TY  - JOUR
AU  - Veličković, Nataša
AU  - Drakulić, Dunja R
AU  - Petrović, Snjezana B
AU  - Grković, Ivana S
AU  - Milosević, Maja S
AU  - Stanojlović, Milos  R
AU  - Horvat, Anica I
PY  - 2012
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/1104
AB  - Recent studies reported that exposure of juvenile rats to cranial irradiation affects hypothalamic-pituitary-adrenal (HPA) axis stability, leading to its activation along with radiation-induced inflammation. In the present study, we hypothesized whether inflammatory reaction in the CNS could be a mediator of HPA axis response to cranial irradiation (CI). Therefore, we analyzed time-course changes of serum corticosterone level, as well IL-1 beta and TNF-alpha level in the serum and hypothalamus of juvenile rats after CI. Protein and gene expression of the glucocorticoid receptor (GR) and nuclear factor kappaB (NF kappa B) were examined in the hippocampus within 24 h postirradiation interval. Cranial irradiation led to rapid induction of both GR and NF kappa B mRNA and protein in the hippocampus at 1 h. The increment in NF kappa B protein persisted for 2 h, therefore NF kappa B/GR protein ratio was turned in favor of NF kappa B. Central inflammation was characterized by increased IL-1 beta in the hypothalamus, with maximum levels at 2 and 4 h after irradiation, while both IL-1 beta and TNF-alpha were undetectable in the serum. Enhanced hypothalamic IL-1 beta probably induced the relocation of hippocampal NF kappa B to the nucleus and decreased NF kappa B mRNA at 6 h, indicating promotion of inflammation in the key tissue for HPA axis regulation. Concomitant increase of corticosterone level and enhanced GR nuclear translocation in the hippocampus at 6 h might represent a compensatory mechanism for observed inflammation. Our results indicate that acute radiation response is characterized by increased central inflammation and concomitant HPA axis activation, most likely having a role in protection of the organism from overwhelming inflammatory reaction.
T2  - Cellular and Molecular Neurobiology
T1  - Time-Course of Hypothalamic-Pituitary-Adrenal Axis Activity and Inflammation in Juvenile Rat Brain After Cranial Irradiation
IS  - 7
VL  - 32
SP  - 129
EP  - 1185
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_1104
ER  - 

@article{
author = "Veličković, Nataša and Drakulić, Dunja R and Petrović, Snjezana B and Grković, Ivana S and Milosević, Maja S and Stanojlović, Milos  R and Horvat, Anica I",
year = "2012",
abstract = "Recent studies reported that exposure of juvenile rats to cranial irradiation affects hypothalamic-pituitary-adrenal (HPA) axis stability, leading to its activation along with radiation-induced inflammation. In the present study, we hypothesized whether inflammatory reaction in the CNS could be a mediator of HPA axis response to cranial irradiation (CI). Therefore, we analyzed time-course changes of serum corticosterone level, as well IL-1 beta and TNF-alpha level in the serum and hypothalamus of juvenile rats after CI. Protein and gene expression of the glucocorticoid receptor (GR) and nuclear factor kappaB (NF kappa B) were examined in the hippocampus within 24 h postirradiation interval. Cranial irradiation led to rapid induction of both GR and NF kappa B mRNA and protein in the hippocampus at 1 h. The increment in NF kappa B protein persisted for 2 h, therefore NF kappa B/GR protein ratio was turned in favor of NF kappa B. Central inflammation was characterized by increased IL-1 beta in the hypothalamus, with maximum levels at 2 and 4 h after irradiation, while both IL-1 beta and TNF-alpha were undetectable in the serum. Enhanced hypothalamic IL-1 beta probably induced the relocation of hippocampal NF kappa B to the nucleus and decreased NF kappa B mRNA at 6 h, indicating promotion of inflammation in the key tissue for HPA axis regulation. Concomitant increase of corticosterone level and enhanced GR nuclear translocation in the hippocampus at 6 h might represent a compensatory mechanism for observed inflammation. Our results indicate that acute radiation response is characterized by increased central inflammation and concomitant HPA axis activation, most likely having a role in protection of the organism from overwhelming inflammatory reaction.",
journal = "Cellular and Molecular Neurobiology",
title = "Time-Course of Hypothalamic-Pituitary-Adrenal Axis Activity and Inflammation in Juvenile Rat Brain After Cranial Irradiation",
number = "7",
volume = "32",
pages = "129-1185",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_1104"
}

Veličković, N., Drakulić, D. R., Petrović, S. B., Grković, I. S., Milosević, M. S., Stanojlović, Milos  R,& Horvat, A. I.. (2012). Time-Course of Hypothalamic-Pituitary-Adrenal Axis Activity and Inflammation in Juvenile Rat Brain After Cranial Irradiation. in Cellular and Molecular Neurobiology, 32(7), 129-1185.
https://hdl.handle.net/21.15107/rcub_ibiss_1104

Veličković N, Drakulić DR, Petrović SB, Grković IS, Milosević MS, Stanojlović, Milos  R, Horvat AI. Time-Course of Hypothalamic-Pituitary-Adrenal Axis Activity and Inflammation in Juvenile Rat Brain After Cranial Irradiation. in Cellular and Molecular Neurobiology. 2012;32(7):129-1185.
https://hdl.handle.net/21.15107/rcub_ibiss_1104 .

Veličković, Nataša, Drakulić, Dunja R, Petrović, Snjezana B, Grković, Ivana S, Milosević, Maja S, Stanojlović, Milos  R, Horvat, Anica I, "Time-Course of Hypothalamic-Pituitary-Adrenal Axis Activity and Inflammation in Juvenile Rat Brain After Cranial Irradiation" in Cellular and Molecular Neurobiology, 32, no. 7 (2012):129-1185,
https://hdl.handle.net/21.15107/rcub_ibiss_1104 .

TY  - JOUR
AU  - Milosević, Maja S
AU  - Petrović, Snjezana B
AU  - Veličković, Nataša
AU  - Grković, Ivana S
AU  - Ignjatović, Marija
AU  - Horvat, Anica I
PY  - 2012
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/1084
AB  - Extracellular nucleotides affect female reproductive functions, fertilization, and pregnancy. The aim of this study was to investigate biochemical characteristics of ATP and ADP hydrolysis and identify E-NTPDases in myometrial cell membranes from Wistar albino rats. The apparent K (m) values were 506.4 +/- A 62.1 and 638.8 +/- A 31.3 mu M, with a calculated V (max) (app) of 3,973.0 +/- A 279.5 and 2,853.9 +/- A 79.8 nmol/min/mg for ATP and ADP, respectively. The enzyme activity described here has common properties characteristic for NTPDases: divalent cation dependence; alkaline pH optimum for both substrates, insensitivity to some of classical ATPase inhibitors (ouabain, oligomycine, theophylline, levamisole) and significant inhibition by suramine and high concentration of sodium azides (5 mM). According to similar apparent K-m values for both substrates, the ATP/ADP hydrolysis ratio, and Chevillard competition plot, NTPDase1 is dominant ATP/ADP hydrolyzing enzyme in myometrial cell membranes. RT-PCR analysis revealed expression of three members of ectonucleoside triphosphate diphosphohydrolase family (NTPDase 1, 2, and 8) in rat uterus. These findings may further elucidate the role of NTPDases and ATP in reproductive physiology.
T2  - Molecular and Cellular Biochemistry
T1  - ATP and ADP hydrolysis in cell membranes from rat myometrium
IS  - 1-2
VL  - 371
SP  - 248
EP  - 208
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_1084
ER  - 

@article{
author = "Milosević, Maja S and Petrović, Snjezana B and Veličković, Nataša and Grković, Ivana S and Ignjatović, Marija and Horvat, Anica I",
year = "2012",
abstract = "Extracellular nucleotides affect female reproductive functions, fertilization, and pregnancy. The aim of this study was to investigate biochemical characteristics of ATP and ADP hydrolysis and identify E-NTPDases in myometrial cell membranes from Wistar albino rats. The apparent K (m) values were 506.4 +/- A 62.1 and 638.8 +/- A 31.3 mu M, with a calculated V (max) (app) of 3,973.0 +/- A 279.5 and 2,853.9 +/- A 79.8 nmol/min/mg for ATP and ADP, respectively. The enzyme activity described here has common properties characteristic for NTPDases: divalent cation dependence; alkaline pH optimum for both substrates, insensitivity to some of classical ATPase inhibitors (ouabain, oligomycine, theophylline, levamisole) and significant inhibition by suramine and high concentration of sodium azides (5 mM). According to similar apparent K-m values for both substrates, the ATP/ADP hydrolysis ratio, and Chevillard competition plot, NTPDase1 is dominant ATP/ADP hydrolyzing enzyme in myometrial cell membranes. RT-PCR analysis revealed expression of three members of ectonucleoside triphosphate diphosphohydrolase family (NTPDase 1, 2, and 8) in rat uterus. These findings may further elucidate the role of NTPDases and ATP in reproductive physiology.",
journal = "Molecular and Cellular Biochemistry",
title = "ATP and ADP hydrolysis in cell membranes from rat myometrium",
number = "1-2",
volume = "371",
pages = "248-208",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_1084"
}

Milosević, M. S., Petrović, S. B., Veličković, N., Grković, I. S., Ignjatović, M.,& Horvat, A. I.. (2012). ATP and ADP hydrolysis in cell membranes from rat myometrium. in Molecular and Cellular Biochemistry, 371(1-2), 248-208.
https://hdl.handle.net/21.15107/rcub_ibiss_1084

Milosević MS, Petrović SB, Veličković N, Grković IS, Ignjatović M, Horvat AI. ATP and ADP hydrolysis in cell membranes from rat myometrium. in Molecular and Cellular Biochemistry. 2012;371(1-2):248-208.
https://hdl.handle.net/21.15107/rcub_ibiss_1084 .

Milosević, Maja S, Petrović, Snjezana B, Veličković, Nataša, Grković, Ivana S, Ignjatović, Marija, Horvat, Anica I, "ATP and ADP hydrolysis in cell membranes from rat myometrium" in Molecular and Cellular Biochemistry, 371, no. 1-2 (2012):248-208,
https://hdl.handle.net/21.15107/rcub_ibiss_1084 .

TY  - JOUR
AU  - Stanojević, Ivana
AU  - Bjelobaba, Ivana
AU  - Nedeljković, Nadežda N.
AU  - Drakulić, Dunja R.
AU  - Petrović, Snježana B.
AU  - Stojiljković, Mirjana B.
AU  - Horvat, Anica I.
PY  - 2011
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/1286
AB  - Ecto-5'-nucleotidase (CD73; EC 3.1.3.5, e-5NT) is regarded as the key enzyme in the extracellular formation of adenosine, which acts as a neuromodulator and important trophic and homeostatic factor in the brain. In the present study, we have investigated e-5NT activity, kinetic properties concerning AMP hydrolysis and the enzyme protein abundance in the purified synaptic plasma membrane (SPM) preparations isolated from whole female rat brain at different ages. We observed pronounced increase in AMP hydrolyzing activity in SPM during maturation, with greatest increment between juvenile (15-day-old) and pre-pubertal (30-day-old) rats. Immunodetection of e-5NT protein in the SPM displayed the reverse pattern of expression, with the maximum relative abundance at juvenile and minimum relative abundance in the adult stage. Negative correlation between the enzyme activity and the enzyme protein abundance in the SPM indicates that e-5NT has additional roles in the synaptic compartment during postnatal brain development, other than those related to AMP hydrolysis. Determination of kinetic parameters, K(m) and V(max), suggested that the increase in the enzyme activity with maturation was entirely due to the increase in the enzyme catalytic efficiency (V(max)/K(m)). Finally, double immunofluorescence staining against e-5NT and presynaptic membrane marker syntaxin provided first direct evidence for the existence of this ecto-enzyme in the presynaptic compartment. The results of the study suggest that e-5NT may be a part of general scheme of brain development and synapse maturation and provide rationale for the previously reported inconsistencies between enzyme immunohistochemical and biochemical studies concerning localization of e-5NT in the brain. (C) 2011 ISDN. Published by Elsevier Ltd. All rights reserved.
PB  - John Wiley and Sons
T2  - International Journal of Developmental Neuroscience
T1  - Ontogenetic profile of ecto-5 '-nucleotidase in rat brain synaptic plasma membranes
IS  - 4
VL  - 29
DO  - 10.1016/j.ijdevneu.2011.03.003
SP  - 397
EP  - 403
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_1286
ER  - 

@article{
author = "Stanojević, Ivana and Bjelobaba, Ivana and Nedeljković, Nadežda N. and Drakulić, Dunja R. and Petrović, Snježana B. and Stojiljković, Mirjana B. and Horvat, Anica I.",
year = "2011",
abstract = "Ecto-5'-nucleotidase (CD73; EC 3.1.3.5, e-5NT) is regarded as the key enzyme in the extracellular formation of adenosine, which acts as a neuromodulator and important trophic and homeostatic factor in the brain. In the present study, we have investigated e-5NT activity, kinetic properties concerning AMP hydrolysis and the enzyme protein abundance in the purified synaptic plasma membrane (SPM) preparations isolated from whole female rat brain at different ages. We observed pronounced increase in AMP hydrolyzing activity in SPM during maturation, with greatest increment between juvenile (15-day-old) and pre-pubertal (30-day-old) rats. Immunodetection of e-5NT protein in the SPM displayed the reverse pattern of expression, with the maximum relative abundance at juvenile and minimum relative abundance in the adult stage. Negative correlation between the enzyme activity and the enzyme protein abundance in the SPM indicates that e-5NT has additional roles in the synaptic compartment during postnatal brain development, other than those related to AMP hydrolysis. Determination of kinetic parameters, K(m) and V(max), suggested that the increase in the enzyme activity with maturation was entirely due to the increase in the enzyme catalytic efficiency (V(max)/K(m)). Finally, double immunofluorescence staining against e-5NT and presynaptic membrane marker syntaxin provided first direct evidence for the existence of this ecto-enzyme in the presynaptic compartment. The results of the study suggest that e-5NT may be a part of general scheme of brain development and synapse maturation and provide rationale for the previously reported inconsistencies between enzyme immunohistochemical and biochemical studies concerning localization of e-5NT in the brain. (C) 2011 ISDN. Published by Elsevier Ltd. All rights reserved.",
publisher = "John Wiley and Sons",
journal = "International Journal of Developmental Neuroscience",
title = "Ontogenetic profile of ecto-5 '-nucleotidase in rat brain synaptic plasma membranes",
number = "4",
volume = "29",
doi = "10.1016/j.ijdevneu.2011.03.003",
pages = "397-403",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_1286"
}

Stanojević, I., Bjelobaba, I., Nedeljković, N. N., Drakulić, D. R., Petrović, S. B., Stojiljković, M. B.,& Horvat, A. I.. (2011). Ontogenetic profile of ecto-5 '-nucleotidase in rat brain synaptic plasma membranes. in International Journal of Developmental Neuroscience
John Wiley and Sons., 29(4), 397-403.
https://doi.org/10.1016/j.ijdevneu.2011.03.003
https://hdl.handle.net/21.15107/rcub_ibiss_1286

Stanojević I, Bjelobaba I, Nedeljković NN, Drakulić DR, Petrović SB, Stojiljković MB, Horvat AI. Ontogenetic profile of ecto-5 '-nucleotidase in rat brain synaptic plasma membranes. in International Journal of Developmental Neuroscience. 2011;29(4):397-403.
doi:10.1016/j.ijdevneu.2011.03.003
https://hdl.handle.net/21.15107/rcub_ibiss_1286 .

Stanojević, Ivana, Bjelobaba, Ivana, Nedeljković, Nadežda N., Drakulić, Dunja R., Petrović, Snježana B., Stojiljković, Mirjana B., Horvat, Anica I., "Ontogenetic profile of ecto-5 '-nucleotidase in rat brain synaptic plasma membranes" in International Journal of Developmental Neuroscience, 29, no. 4 (2011):397-403,
https://doi.org/10.1016/j.ijdevneu.2011.03.003 .,
https://hdl.handle.net/21.15107/rcub_ibiss_1286 .

TY  - JOUR
AU  - Petrović, Snjezana B
AU  - Veličković, Nataša
AU  - Stanojević, Ivana
AU  - Milosević, Maja S
AU  - Drakulić, Dunja R
AU  - Stanojlović, Milos  R
AU  - Horvat, Anica I
PY  - 2011
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/1265
AB  - Our results, as well as those of others, have indicated that 17 beta-estradiol (E2) exerts its nongenomic effects in neuronal cells by affecting plasma membrane Ca2+ flux. In neuronal cells mitochondria possess Ca2+ buffering properties as they both sequester and release Ca2+. The goal of this study was to examine the rapid non-genomic effect of E2 on mitochondria! Ca2+ transport in hippocampal synaptosomes from ovariectomised rats. In addition, we aimed to determine if, and to what extent, E2 receptors participated in mitochondria! Ca2+ transport modulation by E2 in vitro. E2-specific binding and Ca2+ transport was monitored. At physiological E2 concentrations (0.1-1.5 nmol/L), specific E2 binding to mitochondria isolated from hippocampal synaptosomes was detected with a B-max and K-m of 37.6 +/- 2.6 fmol/mg protein and 0.69 +/- 0.14 nmol/L of free E2, respectively. The main mitochondrial Ca2+ influx mechanism is the Ruthenium Red-sensitive uniporter driven by mitochondrial membrane potential. Despite no effect of E2 on Ca2+ influx, a physiological E2 concentration (0.5 nmol/L) protected mitochondrial membrane potential and consequently Ca2+ influx from the uncoupling agent carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (1 mu mol/L). In neuronal cells the predominant mitochondria! Ca2+ efflux mechanism is the Na+/Ca2+ exchanger. E2 caused Ca2+ efflux inhibition (by 46%) coupled with increased affinity of the Na+/Ca2+ exchanger for Na+. Using E2 receptor (ER alpha and ER beta) antagonists and agonists, we confirmed ER beta's involvement in E2-induced mitochondrial membrane potential protection as well as Ca2+ efflux inhibition. In summary, our results indicate that the nongenomic neuromodulatory role of E2 in rat hippocampus is achieved by affecting mitochondria! Ca2+ transport via, in part, mitochondrial ER beta. (C) 2011 IBRO. Published by Elsevier Ltd. All rights reserved.
T2  - Neuroscience
T1  - Inhibition of Mitochondrial Na+-Dependent Ca2+ Efflux By 17 Beta-Estradiol in the Rat Hippocampus
IS  - null
VL  - 192
EP  - 204
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_1265
ER  - 

@article{
author = "Petrović, Snjezana B and Veličković, Nataša and Stanojević, Ivana and Milosević, Maja S and Drakulić, Dunja R and Stanojlović, Milos  R and Horvat, Anica I",
year = "2011",
abstract = "Our results, as well as those of others, have indicated that 17 beta-estradiol (E2) exerts its nongenomic effects in neuronal cells by affecting plasma membrane Ca2+ flux. In neuronal cells mitochondria possess Ca2+ buffering properties as they both sequester and release Ca2+. The goal of this study was to examine the rapid non-genomic effect of E2 on mitochondria! Ca2+ transport in hippocampal synaptosomes from ovariectomised rats. In addition, we aimed to determine if, and to what extent, E2 receptors participated in mitochondria! Ca2+ transport modulation by E2 in vitro. E2-specific binding and Ca2+ transport was monitored. At physiological E2 concentrations (0.1-1.5 nmol/L), specific E2 binding to mitochondria isolated from hippocampal synaptosomes was detected with a B-max and K-m of 37.6 +/- 2.6 fmol/mg protein and 0.69 +/- 0.14 nmol/L of free E2, respectively. The main mitochondrial Ca2+ influx mechanism is the Ruthenium Red-sensitive uniporter driven by mitochondrial membrane potential. Despite no effect of E2 on Ca2+ influx, a physiological E2 concentration (0.5 nmol/L) protected mitochondrial membrane potential and consequently Ca2+ influx from the uncoupling agent carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (1 mu mol/L). In neuronal cells the predominant mitochondria! Ca2+ efflux mechanism is the Na+/Ca2+ exchanger. E2 caused Ca2+ efflux inhibition (by 46%) coupled with increased affinity of the Na+/Ca2+ exchanger for Na+. Using E2 receptor (ER alpha and ER beta) antagonists and agonists, we confirmed ER beta's involvement in E2-induced mitochondrial membrane potential protection as well as Ca2+ efflux inhibition. In summary, our results indicate that the nongenomic neuromodulatory role of E2 in rat hippocampus is achieved by affecting mitochondria! Ca2+ transport via, in part, mitochondrial ER beta. (C) 2011 IBRO. Published by Elsevier Ltd. All rights reserved.",
journal = "Neuroscience",
title = "Inhibition of Mitochondrial Na+-Dependent Ca2+ Efflux By 17 Beta-Estradiol in the Rat Hippocampus",
number = "null",
volume = "192",
pages = "204",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_1265"
}

Petrović, S. B., Veličković, N., Stanojević, I., Milosević, M. S., Drakulić, D. R., Stanojlović, Milos  R,& Horvat, A. I.. (2011). Inhibition of Mitochondrial Na+-Dependent Ca2+ Efflux By 17 Beta-Estradiol in the Rat Hippocampus. in Neuroscience, 192(null).
https://hdl.handle.net/21.15107/rcub_ibiss_1265

Petrović SB, Veličković N, Stanojević I, Milosević MS, Drakulić DR, Stanojlović, Milos  R, Horvat AI. Inhibition of Mitochondrial Na+-Dependent Ca2+ Efflux By 17 Beta-Estradiol in the Rat Hippocampus. in Neuroscience. 2011;192(null):null-204.
https://hdl.handle.net/21.15107/rcub_ibiss_1265 .

Petrović, Snjezana B, Veličković, Nataša, Stanojević, Ivana, Milosević, Maja S, Drakulić, Dunja R, Stanojlović, Milos  R, Horvat, Anica I, "Inhibition of Mitochondrial Na+-Dependent Ca2+ Efflux By 17 Beta-Estradiol in the Rat Hippocampus" in Neuroscience, 192, no. null (2011),
https://hdl.handle.net/21.15107/rcub_ibiss_1265 .