Information Page of SAS Organisation

Centre of Biosciences SAS (Institute of Molecular Physiology and Genetics SAS)

International projects

Mitochondria-endoplasmic reticulum functional interplay in Wolfram Syndrome: emerging role for heart and brain protection
Funkčné prepojenie mitochondrií a endoplazmatického retikula u Wolframovho syndrómu: predpokladaný význam pre ochranu mozgu a srdca
Program: 7RP
Project leader: RNDr. Cagalinec Michal PhD.
Annotation:Wolfram syndrome (WS)is a recessive neurological disorder caused by mutation of the Wfs1 gene. The Wfs1 protein is highly expressed in the brain and heart and is embedded in the endoplasmic reticulum (ER) where it modulates Ca2+ levels and ER stress. Additionally, the main symptoms of the WSare consistent with the ones characteristic for mitochondrial diseases. In fact, our preliminary results showed already that silencing of Wfs1 in mouse neurons decreased the mitochondrial fusion frequency and caused mitochondrial fragmentation, demonstrating strong impact of Wfs1 to mitochondrial function in neurons. Although the high expression of Wfs1 in the heart and cardiac symptoms in WS identified recently emphasize the functional importance of Wfs1 in the heart, the most common causes of morbidity in WS are the neurological manifestations. How is it possible that mutations of Wfs1 causing significant perturbations in the brain functions are not so prominent in the heart? One explanation of this tissue specificity is that a mechanism compensating for loss of Wfs1 protein function is present in the heart but not in neurons.
Project web page:http://www.confolab.sav.sk/indexSASPRO.html
Duration: 1.3.2015 - 31.12.2018

Characterization of novel interaction partners of the N-type (Cav2.2) calcium channel
Charakterizácia nových interakčných partnerov N-typu (Cav2.2) vápnikového kanála
Program: Bilaterálne - iné
Project leader: doc. RNDr. Lacinová Ľubica DrSc.
Annotation:In neurons calcium acts as a second messenger mediating numerous physiological responses including synaptic plasticity, secretion of neurotransmitters, activities of various enzymes, gene expression, cellular differentiation, as well as axonal outgrowth. Precise spatiotemporal regulation of free intracellular [Ca2+]i is vital for the integrity and function of a neuronal system. Well-defined pathways for calcium influx from an extracellular space represent individual members of voltage-gated calcium channels (VGCCs). Spatial diversity of calcium signaling is accomplished by highly specific expression of individual channel types in parts of the neuronal cell body, axon, and/or dendrites. N-type or Cav2.2 calcium channels are expressed solely in neurons. These channels are predominantly located at synaptic nerve terminals and calcium influx through Cav2.2 initiates neurotransmitter secretion ensuring transmission of excitatory stimuli to postsynaptic membranes (Wheeler et al. 1994). These are the only VGCCs capable of generating a large but temporally precise calcium influx (Weber et al. 2010). Integration of VGCCs in complex network structures of protein-interactions is a crucial requirement for precise regulation of channel activity. In the past, we took advantage of the specific properties of the yeast split-ubiquitin system to identify novel interaction partners of the 1 subunit of the Cav2.2 calcium channel. This approach is a completely unbiased strategy to identify novel, so far undescribed calcium channel interacting proteins. In these initial studies we specified a series of so far unknown interaction partners: Tetraspanin-13 (published in Mallmann et al., 2013), RTN1 or reticulon, SLC38A1 a member of the solute carrier family 38, Ptgs or prostaglandin D2 synthase, TMEM 223 an uncharacterized transmembrane protein and GRINA a putative glutamate receptor-associated protein. The major aim of our project is to complete the characterization of each of these individual Cav2.2 interactions with the long-term objective to achieve insights into their physiological context. We will concentrate on the following aspects of the Cav2.2 channel interactome: -Characterization of the nature of the interactions We will verify the interaction by means of co-immunoprecipitation and co-localization studies. Where appropriate we will narrow down the relevant binding structures and analyze the specificity of the interaction for Cav2.2 and possibly other VGCCs. -Regulation of voltage- and time-dependent activity of the Cav2.2 channel itself The main role of Cav2.2 channels is the generation of a large but temporally precise calcium influx into presynaptic neurons. Even minor changes in the channel gating may translate into a significant modulation of synaptic transmission. We will characterize voltage dependence and kinetics of activation and inactivation, cumulative inactivation evoked by high-frequency trains of rectangular depolarizing pulses and/or action-potential-like waveforms in the absence and presence of each putative interacting protein. Further, we will estimate the channel opening probability from a relation between maximal ionic and gating currents. -Modulation of a downstream regulatory pathways We will investigate the modulation of Cav2.2 channels activity by intracellular messengers and/or G-proteins in the absence or presence of each interacting protein.
Duration: 1.1.2016 - 31.12.2017

FWF P28179-B30 - The link between PDS and mitochondria in epileptogenesis
Vzťah medzi PDS a mitochondriami v procese epileptogenézy
Program: Iné
Project leader: RNDr. Cagalinec Michal PhD.
Annotation:PDS are increasingly recognized as epileptogenic factors in acquired forms of epilepsies. Prevention of PDS provides a potential means to decrease the numbers of patients developing epilepsy after brain injuries. It is of crucial importance to understand the pathomechanisms of PDS formation (e.g. common molecular links converging in the aftermaths of hemorrhagic bleeding – both subarachnoidal and intracerebral – or of infection, ischemia,…). Since epilepsy develops only in a certain number of patients, i.e. about 20% after severe brain injury, it is of primary interest to define the factors that predispose to acquisition of an epileptic brain condition. A genetic influence has been noted with respect to this predisposition. The mitochondrial state may represent a critical determinant. Indeed, mitochondrial dysfunction is increasingly understood as a precipitating cause of epilepsy. However, the possibility that formation of PDS plays an essential role therein has not been addressed so far, and thus represents an innovative aspect in targeting epileptogenesis. The vital question is whether supporting mitochondrial function during a critical period after brain injuries can be used to prevent subsequent pathogenesis. Answering this question can be envisaged to provide invaluable insights in viable strategies to counteract acquired forms of epilepsies. Additionally, important information regarding mitochondrial implications in neuronal physiology and pathophysiology can be expected to also arise from our approach.
Duration: 1.1.2016 - 30.6.2018


National projects

BIOGLYCO - Biochips and biosensors for glycorecognition, their development, prepararion and application in cancer research
Biočipy a biosenzory pre glykorozpoznávanie, ich vývoj, príprava a využitie pri výskume rakoviny
Program: APVV
Project leader: Ing. Sulová Zdena DrSc.
Annotation:Due to the rapid development of new technologies is placed a considerable emphasis on their use for improving the quality of life. The most important areas here include the care of human health, such as the fight against cancer. Here is a keen interest in new sensitive diagnostic approaches and technologies to help increase the effectiveness of therapy and cancer research. Development and application of new diagnostic procedures and methods is based in no small measure on the knowledge gained in the basic research in the field of analytical tools based on the biochips and biosensors. The aim of the project is the development and preparation of new analytical glyco-recognizing biochip and biosensor systems based on lectin in different formats in combination with multiple platforms and innovative detection methods, their validation and use in the analysis of real samples in cancer research. The selected design of biochips and used detection platform enable highly sensitive and high-throughput analysis of glycosylation changes, and the involvement of mass spectrometry also accurate identification of glycan structures subject to glycosylation changes. To increase the functionality of biochips will be in the development of analytical systems used also nanotechnology tools, particularly in the preparation of nanostructured surfaces of biochips and for improving the efficiency of detection. Various types of biological samples will be analyzed such as serum, lysates, tissue and isolated glycoproteins from some areas of cancer research (effect of hypoxia, resistance to cytostatics, search of glyco-biomarkers). It is expected that the involvement of the entire spectrum of analytical procedures for glycorecognition developed in this project will help formulate more sophisticated conclusions on the role of glycans in cancer.
Duration: 1.7.2015 - 30.6.2019

CAMYS - Cytoarchitecture of calcium signalling of cardiac myocytes in development of myocardial hypertrophy
Cytoarchitektúra vápnikovej signalizácie srdcových myocytov vo vývoji hypertrofie myokardu
Program: APVV
Project leader: Ing. Zahradníková Alexandra DrSc.
Annotation:In early phases of many cardiovascular diseases, calcium signalling deteriorates and cardiomyocyte hypertrophy is activated. Calcium signalling, in addition to control of contractile function, modulates many of signalling pathways and metabolism of myocytes. Therefore, disruption of calcium signalling in either systolic or diastolic phase may be a consequence, but also the cause of the maladaptive reaction of myocytes to overload. We hypothesize that processes related to hypertrophy lead to changes in distribution of endo/sarcoplasmic reticulum elements responsible for calcium signalling and for proteosynthesis, and to associated changes in expression and distribution of calcium signalling proteins. This study aims to determine differences in the myocyte calcium signalling system of myocardium adapting to pathological or physiological load and to compare them with development of the myocyte calcium signalling system during postnatal maturation of myocardium. The status of the calcium signalling system will be analysed in models of myocardial load in laboratory rats: a/ sedentary model - animals in standard cages, b/ model of physiological load - animals in cages with a running wheel, 3/ model of pressure overload - surgical obstruction of ascending aorta, and d/ growth model of hypertrophy - postnatal development of myocardium. State-of-the-art methods of electrophysiology, molecular biology and ultrastructural microscopy will be used to characterize quality and distribution of calcium signals, the extent and distribution of smooth and rough sarcoplasmic reticulum, and of expression, localization and co-localization of calcium signalling proteins. We expect that the multidisciplinary approach supported by the expertise of team members will allow interpreting cellular and molecular mechanisms of calcium signalling and drawing conclusions relevant to understanding mechanisms of heart failure that will aid development of new therapeutic and preventative procedures.
Project web page:http://www.umfg.sav.sk/ovsb/projektyovsb/aktivne-projekty-v-ovsb/apvv-15-0302/
Duration: 1.7.2016 - 30.6.2019

Diastolic function of the ryanodine receptor and generation of arrhythmogenic calcium waves
Diastolická funkcia ryanodínového receptora a tvorba arytmogénnych vápnikových vĺn
Program: VEGA
Project leader: Ing. Zahradníková Alexandra DrSc.
Annotation:In some acquired and genetic arrhythmias, anomalies in calcium release occur during the diastole, which may result in formation of calcium waves that initiate cardiac arrhythmias. Proper diastolic function of the system of calcium homeostasis involves regulation of diastolic calcium release by ryanodine receptors. Anomalous calcium release from the viewpoint of calcium wave formation is not sufficiently understood. We will concentrate on determination of the relationships between localization of dyads as the sites of calcium release and formation of calcium waves, and on their development during maturation and physiological hypertrophy of myocytes. The outcome of the project will be a better understanding of the factors governing calcium homeostasis in cardiac myocytes and their impairment leading to calcium waves.
Duration: 1.1.2017 - 31.12.2019

Mitochondrial dynamics and morphology in transgenic model of Wolfram syndrome: emerging role for heart protection
Dynamika a morfológia mitochondrií u transgénneho modelu Wolframovho syndrómu: význam pre ochranu srdca
Program: VEGA
Project leader: RNDr. Cagalinec Michal PhD.
Annotation:Wolfram syndrome (WS) is a recessive neurological disorder caused by mutation of the Wfs1 gene. The Wfs1 protein is highly expressed in the brain and heart and is embedded in the endoplasmic reticulum (ER) where it modulates Ca2+ levels and ER stress. Additionally, the main symptoms of the WS are consistent with the ones characteristic for mitochondrial diseases. In fact, our preliminary results showed already that silencing of Wfs1 in mouse neurons decreased the mitochondrial fusion frequency and caused mitochondrial fragmentation, demonstrating strong impact of Wfs1 to mitochondrial function in neurons. Although the high expression of Wfs1 in the heart and cardiac symptoms in WS identified recently emphasize the functional importance of Wfs1 in the heart, the most common causes of morbidity in WS are the neurological manifestations.
Duration: 1.1.2016 - 31.12.2018

Expression and co-localization of dyadic protein complexes in ventricular myocytes in relation to excitation-contraction coupling ontogenesis.
Expresia a kolokalizácia proteínov diadických komplexov komorových myocytov potkana vo vzťahu k ontogenéze väzby excitácie s kontrakciou.
Program: VEGA
Project leader: RNDr. Zahradníková, ml. Alexandra PhD.
Annotation:Excitation-contraction coupling in ventricular myocytes is based on calcium signalling in dyads - specialized junctions of sarcolemma and terminal cisternae of sarcoplasmic reticulum. The relationship between the molecular structure of dyads and the functionality of calcium signalling is not sufficiently understood, since its direct experimental manipulation is not possible. It should be possible to observe correlation between the structure and function of dyads during the postnatal stage of cardiac development, a period of their extensive formation and growth. The aim of the project is to correlate the changes in local calcium signalling to protein composition and interaction in individual dyadic complexes during myocyte ontogenesis. The acquired data will allow us to formulate hypotheses about the relationship between the structure and function of dyads at the molecular level and will help us understand the adaptive processes, occurring during cardiac hypertrophy, at the level of calcium signaling.
Duration: 1.1.2015 - 31.12.2017

Smart MoS2 platform for cancer diagnosis and targeted treatment
Inovatívna MoS2 platforma pre diagnózu a cielenú liečbu rakoviny
Program: APVV
Project leader: doc. Ing. Breier Albert DrSc.
Annotation:The goal of the proposed project is to develop a novel smart 2-dimensional multifunctional nanoplatform based on MoS2 for cancer cell detection and treatment. The MoS2 nanosheets prepared by liquid phase exfoliation and/or Li intercalation and modified for low toxicity and high biocompatibility will be chemically functionalized with antibodies sensitive to specific cancer cells and relevant cytotoxine. In comparison to graphene based nanosheets the MoS2 provides much stronger signal for the advanced laboratory diagnostics such as Raman spectroscopy, X-ray methods, SEM and TEM. Strong Raman signal and photoluminescence of MoS2 nanosheets will allow a label-free in situ tracking of the nanoplatform localization at the cell level. This will be one of the original project contributions to the knowledge of the cell interaction with the functional nanoplatform in general. The new quality of the laboratory testing of the nanoplatform interaction with the cancer cells may bring new knowledge and essential progress in the field of 2D nanoplatform generally. New knowledge is expected also in terms of a smart handling of biocompatibility and toxicity of the nanoplatform which is important for the nanoplatform cell internalization. Newly elaborated technological procedures will have direct implications for tailored 2D materials technology.
Duration: 1.7.2016 - 30.6.2020

Luminal regulation of the cardiac ryanodine receptor and its molecular mechanisms
Luminálna regulácia srdcového ryanodínového receptora a jej molekulárne mechanizmy
Program: VEGA
Project leader: Mgr. Gaburjáková Marta PhD.
Annotation:Ryanodine receptor type 2 (RYR2) plays unchangeable role in the process of excitation-contraction coupling in the heart. The physiologically relevant Ca2+ regulation of the RYR2 channel is predominantly determined by its robust cytosolic domain that harbors binding sites for cytosolic Ca2+. Less characterized channel regulation by luminal Ca2+ could be mediated by two mechanisms namely luminal Ca2+ binding on the luminal RYR2 face and binding of luminal Ca2+ emanating the RYR2 pore to the cytosolic channel face. Our aim is to examine a mutual relationship between these two mechanisms of the luminal regulation. The recent findings about the structure of Ca2+ binding sites on the RYR2 channel allow us to employ non-standard experimental conditions. We expect that trivalent cations and their interaction with the RYR2 channel will contribute to the reconciling of current controversial results and bring a comprehensive picture of the RYR2 Ca2+ regulation that has been implicated in heart diseases.
Duration: 1.1.2017 - 31.12.2019

Possible dual function of P-glycoprotein in leukemia cells: efflux pump and regulatory protein
Možná duálna funkcia P-glykoproteínu pri viacliekovej rezistencii leukemických buniek: efluxná pumpa a regulačný proteín
Program: APVV
Project leader: Ing. Sulová Zdena DrSc.
Annotation:Project is focused on hypothesis that P-glycoprotein besides its generally accepted role as multidrug efflux pump for large groups of lipophilic substances also play additional role as regulatory protein influencing programmed cell death progression. Several lines of evidences indicated, that this regulatory action of P-gp is independent on P-gp transport activity. Therefore, P-gp when express in cells, could depress cell sensitivity also to substances which cannot be effluxed by its transport activity. We studied this trend on mice leukemia cells L1210 in which the P-gp expression was induced either by selection for resistance with vincristine, or by transfection with gene encoding full length P-gp. We found that P-gp overexpressing cells are resistant also to endoplasmic reticulum stressors like tunicamycin (inhibitor of N-glycosylation in endoplasmic reticulum), thapsigargin (inhibitor of Ca (2+)-ATPase from endoplasmic reticulum); but also to cisplatin (chemotherapeutic that is not substrate for P-gp), and lastly to lectin Concanavalin A. However P-gp positive cells were more sensitive than their P-gp negative counterparts to another lectin from wheat germ. These results are associated with massive remodalation of cell surface saccharides in cells with upregulated P-gp. Presence of P-gp in cells seems to induce strong alteration in cell regulatory pathways including either protein phosphorylation and glycosylation or regulation of apoptosis progression. These issues we plane to study by described P-gp cell models, or by using the human leukemia cells, in which the P-gp overexpresion was induced by adaptation with chemotherapeutics like vincristine, mitoxantrone or azacytidine.
Duration: 1.7.2015 - 31.7.2018

Novel pathways of N-type (CaV2.2) calcium channels regulation
Nové spôsoby regulácie N-typu (CaV2.2) vápnikových kanálov
Program: VEGA
Project leader: doc. RNDr. Lacinová Ľubica DrSc.
Annotation:Neurotransmission controlled by CaV2.2 or N-type calcium channels plays a major role in chronic and neuropathic pain which is a major clinical, social and economic problem. Ziconotide is the only one drug specifically targeting CaV2.2 channel. It is delivered intrathecally leaving demand for an oral drug open. Ion channels form macromolecular signaling complexes with a variety of interacting proteins and stabilization or disruption of such interactions may be a base for a novel therapeutic strategy. Modified yeast-two-hybrid assay identified proteins RTN1, SLC38A1, Ptgds, TMEM 223, and Grina as putative interaction partners of the CaV2.2 channels. These proteins are expressed predominantly or solely in neurons and several neuromodulatory functions of these proteins were described. We hypothesize that they may modulate CaV2.2 channels, as well. Our aim is to describe functional mechanisms and/or intracellular pathways involved in the interaction between CaV2.2 channels and listed interacting proteins.
Duration: 1.1.2016 - 31.12.2018

Novel synergistic antitumour properties of nuclear retinoid X receptor (RXR) agonists as a consequence of the conditional RXR-RAR heterodimer formation in human breast cancer cells
Nové synergické protinádorové vlastnosti agonistov nukleárnych retinoidných X receptorov (RXR) ako následok vzniku "conditional" RXR-RAR heterodiméru v ľudských nádorových bunkách prsníka
Program: APVV
Project leader: Ing. Sulová Zdena DrSc.
Annotation:Nuclear retinoid X receptors (RXR) are considered to be important target molecules for treatment of several malignant diseases. Characteristic feature of RXR agonists is their capability to amplify the effect of endogenous ligand of nuclear retinoid receptors (RAR). The objective of the project proposal is the investigation of novel antitumour properties of the biologically active agonists of nuclear retinoid X receptors, which upon binding to RXR molecule function with the "partner" RAR receptors in the presence of all-trans retinoic acid as a "conditional" heterodimer. The object of investigation is a selected group of retinoid X receptors agonists of natural or synthetic origin. For a number of experiments, we will employ two different human breast cancer cell lines (estrogen receptor positive and estrogen receptor negative) in order to find cooperative effects of RXR agonist and RAR ligand at the RXR-RAR heterodimer leading to enhanced tumour cell growth inhibition or induction of apoptosis. The parallel objective of the project is investigation of the effects of the "supramolecular ligand/agonist" formation from a group of RXR agonists of natural or synthetic origin. We expect a number of original results yielding from experimental work that should contribute to the development of novel therapeutical potentialities for breast cancer treatment.
Duration: 1.7.2016 - 30.6.2020

Defense mechanisms of neoplastic cells against chemical stress
Obranné mechanizmy neoplastických buniek proti chemickému stresu
Program: APVV
Project leader: Ing. Sulová Zdena DrSc.
Annotation:Animal cells are disposing with large spectrum of defense mechanisms against damage induced by cytotoxic agents. These mechanisms are based on: i) ability of detoxification enzymes to modify the structure of cytotoxic agents; ii) ability of plasma membrane specialized transporters to eliminate the drugs from cell's intracellular space; iii| changes in expression of cytotoxic agents target proteins; iv| changes in regulatory pathways responsible for initiation of programed cell death mechanisms as response of cell damage induced by cytotoxic agents. Initiation of these mechanisms in neoplastically altered tissue leads to multidrug resistance (MDR) development. Reduced cell sensitivity to large group of structurally unrelated anticancer drugs with different mechanisms of pharmacological action is feature typical for MDR. If MDR is developed in cancer tissue, it represents real obstacle of effective chemotherapy. MDR could develop in cells of alla types of solid malignant tumors and also in blood malignancies. MDR development could be considered as adaptation of neoplastic cells on previously applied chemotherapy cycles. However, MDR could be present in neoplastic cells of patients that were not treated by anticancer drugs. MDR in this case represents specific phenotype of neoplasticaly altered cells. Targeted research on this topic represents aim of this project.
Duration: 1.7.2016 - 30.6.2020

HippoOR - Prenatal and postnatal effects of δ and µ opioid receptor ligands on the hippocampal development and function.
Prenatálne a postnatálne účinky ligandov δ a µ opioidných receptorov na vývoj a funkciu hipokampu.
Program: APVV
Project leader: doc. RNDr. Lacinová Ľubica DrSc.
Annotation:Ligands of opioid receptors δ (DOR) and μ opioid receptors (MOR) are commonly used in treatment of severe acute and chronic pain. DORs are involved also in mood disorders like depression and anxiety, which are related to the hippocampal function. Treatment with DOR ligands does not result in adverse effects including addiction, which are common with MOR ligands. However, much less is known about DOR – activated signaling pathways than about MOR – activated pathways. We will analyze the effect of acute (seconds to minutes) and chronic (hours to days) in in vitro and in vivo (prenatal and postnatal) administration of DOR ligands on the morphological and electrophysiological properties of rat hippocampal neurons and compare them with effects of MOR ligands and with effects of ligands specific for MOR-DOR heteromers. Further, involvement of calcium transporting proteins in signal transduction pathways activated by DORs and MORs ligands will be addressed by molecular biology methods. Possible remodeling of the dendritic spines will be investigated using transmission electron microscopy. Effect of DOR ligands on hippocampal plasticity in control and stressed rats will be examined using behavioral tests and molecular neuroscience techniques. Excitability will be investigated in primary culture of hippocampal neurons by patch clamp and in situ by in vivo electrophysiology. Both models enable to follow effects of acute and chronic drug application as well as possible receptor desensitization and offer complementary advantages. Primary neuronal culture is the possibility to visually identify neurons, characterize in details both action potentials and underlying ionic currents and to correlate electrophysiology and molecular biology on the same batch of neurons. In vivo electrophysiology offers the possibility to measure neuronal activity within its normal environment including all interactions with other brain parts. Finaly, behavioral studies enable investigation of an intact animal.
Duration: 1.7.2016 - 30.6.2020

Study of changes in expression of some regulatory and functional proteins connected with the presence of P-gp protein in leukemic cells
Štúdium zmien expresie niektorých regulačných a štrukturálnych proteínov sprevádzajúcich expresiu P-glykoproteínu v leukemických bunkách
Program: VEGA
Project leader: Ing. Sulová Zdena DrSc.
Annotation:Successful treatment of hematologic malignancies depends on the efficacy of chemotherapy. The P-glycoprotein (P-gp) overexpression is one of the most common causes of multidrug resistance (MDR) in leukemic cells. The presence of this protein in human and murine cell lines leads not only to resistance against P-gp substrates but significant is also its influence on cell response to substrates, which are not P-gp substrates. In our work, we found that P-gp expression in this cells was accompanied by changes in the levels/activities of regulatory or structural proteins. Some of these proteins are involved in the regulation pathway of apoptosis (Bcl-2 family proteins) or they can be the target of leukemia immunotherapy (CD33). The solving of this project have the potential to bring results clinically applicable in therapeutic protocols aimed to appropriate type of neoplastic diseases.
Duration: 1.1.2017 - 31.12.2020

Effects of stress, induced by immune challenge during the gestation in rats, on maternal care behaviour and on hippocampal neuronal excitability in the offsprings
Účinok stresu vyvolaného imunitnou reakciou počas gravidity potkanov na následnú starostlivosť o mláďatá a na hipokampálnu excitabilitu mláďat.
Program: VEGA
Project leader: MMedSc Dremencov Eliyahu PhD
Annotation:There is evidence that children of women which experienced stressful conditions during the pregnancy have higher risk to develop certain brain disorders, such as depression, schizophrenia, and autism. One of the common stressors experienced by pregnant women is acute infective illness. Little is known how prenatal infection influences the functioning of offspring brain. In this study, we plan to investigate the effect of prenatal immune challenge resembling infective illness, on maternal care behaviour of laboratory rats and on hippocampal neuronal activity of their pups. The immune challenge will be induced by the administration of bacterial endotoxin lipopolysaccharide (LPS). Neuronal excitability will be examined in hippocampus, the structure related to depression, schizophrenia and autism, and compared between offsprings of rats treated with LPS and control rats. The results generated by this project will provide better understanding of pathophysiology of brain disorders triggered by prenatal stress.
Duration: 1.1.2015 - 31.12.2017

The role of physiologically relevant metal ions Mg2+ and Zn2+ in the luminal regulation of the cardiac ryanodine receptor.
Úloha fyziologicky aktívnych iónov Mg2+ a Zn2+ v luminálnej regulácii srdcového ryanodínového receptora.
Program: VEGA
Project leader: Mgr. Gaburjáková Jana PhD.
Annotation:The key determinant of cardiac contractility is Ca2+ released from the sarcoplasmic reticulum (SR) via the ryanodine receptor type 2 (RYR2). The major regulation domain of the RYR2 channel is a huge cytosolic head. Smaller luminal domain also contributes to the channel regulation, and importantly, its dysfunction leads to generation of specific ventricular arrhythmias. The main luminal regulator of the RYR2 channel is Ca2+. However, other cations such as Mg2+ and Zn2+ are also present in the SR and can contribute to the luminal regulation by binding to luminal RYR2 sites. These binding sites are localized either on the channel protein or on the associated protein-calsequestrin (CSQ2). Our aim is to examine the luminal regulation of the RYR2 channel in more comprehensive manner in respect to cationic composition of the luminal environment. On the single channel level we will examine the role of Mg2+ and Zn2+ in luminal regulation of the RYR2 channel at diastolic (1 mM) or systolic (0.6 mM) luminal Ca2+. Outcomes of experiments will be correlated to the CSQ2 presence in the RYR2 protein complex.
Duration: 1.1.2015 - 31.12.2017

The effect of endoplasmic reticulum stress inductors and proteaosome inhibitors on leukemia cell line L1210, SKM-1 and MOLM-13 with induced P-glycoprotein overexpression
Vplyv látok vyvolávajúcich stres endoplazmatického retikula a inhibítorov proteozómu na leukemické bunkové línie L1210, SKM-1 a MOLM-13, u ktorých bola vyvolaná nadexpresia P-glykoproteínu
Program: VEGA
Project leader: Mgr. Šereš Mário PhD.
Annotation:The leukemia mouse cells L1210 in which was induced the P-glycoprotein (P-gp) expression of either by selection with vincristine or transfection with the P-gp encoding gene. We have found that cells with P-gp overexpression, have developed resistance not only to substrates of P-gp, but also to cisplatin (antineoplastic agent, which is not P-gp substrate) and endoplasmic reticulum (ER) stress inducing substances. Further, we observed significant differences in interaction with lectins, between P-gp positive and P-gp negative L1210 cells, indicating extensive remodeling of surface carbohydrates, associated with the P-gp expression in these cells. The presence of P-gp in leukemia cells induces large changes of cell regulatory processes involving protein phosphorylations and glycosylations and the regulation of the apoptosis progression. New knowledge of P-gp regulatory role may contribute to a better understanding of the P-gp mediated multidrug resistant mechanism of leukemia cells to cytostatics.
Duration: 1.1.2016 - 31.12.2018

ER stres - myo - Effect of endoplasmic reticulum stress on ultrastructure and metabolism of mammalian cardiomyocytes
Vplyv stresu endoplazmatického retikula na ultraštruktúru a metabolizmus kardiomyocytov cicavcov
Program: APVV
Project leader: RNDr. Novotová Marta CSc.
Annotation:Endoplasmic reticulum (ER) stress has been implicated in many cardiovascular diseases. Currently, the research on ER stress is aimed toward preservation of the beneficial adaptational response and elimination of the deleterious apoptosis in disease. The objective of this project is to study the adaptational changes in the cytoarchitecture of cardiomyocytes under ER stress, with focus on both, the smooth and rough forms of the reticular membrane system. We will characterize remodeling of myocyte cytoarchitecture induced by application of tunicamycine (the reticular stressor) in control mice and in mice with knocked-out sirtuin1 gene (SIRT1 ciKO mice). Sirtuin1 is a general metabolic energy sensor activated in response to cellular stress that protects cells from apoptosis. We hypothesize that SIRT1 may regulate the unfolded protein response and protect the heart against ER stress-induced damages.
Duration: 1.1.2016 - 31.12.2017

The effect of myocardial load on distribution of the endoplasmic membrane reticulum of cardiac myocytes
Vplyv záťaže myokardu na distribúciu vnútorného membránového systému srdcových myocytov
Program: VEGA
Project leader: RNDr. Novotová Marta CSc.
Annotation:In our previous studies, we have shown that cytoarchitecture contributes to intracellular signaling in cardiac myocytes and that cytoarchitectual changes accompany cardiac remodeling. This project aims at characterization of the morphological variability of the sarcolemma and of the sarcoplasmic reticulum with emphasis on the local aspects of excitation-contraction coupling. Changes in the spatial relations of the plasma membrane and of membranes of the sarcoplasmic reticulum will be studied in the mammalian myocardium with the aim to classify and quantify their structural dynamics in the physiologically and pathologically loaded myocardium. Comprehension of the plasticity of the membrane system will be instrumental in understanding of the membrane morphology as the adaptability factor in cardiac myocytes.
Project web page:http://www.confolab.sav.sk/indexVEGMN.html
Duration: 1.1.2015 - 31.12.2017

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Využitie myrozinázy na aktiváciu sulforafanu pre vývoj preparátu s preventívnymi účinkami nádorových ochorení
Program: APVV
Project leader: Ing. Sulová Zdena DrSc.
Duration: 1.7.2017 - 30.6.2021

Changes of leukemia cells drug sensitivity induced by changes of membrane drug transporters expression profile.
Zmeny citlivosti leukemických buniek na chemoterapeutiká vyvolané zmeneným expresným profilom membránových transportérov.
Program: VEGA
Project leader: doc. Ing. Breier Albert DrSc.
Annotation:Multidrug resistance (MDR) of neoplastic cells represents real obstacle in chemotherapy of cancer diseases. Expression of P-glycoprotein (P-gp), a plasma membrane drug transporter represents most often occurred molecular cause of MDR. Several cell processes and regulatory pathways including protein glycosylation and phosphorylation are involved in P-gp mediated MDR development. Study of multi-player cellular mechanisms that may alter regulation of P-gp expression, degradation and transport activity represents highly important topic for intensive research activity. This research has potential to bring data applicable for enlargement of targeted therapeutic protocols effective for concrete type of neoplastic disease of respective individuals with multidrug resistance mediated by P-gp. The project will be focused on hematological malignancies predominantly on leukemia using different human and animal cell models.
Duration: 1.1.2015 - 31.12.2018

Projects total: 23