TY  - JOUR
AU  - Pajić, Tanja
AU  - Stevanović, Katarina
AU  - Todorović, Nataša
AU  - Krmpot, Aleksandar J
AU  - Živić, Miroslav
AU  - Savić-Šević, Svetlana
AU  - Lević, Steva M
AU  - Stanić, Marina
AU  - Pantelić, Dejan
AU  - Jelenković, Brana
AU  - Rabasović, Mihailo D
PY  - 2024
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6666
AB  - Studying the membrane physiology of filamentous fungi is key to understanding their interactions with the environment and crucial for developing new therapeutic strategies for disease-causing pathogens. However, their plasma membrane has been inaccessible for a micron-sized patch-clamp pipette for pA current recordings due to the rigid chitinous cell wall. Here, we report the first femtosecond IR laser nanosurgery of the cell wall of the filamentous fungi, which enabled patch-clamp measurements on protoplasts released from hyphae. A reproducible and highly precise (diffraction-limited, submicron resolution) method for obtaining viable released protoplasts was developed. Protoplast release from the nanosurgery-generated incisions in the cell wall was achieved from different regions of the hyphae. The plasma membrane of the obtained protoplasts formed tight and high-resistance (GΩ) contacts with the recording pipette. The entire nanosurgical procedure followed by the patch-clamp technique could be completed in less than 1 hour. Compared to previous studies using heterologously expressed channels, this technique provides the opportunity to identify new ionic currents and to study the properties of the ion channels in the protoplasts of filamentous fungi in their native environment.
PB  - Springer Nature
T2  - Microsystems & Nanoengineering
T1  - In vivo femtosecond laser nanosurgery of the cell wall enabling patch-clamp measurements on filamentous fungi
VL  - 10
DO  - 10.1038/s41378-024-00664-x
SP  - 47
ER  - 

@article{
author = "Pajić, Tanja and Stevanović, Katarina and Todorović, Nataša and Krmpot, Aleksandar J and Živić, Miroslav and Savić-Šević, Svetlana and Lević, Steva M and Stanić, Marina and Pantelić, Dejan and Jelenković, Brana and Rabasović, Mihailo D",
year = "2024",
abstract = "Studying the membrane physiology of filamentous fungi is key to understanding their interactions with the environment and crucial for developing new therapeutic strategies for disease-causing pathogens. However, their plasma membrane has been inaccessible for a micron-sized patch-clamp pipette for pA current recordings due to the rigid chitinous cell wall. Here, we report the first femtosecond IR laser nanosurgery of the cell wall of the filamentous fungi, which enabled patch-clamp measurements on protoplasts released from hyphae. A reproducible and highly precise (diffraction-limited, submicron resolution) method for obtaining viable released protoplasts was developed. Protoplast release from the nanosurgery-generated incisions in the cell wall was achieved from different regions of the hyphae. The plasma membrane of the obtained protoplasts formed tight and high-resistance (GΩ) contacts with the recording pipette. The entire nanosurgical procedure followed by the patch-clamp technique could be completed in less than 1 hour. Compared to previous studies using heterologously expressed channels, this technique provides the opportunity to identify new ionic currents and to study the properties of the ion channels in the protoplasts of filamentous fungi in their native environment.",
publisher = "Springer Nature",
journal = "Microsystems & Nanoengineering",
title = "In vivo femtosecond laser nanosurgery of the cell wall enabling patch-clamp measurements on filamentous fungi",
volume = "10",
doi = "10.1038/s41378-024-00664-x",
pages = "47"
}

Pajić, T., Stevanović, K., Todorović, N., Krmpot, A. J., Živić, M., Savić-Šević, S., Lević, S. M., Stanić, M., Pantelić, D., Jelenković, B.,& Rabasović, M. D.. (2024). In vivo femtosecond laser nanosurgery of the cell wall enabling patch-clamp measurements on filamentous fungi. in Microsystems & Nanoengineering
Springer Nature., 10, 47.
https://doi.org/10.1038/s41378-024-00664-x

Pajić T, Stevanović K, Todorović N, Krmpot AJ, Živić M, Savić-Šević S, Lević SM, Stanić M, Pantelić D, Jelenković B, Rabasović MD. In vivo femtosecond laser nanosurgery of the cell wall enabling patch-clamp measurements on filamentous fungi. in Microsystems & Nanoengineering. 2024;10:47.
doi:10.1038/s41378-024-00664-x .

Pajić, Tanja, Stevanović, Katarina, Todorović, Nataša, Krmpot, Aleksandar J, Živić, Miroslav, Savić-Šević, Svetlana, Lević, Steva M, Stanić, Marina, Pantelić, Dejan, Jelenković, Brana, Rabasović, Mihailo D, "In vivo femtosecond laser nanosurgery of the cell wall enabling patch-clamp measurements on filamentous fungi" in Microsystems & Nanoengineering, 10 (2024):47,
https://doi.org/10.1038/s41378-024-00664-x . .

TY  - CONF
AU  - Vitkovac, Aleksandra
AU  - Pajić, Tanja
AU  - Bukumira, Marta
AU  - Stanić, Marina
AU  - Rabasović, Mihailo
AU  - Todorović, Nataša
PY  - 2023
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6247
AB  - Adaptive changes in mitochondrial morphology are associated with changes in the mitochondrial function and metabolic fitness of eukaryotic cells. We previously described in young hyphae of the filamentous fungus Phycomyces blakesleeanus a dramatic effect of an increase in ambient temperature during growth: a 3°C warmer environment compared with a control temperature of 22°C resulted in the appearance of long elongated (“tubular”) mitochondria accompanied by an increase in lipid droplet density. Here, we examined how cooler ambient temperature (18°C) during growth affects mitochondrial morphology in P. blakesleeanus compared with the control grown at 22°C. We used two-photon fluorescence imaging (TPEF) of live hyphae stained with the vital mitochondrial dye rhodamine 123. Extraction of relevant parameters (number, size, and shape of mitochondria) from TPEF images was performed using the Ilastik machine learning-based software. The suitability of the Ilastik analysis was compared with the Particle Analysis (ImageJ). Cold treatment resulted in the appearance of tubular mitochondrial morphology that was absent in the control group. Tubular mitochondrial morphology appears to be an adaptive feature that occurs in both warmer and colder conditions and is likely part of the stress response.
PB  - Kragujevac: Institute for Information Technologies, University of Belgrade
C3  - Book of Proceedings: 2nd International Conference on Chemo and Bioinformatics ICCBIKG 2023; 2023 Sep 28-29; Kragujevac, Serbia
T1  - Slight cooling during growth induced changes in filamentous fungi hypha mitochondrial morphology
DO  - 10.46793/ICCBI23.334V
SP  - 334
EP  - 337
ER  - 

@conference{
author = "Vitkovac, Aleksandra and Pajić, Tanja and Bukumira, Marta and Stanić, Marina and Rabasović, Mihailo and Todorović, Nataša",
year = "2023",
abstract = "Adaptive changes in mitochondrial morphology are associated with changes in the mitochondrial function and metabolic fitness of eukaryotic cells. We previously described in young hyphae of the filamentous fungus Phycomyces blakesleeanus a dramatic effect of an increase in ambient temperature during growth: a 3°C warmer environment compared with a control temperature of 22°C resulted in the appearance of long elongated (“tubular”) mitochondria accompanied by an increase in lipid droplet density. Here, we examined how cooler ambient temperature (18°C) during growth affects mitochondrial morphology in P. blakesleeanus compared with the control grown at 22°C. We used two-photon fluorescence imaging (TPEF) of live hyphae stained with the vital mitochondrial dye rhodamine 123. Extraction of relevant parameters (number, size, and shape of mitochondria) from TPEF images was performed using the Ilastik machine learning-based software. The suitability of the Ilastik analysis was compared with the Particle Analysis (ImageJ). Cold treatment resulted in the appearance of tubular mitochondrial morphology that was absent in the control group. Tubular mitochondrial morphology appears to be an adaptive feature that occurs in both warmer and colder conditions and is likely part of the stress response.",
publisher = "Kragujevac: Institute for Information Technologies, University of Belgrade",
journal = "Book of Proceedings: 2nd International Conference on Chemo and Bioinformatics ICCBIKG 2023; 2023 Sep 28-29; Kragujevac, Serbia",
title = "Slight cooling during growth induced changes in filamentous fungi hypha mitochondrial morphology",
doi = "10.46793/ICCBI23.334V",
pages = "334-337"
}

Vitkovac, A., Pajić, T., Bukumira, M., Stanić, M., Rabasović, M.,& Todorović, N.. (2023). Slight cooling during growth induced changes in filamentous fungi hypha mitochondrial morphology. in Book of Proceedings: 2nd International Conference on Chemo and Bioinformatics ICCBIKG 2023; 2023 Sep 28-29; Kragujevac, Serbia
Kragujevac: Institute for Information Technologies, University of Belgrade., 334-337.
https://doi.org/10.46793/ICCBI23.334V

Vitkovac A, Pajić T, Bukumira M, Stanić M, Rabasović M, Todorović N. Slight cooling during growth induced changes in filamentous fungi hypha mitochondrial morphology. in Book of Proceedings: 2nd International Conference on Chemo and Bioinformatics ICCBIKG 2023; 2023 Sep 28-29; Kragujevac, Serbia. 2023;:334-337.
doi:10.46793/ICCBI23.334V .

Vitkovac, Aleksandra, Pajić, Tanja, Bukumira, Marta, Stanić, Marina, Rabasović, Mihailo, Todorović, Nataša, "Slight cooling during growth induced changes in filamentous fungi hypha mitochondrial morphology" in Book of Proceedings: 2nd International Conference on Chemo and Bioinformatics ICCBIKG 2023; 2023 Sep 28-29; Kragujevac, Serbia (2023):334-337,
https://doi.org/10.46793/ICCBI23.334V . .

TY  - CONF
AU  - Žižić, Milan
AU  - Stanić, Marina
AU  - Aquilanti, Giuliana
AU  - Cvetić Antić, Tijana
AU  - Todorović, Nataša
AU  - Živić, Miroslav
AU  - Zakrzewska, Joanna
PY  - 2022
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5733
AB  - In this study, mycelium of fungus Phycomyces blakesleeanus was exposed to soluble toxic form of selenium, selenite (Se+4), with the aim of determining the flow and products of its biotransformation. Selenite is reduced to Se0 in the form selenium nanoparticles (SeNPs) and Se-2 in the form of methylated volatile selenides. Low concentrations of Se+4 in the mycelium form RSSeSR type compounds, which could be a step in the process of SeNPs formation, or incorporation of Se into metabolites such as Se-amino acids.
PB  - Belgrade: Society of Physical Chemists of Serbia
C3  - Proceedings: 16th International Conference on Fundamental and Applied Aspects of Physical Chemistry: Physical Chemistry 2022, Vol. 1; 2022 Sep 26-30; Belgrade, Serbia
T1  - Selenite metabolism in the mycelium of the fungus Phycomyces blakesleeanus
SP  - 93
EP  - 96
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_5733
ER  - 

@conference{
author = "Žižić, Milan and Stanić, Marina and Aquilanti, Giuliana and Cvetić Antić, Tijana and Todorović, Nataša and Živić, Miroslav and Zakrzewska, Joanna",
year = "2022",
abstract = "In this study, mycelium of fungus Phycomyces blakesleeanus was exposed to soluble toxic form of selenium, selenite (Se+4), with the aim of determining the flow and products of its biotransformation. Selenite is reduced to Se0 in the form selenium nanoparticles (SeNPs) and Se-2 in the form of methylated volatile selenides. Low concentrations of Se+4 in the mycelium form RSSeSR type compounds, which could be a step in the process of SeNPs formation, or incorporation of Se into metabolites such as Se-amino acids.",
publisher = "Belgrade: Society of Physical Chemists of Serbia",
journal = "Proceedings: 16th International Conference on Fundamental and Applied Aspects of Physical Chemistry: Physical Chemistry 2022, Vol. 1; 2022 Sep 26-30; Belgrade, Serbia",
title = "Selenite metabolism in the mycelium of the fungus Phycomyces blakesleeanus",
pages = "93-96",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_5733"
}

Žižić, M., Stanić, M., Aquilanti, G., Cvetić Antić, T., Todorović, N., Živić, M.,& Zakrzewska, J.. (2022). Selenite metabolism in the mycelium of the fungus Phycomyces blakesleeanus. in Proceedings: 16th International Conference on Fundamental and Applied Aspects of Physical Chemistry: Physical Chemistry 2022, Vol. 1; 2022 Sep 26-30; Belgrade, Serbia
Belgrade: Society of Physical Chemists of Serbia., 93-96.
https://hdl.handle.net/21.15107/rcub_ibiss_5733

Žižić M, Stanić M, Aquilanti G, Cvetić Antić T, Todorović N, Živić M, Zakrzewska J. Selenite metabolism in the mycelium of the fungus Phycomyces blakesleeanus. in Proceedings: 16th International Conference on Fundamental and Applied Aspects of Physical Chemistry: Physical Chemistry 2022, Vol. 1; 2022 Sep 26-30; Belgrade, Serbia. 2022;:93-96.
https://hdl.handle.net/21.15107/rcub_ibiss_5733 .

Žižić, Milan, Stanić, Marina, Aquilanti, Giuliana, Cvetić Antić, Tijana, Todorović, Nataša, Živić, Miroslav, Zakrzewska, Joanna, "Selenite metabolism in the mycelium of the fungus Phycomyces blakesleeanus" in Proceedings: 16th International Conference on Fundamental and Applied Aspects of Physical Chemistry: Physical Chemistry 2022, Vol. 1; 2022 Sep 26-30; Belgrade, Serbia (2022):93-96,
https://hdl.handle.net/21.15107/rcub_ibiss_5733 .

TY  - JOUR
AU  - Nikolić, Ljiljana
AU  - Todorović, Nataša
AU  - Zakrzewska, Joanna S
AU  - Stanić, Marina B
AU  - Rauš Balind, Snežana
AU  - Kalauzi, Aleksandar
AU  - Petković, Branka
PY  - 2012
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/1168
UR  - https://link.springer.com/article/10.1007/s00359-012-0727-0
AB  - The spontaneously active Br neuron from the brain-subesophageal ganglion complex of the garden snail Helix pomatia rhythmically generates regular bursts of action potentials with quiescent intervals accompanied by slow oscillations of membrane potential. We examined the involvement of the Na+/K+ pump in modulating its bursting activity by applying a static magnetic field. Whole snail brains and Br neuron were exposed to the 10-mT static magnetic field for 15 min. Biochemical data showed that Na+/K+-ATPase activity increased almost twofold after exposure of snail brains to the static magnetic field. Similarly, P-31 NMR data revealed a trend of increasing ATP consumption and increase in intracellular pH mediated by the Na+/H+ exchanger in snail brains exposed to the static magnetic field. Importantly, current clamp recordings from the Br neuron confirmed the increase in activity of the Na+/K+ pump after exposure to the static magnetic field, as the magnitude of ouabain's effect measured on the membrane resting potential, action potential, and interspike interval duration was higher in neurons exposed to the magnetic field. Metabolic pathways through which the magnetic field influenced the Na+/K+ pump could involve phosphorylation and dephosphorylation, as blocking these processes abolished the effect of the static magnetic field.
PB  - Springer
T2  - Journal of Comparative Physiology A-Neuroethology Sensory Neural and Behavioral Physiology
T1  - Involvement of Na+/K+ pump in fine modulation of bursting activity of the snail Br neuron by 10 mT static magnetic field
IS  - 7
VL  - 198
DO  - 10.1007/s00359-012-0727-0
SP  - 525
EP  - 540
ER  - 

@article{
author = "Nikolić, Ljiljana and Todorović, Nataša and Zakrzewska, Joanna S and Stanić, Marina B and Rauš Balind, Snežana and Kalauzi, Aleksandar and Petković, Branka",
year = "2012",
abstract = "The spontaneously active Br neuron from the brain-subesophageal ganglion complex of the garden snail Helix pomatia rhythmically generates regular bursts of action potentials with quiescent intervals accompanied by slow oscillations of membrane potential. We examined the involvement of the Na+/K+ pump in modulating its bursting activity by applying a static magnetic field. Whole snail brains and Br neuron were exposed to the 10-mT static magnetic field for 15 min. Biochemical data showed that Na+/K+-ATPase activity increased almost twofold after exposure of snail brains to the static magnetic field. Similarly, P-31 NMR data revealed a trend of increasing ATP consumption and increase in intracellular pH mediated by the Na+/H+ exchanger in snail brains exposed to the static magnetic field. Importantly, current clamp recordings from the Br neuron confirmed the increase in activity of the Na+/K+ pump after exposure to the static magnetic field, as the magnitude of ouabain's effect measured on the membrane resting potential, action potential, and interspike interval duration was higher in neurons exposed to the magnetic field. Metabolic pathways through which the magnetic field influenced the Na+/K+ pump could involve phosphorylation and dephosphorylation, as blocking these processes abolished the effect of the static magnetic field.",
publisher = "Springer",
journal = "Journal of Comparative Physiology A-Neuroethology Sensory Neural and Behavioral Physiology",
title = "Involvement of Na+/K+ pump in fine modulation of bursting activity of the snail Br neuron by 10 mT static magnetic field",
number = "7",
volume = "198",
doi = "10.1007/s00359-012-0727-0",
pages = "525-540"
}

Nikolić, L., Todorović, N., Zakrzewska, J. S., Stanić, M. B., Rauš Balind, S., Kalauzi, A.,& Petković, B.. (2012). Involvement of Na+/K+ pump in fine modulation of bursting activity of the snail Br neuron by 10 mT static magnetic field. in Journal of Comparative Physiology A-Neuroethology Sensory Neural and Behavioral Physiology
Springer., 198(7), 525-540.
https://doi.org/10.1007/s00359-012-0727-0

Nikolić L, Todorović N, Zakrzewska JS, Stanić MB, Rauš Balind S, Kalauzi A, Petković B. Involvement of Na+/K+ pump in fine modulation of bursting activity of the snail Br neuron by 10 mT static magnetic field. in Journal of Comparative Physiology A-Neuroethology Sensory Neural and Behavioral Physiology. 2012;198(7):525-540.
doi:10.1007/s00359-012-0727-0 .

Nikolić, Ljiljana, Todorović, Nataša, Zakrzewska, Joanna S, Stanić, Marina B, Rauš Balind, Snežana, Kalauzi, Aleksandar, Petković, Branka, "Involvement of Na+/K+ pump in fine modulation of bursting activity of the snail Br neuron by 10 mT static magnetic field" in Journal of Comparative Physiology A-Neuroethology Sensory Neural and Behavioral Physiology, 198, no. 7 (2012):525-540,
https://doi.org/10.1007/s00359-012-0727-0 . .