TY  - JOUR
AU  - Perić, Mina
AU  - Bataveljić, Danijela
AU  - Bijelić, Dunja
AU  - Milićević, Katarina
AU  - Andjus, Pavle R.
AU  - Bogdanović Pristov, Jelena
AU  - Nikolić, Ljiljana
PY  - 2022
UR  - https://onlinelibrary.wiley.com/doi/10.1002/jemt.24066
UR  - http://www.ncbi.nlm.nih.gov/pubmed/35088507
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/4790
AB  - We describe an approach for studying the physiology of single live cells using the conceptionally novel upright microscope/patch-clamp configuration. Electrophysiology experiments typically require a microscope with the fixed stage position and the motion control of the microscope objective. Here, we demonstrate that a microscope with a z-axis movable stage and a fixed objective can also be efficiently used in combination with the patch-clamp technique. We define a set of underlying principles governing the operation of this microscope/patch-clamp configuration and demonstrate its performance in practice using cultured astrocytes, microglia, and oligodendrocytes. Experimental results show that our custom configuration provides stable recordings, has a high success rate of the whole-cell patch-clamp trials, can be effectively applied to study cellular physiology of glial cells, and provides comparable performance and usability to the commercially available systems. Our system can be easily replicated or adapted to suit the needs of the research groups and can be cost-effective in reducing the investments in purchasing additional equipment. We provide step-by-step instructions on implementing an upright microscope with z-axis movable stage as a routine workhorse for patch-clamping.
PB  - Hoboken: John Wiley and Sons Inc.
T2  - Microscopy Research and Technique
T1  - Approach for patch-clamping using an upright microscope with z-axis movable stage.
IS  - 6
VL  - 85
DO  - 10.1002/jemt.24066
SP  - 2095
EP  - 2104
ER  - 

@article{
author = "Perić, Mina and Bataveljić, Danijela and Bijelić, Dunja and Milićević, Katarina and Andjus, Pavle R. and Bogdanović Pristov, Jelena and Nikolić, Ljiljana",
year = "2022",
abstract = "We describe an approach for studying the physiology of single live cells using the conceptionally novel upright microscope/patch-clamp configuration. Electrophysiology experiments typically require a microscope with the fixed stage position and the motion control of the microscope objective. Here, we demonstrate that a microscope with a z-axis movable stage and a fixed objective can also be efficiently used in combination with the patch-clamp technique. We define a set of underlying principles governing the operation of this microscope/patch-clamp configuration and demonstrate its performance in practice using cultured astrocytes, microglia, and oligodendrocytes. Experimental results show that our custom configuration provides stable recordings, has a high success rate of the whole-cell patch-clamp trials, can be effectively applied to study cellular physiology of glial cells, and provides comparable performance and usability to the commercially available systems. Our system can be easily replicated or adapted to suit the needs of the research groups and can be cost-effective in reducing the investments in purchasing additional equipment. We provide step-by-step instructions on implementing an upright microscope with z-axis movable stage as a routine workhorse for patch-clamping.",
publisher = "Hoboken: John Wiley and Sons Inc.",
journal = "Microscopy Research and Technique",
title = "Approach for patch-clamping using an upright microscope with z-axis movable stage.",
number = "6",
volume = "85",
doi = "10.1002/jemt.24066",
pages = "2095-2104"
}

Perić, M., Bataveljić, D., Bijelić, D., Milićević, K., Andjus, P. R., Bogdanović Pristov, J.,& Nikolić, L.. (2022). Approach for patch-clamping using an upright microscope with z-axis movable stage.. in Microscopy Research and Technique
Hoboken: John Wiley and Sons Inc.., 85(6), 2095-2104.
https://doi.org/10.1002/jemt.24066

Perić M, Bataveljić D, Bijelić D, Milićević K, Andjus PR, Bogdanović Pristov J, Nikolić L. Approach for patch-clamping using an upright microscope with z-axis movable stage.. in Microscopy Research and Technique. 2022;85(6):2095-2104.
doi:10.1002/jemt.24066 .

Perić, Mina, Bataveljić, Danijela, Bijelić, Dunja, Milićević, Katarina, Andjus, Pavle R., Bogdanović Pristov, Jelena, Nikolić, Ljiljana, "Approach for patch-clamping using an upright microscope with z-axis movable stage." in Microscopy Research and Technique, 85, no. 6 (2022):2095-2104,
https://doi.org/10.1002/jemt.24066 . .

TY  - CONF
AU  - Perić, Mina
AU  - Nikolić, Ljiljana
AU  - Anđus, Pavle R.
AU  - Bataveljić, Danijela
PY  - 2020
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5499
AB  - Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease affecting lower
and upper motor neurons, leading to muscle atrophy, paralysis and death. Although
ALS is characterized by motoneuron degeneration, non-neuronal cells such as
oligodendrocytes play an essential role in disease onset and progression. The aim
of this study was to investigate functional properties of oligodendrocytes in the ALS
with particular focus on their role in the regulation of extracellular K+ through inwardly
rectifying potassium channel Kir4.1 abundantly expressed in these glial cells. We
employed immunolabeling of Kir4.1, and oligodendrocyte-specific marker CNPase,
to examine the expression of Kir4.1 in oligodendrocytes of the lumbar spinal cord of
SOD1G93A rat model of ALS. Furthermore, whole-cell patch-clamp recordings were
performed on primary oligodendrocyte cultures from ALS and control rats to
investigate functional properties of these cells. Our data show reduced expression
of Kir4.1 in the oligodendrocytes in the lumbar spinal cord of SOD1G93A rat compared
to control. Moreover, we found elevated number of dysmorphic oligodendrocytes in
the ALS spinal cord, indicative of a degenerative phenotype. Oligodendrocytes
isolated from SOD1G93A spinal cord display similar processes ramification as the
control, however expressing a lower level of Kir4.1. Electrophysiological examination
of cultured SOD1G93A oligodendrocytes revealed compromised membrane
properties and diminished inward currents in comparison to control. In addition, the
Ba2+-sensitive Kir current is decreased in ALS oligodendrocytes. Altogether, our
findings provide the evidence of impaired Kir4.1 expression and function in
SOD1G93A spinal cord oligodendrocytes suggesting a critical role of oligodendrocyte
Kir4.1 channel in ALS pathophysiology.
PB  - Querétaro, México: Instituto de neurobiologia
C3  - Proceedings: 3rd Symposium on Physiology and pathology of neuroglia; 2020 Noc 24-25; Virtual
T1  - Dysfunction of oligodendrocyte inwardly rectifying potassium channel in a rat model of amyotrophic lateral sclerosis
SP  - 75
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_5499
ER  - 

@conference{
author = "Perić, Mina and Nikolić, Ljiljana and Anđus, Pavle R. and Bataveljić, Danijela",
year = "2020",
abstract = "Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease affecting lower
and upper motor neurons, leading to muscle atrophy, paralysis and death. Although
ALS is characterized by motoneuron degeneration, non-neuronal cells such as
oligodendrocytes play an essential role in disease onset and progression. The aim
of this study was to investigate functional properties of oligodendrocytes in the ALS
with particular focus on their role in the regulation of extracellular K+ through inwardly
rectifying potassium channel Kir4.1 abundantly expressed in these glial cells. We
employed immunolabeling of Kir4.1, and oligodendrocyte-specific marker CNPase,
to examine the expression of Kir4.1 in oligodendrocytes of the lumbar spinal cord of
SOD1G93A rat model of ALS. Furthermore, whole-cell patch-clamp recordings were
performed on primary oligodendrocyte cultures from ALS and control rats to
investigate functional properties of these cells. Our data show reduced expression
of Kir4.1 in the oligodendrocytes in the lumbar spinal cord of SOD1G93A rat compared
to control. Moreover, we found elevated number of dysmorphic oligodendrocytes in
the ALS spinal cord, indicative of a degenerative phenotype. Oligodendrocytes
isolated from SOD1G93A spinal cord display similar processes ramification as the
control, however expressing a lower level of Kir4.1. Electrophysiological examination
of cultured SOD1G93A oligodendrocytes revealed compromised membrane
properties and diminished inward currents in comparison to control. In addition, the
Ba2+-sensitive Kir current is decreased in ALS oligodendrocytes. Altogether, our
findings provide the evidence of impaired Kir4.1 expression and function in
SOD1G93A spinal cord oligodendrocytes suggesting a critical role of oligodendrocyte
Kir4.1 channel in ALS pathophysiology.",
publisher = "Querétaro, México: Instituto de neurobiologia",
journal = "Proceedings: 3rd Symposium on Physiology and pathology of neuroglia; 2020 Noc 24-25; Virtual",
title = "Dysfunction of oligodendrocyte inwardly rectifying potassium channel in a rat model of amyotrophic lateral sclerosis",
pages = "75",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_5499"
}

Perić, M., Nikolić, L., Anđus, P. R.,& Bataveljić, D.. (2020). Dysfunction of oligodendrocyte inwardly rectifying potassium channel in a rat model of amyotrophic lateral sclerosis. in Proceedings: 3rd Symposium on Physiology and pathology of neuroglia; 2020 Noc 24-25; Virtual
Querétaro, México: Instituto de neurobiologia., 75.
https://hdl.handle.net/21.15107/rcub_ibiss_5499

Perić M, Nikolić L, Anđus PR, Bataveljić D. Dysfunction of oligodendrocyte inwardly rectifying potassium channel in a rat model of amyotrophic lateral sclerosis. in Proceedings: 3rd Symposium on Physiology and pathology of neuroglia; 2020 Noc 24-25; Virtual. 2020;:75.
https://hdl.handle.net/21.15107/rcub_ibiss_5499 .

Perić, Mina, Nikolić, Ljiljana, Anđus, Pavle R., Bataveljić, Danijela, "Dysfunction of oligodendrocyte inwardly rectifying potassium channel in a rat model of amyotrophic lateral sclerosis" in Proceedings: 3rd Symposium on Physiology and pathology of neuroglia; 2020 Noc 24-25; Virtual (2020):75,
https://hdl.handle.net/21.15107/rcub_ibiss_5499 .