Influence of valproic acid application during breeding and gestation in the mouse animal model - effect on psychomotor development in offspring

Osnovni cilj ove doktorske disertacije je bio da se ispitaju posledice intrauterinog izlaganja različitim dozama valproata na tok fizičkog i psihomotornog razvoja jedinki miša. Ispitivani su efekti valproata na CNS-u u subterapijskim dozama i dozno- zavisni efekat. Testovi primenjeni na potomstvu su u literaturi okarakterisani kao odgovarajući za procenu stanja motornog i senzornog sistema u određenoj fazi postnatalnog razvoja. Određeni testovi, pored opšte motorne sposobnosti, omogućavali su i procenu ponašanja tipa anksioznosti / depresije, hiper- i hipoaktivnosti jedinki. Metode. U eksperimentima su korišćene ženke i mužjaci zdravih laboratorijskih miševa soja NMRI, odabrane metodom slučajnog izbora iz okota na Vojnomedicinskoj akademiji u Beogradu. Eksperiment je počinjao sparivanjem dve ženke i jednog mužjaka u jednom kavezu. Shodno tretmanu, oformljeno je 5 eksperimentalnih grupa i to jedna kontrolna (n=9 ženki), koja je bila tretirana fiziološkim rastvorom (Natrii Chloridi Infundibule 0,9%, HemofarmhospitalLogica), i 4 grupe koje su tretirane valproatom (Valproic acid sodium salt – 2 propylpentanoic sodium, ≥98, P4543 Sigma Aldrich) u dozi od 50 mg/kg, 100 mg/kg, 200 mg/kg ili 400 mg/kg (n=10-11 ženki po grupi). Natrijumova so valproinske kiseline rastvarana je u fiziološkom rastvoru. Injeciranje je obavljano svakodnevno, od momenta sparivanja, tokom čitavog perioda priploda i gestacije. Nakon rađanja potomstva tretman je prestajao i ženke su sa svojim potomstvom odvajane u zasebne kaveze. Pristup hrani i vodi je bio neograničen. Procenjivan je odgovor potomstva u sledećim testovima: (1) test refleksa uspravljanja , (2)test kačenja o rep, (3) test izlaganja vrućoj ploči, (4) test uzdignutog krstastog lavirinta, (5) test otvorenog polja Rezultati. Kontinuirana primena različitih doza valproata (50, 100, 200 i 400 mg/kg) na ženkama miša NMRI soja u periodu priploda ima negativan dozno-zavisni uticaj na fertilitet, a kontinuirana primena u periodu priploda i gestacije na smrtnost ženki (akcenat na visokim dozama). Tretman nema značajnog uticaja na telesnu težinu ženki. Intrauterino izlaganje valproatu ima negativan uticaj na telesnu težinu jedinki tokom postnatalnog razvoja, uključujući period adolescencije. Tretman je povezan i sa kašnjenjem u otvaranju očiju jedinki u periodu do 15. dana postanatalnog razvoja. Intrauterino izlaganje različitim dozama valproata značajno utiče na ponašanje jedinki u testu uspravljanja. Shodno starosti jedinki efekat tretmana se ogleda kako u kvalitetu (tipu) odgovora i u vremenu koje je potrebno jedinki da se uspravi. Tretman valproatom tokom prenatalnog perioda utiče na ishod testa kačenja o rep kada se jedinke testiraju u periodu od 5. do 10. postnatalnog dana (sa akcentom na doze 200 mg/kg i 400 mg/kg). Kod jedinki starih 15 postnatalnih dana posledice tretmana nisu uočene. Intrauterino izlaganje valproatu značajno utiče na ishod u testu izlaganja vrućoj ploči jedinki starih 25 i 32 postnatalna dana. Efekat se ogleda u povećanju broja jedinki koje neadekvatno odgovaraju na test, kao i u produženoj latenci odgovora na stimulus kod jedinki koje adekvatno odgovore na test. Izlaganje majki tokom trudnoće valproatu značajno utiče na ponašanje adolescentnog potomstva (PND35) u testu uzdignutog krstastog lavirinta, a posledice su dozno- i polno-specifične. U testu otvorenog polja (PND40), izlaganje majki tokom trudnoće valproatu značajno utiče na ponašanje adolescentnog potomstva oba pola. Shodno primenjenoj dozi, efekat se ogleda u finom narušavanju analiziranih parametara (lokomotorna, stereotipna ili vertikalna aktivnost) u određenom periodu boravka u otvorenom polju. Zaključak. Primenjeni eksperimentalni model kontirnuirane aplikacije valproata tokom priploda i gestacije na jedinkama miša je u korelaciji sa njegovom terapijskom primenom u tretmanu epilepsija tokom trudnoće u humanoj populaciji. Rezultati dobijeni u našim istraživanjima su jasno ukazali da posledice intrauterinog izlaganja valproatu zavise kako od primenjene doze tako i od pola potomstva. Od posebnog značaja su pokazatelji da subterapijske doze valproata primenjene tokom gestacije značajno utiču na psihomotorni razvoj jedinki. U metodološkom smislu naši eksperimenti su pokazali da za pravilno sagledavanje posledica intrauterinog izlaganja valproatu mora da postoji adekvatan opseg postnatalnih starosti jedinki. Shodno činjenici da je metabolizam valproata kod jedinki miševa ubrzan u odnosu na metabolizam ovog leka u humanoj populaciji, kritički pristup u aproksimaciji podataka dobijenih iz animalnog modela miša na ljudsku populaciju je neophodan. Cilj ove doktorske disertacije je bio da se ispitaju posledice intrauterinog izlaganja različitim dozama valproata (sa akcentom na subterapijske doze) na psihomotorni razvoj jedinki miša, koje su testirane u različitim fazama postnatalnog razvoja. Baterija testova za ispitivanje ponašanja koja je upotrebljena tokom ove studije uključivala je sledeće testove: test refleksa uspravljanja i test kačenja o rep – tokom prve dve postnatalne nedelje, test izlaganja vrućoj ploči – u periadolescentnoj fazi, test uzdignutog krstastog lavirinta i test otvorenog polja – tokom adolescencije. Podaci dobijeni u testu refleksa uspravljanja i testu kačenja jedinke o rep su jasno ukazali na kašnjenje u razvoju senzornog i motornog sistema jedinki koje su tokom gestacije bile izložene različitim dozama valproata, koje je uočljivo kroz kvalitativno i kvantitativno drugačiji odgovor jedinki na date testove u periodu ranog postnatalnog razvoja. Velike fizičke anomalije nisu detektovane, ali je jasno pokazan negativan uticaj intrauterinog izlaganja valproatu na telesnu težinu i otvaranje očiju. Podaci dobijeni u testu vruće ploče su dodatno ukazali na kašnjenje u sazrevanju senzornog sistema značajnog za percepciju bolnog stimulusa, sa posebnim akcentom na uticaj malih 5 doza valproata. Testovi primenjeni na jedinkama u periodu adolescencije su ukazali da intrauterino izlaganje valproatu izaziva polno- i dozno-specifične promene u ponašanju, u smislu anksiozno/depresivnih stanja kod jedinki muškog pola usled primene velikih doza valproata i hiperaktivnih stanja kod jedinki ženskog pola usled primene malih doza valproata. Rezultati dobijeni u ovom istraživanju pružaju mogućnost sagledavanja posledica primene valproata tokom trudnoće na funkcionisanje centralnog nervnog sistema potomstva, sa aspekta ponašanja. U skladu sa literaturom, naši eksperimenti su ukazali da intrauterino izlaganje valproatu izaziva kompleksne promene u ponašanju potomstva, usled morfo-funkcionalnih poremećaja u određenim moždanim regionima. Ipak, ostaje da se precizno utvrde neuralni korelati bihejvioralnih efekata opisanih u našoj studiji koja je za model imala kontinuiranu primenu valproata tokom čitave gestacije, što je eksperimentalno za sada nedovoljno ispitano.The aim of this study was to examine the effects of intrauterine exposure to different doses of valproate on the course of physical and psychomotor development of mouse. The effects of valproate in the CNS was evaluated at subtherapeutic doses and dose-dependent effect. The tests applied to the offspring in the literature are labeled as suitable for the assessment of motor and sensory system at a certain stage of postnatal development. Certain tests, in addition to general motor skills, allowed the assessment of behavioral state as anxiety / depression, hyper-and hypo-activity of offspring. The study was done on adult female NMRI mice. Four experimental and one control group was formed. Females were treated with subcutaneous injection of 50, 100, 200 and 400 mg/kg VPA (Valproic acid sodium salt – 2 propylpentanoic sodium, ≥98, P4543 Sigma Aldrich) or with saline (Natrii Chloridi Infundibule 0,9%, HemofarmhospitalLogica) (control group). All treatments were performed during breeding and whole gestation period. Mice were given ad libitum access to food and water. The offspring was examined in tests: (1) Righting test, (2) Tail suspension test, (3) Hot plate test, (4) Elevated plus maze test and (5) Open field test Results: Different doses of VPA (50, 100, 200 and 400 mg/kg) continuously applied on female NMRI mice during breeding and whole gestation caused female mortality and lower fertility dose-dependent effect. Treatment had no influence on female body weight. In utero exposure to valproate has a negative impact on body weight of individuals during postnatal development, including the adolescence. Treatment is associated with a delay in eye-opening at the 15th postnatal day (PND). Prenatal exposition to different doses of VPA significantly influence on offspring behaviour in righting reflex. Regarding the critical developmental postnatal points, treatment influence on type response and latency during righting reflex testing. VPA treatment during gestation affect outcome of tail suspension test at the 5th and 10th PND (regarding the doses of 200 and 400mg/kg). There are no concequences in treated groups in tst at 15th PND. VPA exposition in utero distinctly influence on outcome during hot plate testing at 25th and 32nd PND. Increased procent of inadequately response and extended latency in adequate response of treated mice was obtained in hot plate test. Offspring which mothers were treated with VPA shown different pattern of behaviour (dose- and gender- dependent) during adolescence in elevated plus maze test (PND 35). Mice prenataly exposed to VPA during adolescence (PND40), represent significant differences during locomotor, stereotipic and vertical activity in the open field test. Conclusion: Experimental model of continuous application of VPA during breeding and gestation on mice is congruent with therapeutic application in epilepsy treatment during pregnancy in human population. Obtained results in this study indicate that exposure to VPA induce the effect which is dose- and gender-dependent. A subtherapeutic doses applied during gestation significantly influence on the course of psychomotor development. Methodolically, this experiments had shown that the consequences of exposition to valproate in-utero must be examined during adequate postnatal period. Mice has higher rate of VPA metabolism compared with the human rate. Regarding this fact, critical approach must be considered in approximation of the results obtained from animal model of mice to human population

Influence of valproic acid application during breeding and gestation in the mouse animal model - effect on psychomotor development in offspring

Osnovni cilj ove doktorske disertacije je bio da se ispitaju posledice intrauterinog izlaganja različitim dozama valproata na tok fizičkog i psihomotornog razvoja jedinki miša. Ispitivani su efekti valproata na CNS-u u subterapijskim dozama i dozno- zavisni efekat. Testovi primenjeni na potomstvu su u literaturi okarakterisani kao odgovarajući za procenu stanja motornog i senzornog sistema u određenoj fazi postnatalnog razvoja. Određeni testovi, pored opšte motorne sposobnosti, omogućavali su i procenu ponašanja tipa anksioznosti / depresije, hiper- i hipoaktivnosti jedinki. Metode. U eksperimentima su korišćene ženke i mužjaci zdravih laboratorijskih miševa soja NMRI, odabrane metodom slučajnog izbora iz okota na Vojnomedicinskoj akademiji u Beogradu. Eksperiment je počinjao sparivanjem dve ženke i jednog mužjaka u jednom kavezu. Shodno tretmanu, oformljeno je 5 eksperimentalnih grupa i to jedna kontrolna (n=9 ženki), koja je bila tretirana fiziološkim rastvorom (Natrii Chloridi Infundibule 0,9%, HemofarmhospitalLogica), i 4 grupe koje su tretirane valproatom (Valproic acid sodium salt – 2 propylpentanoic sodium, ≥98, P4543 Sigma Aldrich) u dozi od 50 mg/kg, 100 mg/kg, 200 mg/kg ili 400 mg/kg (n=10-11 ženki po grupi). Natrijumova so valproinske kiseline rastvarana je u fiziološkom rastvoru. Injeciranje je obavljano svakodnevno, od momenta sparivanja, tokom čitavog perioda priploda i gestacije. Nakon rađanja potomstva tretman je prestajao i ženke su sa svojim potomstvom odvajane u zasebne kaveze. Pristup hrani i vodi je bio neograničen. Procenjivan je odgovor potomstva u sledećim testovima: (1) test refleksa uspravljanja , (2)test kačenja o rep, (3) test izlaganja vrućoj ploči, (4) test uzdignutog krstastog lavirinta, (5) test otvorenog polja Rezultati. Kontinuirana primena različitih doza valproata (50, 100, 200 i 400 mg/kg) na ženkama miša NMRI soja u periodu priploda ima negativan dozno-zavisni uticaj na fertilitet, a kontinuirana primena u periodu priploda i gestacije na smrtnost ženki (akcenat na visokim dozama). Tretman nema značajnog uticaja na telesnu težinu ženki. Intrauterino izlaganje valproatu ima negativan uticaj na telesnu težinu jedinki tokom postnatalnog razvoja, uključujući period adolescencije. Tretman je povezan i sa kašnjenjem u otvaranju očiju jedinki u periodu do 15. dana postanatalnog razvoja. Intrauterino izlaganje različitim dozama valproata značajno utiče na ponašanje jedinki u testu uspravljanja. Shodno starosti jedinki efekat tretmana se ogleda kako u kvalitetu (tipu) odgovora i u vremenu koje je potrebno jedinki da se uspravi. Tretman valproatom tokom prenatalnog perioda utiče na ishod testa kačenja o rep kada se jedinke testiraju u periodu od 5. do 10. postnatalnog dana (sa akcentom na doze 200 mg/kg i 400 mg/kg). Kod jedinki starih 15 postnatalnih dana posledice tretmana nisu uočene. Intrauterino izlaganje valproatu značajno utiče na ishod u testu izlaganja vrućoj ploči jedinki starih 25 i 32 postnatalna dana. Efekat se ogleda u povećanju broja jedinki koje neadekvatno odgovaraju na test, kao i u produženoj latenci odgovora na stimulus kod jedinki koje adekvatno odgovore na test. Izlaganje majki tokom trudnoće valproatu značajno utiče na ponašanje adolescentnog potomstva (PND35) u testu uzdignutog krstastog lavirinta, a posledice su dozno- i polno-specifične. U testu otvorenog polja (PND40), izlaganje majki tokom trudnoće valproatu značajno utiče na ponašanje adolescentnog potomstva oba pola. Shodno primenjenoj dozi, efekat se ogleda u finom narušavanju analiziranih parametara (lokomotorna, stereotipna ili vertikalna aktivnost) u određenom periodu boravka u otvorenom polju. Zaključak. Primenjeni eksperimentalni model kontirnuirane aplikacije valproata tokom priploda i gestacije na jedinkama miša je u korelaciji sa njegovom terapijskom primenom u tretmanu epilepsija tokom trudnoće u humanoj populaciji. Rezultati dobijeni u našim istraživanjima su jasno ukazali da posledice intrauterinog izlaganja valproatu zavise kako od primenjene doze tako i od pola potomstva. Od posebnog značaja su pokazatelji da subterapijske doze valproata primenjene tokom gestacije značajno utiču na psihomotorni razvoj jedinki. U metodološkom smislu naši eksperimenti su pokazali da za pravilno sagledavanje posledica intrauterinog izlaganja valproatu mora da postoji adekvatan opseg postnatalnih starosti jedinki. Shodno činjenici da je metabolizam valproata kod jedinki miševa ubrzan u odnosu na metabolizam ovog leka u humanoj populaciji, kritički pristup u aproksimaciji podataka dobijenih iz animalnog modela miša na ljudsku populaciju je neophodan. Cilj ove doktorske disertacije je bio da se ispitaju posledice intrauterinog izlaganja različitim dozama valproata (sa akcentom na subterapijske doze) na psihomotorni razvoj jedinki miša, koje su testirane u različitim fazama postnatalnog razvoja. Baterija testova za ispitivanje ponašanja koja je upotrebljena tokom ove studije uključivala je sledeće testove: test refleksa uspravljanja i test kačenja o rep – tokom prve dve postnatalne nedelje, test izlaganja vrućoj ploči – u periadolescentnoj fazi, test uzdignutog krstastog lavirinta i test otvorenog polja – tokom adolescencije. Podaci dobijeni u testu refleksa uspravljanja i testu kačenja jedinke o rep su jasno ukazali na kašnjenje u razvoju senzornog i motornog sistema jedinki koje su tokom gestacije bile izložene različitim dozama valproata, koje je uočljivo kroz kvalitativno i kvantitativno drugačiji odgovor jedinki na date testove u periodu ranog postnatalnog razvoja. Velike fizičke anomalije nisu detektovane, ali je jasno pokazan negativan uticaj intrauterinog izlaganja valproatu na telesnu težinu i otvaranje očiju. Podaci dobijeni u testu vruće ploče su dodatno ukazali na kašnjenje u sazrevanju senzornog sistema značajnog za percepciju bolnog stimulusa, sa posebnim akcentom na uticaj malih 5 doza valproata. Testovi primenjeni na jedinkama u periodu adolescencije su ukazali da intrauterino izlaganje valproatu izaziva polno- i dozno-specifične promene u ponašanju, u smislu anksiozno/depresivnih stanja kod jedinki muškog pola usled primene velikih doza valproata i hiperaktivnih stanja kod jedinki ženskog pola usled primene malih doza valproata. Rezultati dobijeni u ovom istraživanju pružaju mogućnost sagledavanja posledica primene valproata tokom trudnoće na funkcionisanje centralnog nervnog sistema potomstva, sa aspekta ponašanja. U skladu sa literaturom, naši eksperimenti su ukazali da intrauterino izlaganje valproatu izaziva kompleksne promene u ponašanju potomstva, usled morfo-funkcionalnih poremećaja u određenim moždanim regionima. Ipak, ostaje da se precizno utvrde neuralni korelati bihejvioralnih efekata opisanih u našoj studiji koja je za model imala kontinuiranu primenu valproata tokom čitave gestacije, što je eksperimentalno za sada nedovoljno ispitano.The aim of this study was to examine the effects of intrauterine exposure to different doses of valproate on the course of physical and psychomotor development of mouse. The effects of valproate in the CNS was evaluated at subtherapeutic doses and dose-dependent effect. The tests applied to the offspring in the literature are labeled as suitable for the assessment of motor and sensory system at a certain stage of postnatal development. Certain tests, in addition to general motor skills, allowed the assessment of behavioral state as anxiety / depression, hyper-and hypo-activity of offspring. The study was done on adult female NMRI mice. Four experimental and one control group was formed. Females were treated with subcutaneous injection of 50, 100, 200 and 400 mg/kg VPA (Valproic acid sodium salt – 2 propylpentanoic sodium, ≥98, P4543 Sigma Aldrich) or with saline (Natrii Chloridi Infundibule 0,9%, HemofarmhospitalLogica) (control group). All treatments were performed during breeding and whole gestation period. Mice were given ad libitum access to food and water. The offspring was examined in tests: (1) Righting test, (2) Tail suspension test, (3) Hot plate test, (4) Elevated plus maze test and (5) Open field test Results: Different doses of VPA (50, 100, 200 and 400 mg/kg) continuously applied on female NMRI mice during breeding and whole gestation caused female mortality and lower fertility dose-dependent effect. Treatment had no influence on female body weight. In utero exposure to valproate has a negative impact on body weight of individuals during postnatal development, including the adolescence. Treatment is associated with a delay in eye-opening at the 15th postnatal day (PND). Prenatal exposition to different doses of VPA significantly influence on offspring behaviour in righting reflex. Regarding the critical developmental postnatal points, treatment influence on type response and latency during righting reflex testing. VPA treatment during gestation affect outcome of tail suspension test at the 5th and 10th PND (regarding the doses of 200 and 400mg/kg). There are no concequences in treated groups in tst at 15th PND. VPA exposition in utero distinctly influence on outcome during hot plate testing at 25th and 32nd PND. Increased procent of inadequately response and extended latency in adequate response of treated mice was obtained in hot plate test. Offspring which mothers were treated with VPA shown different pattern of behaviour (dose- and gender- dependent) during adolescence in elevated plus maze test (PND 35). Mice prenataly exposed to VPA during adolescence (PND40), represent significant differences during locomotor, stereotipic and vertical activity in the open field test. Conclusion: Experimental model of continuous application of VPA during breeding and gestation on mice is congruent with therapeutic application in epilepsy treatment during pregnancy in human population. Obtained results in this study indicate that exposure to VPA induce the effect which is dose- and gender-dependent. A subtherapeutic doses applied during gestation significantly influence on the course of psychomotor development. Methodolically, this experiments had shown that the consequences of exposition to valproate in-utero must be examined during adequate postnatal period. Mice has higher rate of VPA metabolism compared with the human rate. Regarding this fact, critical approach must be considered in approximation of the results obtained from animal model of mice to human population

Psychomotor activity and body weight gain after exposure to low ribavirin doses in rats: role of treatment duration

Clinically-related basic studies on the behavioral effects of ribavirin treatment are still lacking despite its wide use as an antiviral medication. This paper considers the effects of low ribavirin doses (10, 20 and 30 mg/kg/day) on psychomotor activity (novelty-induced exploratory behavior, d-amphetamine (AMPH, 1.5 mg/kg, intraperitoneal)-induced motor activity), and body weight gain in socially undisturbed adult male Wistar rats 24 h after the first, seventh and fourteenth once-a-day injection. Low doses of ribavirin were tested in an attempt to avoid the recognized systemic side effects related to high-dose usage. None of the singly applied ribavirin doses affected exploratory/spontaneous and AMPH-induced motor behavior (locomotion, stereotypy-like and vertical activity), however, body weight gain was significantly lower after treatment with 30 mg/kg of ribavirin. The 7- and 14-day treatments with 10 and 30 mg/kg/day of ribavirin significantly suppressed novelty-induced locomotion and body weight gain; the 14-day treatment with ribavirin at a dose of 30 mg/kg/ day decreased AMPH-induced stereotypy. These findings indicate that repeated application (up to 14 days) of low ribavirin doses results in low novelty-induced locomotion along with reduced weight gain, accentuating the existence of a U-shaped dose-response relationship with a prolonged duration of ribavirin treatment

Monitoring the progressive increase of the longest episode of spontaneous movements in Guinea pig fetus

The aim of this work was to determine the changes in the duration of spontaneous movements in the guinea pig fetus after the appearance of its first movements. Every day from the 25th to the 35th gestation day, one fetus from each of twenty pregnant animals was examined by ultrasound. Fetal movements were observed for 5 min. The episode with the longest period of movement was taken into consideration and was recorded as: 3 s. Days 25 and 26 were characterized by episodes lasting 3 s (χ2 = 140.51 p <0.05). Tracking the dynamics of progressive increases in the longest episode of spontaneous movement could be a useful factor in estimating the maturity and condition of a fetus.Projekat ministarstva br. 175006/201

Influence of valproic acid application during breeding and gestation in the mouse animal model - effect on psychomotor development in offspring

Osnovni cilj ove doktorske disertacije je bio da se ispitaju posledice intrauterinog izlaganja različitim dozama valproata na tok fizičkog i psihomotornog razvoja jedinki miša. Ispitivani su efekti valproata na CNS-u u subterapijskim dozama i dozno- zavisni efekat. Testovi primenjeni na potomstvu su u literaturi okarakterisani kao odgovarajući za procenu stanja motornog i senzornog sistema u određenoj fazi postnatalnog razvoja. Određeni testovi, pored opšte motorne sposobnosti, omogućavali su i procenu ponašanja tipa anksioznosti / depresije, hiper- i hipoaktivnosti jedinki. Metode. U eksperimentima su korišćene ženke i mužjaci zdravih laboratorijskih miševa soja NMRI, odabrane metodom slučajnog izbora iz okota na Vojnomedicinskoj akademiji u Beogradu. Eksperiment je počinjao sparivanjem dve ženke i jednog mužjaka u jednom kavezu. Shodno tretmanu, oformljeno je 5 eksperimentalnih grupa i to jedna kontrolna (n=9 ženki), koja je bila tretirana fiziološkim rastvorom (Natrii Chloridi Infundibule 0,9%, HemofarmhospitalLogica), i 4 grupe koje su tretirane valproatom (Valproic acid sodium salt – 2 propylpentanoic sodium, ≥98, P4543 Sigma Aldrich) u dozi od 50 mg/kg, 100 mg/kg, 200 mg/kg ili 400 mg/kg (n=10-11 ženki po grupi). Natrijumova so valproinske kiseline rastvarana je u fiziološkom rastvoru. Injeciranje je obavljano svakodnevno, od momenta sparivanja, tokom čitavog perioda priploda i gestacije. 137 Nakon rađanja potomstva tretman je prestajao i ženke su sa svojim potomstvom odvajane u zasebne kaveze. Pristup hrani i vodi je bio neograničen. Procenjivan je odgovor potomstva u sledećim testovima: (1) test refleksa uspravljanja , (2)test kačenja o rep, (3) test izlaganja vrućoj ploči, (4) test uzdignutog krstastog lavirinta, (5) test otvorenog polja Rezultati. Kontinuirana primena različitih doza valproata (50, 100, 200 i 400 mg/kg) na ženkama miša NMRI soja u periodu priploda ima negativan dozno-zavisni uticaj na fertilitet, a kontinuirana primena u periodu priploda i gestacije na smrtnost ženki (akcenat na visokim dozama). Tretman nema značajnog uticaja na telesnu težinu ženki. Intrauterino izlaganje valproatu ima negativan uticaj na telesnu težinu jedinki tokom postnatalnog razvoja, uključujući period adolescencije. Tretman je povezan i sa kašnjenjem u otvaranju očiju jedinki u periodu do 15. dana postanatalnog razvoja. Intrauterino izlaganje različitim dozama valproata značajno utiče na ponašanje jedinki u testu uspravljanja. Shodno starosti jedinki efekat tretmana se ogleda kako u kvalitetu (tipu) odgovora i u vremenu koje je potrebno jedinki da se uspravi. Tretman valproatom tokom prenatalnog perioda utiče na ishod testa kačenja o rep kada se jedinke testiraju u periodu od 5. do 10. postnatalnog dana (sa akcentom na doze 200 mg/kg i 400 mg/kg). Kod jedinki starih 15 postnatalnih dana posledice tretmana nisu uočene. Intrauterino izlaganje valproatu značajno utiče na ishod u testu izlaganja vrućoj ploči jedinki starih 25 i 32 postnatalna 138 dana. Efekat se ogleda u povećanju broja jedinki koje neadekvatno odgovaraju na test, kao i u produženoj latenci odgovora na stimulus kod jedinki koje adekvatno odgovore na test. Izlaganje majki tokom trudnoće valproatu značajno utiče na ponašanje adolescentnog potomstva (PND35) u testu uzdignutog krstastog lavirinta, a posledice su dozno- i polno-specifične. U testu otvorenog polja (PND40), izlaganje majki tokom trudnoće valproatu značajno utiče na ponašanje adolescentnog potomstva oba pola. Shodno primenjenoj dozi, efekat se ogleda u finom narušavanju analiziranih parametara (lokomotorna, stereotipna ili vertikalna aktivnost) u određenom periodu boravka u otvorenom polju. Zaključak. Primenjeni eksperimentalni model kontirnuirane aplikacije valproata tokom priploda i gestacije na jedinkama miša je u korelaciji sa njegovom terapijskom primenom u tretmanu epilepsija tokom trudnoće u humanoj populaciji. Rezultati dobijeni u našim istraživanjima su jasno ukazali da posledice intrauterinog izlaganja valproatu zavise kako od primenjene doze tako i od pola potomstva. Od posebnog značaja su pokazatelji da subterapijske doze valproata primenjene tokom gestacije značajno utiču na psihomotorni razvoj jedinki. U metodološkom smislu naši eksperimenti su pokazali da za pravilno sagledavanje posledica intrauterinog izlaganja valproatu mora da postoji adekvatan opseg postnatalnih starosti jedinki. Shodno činjenici da je metabolizam valproata kod jedinki miševa ubrzan u odnosu na metabolizam ovog leka u humanoj populaciji, kritički pristup u aproksimaciji podataka dobijenih iz animalnog modela miša na ljudsku populaciju je neophodna.The aim of this study was to examine the effects of intrauterine exposure to different doses of valproate on the course of physical and psychomotor development of mouse. The effects of valproate in the CNS was evaluated at subtherapeutic doses and dose-dependent effect. The tests applied to the offspring in the literature are labeled as suitable for the assessment of motor and sensory system at a certain stage of postnatal development. Certain tests, in addition to general motor skills, allowed the assessment of behavioral state as anxiety / depression, hyper-and hypo-activity of offspring. The study was done on adult female NMRI mice. Four experimental and one control group was formed. Females were treated with subcutaneous injection of 50, 100, 200 and 400 mg/kg VPA (Valproic acid sodium salt – 2 propylpentanoic sodium, ≥98, P4543 Sigma Aldrich) or with saline (Natrii Chloridi Infundibule 0,9%, HemofarmhospitalLogica) (control group). All treatments were performed during breeding and whole gestation period. Mice were given ad libitum access to food and water. The offspring was examined in tests: (1) Righting test, (2) Tail suspension test, (3) Hot plate test, (4) Elevated plus maze test and (5) Open field test Results: Different doses of VPA (50, 100, 200 and 400 mg/kg) continuously applied on female NMRI mice during breeding and whole 142 gestation caused female mortality and lower fertility dose-dependent effect. Treatment had no influence on female body weight. In utero exposure to valproate has a negative impact on body weight of individuals during postnatal development, including the adolescence. Treatment is associated with a delay in eye-opening at the 15th postnatal day (PND). Prenatal exposition to different doses of VPA significantly influence on offspring behaviour in righting reflex. Regarding the critical developmental postnatal points, treatment influence on type response and latency during righting reflex testing. VPA treatment during gestation affect outcome of tail suspension test at the 5th and 10th PND (regarding the doses of 200 and 400mg/kg). There are no concequences in treated groups in tst at 15th PND. VPA exposition in utero distinctly influence on outcome during hot plate testing at 25th and 32nd PND. Increased procent of inadequately response and extended latency in adequate response of treated mice was obtained in hot plate test. Offspring which mothers were treated with VPA shown different pattern of behaviour (dose- and gender- dependent) during adolescence in elevated plus maze test (PND 35). Mice prenataly exposed to VPA during adolescence (PND40), represent significant differences during locomotor, stereotipic and vertical activity in the open field test. Conclusion: Experimental model of continuous application of VPA during breeding and gestation on mice is congruent with therapeutic application in epilepsy treatment during pregnancy in human population. Obtained results in this study indicate that exposure to VPA induce the effect which is dose- and 143 gender-dependent. A subtherapeutic doses applied during gestation significantly influence on the course of psychomotor development. Methodolically, this experiments had shown that the consequences of exposition to valproate in-utero must be examined during adequate postnatal period. Mice has higher rate of VPA metabolism compared with the human rate. Regarding this fact, critical approach must be considered in approximation of the results obtained from animal model of mice to human population

Function of serotonin receptors and its role in the behavior

Serotonin is, evolutionary, the oldest neurotransmitter. Ubiquitous distribution of serotonin in central nervous system, also implication in many physiological functions indicates significance of serotonergic system. Expression of the serotonin function is the most conditioned by the interaction with postsynaptic receptors. In this article, the classification of the serotonergic receptors and its role is reviewed. The expectations of future research are detection of new subpopulations of receptors, their functions and summation of knowledge in order to find new therapy and new manner in treatment of central nervous system disturbances, which imply serotonergic dysfunction.Serotonin predstavlja jedan od evolutivno najstarijih neurotransmitrera. Njegova široka distribucija u centralnom nervnom sistemu i povezanost sa mnogobrojnim fiziološkim funkcijama ukazuje na značaj serotoninskog sistema. Karakter ispoljavanja serotonina u najvećoj meri određuje postsinaptički receptor sa kojim stupa u kontakt. U radu su prikazani do sada klasifikovani receptori i njihova uloga. Od budućih istraživanja očekuje se otkrivanje novih subpopulacija receptora, njihove uloge i sumacija saznanja u cilju pronalaženja novih lekova i načina lečenja uzrokovanih poremećajima serotinskog sistema CNS-a.nul

Linear Analysis of Epileptic Forms in the Electroencephalogram

In our study, we investigated the properties of electroencephalogram (EEG) of humans experiencing epileptic seizures. By applying linear methods, such as spectral analysis and wavelet transformation, we compared characteristics of EEGs of patients suffering from epilepsia with the characteristics of EEGs of healthy individuals. Our goal was to establish the real effect of epileptic attacks in frequency domains, distinguishing them from various types of artifacts caused by mental and physical activities in humans. The frequency range of 1 to 2 Hz was found to be interesting, where we have detected a power spectral density peak typical of epileptic seizures. Also, we have shown that there exists an anomalous distribution of the relative wavelet energy of the signal within this frequency range. We also showed that the frequency range 7.97 to 15.94 Hz can serve as an indicator of preictal activity in the EEG signal.Ministry of Education and Science of Republic of Serbia [175006

Linear Analysis of Epileptic Forms in the Electroencephalogram

In our study, we investigated the properties of electroencephalogram (EEG) of humans experiencing epileptic seizures. By applying linear methods, such as spectral analysis and wavelet transformation, we compared characteristics of EEGs of patients suffering from epilepsia with the characteristics of EEGs of healthy individuals. Our goal was to establish the real effect of epileptic attacks in frequency domains, distinguishing them from various types of artifacts caused by mental and physical activities in humans. The frequency range of 1 to 2 Hz was found to be interesting, where we have detected a power spectral density peak typical of epileptic seizures. Also, we have shown that there exists an anomalous distribution of the relative wavelet energy of the signal within this frequency range. We also showed that the frequency range 7.97 to 15.94 Hz can serve as an indicator of preictal activity in the EEG signal.Ministry of Education and Science of Republic of Serbia [175006

Hydraulic Jump in Trapezoidal Channel (Wassersprung im Trapezkanal)

The features of rat cerebral and cerebellar electrocortical activity (ECoG) under different types of anaesthesia (nembutal. ketamine or zoletil) were examined by the distribution of spectral entropy across frequency bands of ECoG and by calculation of fractal dimension determined on the basis of Higuchi's algorithm. Spectral entropy, as a measure of activity, in the case of cerebrum had greater values than the spectral entropy of cerebellum in low frequency ranges, regardless of the type or applied anesthetic. Various anesthetics evoked different effects on spectral entropy of electrocortical activity: spectral entropy of delta range greatly dominated under nembutal anesthesia, while ketamine or zoletil appeared to affect the spectral entropy of higher frequency ranges. The pronounced effect of ketamine or zoletil anesthesia on spectral entropy of higher frequency was confirmed by the higher values of Higucihi's fractal dimension (ED) of ECoGs, with a tendency of higher ED values in cerebellar activity than cerebral activity.Ministry of Science and Technological Development of Republic of Serbia [143021

Function of serotonin receptors and its role in the behavior

Serotonin is, evolutionary, the oldest neurotransmitter. Ubiquitous distribution of serotonin in central nervous system, also implication in many physiological functions indicates significance of serotonergic system. Expression of the serotonin function is the most conditioned by the interaction with postsynaptic receptors. In this article, the classification of the serotonergic receptors and its role is reviewed. The expectations of future research are detection of new subpopulations of receptors, their functions and summation of knowledge in order to find new therapy and new manner in treatment of central nervous system disturbances, which imply serotonergic dysfunction.Serotonin predstavlja jedan od evolutivno najstarijih neurotransmitrera. Njegova široka distribucija u centralnom nervnom sistemu i povezanost sa mnogobrojnim fiziološkim funkcijama ukazuje na značaj serotoninskog sistema. Karakter ispoljavanja serotonina u najvećoj meri određuje postsinaptički receptor sa kojim stupa u kontakt. U radu su prikazani do sada klasifikovani receptori i njihova uloga. Od budućih istraživanja očekuje se otkrivanje novih subpopulacija receptora, njihove uloge i sumacija saznanja u cilju pronalaženja novih lekova i načina lečenja uzrokovanih poremećajima serotinskog sistema CNS-a.nul