TY  - JOUR
AU  - Milošević, Milena
AU  - Stenovec, Matjaz
AU  - Kreft, Marko
AU  - Petrušić, Vladimir
AU  - Stević, Zorica
AU  - Trkov, Saša
AU  - Anđus, Pavle
AU  - Zorec, Robert
PY  - 2013
UR  - http://intor.torlakinstitut.com/handle/123456789/378
AB  - Astrocytes are considered essential in the etiopathogenesis of amyotrophic lateral sclerosis (ALS). We have demonstrated previously that immunoglobulins G (IgG) isolated from patients with ALS enhance the mobility of acidic vesicles in cultured astrocytes in a Ca2+-dependent manner. Here we directly examined the impact of purified sporadic ALS IgG on cytosolic [Ca2+] ([Ca2+](i)) in astrocytes. Confocal time-lapse images were acquired and fluorescence of a non-ratiometric Ca2+ indicator was recorded before and after the application of IgG. ALS IgG (0.1 mg/ml) from 7 patients evoked transient increases in [Ca2+](i) in similar to 50% of tested astrocytes. The probability of observing a response was independent of extracellular Ca2+. The peak increase in [Ca2+](i) developed similar to 3 times faster and the time integral of evoked transients was similar to 2-fold larger; the peak amplitude itself was not affected by extracellular Ca2+. Application of pharmacological inhibitors revealed that activation of inositol-1,4,5-triphosphate receptors is necessary and sufficient to initiate transients in [Ca2+](i) the Ca2+ influx through store-operated calcium entry prolongs the transient increase in [Ca2+](i). Thus, ALS IgG acutely affect [Ca2+](i) by mobilizing both, intra- and extracellular Ca2+ into the cytosol of cultured astrocytes. (C) 2013 Elsevier Ltd. All rights reserved.
PB  - Elsevier Sci Ltd, Oxford
T2  - Cell Calcium
T1  - Immunoglobulins G from patients with sporadic amyotrophic lateral sclerosis affects cytosolic Ca2+ homeostasis in cultured rat astrocytes
EP  - 25
IS  - 1
SP  - 17
VL  - 54
DO  - 10.1016/j.ceca.2013.03.005
ER  - 

@article{
author = "Milošević, Milena and Stenovec, Matjaz and Kreft, Marko and Petrušić, Vladimir and Stević, Zorica and Trkov, Saša and Anđus, Pavle and Zorec, Robert",
year = "2013",
abstract = "Astrocytes are considered essential in the etiopathogenesis of amyotrophic lateral sclerosis (ALS). We have demonstrated previously that immunoglobulins G (IgG) isolated from patients with ALS enhance the mobility of acidic vesicles in cultured astrocytes in a Ca2+-dependent manner. Here we directly examined the impact of purified sporadic ALS IgG on cytosolic [Ca2+] ([Ca2+](i)) in astrocytes. Confocal time-lapse images were acquired and fluorescence of a non-ratiometric Ca2+ indicator was recorded before and after the application of IgG. ALS IgG (0.1 mg/ml) from 7 patients evoked transient increases in [Ca2+](i) in similar to 50% of tested astrocytes. The probability of observing a response was independent of extracellular Ca2+. The peak increase in [Ca2+](i) developed similar to 3 times faster and the time integral of evoked transients was similar to 2-fold larger; the peak amplitude itself was not affected by extracellular Ca2+. Application of pharmacological inhibitors revealed that activation of inositol-1,4,5-triphosphate receptors is necessary and sufficient to initiate transients in [Ca2+](i) the Ca2+ influx through store-operated calcium entry prolongs the transient increase in [Ca2+](i). Thus, ALS IgG acutely affect [Ca2+](i) by mobilizing both, intra- and extracellular Ca2+ into the cytosol of cultured astrocytes. (C) 2013 Elsevier Ltd. All rights reserved.",
publisher = "Elsevier Sci Ltd, Oxford",
journal = "Cell Calcium",
title = "Immunoglobulins G from patients with sporadic amyotrophic lateral sclerosis affects cytosolic Ca2+ homeostasis in cultured rat astrocytes",
pages = "25-17",
number = "1",
volume = "54",
doi = "10.1016/j.ceca.2013.03.005"
}

Milošević, M., Stenovec, M., Kreft, M., Petrušić, V., Stević, Z., Trkov, S., Anđus, P.,& Zorec, R.. (2013). Immunoglobulins G from patients with sporadic amyotrophic lateral sclerosis affects cytosolic Ca2+ homeostasis in cultured rat astrocytes. in Cell Calcium
Elsevier Sci Ltd, Oxford., 54(1), 17-25.
https://doi.org/10.1016/j.ceca.2013.03.005

Milošević M, Stenovec M, Kreft M, Petrušić V, Stević Z, Trkov S, Anđus P, Zorec R. Immunoglobulins G from patients with sporadic amyotrophic lateral sclerosis affects cytosolic Ca2+ homeostasis in cultured rat astrocytes. in Cell Calcium. 2013;54(1):17-25.
doi:10.1016/j.ceca.2013.03.005 .

Milošević, Milena, Stenovec, Matjaz, Kreft, Marko, Petrušić, Vladimir, Stević, Zorica, Trkov, Saša, Anđus, Pavle, Zorec, Robert, "Immunoglobulins G from patients with sporadic amyotrophic lateral sclerosis affects cytosolic Ca2+ homeostasis in cultured rat astrocytes" in Cell Calcium, 54, no. 1 (2013):17-25,
https://doi.org/10.1016/j.ceca.2013.03.005 . .

TY  - CONF
AU  - Anđus, Pavle
AU  - Stenovec, Matjaz
AU  - Milošević, Milena
AU  - Petrušić, Vladimir
AU  - Potokar, Maja
AU  - Stević, Zorica
AU  - Prebil, Mateja
AU  - Kreft, Marko
AU  - Trkov, Saša
AU  - Zorec, Robert
PY  - 2012
UR  - http://intor.torlakinstitut.com/handle/123456789/355
PB  - Cell Press, Cambridge
C3  - Biophysical Journal
T1  - Amyotrophic Lateral Sclerosis IgGs Enhance the Mobility of Lysotracker-Labelled Vesicles in Cultured Rat Astrocytes
EP  - 380A
IS  - 3
SP  - 380A
VL  - 102
DO  - 10.1016/j.bpj.2011.11.2079
ER  - 

@conference{
author = "Anđus, Pavle and Stenovec, Matjaz and Milošević, Milena and Petrušić, Vladimir and Potokar, Maja and Stević, Zorica and Prebil, Mateja and Kreft, Marko and Trkov, Saša and Zorec, Robert",
year = "2012",
publisher = "Cell Press, Cambridge",
journal = "Biophysical Journal",
title = "Amyotrophic Lateral Sclerosis IgGs Enhance the Mobility of Lysotracker-Labelled Vesicles in Cultured Rat Astrocytes",
pages = "380A-380A",
number = "3",
volume = "102",
doi = "10.1016/j.bpj.2011.11.2079"
}

Anđus, P., Stenovec, M., Milošević, M., Petrušić, V., Potokar, M., Stević, Z., Prebil, M., Kreft, M., Trkov, S.,& Zorec, R.. (2012). Amyotrophic Lateral Sclerosis IgGs Enhance the Mobility of Lysotracker-Labelled Vesicles in Cultured Rat Astrocytes. in Biophysical Journal
Cell Press, Cambridge., 102(3), 380A-380A.
https://doi.org/10.1016/j.bpj.2011.11.2079

Anđus P, Stenovec M, Milošević M, Petrušić V, Potokar M, Stević Z, Prebil M, Kreft M, Trkov S, Zorec R. Amyotrophic Lateral Sclerosis IgGs Enhance the Mobility of Lysotracker-Labelled Vesicles in Cultured Rat Astrocytes. in Biophysical Journal. 2012;102(3):380A-380A.
doi:10.1016/j.bpj.2011.11.2079 .

Anđus, Pavle, Stenovec, Matjaz, Milošević, Milena, Petrušić, Vladimir, Potokar, Maja, Stević, Zorica, Prebil, Mateja, Kreft, Marko, Trkov, Saša, Zorec, Robert, "Amyotrophic Lateral Sclerosis IgGs Enhance the Mobility of Lysotracker-Labelled Vesicles in Cultured Rat Astrocytes" in Biophysical Journal, 102, no. 3 (2012):380A-380A,
https://doi.org/10.1016/j.bpj.2011.11.2079 . .

TY  - JOUR
AU  - Stenovec, Matjaz
AU  - Milošević, Milena
AU  - Petrušić, Vladimir
AU  - Potokar, Maja
AU  - Stević, Zorica
AU  - Prebil, Mateja
AU  - Kreft, Marko
AU  - Trkov, Saša
AU  - Anđus, Pavle
AU  - Zorec, Robert
PY  - 2011
UR  - http://intor.torlakinstitut.com/handle/123456789/331
AB  - Aim: We examined the effect of purified immunoglobulins G (IgG) from patients with amyotrophic lateral sclerosis (ALS) on the mobility and exocytotic release from Lysotracker-stained vesicles in cultured rat astrocytes. Methods: Time-lapse confocal images were acquired, and vesicle mobility was analysed before and after the application of ALS IgG. The vesicle counts were obtained to assess cargo exocytosis from stained organelles. Results: At rest, when mobility was monitored for 2 min in bath with Ca2+, two vesicle populations were discovered: (1) non-mobile vesicles (6.1%) with total track length (TL)  lt  1 mu m, averaging at 0.33 +/- 0.01 mu m (n = 1305) and (2) mobile vesicles (93.9%) with TL  gt  1 mu m, averaging at 3.03 +/- 0.01 mu m (n = 20 200). ALS IgG (0.1 mg mL(-1)) from 12 of 13 patients increased the TL of mobile vesicles by approx. 24% and maximal displacement (MD) by approx. 26% within 4 min, while the IgG from control group did not alter the vesicle mobility. The mobility enhancement by ALS IgG was reduced in extracellular solution devoid of Ca2+, indicating that ALS IgG vesicle mobility enhancement involves changes in Ca2+ homeostasis. To examine whether enhanced mobility relates to elevated Ca2+ activity, cells were stimulated by 1 mm ATP, a cytosolic Ca2+ increasing agent, in the presence (2 mm) and in the absence of extracellular Ca2+. ATP stimulation triggered an increase in TL by approx. 7% and 12% and a decrease in MD by approx. 11% and 1%, within 4 min respectively. Interestingly, none of the stimuli triggered the release of vesicle cargo. Conclusion: Amyotrophic lateral sclerosis-IgG-enhanced vesicle mobility in astrocytes engages changes in calcium homeostasis.
PB  - Wiley, Hoboken
T2  - Acta Physiologica
T1  - Amyotrophic lateral sclerosis immunoglobulins G enhance the mobility of Lysotracker-labelled vesicles in cultured rat astrocytes
EP  - 471
IS  - 4
SP  - 457
VL  - 203
DO  - 10.1111/j.1748-1716.2011.02337.x
ER  - 

@article{
author = "Stenovec, Matjaz and Milošević, Milena and Petrušić, Vladimir and Potokar, Maja and Stević, Zorica and Prebil, Mateja and Kreft, Marko and Trkov, Saša and Anđus, Pavle and Zorec, Robert",
year = "2011",
abstract = "Aim: We examined the effect of purified immunoglobulins G (IgG) from patients with amyotrophic lateral sclerosis (ALS) on the mobility and exocytotic release from Lysotracker-stained vesicles in cultured rat astrocytes. Methods: Time-lapse confocal images were acquired, and vesicle mobility was analysed before and after the application of ALS IgG. The vesicle counts were obtained to assess cargo exocytosis from stained organelles. Results: At rest, when mobility was monitored for 2 min in bath with Ca2+, two vesicle populations were discovered: (1) non-mobile vesicles (6.1%) with total track length (TL)  lt  1 mu m, averaging at 0.33 +/- 0.01 mu m (n = 1305) and (2) mobile vesicles (93.9%) with TL  gt  1 mu m, averaging at 3.03 +/- 0.01 mu m (n = 20 200). ALS IgG (0.1 mg mL(-1)) from 12 of 13 patients increased the TL of mobile vesicles by approx. 24% and maximal displacement (MD) by approx. 26% within 4 min, while the IgG from control group did not alter the vesicle mobility. The mobility enhancement by ALS IgG was reduced in extracellular solution devoid of Ca2+, indicating that ALS IgG vesicle mobility enhancement involves changes in Ca2+ homeostasis. To examine whether enhanced mobility relates to elevated Ca2+ activity, cells were stimulated by 1 mm ATP, a cytosolic Ca2+ increasing agent, in the presence (2 mm) and in the absence of extracellular Ca2+. ATP stimulation triggered an increase in TL by approx. 7% and 12% and a decrease in MD by approx. 11% and 1%, within 4 min respectively. Interestingly, none of the stimuli triggered the release of vesicle cargo. Conclusion: Amyotrophic lateral sclerosis-IgG-enhanced vesicle mobility in astrocytes engages changes in calcium homeostasis.",
publisher = "Wiley, Hoboken",
journal = "Acta Physiologica",
title = "Amyotrophic lateral sclerosis immunoglobulins G enhance the mobility of Lysotracker-labelled vesicles in cultured rat astrocytes",
pages = "471-457",
number = "4",
volume = "203",
doi = "10.1111/j.1748-1716.2011.02337.x"
}

Stenovec, M., Milošević, M., Petrušić, V., Potokar, M., Stević, Z., Prebil, M., Kreft, M., Trkov, S., Anđus, P.,& Zorec, R.. (2011). Amyotrophic lateral sclerosis immunoglobulins G enhance the mobility of Lysotracker-labelled vesicles in cultured rat astrocytes. in Acta Physiologica
Wiley, Hoboken., 203(4), 457-471.
https://doi.org/10.1111/j.1748-1716.2011.02337.x

Stenovec M, Milošević M, Petrušić V, Potokar M, Stević Z, Prebil M, Kreft M, Trkov S, Anđus P, Zorec R. Amyotrophic lateral sclerosis immunoglobulins G enhance the mobility of Lysotracker-labelled vesicles in cultured rat astrocytes. in Acta Physiologica. 2011;203(4):457-471.
doi:10.1111/j.1748-1716.2011.02337.x .

Stenovec, Matjaz, Milošević, Milena, Petrušić, Vladimir, Potokar, Maja, Stević, Zorica, Prebil, Mateja, Kreft, Marko, Trkov, Saša, Anđus, Pavle, Zorec, Robert, "Amyotrophic lateral sclerosis immunoglobulins G enhance the mobility of Lysotracker-labelled vesicles in cultured rat astrocytes" in Acta Physiologica, 203, no. 4 (2011):457-471,
https://doi.org/10.1111/j.1748-1716.2011.02337.x . .