A, Kasymov V, Karagiannis A, Hosford PS, Mosienko V,Teschemacher AG, Vihko P, Paton JF, Kasparov S, Gourine AV. “Brainstem hypoxia contributes to the development of hypertension in the spontaneously hypertensive rat.” Hypertension. 2015 Apr;65(4):775-83.

3. Ackland GL, Kasymov V, Marina N, Singer M, Gourine AV. “Peripheral neural detection of danger-associated and pathogen-associated molecular patterns.” Crit Care Med. 2013 Jun;41(6)

4. Bogdanov E, Dominova I, Shusharina N, Botman S, Kasymov V, Patrushev M. “Silicon dioxide thin film mediated single cell nucleic acid isolation.” PLoS One. 2013 Jul 10;8(7):

5. Kasymov V, Larina O, Castaldo C, Marina N, Patrushev M, Kasparov S, Gourine AV.“Differential sensitivity of brainstem versus cortical astrocytes to changes in pH reveals

functional regional specialization of astroglia.” J Neurosci. 2013 Jan 9;33(2)

6. Figueiredo M, Lane S, Tang F, Liu BH, Hewinson J, Marina N, Kasymov V, Souslova EA, Chudakov DM, Gourine AV, Teschemacher AG, Kasparov S. “Optogenetic experimentation on astrocytes.” Exp Physiol. 2011 Jan;96(1)

7. Gourine AV, Kasymov V, Marina N, Tang F, Figueiredo MF, Lane S, Teschemacher AG, Spyer KM, Deisseroth K, Kasparov S. “Astrocytes control breathing through pH-dependent release of ATP”, Science. 2010 Jul 30.

8. Paschalis A, Churchill L, Marina N, Kasymov V, Gourine A, Ackland G. “beta1- Adrenoceptor distribution in the rat brain: an immunohistochemical study” Neurosci Lett. 2009 Jul 17;458(2):84-8. Epub 2009 Apr 15

9. Zinchenko VP, Goncharov NV, Teplova VV, Kasymov VA, Petrova OI, Berezhnov AV, Senchenkov EV, Mindukshev IV, Jenkins RO, Radilov AS. “Studies of interaction of intracellular signalling and metabolic pathways under inhibition of mitochondrial aconitase with fluoroacetate.” Tsitologiia. 2007;49(12):1023-31. Russian

10. Ackland GL, Kasymov V, Gourine AV. “Physiological and pathophysiological roles of extracellular ATP in chemosensory control of breathing.” Biochem Soc Trans. 2007 Nov;35(Pt5):1264-8. Review.

11. Patrushev M., Kasymov V., Patrusheva V., Ushakova T., Gogvadze V., Gaziev A. “Release of mitochondrial DNA fragments from brain mitochondria of irradiated mice.” Mitochondrion, 2006 Jan 11; V. 6(1): 43-7

12. Kasymov V.A., Li V.V., Kaimachnikov N.P., Zinchenko V.P. “Role of calmodulin and phospholipase A 2 in the induction of non-specific Ca 2+ -channel in cell plasma membrane.” Biologocal Membranes (Russian). 2006 Jan; Vol. 23(2): 182-189.

13. V.P. Zinchenko, V.A. Kasymov, V.V. Li, N.P. Kaimachnikov “Calmodulin inhibitor R24571 induces the short-term Са 2+ entry and the pulse-like secretion of ATP in Ehrlich ascites tumor cells.” Biophyzika (Russian). 2005 Nov-Dec;Vol. 50(6): 1055-69

14. Patrushev M., Kasymov V., Patrusheva V., Ushakova T., Gogvadze V., Gaziev A.

“Mitochondrial permeability transition triggers the release of mtDNA fragments.”

Cell Mol Life Sci. 2004 Dec;61(24):3100-3.

15. V.A. Stupin, E.V. Silina, R.G. Oganov, Y.A. Bogdanov and N.N. Shusharina. Development of An Invasive Device for Long-term Remote Monitoring of Cardiovascular System Parameters, Including Blood Pressure, in Patients with Comorbid Conditions. Biosciences Biotechnology Research Asia. 2015 Vol. 12(2). - 1255-1263.

16. S. Botman, D. Borchevkin, V. Petrov, E. Bogdanov, M. Patrushev, N. Shusharina. Photoplethysmography-Based Device Designing for Cardiovascular System Diagnostics. World Academy of Science, Engineering and Technology, International Journal of Medical, Health, Biomedical, Bioengineering and Pharmaceutical Engineering. 2015 Т. 9. – №. 9. – С. 685-689.

17. Petrov V, Shestakov A, Borchevkin D, Botman S, Bogdanov E, Patrushev M., Shusharina N. Device for Ambulatory Diagnosis of Pet’s Cardiovascular System Functional State. J Veterinar Sci Technol. 2015 6: 263. doi:10.4172/2157-7579.1000263

18. Rumyantseva S. A., Stupin V. A., Oganov R. G., Silina E. V., Kolesnikova E. A., Petrov V.A., Kasymov V. A., Shusharina N. N., Ladaniy D. V., Bogdanov E. A The ways of long-term blood pressure control systems development. Cardiovascular Therapy and Prevention 201 14(1): 78–82

19. O. Tuchina, S. Koczan, S. Harzsch, J. Rybak, G. Wolff, N.J. Strausfeld, B.S. Hansson.Central projections of antennular chemosensory and mechanosensory afferents in the brain of the terrestrial hermit crab (Coenobita clypeatus; Coenobitidae, Anomura). Frontiers in neuroanatomy 2015 Т. 9. - pp. 94. doi: 10.3389/fnana.2015.00094. eCollection 2015.