TY  - JOUR
AU  - Milosavljević, Filip
AU  - Brusini, Irene
AU  - Atanasov, Andrea
AU  - Manojlović, Marina
AU  - Vučić, Marija
AU  - Oreščanin-Dušić, Zorana
AU  - Brkljačić, Jelena
AU  - Miljević, Čedo
AU  - Nikolić-Kokić, Aleksandra
AU  - Blagojević, Duško
AU  - Wang, Chunliang
AU  - Damberg, Peter
AU  - Pešić, Vesna
AU  - Tyndale, Rachel F.
AU  - Ingelman-Sundberg, Magnus
AU  - Jukić, Marin
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4516
AB  - Aims: CYP2C19 transgenic mouse expresses the human CYP2C19 gene in the liver and developing brain, and it exhibits altered neurodevelopment associated with impairments in emotionality and locomotion. Because the validation of new animal models is essential for the understanding of the aetiology and pathophysiology of movement disorders, the objective was to characterise motoric phenotype in CYP2C19 transgenic mice and to investigate its validity as a new animal model of ataxia. Methods: The rotarod, paw-print and beam-walking tests were utilised to characterise the motoric phenotype. The volumes of 20 brain regions in CYP2C19 transgenic and wild-type mice were quantified by 9.4T gadolinium-enhanced post-mortem structural neuroimaging. Antioxidative enzymatic activity was quantified biochemically. Dopaminergic alterations were characterised by chromatographic quantification of concentrations of dopamine and its metabolites and by subsequent immunohistochemical analyses. The beam-walking test was repeated after the treatment with dopamine receptor antagonists ecopipam and raclopride. Results: CYP2C19 transgenic mice exhibit abnormal, unilateral ataxia-like gait, clasping reflex and 5.6-fold more paw-slips in the beam-walking test; the motoric phenotype was more pronounced in youth. Transgenic mice exhibited a profound reduction of 12% in cerebellar volume and a moderate reduction of 4% in hippocampal volume; both regions exhibited an increased antioxidative enzyme activity. CYP2C19 mice were hyperdopaminergic; however, the motoric impairment was not ameliorated by dopamine receptor antagonists, and there was no alteration in the number of midbrain dopaminergic neurons in CYP2C19 mice. Conclusions: Humanised CYP2C19 transgenic mice exhibit altered gait and functional motoric impairments; this phenotype is likely caused by an aberrant cerebellar development.
PB  - John Wiley and Sons Inc
T2  - Neuropathology and Applied Neurobiology
T1  - The humanised CYP2C19 transgenic mouse exhibits cerebellar atrophy and movement impairment reminiscent of ataxia
VL  - 49
IS  - 1
DO  - 10.1111/nan.12867
ER  - 

@article{
author = "Milosavljević, Filip and Brusini, Irene and Atanasov, Andrea and Manojlović, Marina and Vučić, Marija and Oreščanin-Dušić, Zorana and Brkljačić, Jelena and Miljević, Čedo and Nikolić-Kokić, Aleksandra and Blagojević, Duško and Wang, Chunliang and Damberg, Peter and Pešić, Vesna and Tyndale, Rachel F. and Ingelman-Sundberg, Magnus and Jukić, Marin",
year = "2023",
abstract = "Aims: CYP2C19 transgenic mouse expresses the human CYP2C19 gene in the liver and developing brain, and it exhibits altered neurodevelopment associated with impairments in emotionality and locomotion. Because the validation of new animal models is essential for the understanding of the aetiology and pathophysiology of movement disorders, the objective was to characterise motoric phenotype in CYP2C19 transgenic mice and to investigate its validity as a new animal model of ataxia. Methods: The rotarod, paw-print and beam-walking tests were utilised to characterise the motoric phenotype. The volumes of 20 brain regions in CYP2C19 transgenic and wild-type mice were quantified by 9.4T gadolinium-enhanced post-mortem structural neuroimaging. Antioxidative enzymatic activity was quantified biochemically. Dopaminergic alterations were characterised by chromatographic quantification of concentrations of dopamine and its metabolites and by subsequent immunohistochemical analyses. The beam-walking test was repeated after the treatment with dopamine receptor antagonists ecopipam and raclopride. Results: CYP2C19 transgenic mice exhibit abnormal, unilateral ataxia-like gait, clasping reflex and 5.6-fold more paw-slips in the beam-walking test; the motoric phenotype was more pronounced in youth. Transgenic mice exhibited a profound reduction of 12% in cerebellar volume and a moderate reduction of 4% in hippocampal volume; both regions exhibited an increased antioxidative enzyme activity. CYP2C19 mice were hyperdopaminergic; however, the motoric impairment was not ameliorated by dopamine receptor antagonists, and there was no alteration in the number of midbrain dopaminergic neurons in CYP2C19 mice. Conclusions: Humanised CYP2C19 transgenic mice exhibit altered gait and functional motoric impairments; this phenotype is likely caused by an aberrant cerebellar development.",
publisher = "John Wiley and Sons Inc",
journal = "Neuropathology and Applied Neurobiology",
title = "The humanised CYP2C19 transgenic mouse exhibits cerebellar atrophy and movement impairment reminiscent of ataxia",
volume = "49",
number = "1",
doi = "10.1111/nan.12867"
}

Milosavljević, F., Brusini, I., Atanasov, A., Manojlović, M., Vučić, M., Oreščanin-Dušić, Z., Brkljačić, J., Miljević, Č., Nikolić-Kokić, A., Blagojević, D., Wang, C., Damberg, P., Pešić, V., Tyndale, R. F., Ingelman-Sundberg, M.,& Jukić, M.. (2023). The humanised CYP2C19 transgenic mouse exhibits cerebellar atrophy and movement impairment reminiscent of ataxia. in Neuropathology and Applied Neurobiology
John Wiley and Sons Inc., 49(1).
https://doi.org/10.1111/nan.12867

Milosavljević F, Brusini I, Atanasov A, Manojlović M, Vučić M, Oreščanin-Dušić Z, Brkljačić J, Miljević Č, Nikolić-Kokić A, Blagojević D, Wang C, Damberg P, Pešić V, Tyndale RF, Ingelman-Sundberg M, Jukić M. The humanised CYP2C19 transgenic mouse exhibits cerebellar atrophy and movement impairment reminiscent of ataxia. in Neuropathology and Applied Neurobiology. 2023;49(1).
doi:10.1111/nan.12867 .

Milosavljević, Filip, Brusini, Irene, Atanasov, Andrea, Manojlović, Marina, Vučić, Marija, Oreščanin-Dušić, Zorana, Brkljačić, Jelena, Miljević, Čedo, Nikolić-Kokić, Aleksandra, Blagojević, Duško, Wang, Chunliang, Damberg, Peter, Pešić, Vesna, Tyndale, Rachel F., Ingelman-Sundberg, Magnus, Jukić, Marin, "The humanised CYP2C19 transgenic mouse exhibits cerebellar atrophy and movement impairment reminiscent of ataxia" in Neuropathology and Applied Neurobiology, 49, no. 1 (2023),
https://doi.org/10.1111/nan.12867 . .

TY  - JOUR
AU  - Pridgeon, Chris
AU  - Bolhuis, Dian
AU  - Milosavljević, Filip
AU  - Manojlović, Marina
AU  - Végvári, Ákos
AU  - Gaetani, Massimiliano
AU  - Jukić, Marin
AU  - Ingelman-Sundberg, Magnus
PY  - 2022
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4111
AB  - The in vivo-relevant phenotype of 3D liver spheroids allows for long-term studies of, e.g., novel mechanisms of chronic drug-induced liver toxicity. Using this system, we present a novel drug-induced stress response in human and murine hepatocyte spheroids, wherein long slender filaments form after chronic treatment with four different drugs, of which three are PPARα antagonists. The morphology of the thorns varies between donors and the compounds used. They are mainly composed of diverse protein fibres, which are glycosylated. Their formation is inhibited by treatment with fatty acids or antioxidants. Treatment of mice with GW6471 revealed changes in gene and protein expression, such as those in the spheroids. In addition, similar changes in keratin expression were seen following the treatment of hepatotoxic drugs, including aflatoxin B1, paracetamol, chlorpromazine, cyclosporine, and ketoconazole. We suggest that thorn formation may be indicative of hepatocyte metaplasia in response to toxicity and that more focus should be placed on alterations of ECM-derived protein expression as biomarkers of liver disease and chronic drug-induced hepatotoxicity, changes that can be studied in stable in vivo-like hepatic cell systems, such as the spheroids.
PB  - MDPI
T2  - Cells
T1  - Hepatocyte Thorns, A Novel Drug-Induced Stress Response in Human and Mouse Liver Spheroids
VL  - 11
IS  - 10
DO  - 10.3390/cells11101597
ER  - 

@article{
author = "Pridgeon, Chris and Bolhuis, Dian and Milosavljević, Filip and Manojlović, Marina and Végvári, Ákos and Gaetani, Massimiliano and Jukić, Marin and Ingelman-Sundberg, Magnus",
year = "2022",
abstract = "The in vivo-relevant phenotype of 3D liver spheroids allows for long-term studies of, e.g., novel mechanisms of chronic drug-induced liver toxicity. Using this system, we present a novel drug-induced stress response in human and murine hepatocyte spheroids, wherein long slender filaments form after chronic treatment with four different drugs, of which three are PPARα antagonists. The morphology of the thorns varies between donors and the compounds used. They are mainly composed of diverse protein fibres, which are glycosylated. Their formation is inhibited by treatment with fatty acids or antioxidants. Treatment of mice with GW6471 revealed changes in gene and protein expression, such as those in the spheroids. In addition, similar changes in keratin expression were seen following the treatment of hepatotoxic drugs, including aflatoxin B1, paracetamol, chlorpromazine, cyclosporine, and ketoconazole. We suggest that thorn formation may be indicative of hepatocyte metaplasia in response to toxicity and that more focus should be placed on alterations of ECM-derived protein expression as biomarkers of liver disease and chronic drug-induced hepatotoxicity, changes that can be studied in stable in vivo-like hepatic cell systems, such as the spheroids.",
publisher = "MDPI",
journal = "Cells",
title = "Hepatocyte Thorns, A Novel Drug-Induced Stress Response in Human and Mouse Liver Spheroids",
volume = "11",
number = "10",
doi = "10.3390/cells11101597"
}

Pridgeon, C., Bolhuis, D., Milosavljević, F., Manojlović, M., Végvári, Á., Gaetani, M., Jukić, M.,& Ingelman-Sundberg, M.. (2022). Hepatocyte Thorns, A Novel Drug-Induced Stress Response in Human and Mouse Liver Spheroids. in Cells
MDPI., 11(10).
https://doi.org/10.3390/cells11101597

Pridgeon C, Bolhuis D, Milosavljević F, Manojlović M, Végvári Á, Gaetani M, Jukić M, Ingelman-Sundberg M. Hepatocyte Thorns, A Novel Drug-Induced Stress Response in Human and Mouse Liver Spheroids. in Cells. 2022;11(10).
doi:10.3390/cells11101597 .

Pridgeon, Chris, Bolhuis, Dian, Milosavljević, Filip, Manojlović, Marina, Végvári, Ákos, Gaetani, Massimiliano, Jukić, Marin, Ingelman-Sundberg, Magnus, "Hepatocyte Thorns, A Novel Drug-Induced Stress Response in Human and Mouse Liver Spheroids" in Cells, 11, no. 10 (2022),
https://doi.org/10.3390/cells11101597 . .

TY  - CONF
AU  - Manojlović, Marina
AU  - Milosavljević, Filip
AU  - Atanasov, Andrea
AU  - Batinić, Bojan
AU  - Sitarica, Pavle
AU  - Jukić, Marin
PY  - 2022
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4728
AB  - Background: Due to the COVID-19 pandemic, the prevalence of anxiety and
depression disorders increased by approximately 25-30% [1]. Reasons behind
this are likely associated with the increased average daily level of stress, especially during social isolation. Since one of the main adolescence hallmarks is
formation of meaningful social and love relationships, it represents a particularly
vulnerable period for social stress [2]. The aim of this study was to evaluate
whether isolation stress during the adolescence causes general and social anxiety
in rats.
Methods: Sprague-Dawley rats of both sexes were used for the adolescence social
isolation experiment. From the fourth postnatal week, 24 rats were single caged
and therefore subjected to the social isolation stress, while 24 non-stressed
control rats remained in groups of four per cage and were handled three times a
week. All rats were handled daily for one week and weighted before behavioural
tests. From eleventh week, rats were subjected to the open field, elevated plus
maze, and three-chamber sociability tests, with two-day gap between different
tests. In open field test, time spent in the centre was measured, while in the
elevated plus maze test, the time spent in open arms, normalized to time spent in
all arms was measured; reduction in values of these parameters was interpreted
as general anxiety. In the social preference test, social preference ratio was
calculated for each animal, according to the previously published protocol [3],
and reduction in this ratio was interpreted as social anxiety. 2-way ANOVA, with
sex and isolation stress as factors, was used for parametric data analysis, and
Fischer’s post hoc test was used to detect statistically significant between group
differences. Kruskal-Wallis test was used for non-parametric data analysis, and
Mann-Whitney post hoc test with multiple comparison correction was used to
detect statistically significant between group differences.
Results: Compared to control rats, time spent in the centre of open field (-44%
[95%CI: -76%, -12%] p¼0,008) and total distance travelled (-17% [95%CI:
-32%, -2%], p¼0.028) were decreased in isolated males, but not in females
(p>0.1). In elevated plus maze test, time spent in the open arms was significantly
decreased in male isolated rats (isolated: [median: 0.76, IQR: 0.00 – 2.06] vs
control: [median: 4.96, IQR: 2.04 – 17.75], p¼0.034); however, this change did
not remain significant after multiple comparisons corrections. In sociability tests,
both male and female isolated rats exhibited increase in social preference ratio
(+32% [95%CI: 12%, 51%], p¼0.002) and preference for novel animal over
familiar one (+48% [95%CI: 10%, 86%], p¼0.015), compared to control rats.
Body weight, measured after six weeks of social isolation at week ten, was
increased in isolated male compared to control rats (+19% [95%CI: 14%, 24%],
p<0.001), while there was no such difference observed in female rats.
Conclusion: Social isolation caused increase in general anxiety in male, but not
female rats. In addition, both male and female rats exhibited robust increase in
preference for social interaction and novel social stimulus, which is the result
opposite from the expected social anxiety as initially hypothesized.
PB  - Elsevier
C3  - Neuroscience Applied
T1  - Social adolescent stress causes increased general anxiety in male rats and reduced social anxiety in both male and female rats
VL  - 1
IS  - Supplement 2
SP  - 19
EP  - 19
DO  - 10.1016/j.nsa.2022.100149
ER  - 

@conference{
author = "Manojlović, Marina and Milosavljević, Filip and Atanasov, Andrea and Batinić, Bojan and Sitarica, Pavle and Jukić, Marin",
year = "2022",
abstract = "Background: Due to the COVID-19 pandemic, the prevalence of anxiety and
depression disorders increased by approximately 25-30% [1]. Reasons behind
this are likely associated with the increased average daily level of stress, especially during social isolation. Since one of the main adolescence hallmarks is
formation of meaningful social and love relationships, it represents a particularly
vulnerable period for social stress [2]. The aim of this study was to evaluate
whether isolation stress during the adolescence causes general and social anxiety
in rats.
Methods: Sprague-Dawley rats of both sexes were used for the adolescence social
isolation experiment. From the fourth postnatal week, 24 rats were single caged
and therefore subjected to the social isolation stress, while 24 non-stressed
control rats remained in groups of four per cage and were handled three times a
week. All rats were handled daily for one week and weighted before behavioural
tests. From eleventh week, rats were subjected to the open field, elevated plus
maze, and three-chamber sociability tests, with two-day gap between different
tests. In open field test, time spent in the centre was measured, while in the
elevated plus maze test, the time spent in open arms, normalized to time spent in
all arms was measured; reduction in values of these parameters was interpreted
as general anxiety. In the social preference test, social preference ratio was
calculated for each animal, according to the previously published protocol [3],
and reduction in this ratio was interpreted as social anxiety. 2-way ANOVA, with
sex and isolation stress as factors, was used for parametric data analysis, and
Fischer’s post hoc test was used to detect statistically significant between group
differences. Kruskal-Wallis test was used for non-parametric data analysis, and
Mann-Whitney post hoc test with multiple comparison correction was used to
detect statistically significant between group differences.
Results: Compared to control rats, time spent in the centre of open field (-44%
[95%CI: -76%, -12%] p¼0,008) and total distance travelled (-17% [95%CI:
-32%, -2%], p¼0.028) were decreased in isolated males, but not in females
(p>0.1). In elevated plus maze test, time spent in the open arms was significantly
decreased in male isolated rats (isolated: [median: 0.76, IQR: 0.00 – 2.06] vs
control: [median: 4.96, IQR: 2.04 – 17.75], p¼0.034); however, this change did
not remain significant after multiple comparisons corrections. In sociability tests,
both male and female isolated rats exhibited increase in social preference ratio
(+32% [95%CI: 12%, 51%], p¼0.002) and preference for novel animal over
familiar one (+48% [95%CI: 10%, 86%], p¼0.015), compared to control rats.
Body weight, measured after six weeks of social isolation at week ten, was
increased in isolated male compared to control rats (+19% [95%CI: 14%, 24%],
p<0.001), while there was no such difference observed in female rats.
Conclusion: Social isolation caused increase in general anxiety in male, but not
female rats. In addition, both male and female rats exhibited robust increase in
preference for social interaction and novel social stimulus, which is the result
opposite from the expected social anxiety as initially hypothesized.",
publisher = "Elsevier",
journal = "Neuroscience Applied",
title = "Social adolescent stress causes increased general anxiety in male rats and reduced social anxiety in both male and female rats",
volume = "1",
number = "Supplement 2",
pages = "19-19",
doi = "10.1016/j.nsa.2022.100149"
}

Manojlović, M., Milosavljević, F., Atanasov, A., Batinić, B., Sitarica, P.,& Jukić, M.. (2022). Social adolescent stress causes increased general anxiety in male rats and reduced social anxiety in both male and female rats. in Neuroscience Applied
Elsevier., 1(Supplement 2), 19-19.
https://doi.org/10.1016/j.nsa.2022.100149

Manojlović M, Milosavljević F, Atanasov A, Batinić B, Sitarica P, Jukić M. Social adolescent stress causes increased general anxiety in male rats and reduced social anxiety in both male and female rats. in Neuroscience Applied. 2022;1(Supplement 2):19-19.
doi:10.1016/j.nsa.2022.100149 .

Manojlović, Marina, Milosavljević, Filip, Atanasov, Andrea, Batinić, Bojan, Sitarica, Pavle, Jukić, Marin, "Social adolescent stress causes increased general anxiety in male rats and reduced social anxiety in both male and female rats" in Neuroscience Applied, 1, no. Supplement 2 (2022):19-19,
https://doi.org/10.1016/j.nsa.2022.100149 . .

TY  - CONF
AU  - Milosavljević, Filip
AU  - Brusini, Irene
AU  - Atanasov, Andrea
AU  - Manojlović, Marina
AU  - Vučić, Marija
AU  - Oreščanin-Dušić, Zorana
AU  - Brkljačić, Jelena
AU  - Sitarica, Pavle
AU  - Miljević, Čedo
AU  - Nikolić-Kokić, Aleksandra
AU  - Blagojević, Duško
AU  - Wang, Chunliang
AU  - Damberg, Peter
AU  - Pešić, Vesna
AU  - Tyndale, Rachel
AU  - Ingelman-Sundberg, Magnus
AU  - Jukić, Marin
PY  - 2022
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4729
AB  - Background: Animal models are essential for understanding aetiology and pathophysiology of movement disorders. Previously, it had been found that mice transgenic for the human CYP2C19 gene, which is expressed in the liver and developing brain, exhibit altered neurodevelopment associated with impairments of their motor function and emotionality [1, 2]. The aim of this study was to characterize motoric phenotype of the CYP2C19 transgenic mouse and validate its potential usefulness as an animal model for ataxia.

Methods: Experiments were performed on CYP2C19 transgenic mice and control wild-type littermate mice. Body weight of mice was measured between the 21st and 42nd postnatal day. Mouse gait was analysed with footprint test [3] in young animals at four time points and once in adult mice. The maximal height of hindpaw elevation while walking was measured offline from the video footage of the footprint test. Motoric function was quantified by the rotarod and beam-walking tests. Structural differences in 20 brain regions of wild-type and transgenic mice were investigated with 9.4T gadolinium-enhanced postmortem neuroimaging. Antioxidative enzyme status was determined in the brain tissue in order to assess potential differences in the brain oxidative-antioxidative balance between wild-type and transgenic mice. When multiple brain regions or multiple antioxidant enzyme activities were analysed, p-values were FDR corrected for multiple comparisons.

Results: CYP2C19 transgenic (TG) animals exhibited approximately 5-10% reduced body weight (p=0.015) during 3rd and 4th postnatal week, while after postnatal day 31, the differences in the body weight were no longer statistically significant. The TG animals exhibited approximately two fold higher maximal hindpaw elevation in young (2.1-fold [CI95%: 2.0, 2.2], p<0.0001) and adult mice (1.9-fold [CI95%: 1.8, 2.1], p<0.0001), compared to wild-types. In the 5th postnatal week, all transgenic mice exhibited increase in elevation of both hindpaws, while after this point they gradually started exhibiting unilateral phenotype until almost all (49 of 51) animals became unilaterally affected in the adulthood. Footprint analysis and rotarod test did not detect significant differences (p>0.1) between TG and control mice in any of the analysed parameters, accounting for all examined time points. CYP2C19 transgenic mice exhibited 14% increase in beam crossing time (14%, [95%CI: 6.4, 22], p=0.0014) and 5.6-fold more paw-slips (p<0.0001, n=89) in the beam-walking test. CYP2C19 transgenic mice exhibited profound reduction in cerebellar volume (-11.8% [95%CI: -14.7, -9.0], q<0.0001, n=59) and moderate reduction in hippocampal volume (-4.2% [95%CI: -6.4%, -1.9%], q=0.015, n=59); compared to the corresponding volumes measured in WT mice. Superoxide dismutase activity was slightly increased (1.14-fold [CI95%: 1.06, 1.23], p=0.0010, q=0.023) in the cerebelli and moderately increased (1.3-fold, [CI95%: 1.18, 1.47], p<0.0001, q=0.0013) in the hippocampi of transgenic mice compared to wild-types, while glutathione reductase activity was significantly increased (1.2-fold [CI95%: 1.13, 1.35], p<0.0001, q=0.0021) in the hippocampi of TG mice.

Conclusions: Humanized CYP2C19 transgenic mice exhibit altered motoric function, functional motoric impairments and reduced cerebellar volume. CYP2C19 transgenic mice can be a useful tool for the studies focused on understanding the physiology of cerebellar development, as well as aetiology and pathophysiology of cerebellum-related disorders.
PB  - Elsevier
C3  - Neuroscience Applied
T1  - Humanized CYP2C19 transgenic mouse as an animal model of cerebellar ataxia
VL  - 1
IS  - Supplement 2
SP  - 64
EP  - 64
DO  - 10.1016/j.nsa.2022.100236
ER  - 

@conference{
author = "Milosavljević, Filip and Brusini, Irene and Atanasov, Andrea and Manojlović, Marina and Vučić, Marija and Oreščanin-Dušić, Zorana and Brkljačić, Jelena and Sitarica, Pavle and Miljević, Čedo and Nikolić-Kokić, Aleksandra and Blagojević, Duško and Wang, Chunliang and Damberg, Peter and Pešić, Vesna and Tyndale, Rachel and Ingelman-Sundberg, Magnus and Jukić, Marin",
year = "2022",
abstract = "Background: Animal models are essential for understanding aetiology and pathophysiology of movement disorders. Previously, it had been found that mice transgenic for the human CYP2C19 gene, which is expressed in the liver and developing brain, exhibit altered neurodevelopment associated with impairments of their motor function and emotionality [1, 2]. The aim of this study was to characterize motoric phenotype of the CYP2C19 transgenic mouse and validate its potential usefulness as an animal model for ataxia.

Methods: Experiments were performed on CYP2C19 transgenic mice and control wild-type littermate mice. Body weight of mice was measured between the 21st and 42nd postnatal day. Mouse gait was analysed with footprint test [3] in young animals at four time points and once in adult mice. The maximal height of hindpaw elevation while walking was measured offline from the video footage of the footprint test. Motoric function was quantified by the rotarod and beam-walking tests. Structural differences in 20 brain regions of wild-type and transgenic mice were investigated with 9.4T gadolinium-enhanced postmortem neuroimaging. Antioxidative enzyme status was determined in the brain tissue in order to assess potential differences in the brain oxidative-antioxidative balance between wild-type and transgenic mice. When multiple brain regions or multiple antioxidant enzyme activities were analysed, p-values were FDR corrected for multiple comparisons.

Results: CYP2C19 transgenic (TG) animals exhibited approximately 5-10% reduced body weight (p=0.015) during 3rd and 4th postnatal week, while after postnatal day 31, the differences in the body weight were no longer statistically significant. The TG animals exhibited approximately two fold higher maximal hindpaw elevation in young (2.1-fold [CI95%: 2.0, 2.2], p<0.0001) and adult mice (1.9-fold [CI95%: 1.8, 2.1], p<0.0001), compared to wild-types. In the 5th postnatal week, all transgenic mice exhibited increase in elevation of both hindpaws, while after this point they gradually started exhibiting unilateral phenotype until almost all (49 of 51) animals became unilaterally affected in the adulthood. Footprint analysis and rotarod test did not detect significant differences (p>0.1) between TG and control mice in any of the analysed parameters, accounting for all examined time points. CYP2C19 transgenic mice exhibited 14% increase in beam crossing time (14%, [95%CI: 6.4, 22], p=0.0014) and 5.6-fold more paw-slips (p<0.0001, n=89) in the beam-walking test. CYP2C19 transgenic mice exhibited profound reduction in cerebellar volume (-11.8% [95%CI: -14.7, -9.0], q<0.0001, n=59) and moderate reduction in hippocampal volume (-4.2% [95%CI: -6.4%, -1.9%], q=0.015, n=59); compared to the corresponding volumes measured in WT mice. Superoxide dismutase activity was slightly increased (1.14-fold [CI95%: 1.06, 1.23], p=0.0010, q=0.023) in the cerebelli and moderately increased (1.3-fold, [CI95%: 1.18, 1.47], p<0.0001, q=0.0013) in the hippocampi of transgenic mice compared to wild-types, while glutathione reductase activity was significantly increased (1.2-fold [CI95%: 1.13, 1.35], p<0.0001, q=0.0021) in the hippocampi of TG mice.

Conclusions: Humanized CYP2C19 transgenic mice exhibit altered motoric function, functional motoric impairments and reduced cerebellar volume. CYP2C19 transgenic mice can be a useful tool for the studies focused on understanding the physiology of cerebellar development, as well as aetiology and pathophysiology of cerebellum-related disorders.",
publisher = "Elsevier",
journal = "Neuroscience Applied",
title = "Humanized CYP2C19 transgenic mouse as an animal model of cerebellar ataxia",
volume = "1",
number = "Supplement 2",
pages = "64-64",
doi = "10.1016/j.nsa.2022.100236"
}

Milosavljević, F., Brusini, I., Atanasov, A., Manojlović, M., Vučić, M., Oreščanin-Dušić, Z., Brkljačić, J., Sitarica, P., Miljević, Č., Nikolić-Kokić, A., Blagojević, D., Wang, C., Damberg, P., Pešić, V., Tyndale, R., Ingelman-Sundberg, M.,& Jukić, M.. (2022). Humanized CYP2C19 transgenic mouse as an animal model of cerebellar ataxia. in Neuroscience Applied
Elsevier., 1(Supplement 2), 64-64.
https://doi.org/10.1016/j.nsa.2022.100236

Milosavljević F, Brusini I, Atanasov A, Manojlović M, Vučić M, Oreščanin-Dušić Z, Brkljačić J, Sitarica P, Miljević Č, Nikolić-Kokić A, Blagojević D, Wang C, Damberg P, Pešić V, Tyndale R, Ingelman-Sundberg M, Jukić M. Humanized CYP2C19 transgenic mouse as an animal model of cerebellar ataxia. in Neuroscience Applied. 2022;1(Supplement 2):64-64.
doi:10.1016/j.nsa.2022.100236 .

Milosavljević, Filip, Brusini, Irene, Atanasov, Andrea, Manojlović, Marina, Vučić, Marija, Oreščanin-Dušić, Zorana, Brkljačić, Jelena, Sitarica, Pavle, Miljević, Čedo, Nikolić-Kokić, Aleksandra, Blagojević, Duško, Wang, Chunliang, Damberg, Peter, Pešić, Vesna, Tyndale, Rachel, Ingelman-Sundberg, Magnus, Jukić, Marin, "Humanized CYP2C19 transgenic mouse as an animal model of cerebellar ataxia" in Neuroscience Applied, 1, no. Supplement 2 (2022):64-64,
https://doi.org/10.1016/j.nsa.2022.100236 . .

TY  - CONF
AU  - Manojlović, Marina
AU  - Miloševski, Teodora
PY  - 2021
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/3831
AB  - Увод: Трансгени мишеви, носиоци хуманог гена за CYP2C19, показују понашање слично депресији, повећану осетљивост на стрес и карактеристичан моторни фенотип. Магнетном резонанцом високе резолуције могућа је тродимензионална визуализација унутрашњих структура мозга и мерење запремина регија од интереса које су повезане са оваквим фенотипом.
Циљ рада: Циљ овог рада био је квантификација промена у запремини структура мозга код CYP2C19 трансгених мишева у односу на немутиране контроле, узимајући у обзир и утицај пола.
Материјал и методе: Kоришћено је 29 CYP2C19 трансгених и 30 контролних мишева. Како би се појачао сигнал на магнетној резонанци, животиње су пре жртвовања транскардијално перфундоване контрастним средством које садржи гадолинијум. Узорци су анализирани помоћу анималног MRI Siemens скенера јачине 9,4 Тesla и мерене су запремине 21 регије од интереса.
Резултати: Приликом поређења на основу генотипа, статистички значајне промене у запремини запажене су код амигдала, малог мозга, коре великог мозга, корпус стријатума, глобус палидуса, хипокампусне формације, хипоталамуса, нуклеус акумбенса, олфакторног булбуса, септалног једра и супериорног коликулуса, док је пол показао утицај само код хипотaламуса и олфакторног булбуса.
Закључак: Повећање запремине амигдала и смањење запремине хипокампуса код ових животиња може бити објашњење за понашање слично депресији и повећану анксиозност, док је смањење запремине малог мозга највероватније узрок карактеристичног моторног фенотипа сличног атаксији.
AB  - Introduction: Transgenic mice, carriers of the human CYP2C19 gene, exhibit depression-like behavior, increased sensitivity to stress and altered motoric function. Using high-resolution magnetic resonance imaging, it is possible to visualize the internal structures of the brain in 3D and to measure the volumes of regions associated with the observed phenotype.
The Aim: The aim of this study was to quantify the change in the brain structure volume in CYP2C19 transgenic mice, considering also the influence of sex.
Material and Methods: Twenty nine CYP2C19Tg and 30 control mice were used. Prior to sacrificing, mice were transcardially perfused with fixating solution which contains gadolinium-based MRI contrast agent in order to enhance the contrast. Volumetric changes were assessed from the animal 9.4 Tesla Siemens scanner readouts.
Results: Statistically significant changes in the volumes were found in: amygdala, cerebellum, cerebral cortex, corpus striatum, globus pallidus, hippocampal formation, hypothalamus, nucleus accumbens, olfactory bulb, septal nucleus and superior colliculus, whereas sex was factor only in hypothalamus and olfactory bulb.
Conclusion: Increased amygdala and decreased hippocampal volume in transgenic mice may explain the depression-like behavior and increased susceptibility to anxiety, while decreased cerebellum volume may explain altered motoric function.
T1  - Волуметријске промене структура мозга код CYP2C19 трансгеног миша – post-mortem структурна магнетна резонанца на 9,4 Tesla скенеру
T1  - Volumetric brain structure alterations in CYP2C19 transgenic mouse – post-mortem structural magnetic resonance on 9.4 Tesla scanner
UR  - https://hdl.handle.net/21.15107/rcub_farfar_3831
ER  - 

@conference{
author = "Manojlović, Marina and Miloševski, Teodora",
year = "2021",
abstract = "Увод: Трансгени мишеви, носиоци хуманог гена за CYP2C19, показују понашање слично депресији, повећану осетљивост на стрес и карактеристичан моторни фенотип. Магнетном резонанцом високе резолуције могућа је тродимензионална визуализација унутрашњих структура мозга и мерење запремина регија од интереса које су повезане са оваквим фенотипом.
Циљ рада: Циљ овог рада био је квантификација промена у запремини структура мозга код CYP2C19 трансгених мишева у односу на немутиране контроле, узимајући у обзир и утицај пола.
Материјал и методе: Kоришћено је 29 CYP2C19 трансгених и 30 контролних мишева. Како би се појачао сигнал на магнетној резонанци, животиње су пре жртвовања транскардијално перфундоване контрастним средством које садржи гадолинијум. Узорци су анализирани помоћу анималног MRI Siemens скенера јачине 9,4 Тesla и мерене су запремине 21 регије од интереса.
Резултати: Приликом поређења на основу генотипа, статистички значајне промене у запремини запажене су код амигдала, малог мозга, коре великог мозга, корпус стријатума, глобус палидуса, хипокампусне формације, хипоталамуса, нуклеус акумбенса, олфакторног булбуса, септалног једра и супериорног коликулуса, док је пол показао утицај само код хипотaламуса и олфакторног булбуса.
Закључак: Повећање запремине амигдала и смањење запремине хипокампуса код ових животиња може бити објашњење за понашање слично депресији и повећану анксиозност, док је смањење запремине малог мозга највероватније узрок карактеристичног моторног фенотипа сличног атаксији., Introduction: Transgenic mice, carriers of the human CYP2C19 gene, exhibit depression-like behavior, increased sensitivity to stress and altered motoric function. Using high-resolution magnetic resonance imaging, it is possible to visualize the internal structures of the brain in 3D and to measure the volumes of regions associated with the observed phenotype.
The Aim: The aim of this study was to quantify the change in the brain structure volume in CYP2C19 transgenic mice, considering also the influence of sex.
Material and Methods: Twenty nine CYP2C19Tg and 30 control mice were used. Prior to sacrificing, mice were transcardially perfused with fixating solution which contains gadolinium-based MRI contrast agent in order to enhance the contrast. Volumetric changes were assessed from the animal 9.4 Tesla Siemens scanner readouts.
Results: Statistically significant changes in the volumes were found in: amygdala, cerebellum, cerebral cortex, corpus striatum, globus pallidus, hippocampal formation, hypothalamus, nucleus accumbens, olfactory bulb, septal nucleus and superior colliculus, whereas sex was factor only in hypothalamus and olfactory bulb.
Conclusion: Increased amygdala and decreased hippocampal volume in transgenic mice may explain the depression-like behavior and increased susceptibility to anxiety, while decreased cerebellum volume may explain altered motoric function.",
title = "Волуметријске промене структура мозга код CYP2C19 трансгеног миша – post-mortem структурна магнетна резонанца на 9,4 Tesla скенеру, Volumetric brain structure alterations in CYP2C19 transgenic mouse – post-mortem structural magnetic resonance on 9.4 Tesla scanner",
url = "https://hdl.handle.net/21.15107/rcub_farfar_3831"
}

Manojlović, M.,& Miloševski, T.. (2021). Волуметријске промене структура мозга код CYP2C19 трансгеног миша – post-mortem структурна магнетна резонанца на 9,4 Tesla скенеру. .
https://hdl.handle.net/21.15107/rcub_farfar_3831

Manojlović M, Miloševski T. Волуметријске промене структура мозга код CYP2C19 трансгеног миша – post-mortem структурна магнетна резонанца на 9,4 Tesla скенеру. 2021;.
https://hdl.handle.net/21.15107/rcub_farfar_3831 .

Manojlović, Marina, Miloševski, Teodora, "Волуметријске промене структура мозга код CYP2C19 трансгеног миша – post-mortem структурна магнетна резонанца на 9,4 Tesla скенеру" (2021),
https://hdl.handle.net/21.15107/rcub_farfar_3831 .

TY  - CONF
AU  - Milosavljević, Filip
AU  - Vučić, Marija
AU  - Manojlović, Marina
AU  - Miloševski, Teodora
AU  - Batinić, Bojan
AU  - Novalen, Maria
AU  - Miksys, Sharon
AU  - Tyndale, Rachel
AU  - Ingelman-Sundberg, Magnus
AU  - Pešić, Vesna
AU  - Jukić, Marin
PY  - 2020
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4753
AB  - Introduction: CYP2C19 transgenic mouse (2C19TG) is generated by the insertion of 12 copies of the human CYP2C19 gene into mouse genome. This animal model is a tool to study the neurodevelopmental role of CYP2C19 in vivo, since this enzyme is expressed in the foetal brains of 2C19TG mice and humans. Previous studies [1,2] showed anxiety and depression-like behaviour in these mice, while the aim of this study was to characterize the motoric function of the 2C19TG mouse.
Methods: Whole brain dopamine concentration was measured in the brain homogenate of 23 adult mice by the HPLC-MS method. Motoric function in 50 mutant and 43 control mice of both genders was tested by the rotarod and beam walking tests. Beam walking test was repeated after treatment with dopaminergic receptor antagonists, Ecopipam (0.1 mg/kg) and Raclopride (0.25 mg/kg) as a follow-up. The sections of 10 6-month old and 8 15-months old mice were stained with anti-tyrosine hydroxylase antibody and the number of dopaminergic neurons was counted on histological slides under microscope. Next, after transcardial perfusion of 30 2C19TG and 30 control mice of both genders with contrast agent (4% Paraformaldehyde, 0.05M Gadoteridole, 0.01M Phosphate buffered saline, pH=7.4), cranium containing the whole brain was scanned overnight by the 9.4T MRI scanner. The volumes of 39 brain regions were quantified according to the mouse brain atlas [3]. Student's t-test and two-way ANOVA were used to evaluate statistical significance of between-group differences.
Results: Adult 2C19TG mice are hyperdopaminergic, as they exhibit 15% increased dopamine concentration (p<0.001). They also show hyperkinetic motoric phenotype with the ataxia-like walking pattern and pathological clasping reflex. In the beam walking test 2C19TG mice had 20% longer beam crossing time (p=0.007) and 60% more paw slips (p<0.001) then the controls, and this motoric impairment could not be improved with antidopaminergic drugs. Both younger and older 2C19TG mice exhibited only a marginal reduction in the number of dopaminergic neurons of both substantia nigra and ventral tegmental area in a subset of coronal sections. This was confirmed by the gadolinium-enhanced neuroimaging that showed no change in substantia nigra volume in 2C19TG mice. On the other hand, significant differences in volume were identified in 11 regions, including cerebellum (-8.3% p<0.001) and striatum (+3.0%, p<0.001), which are the regions connected with the motoric function. The volumetric changes were detected in the hippocampus (-1.3%, p=0.027), amygdala (+2.8%, p<0.001), septum (+3.3%, p=0.014) and nucleus accumbens (+3.5, p=0.004) of 2C19TG mice. These brain regions are involved in emotional and motivational functions.
Conclusion: Ataxia-like motoric phenotype in 2C19TG transgenic mice is probably caused by changes in cerebellum, while hyperdopaminergism is most likely the compensatory adaptation, whereas the changes in the hippocampus, amygdala, septum, and nucleus accumbens may be connected with the mutants’ depression-like phenotype and susceptibility to stress. Therefore, CYP2C19 transgenic mouse in potentially useful model of hyperkinetic disorders, and our findings hint at the possible impact of CYP2C19 enzyme on the development of the several brain regions involved in motor and emotional functioning.
PB  - Elsevier
C3  - European Neuropsychopharmacology
T1  - Reduced cerebellum volume and ataxia-like motoric phenotype in transgenic mouse, carrier of human CYP2C19 gene
VL  - 40
IS  - 1
SP  - S207
EP  - S208
DO  - 10.1016/j.euroneuro.2020.09.271
ER  - 

@conference{
author = "Milosavljević, Filip and Vučić, Marija and Manojlović, Marina and Miloševski, Teodora and Batinić, Bojan and Novalen, Maria and Miksys, Sharon and Tyndale, Rachel and Ingelman-Sundberg, Magnus and Pešić, Vesna and Jukić, Marin",
year = "2020",
abstract = "Introduction: CYP2C19 transgenic mouse (2C19TG) is generated by the insertion of 12 copies of the human CYP2C19 gene into mouse genome. This animal model is a tool to study the neurodevelopmental role of CYP2C19 in vivo, since this enzyme is expressed in the foetal brains of 2C19TG mice and humans. Previous studies [1,2] showed anxiety and depression-like behaviour in these mice, while the aim of this study was to characterize the motoric function of the 2C19TG mouse.
Methods: Whole brain dopamine concentration was measured in the brain homogenate of 23 adult mice by the HPLC-MS method. Motoric function in 50 mutant and 43 control mice of both genders was tested by the rotarod and beam walking tests. Beam walking test was repeated after treatment with dopaminergic receptor antagonists, Ecopipam (0.1 mg/kg) and Raclopride (0.25 mg/kg) as a follow-up. The sections of 10 6-month old and 8 15-months old mice were stained with anti-tyrosine hydroxylase antibody and the number of dopaminergic neurons was counted on histological slides under microscope. Next, after transcardial perfusion of 30 2C19TG and 30 control mice of both genders with contrast agent (4% Paraformaldehyde, 0.05M Gadoteridole, 0.01M Phosphate buffered saline, pH=7.4), cranium containing the whole brain was scanned overnight by the 9.4T MRI scanner. The volumes of 39 brain regions were quantified according to the mouse brain atlas [3]. Student's t-test and two-way ANOVA were used to evaluate statistical significance of between-group differences.
Results: Adult 2C19TG mice are hyperdopaminergic, as they exhibit 15% increased dopamine concentration (p<0.001). They also show hyperkinetic motoric phenotype with the ataxia-like walking pattern and pathological clasping reflex. In the beam walking test 2C19TG mice had 20% longer beam crossing time (p=0.007) and 60% more paw slips (p<0.001) then the controls, and this motoric impairment could not be improved with antidopaminergic drugs. Both younger and older 2C19TG mice exhibited only a marginal reduction in the number of dopaminergic neurons of both substantia nigra and ventral tegmental area in a subset of coronal sections. This was confirmed by the gadolinium-enhanced neuroimaging that showed no change in substantia nigra volume in 2C19TG mice. On the other hand, significant differences in volume were identified in 11 regions, including cerebellum (-8.3% p<0.001) and striatum (+3.0%, p<0.001), which are the regions connected with the motoric function. The volumetric changes were detected in the hippocampus (-1.3%, p=0.027), amygdala (+2.8%, p<0.001), septum (+3.3%, p=0.014) and nucleus accumbens (+3.5, p=0.004) of 2C19TG mice. These brain regions are involved in emotional and motivational functions.
Conclusion: Ataxia-like motoric phenotype in 2C19TG transgenic mice is probably caused by changes in cerebellum, while hyperdopaminergism is most likely the compensatory adaptation, whereas the changes in the hippocampus, amygdala, septum, and nucleus accumbens may be connected with the mutants’ depression-like phenotype and susceptibility to stress. Therefore, CYP2C19 transgenic mouse in potentially useful model of hyperkinetic disorders, and our findings hint at the possible impact of CYP2C19 enzyme on the development of the several brain regions involved in motor and emotional functioning.",
publisher = "Elsevier",
journal = "European Neuropsychopharmacology",
title = "Reduced cerebellum volume and ataxia-like motoric phenotype in transgenic mouse, carrier of human CYP2C19 gene",
volume = "40",
number = "1",
pages = "S207-S208",
doi = "10.1016/j.euroneuro.2020.09.271"
}

Milosavljević, F., Vučić, M., Manojlović, M., Miloševski, T., Batinić, B., Novalen, M., Miksys, S., Tyndale, R., Ingelman-Sundberg, M., Pešić, V.,& Jukić, M.. (2020). Reduced cerebellum volume and ataxia-like motoric phenotype in transgenic mouse, carrier of human CYP2C19 gene. in European Neuropsychopharmacology
Elsevier., 40(1), S207-S208.
https://doi.org/10.1016/j.euroneuro.2020.09.271

Milosavljević F, Vučić M, Manojlović M, Miloševski T, Batinić B, Novalen M, Miksys S, Tyndale R, Ingelman-Sundberg M, Pešić V, Jukić M. Reduced cerebellum volume and ataxia-like motoric phenotype in transgenic mouse, carrier of human CYP2C19 gene. in European Neuropsychopharmacology. 2020;40(1):S207-S208.
doi:10.1016/j.euroneuro.2020.09.271 .

Milosavljević, Filip, Vučić, Marija, Manojlović, Marina, Miloševski, Teodora, Batinić, Bojan, Novalen, Maria, Miksys, Sharon, Tyndale, Rachel, Ingelman-Sundberg, Magnus, Pešić, Vesna, Jukić, Marin, "Reduced cerebellum volume and ataxia-like motoric phenotype in transgenic mouse, carrier of human CYP2C19 gene" in European Neuropsychopharmacology, 40, no. 1 (2020):S207-S208,
https://doi.org/10.1016/j.euroneuro.2020.09.271 . .

TY  - CONF
AU  - Milosavljević, Filip
AU  - Vučić, M.
AU  - Manojlović, Marina
AU  - Ašujić, N.
AU  - Batinić, Bojan
AU  - Novalen, M.
AU  - Miksys, S.
AU  - Tyndale, R.F.
AU  - Ingelman-Sundberg, Magnus
AU  - Pešić, Vesna
AU  - Jukić, Marin
PY  - 2019
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/3297
PB  - Elsevier Science BV, Amsterdam
C3  - European Neuropsychopharmacology
T1  - Transgenic mouse, carrier of human CYP2C19 gene, as an animal model for hyperdopaminergism-induced hyperkinesia
VL  - 29
IS  - Supplement 2
SP  - S672
EP  - S673
DO  - 10.1016/j.euroneuro.2019.01.056
ER  - 

@conference{
author = "Milosavljević, Filip and Vučić, M. and Manojlović, Marina and Ašujić, N. and Batinić, Bojan and Novalen, M. and Miksys, S. and Tyndale, R.F. and Ingelman-Sundberg, Magnus and Pešić, Vesna and Jukić, Marin",
year = "2019",
publisher = "Elsevier Science BV, Amsterdam",
journal = "European Neuropsychopharmacology",
title = "Transgenic mouse, carrier of human CYP2C19 gene, as an animal model for hyperdopaminergism-induced hyperkinesia",
volume = "29",
number = "Supplement 2",
pages = "S672-S673",
doi = "10.1016/j.euroneuro.2019.01.056"
}

Milosavljević, F., Vučić, M., Manojlović, M., Ašujić, N., Batinić, B., Novalen, M., Miksys, S., Tyndale, R.F., Ingelman-Sundberg, M., Pešić, V.,& Jukić, M.. (2019). Transgenic mouse, carrier of human CYP2C19 gene, as an animal model for hyperdopaminergism-induced hyperkinesia. in European Neuropsychopharmacology
Elsevier Science BV, Amsterdam., 29(Supplement 2), S672-S673.
https://doi.org/10.1016/j.euroneuro.2019.01.056

Milosavljević F, Vučić M, Manojlović M, Ašujić N, Batinić B, Novalen M, Miksys S, Tyndale R, Ingelman-Sundberg M, Pešić V, Jukić M. Transgenic mouse, carrier of human CYP2C19 gene, as an animal model for hyperdopaminergism-induced hyperkinesia. in European Neuropsychopharmacology. 2019;29(Supplement 2):S672-S673.
doi:10.1016/j.euroneuro.2019.01.056 .

Milosavljević, Filip, Vučić, M., Manojlović, Marina, Ašujić, N., Batinić, Bojan, Novalen, M., Miksys, S., Tyndale, R.F., Ingelman-Sundberg, Magnus, Pešić, Vesna, Jukić, Marin, "Transgenic mouse, carrier of human CYP2C19 gene, as an animal model for hyperdopaminergism-induced hyperkinesia" in European Neuropsychopharmacology, 29, no. Supplement 2 (2019):S672-S673,
https://doi.org/10.1016/j.euroneuro.2019.01.056 . .