Significant scientific results 2017

Evaluation of protein phosphorylation of bull spermatozoa during maturation in the testis

Jana Jankovičová, Katarína Michalková, Petra Sečová, Ľubica Horovská, Jana Antalíková

Protein phosphorylation is an extremely important post-translational modification regulating the function of mature mammalian sperm. Post-testicular maturation of spermatozoa during passage through the testis is a process common to all mammals, but the mechanism of protein modification is species-specific. By fluorescence and phospho-proteomic analysis of bull spermatozoa, we documented an increasing trend of protein phosphorylation during their passage through the epididymis, concomitant with an altered reaction pattern detected on spermatozoa within the tissue as well as spermatozoa isolated from different parts of the epididymis.

Cryosomes of bull testes (a-d) and epididymis (caput, e-h; corpus, i-l; cauda, m-p) labeled with antiphosphotyrosine antibody P Tyr-01 (green), sperm acrosomes labeled with PNA (red), nuclear DNA (DAPI, blue).

JANKOVIČOVÁ, JanaMICHALKOVÁ, KatarínaSEČOVÁ, PetraHOROVSKÁ, Ľubica – MAŇÁSKOVÁ-POSTLEROVÁ, Pavla – ANTALÍKOVÁ, Jana. Evaluation of protein phosphorylation in bull sperm during their maturation in the epididymis. In Cell and Tissue Research, 2017, Accepted: 15 September 2017, Published on line 23.october 2017. (2.787 – IF2016). (2017 – Current Contents). ISSN 0302-766X.

Neural mechanisms of the antidepressant effect of physical exercise

Eliyahu Dremencov, Kristína Csatlósová, Barbora Ďurišová, Lucia Moravčíková, Ľubica Lacinová

The aim of the work was to investigate the effect of exercise or exercise combined with an antidepressant on monoamine neurons in the brain. We found that in rats, voluntary wheel running (VWR) activates serotonin (5-HT) neurons and reduces the inhibitory effect of acute administration of escitalopram on the activity of 5-HT neurons (Fig 1). From our results, we conclude that the antidepressant-like effect of exercise is mediated by stimulation of 5-HT neurons, and that exercise may potentiate and facilitate the response to antidepressants in patients with depression.

Účinky VWR a escitalopramu na excitabilitu 5-HT neurónov u potkanov

DREMENCOV, EliyahuCSATLÓSOVÁ, KristínaĎURIŠOVÁ, BarboraMORAVČÍKOVÁ, LuciaLACINOVÁ, Ľubica – JEŽOVÁ, Daniela. Effect of physical exercise and acute escitalopram on the excitability of brain monoamine neurons: in vivo electrophysiological study in rats. In International Journal of Neuropsychopharmacology, 2017, vol. 20, no 7, p. 585-592. (4.712 IF2016). (2017 – Current Contents). ISSN 1461-1457.

The use of aptasensors for cancer diagnosis

Alexandra Poturnayová

Nowadays, the advances in modern medicine and the increased interest in nanoncology are revolutionizing the diagnosis and therapy of cancer. Nano-oncology is contributing to the development of more sensitive biosensors for the detection of early stages of disease, thanks to the discovery of new biomarkers. New diagnostic approaches capable of improving the accurate detection of cancer cells are crucial in cancer diagnosis. The development of sensitive biosensors based on DNA aptamers is proving to be a suitable tool for their early detection. Our results show that DNA aptamers are suitable for the detection of proteins embedded in model cell membranes or membranes of living cells.

LEITNER, Míchael – POTURNAYOVÁ, Alexandra – LAMPRECHT, Constanze – WEICH, Sabine ŠNEJDÁRKOVÁ, Maja – KARPIŠOVÁ, Ivana – HIANIK, Tibor – EBNER, Andreas. Characterization of the specific interaction between the DNA aptamer sgc8c and protein tyrosine kinase-7 receptors at the surface of T-cells by biosensing AFM. In Analytical and Bioanalytical Biochemistry, 2017, vol. 409, iss. 11, p. 2767-2776. (3.431 – IF2016). (2017 – Current Contents). ISSN 1618-2642.

POTURNAYOVÁ, Alexandra – CASTILLO, Gabriela – SUBJAKOVÁ, Veronika – TATARKO, Marek – ŠNEJDÁRKOVÁ, Maja – HIANIK, Tibor. Optimization of cytochrome c detection by acoustic and electrochemical methods based on aptamer sensors. In Sensors and Actuators B: Chemical, 2017, vol. 238, p. 817-827. (5.401 – IF2016). (2017 – Current Contents). ISSN 0925-4005.

Accurate measurement of membrane capacitance by deconvolution of current responses

Matej Hoťka, Ivan Zahradník

The membrane capacity of all living cells changes as a result of cellular processes associated with growth, transport of substances, changes in membrane composition, as well as during cell signalling. To date, accurate methods for measuring membrane capacitance require special instrumentation and considerable expertise due to the artefacts present in the measurements. We have developed a software method to remove a major source of artifacts based on reconstruction of membrane current responses to voltage step stimuli.

Time course of the current response of a cell (IM) to a voltage step change (VStim) before and after reconstruction using a deconvolution procedure

By using deconvolution in the frequency domain, we remove the influence of the transfer function of the measurement system. Thus, we obtain current records without major measurement artefacts, suitable for analysis with a model of the measured system. The actual analysis then provides impedance parameters without crosstalk, with high absolute accuracy and measurement reliability. The software developed for automatic membrane current collection and analysis, as well as for the evaluation of instantaneous membrane capacitance values, is freely accessible via a public web server. Using experiments on isolated cardiac muscle cells, we have demonstrated that the new analysis method allows membrane capacitance to be measured at the physical limit, i.e. with an amplitude resolution of one femtofarad up to one thousand times per second.

HOŤKA, MatejZAHRADNÍK, Ivan. Reconstruction of membrane current by deconvolution and its application to membrane capacitance measurements in cardiac myocytes. In PLoS ONE, 2017, vol. 12., iss. 11, e0188452. (2.806 – IF2016). ISSN 1932-6203.

Mapping livestock welfar research in Europe

Ľubor Košťál, Boris Bilčík

Following the successful completion of the FP7 AWARE project , we published in 2017, in collaboration with Austrian colleagues, the results of a mapping of the state of livestock welfarist research in Europe, building on the results of a similar mapping of livestock welfarist education in Europe (Illman et al 2014). The research mapping showed that in Western and Northern European countries there are more researchers active in this field, that in Eastern and Southern Europe this research is less funded or not funded at all. Respondents from Western and Northern Europe were significantly more involved in EU-funded schemes such as FP7 and ERA-NET projects. Associations were found between scientific output in this field and the economic performance of countries, resulting in lower ‘visibility’ and influence of researchers in Eastern and South-Eastern Europe.

Participation in international projects in the field of welfar by country.

ILLMANN, Gudrun – KEELING, Linda – MELIŠOVÁ, Dana – ŠIMEČKOVÁ, M. – ILIESKI, Vlatko – WINCKLER, Christoph – KOŠŤÁL, Ľubor. Mapping farm animal welfare education at university level in Europe. In Animal Welfare, 2014, vol. 23, p. 401-410. (1.228 – IF2013).

KIRCHNER, Marlene K. – KOŠŤÁL, ĽuborBILČÍK, Boris – WINCKLER, Christoph. Mapping farm animal welfare research in an enlarged Europe: international collaboration, bibliometric output, research resources and relation to economic indices. In Scientometrics, 2017, vol. 113, iss. 2, p. 909-922. (2.147 – IF2016).

Regulation of heart muscle growth after birth

Marta Novotová

The heart muscle of newborn mammals initially develops rapidly. Its muscle cells multiply, grow and gradually refine their structure so that the heart can meet the growing demands of the organism. This process gradually slows down. Organ growth is controlled by general mechanisms. In the case of cardiac muscle cells, the process is specific. We studied the role of the Fat4 protein, which is part of the signaling mechanisms controlling cell growth, because its damage leads to death of newborn mice due to disorganization of the heart muscle. Using molecular biology, genetics, cytology, and electron microscopy (our contribution), we characterized the molecular signaling mechanisms underlying this regulatory process. We found that Fat4 regulates cardiomyocyte growth in concert with Amotl1 and Yap1 through the Hippo pathway. This finding has implications for understanding regenerative processes and developing tools to control them in the treatment of injured and failing myocardium.

RAGNI, Chiara V. – DIGUET, Nicolas – LE GARREC, Jean-Francois – NOVOTOVÁ, Marta – RESENDE, Tatiana P. – POP, Sorin – CHARON, Nicolas – GUILLEMOT, Laurent – KITASATO, Lisa – BADOUEL, Caroline – DUFOUR, Alexandre – OLIVO-MARIN, Jean-Christophe – TROUVE, Alain – MCNEILL, Helen – MEILHAC, Sigolene M. Amotl1 mediates sequestration of the Hippo effector Yap1 downstream of Fat4 to restrict heart growth. In Nature Communications, 2017, vol. 8, p. 14582. (12.124 – IF2016). (2017 – Current Contents). ISSN 2041-1723.