Serotonergní systém je jedním z klíčových neurotransmiterových systémů ovlivňovaný antipsychotiky. Některá antipsychotika (clozapin, olanzapin) vykazují vysokou afinitu k serotoninovým receptorům typu 5-HT2A a 5-HT2C. Cílem této práce bylo zjistit roli těchto receptorů ve zpracování informací v animálním farmakologickém modelu schizofrenii podobného chování vyvolaném akutním podáním nekompetitivního antagonisty N-metyl-D-aspartátového (NMDA) receptoru – dizocilpinu (MK-801). Použili jsme antagonisty serotoninových 5-HT2A/2C (ritanserin) a 5-HT2C (SB 242084) receptorů a sledovali jsme jejich vliv na deficit ve zpracování informací po podání MK-801. Úroveň zpracování informací byla měřena testem prepulzní inhibice úlekové reakce (PPI) u potkana. Podání MK-801 vedlo v našem modelu k deficitu v PPI. Ritanserin zlepšoval deficit v PPI způsobený MK-801, zatímco SB 242084 na deficit vliv nemělo. Na rozdíl od ritanserinu, který neměnil spontánní PPI (bez MK-801), antagonista serotoninového 5-HT2C receptoru zvětšoval úroveň spontánní PPI. Výsledky ukazují, že serotoninové 5-HT2A receptory hrají významnou úlohu ve zpracování informací v animálním modelu schizofrenii podobného chování.

The serotonergic system is one of the key neurotransmitter systems affected by the effects of antipsychotic drugs. Some antipsychotics (clozapine, olanzapine) show high affinities at serotonin 5-HT2A a 5-HT2C receptors. The aim of this study was to evaluate the role of these receptor subtypes in information processing in pharmacological animal model of schizophrenia-like behavior induced by acute administration of non-competitive N-methyl- D-aspartate (NMDA) receptor antagonist – dizocilpine (MK-801). We used a serotonin 5-HT2A/2C receptor antagonist (ritanserin) and a selective, brain penetrant, serotonin 5-HT2C receptor antagonist (SB 242084). We have concentrated on how they affect deficits in information processing induced by MK-801. The extent of information processing was measured by the Prepulse inhibition of the acoustic startle response test (PPI) in rats. Administration of MK-801 induced severe deficits in PPI. Ritanserin ameliorated this deficit induced by MK-801, while SB 242084 had no effects. On the other hand, compared to ritanserin that had no effect on spontaneous PPI, SB 242084 markedly increased PPI rate in the control animals. Our results show that serotonin 5-HT2A receptors play an important role in information processing in animal models of schizophrenia-like behavior.

• MeSH
• antagonisté serotoninu farmakologie   terapeutické užití   MeSH
• dizocilpinmaleát farmakologie   MeSH
• potkani Wistar MeSH
• ritanserin farmakologie   terapeutické užití   MeSH
• schizofrenie chemicky indukované   MeSH
• úleková reakce účinky léků   MeSH

Levels of Rb2/p130 protein are increased 5-10-fold following all-trans-retinoic acid (ATRA) treatment of the retinoid-sensitive ovarian adenocarcinoma cell line CAOV3, but not the retinoid-resistant adenocarcinoma cell line SKOV3. We found that this increase in Rb2/p130 protein levels in ATRA-treated CAOV3 cells was the result of an increased protein stability. Moreover, Rb2/p130 exhibited a decreased ubiquitination following ATRA treatment. Because phosphorylation frequently mediates ubiquitination of proteins, we examined the serine/threonine phosphatase activity in our CAOV3 cells following ATRA treatment. A significant increase in Ser/Thr phosphatase activity was found, which correlated with a rise in the level of protein phosphatase 2A (PP2A) catalytic subunit-alpha. In addition, co-immunoprecipitation and glutathione S-transferase pull-down studies demonstrated that PP2A and Rb2/p130 associate. We have made use of a battery of Rb2/p130 mutants to determine the sites dephosphorylated in response to ATRA treatment of CAOV3 cells. Obligate CDK4 phosphorylation sites seemed most important to the stability of the protein and are among the candidate sites that are dephosphorylated by PP2A following ATRA treatment. Finally, using both small interfering RNA specific to the catalytic subunit of PP2A and a variant of the SKOV3 cell line that overexpresses PP2A, we have shown that modulation of PP2A protein levels correlates with the ability of ATRA to inhibit growth of ovarian carcinoma cells. Our data suggest that ATRA mediates growth inhibition by stabilizing Rb2/p130 via a mechanism that involves induction of PP2A, an enzyme that can potentially dephosphorylate Rb2/p130, thereby protecting it from degradation by the proteasome.

• MeSH
• buněčné dělení účinky léků   MeSH
• endopeptidasy metabolismus   MeSH
• fosfoproteiny metabolismus   účinky léků   MeSH
• fosforylace MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• nádory vaječníků metabolismus   patologie   MeSH
• protein p130 podobný retinoblastomu MeSH
• proteinfosfatasa 2 MeSH
• proteinfosfatasy * fyziologie   metabolismus   MeSH
• proteiny * MeSH
• tretinoin * farmakologie   MeSH
• ubikvitiny metabolismus   MeSH
• upregulace účinky léků   MeSH
• vazba proteinů fyziologie   účinky léků   MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH

• MeSH
• akutní lymfatická leukemie * genetika   MeSH
• genová přestavba MeSH
• lidé MeSH
• pre-B-buněčná leukemie * genetika   MeSH
• retrospektivní studie MeSH
• transkripční faktory MEF2 MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH

Ankyrin repeat proteins (ARPs) appear to be abundant in organisms from all phyla, and play critical regulatory roles, mediating specific interactions with target biomolecules and thus ordering the sequence of events in diverse cellular processes. ARPs possess a non-globular scaffold consisting of repeating motifs named ankyrin (ANK) repeats, which stack on each other. The modular architecture of ARPs provides a new paradigm for understanding protein stability and folding mechanisms. In the present study, the stability of various C-terminal fragments of the ARP p18(INK4c) was investigated by all-atomic 450 ns molecular dynamics (MD) simulations in explicit water solvent. Only motifs with at least two ANK repeats made stable systems in the available timescale. All smaller fragments were unstable, readily losing their native fold and alpha-helical content. Since each non-terminal ANK repeat has two hydrophobic sides, we may hypothesize that at least one hydrophobic side must be fully covered and shielded from the water as a necessary, but not sufficient, condition to maintain ANK repeat stability. Consequently, at least two ANK repeats are required to make a stable ARP.

• MeSH
• aminokyselinové motivy MeSH
• ankyrinová repetice MeSH
• financování organizované MeSH
• inhibitor p18 cyklin-dependentní kinasy chemie   MeSH
• krystalografie rentgenová MeSH
• voda chemie   MeSH

Primary cilium projects from cells to provide a communication platform with neighboring cells and the surrounding environment. This is ensured by the selective entry of membrane receptors and signaling molecules, producing fine-tuned and effective responses to the extracellular cues. In this study, we focused on one family of signaling molecules, the fibroblast growth factor receptors (FGFRs), their residence within cilia, and its role in FGFR signaling. We show that FGFR1 and FGFR2, but not FGFR3 and FGFR4, localize to primary cilia of the developing mouse tissues and in vitro cells. For FGFR2, we demonstrate that the ciliary residence is necessary for its signaling and expression of target morphogenic genes. We also show that the pathogenic FGFR2 variants have minimal cilium presence, which can be rescued for the p.P253R variant associated with the Apert syndrome by using the RLY-4008 kinase inhibitor. Finally, we determine the molecular regulators of FGFR2 trafficking to cilia, including IFT144, BBS1, and the conserved T429V430 motif within FGFR2.

• MeSH
• cilie * metabolismus   genetika   MeSH
• epitelové buňky * metabolismus   MeSH
• lidé MeSH
• myši MeSH
• receptor fibroblastových růstových faktorů, typ 1 metabolismus   genetika   MeSH
• receptor fibroblastových růstových faktorů, typ 2 * metabolismus   genetika   MeSH
• signální transdukce * MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Fibroblast growth factor (FGF) signaling is crucial for mammary gland development. Although multiple roles for FGF signaling in the epithelium have been described, the function of FGF signaling in mammary stroma has not been elucidated. In this study, we investigated FGF signaling in mammary fibroblasts. We found that murine mammary fibroblasts express FGF receptors FGFR1 and FGFR2 and respond to FGF ligands. In particular, FGF2 and FGF9 induce sustained ERK1/2 signaling and promote fibroblast proliferation and migration in 2D cultures. Intriguingly, only FGF2 induces fibroblast migration in 3D extracellular matrix (ECM) through regulation of actomyosin cytoskeleton and promotes force-mediated collagen remodeling by mammary fibroblasts. Moreover, FGF2 regulates production of ECM proteins by mammary fibroblasts, including collagens, fibronectin, osteopontin and matrix metalloproteinases. Finally, using organotypic 3D co-cultures we show that FGF2 and FGF9 signaling in mammary fibroblasts enhances fibroblast-induced branching of mammary epithelium by modulating paracrine signaling, and that knockdown of Fgfr1 and Fgfr2 in mammary fibroblasts reduces branching of mammary epithelium. Our results demonstrate a pleiotropic role for FGF signaling in mammary fibroblasts, with implications for regulation of mammary stromal functions and epithelial branching morphogenesis.

• MeSH
• fibroblastový růstový faktor 2 metabolismus   MeSH
• fibroblastový růstový faktor 9 metabolismus   MeSH
• fibroblasty cytologie   metabolismus   MeSH
• MAP kinasový signální systém * MeSH
• mléčné žlázy zvířat cytologie   embryologie   MeSH
• myši inbrední ICR MeSH
• myši MeSH
• parakrinní signalizace * MeSH
• receptor fibroblastových růstových faktorů, typ 1 metabolismus   MeSH
• receptor fibroblastových růstových faktorů, typ 2 metabolismus   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Genome size and chromosome number of five Cimicidae species were compared with the similar data recently received from Cimex lectularius parasitizing human. The average nuclear DNA content (males) was 2C = 1.47 pg in C. hemipterus, 2C = 1.61 pg in C. hirundinis, 2C = 1.80 pg in C. lectularius from bats, 2C = 1.68 pg in C. pipistrelli, and 2C = 1.22 pg in Paracimex cf. chaeturus. In the genomes of all cimicid species analyzed, the average GC content ranged from 32.74% in C. pipistrelli to 35.87% in P. cf. chaeturus. Chromosome variability with two male cytotypes, 2n = 28 + X1 X2 Y and 28 + X1 X2 X3 Y, was confirmed in C. pipistrelli. In addition, intraspecific variability in chromosome number was revealed in C. lectularius from bats with 2n = 26 + X1 X2 Y and 26 + X1 X2 X3 Y. We suggest that the origin of intraspecific variability in chromosome number of C. lectularius from bats and C. pipistrelli is not only the result of simple fragmentation, but additive rearrangements like duplications are probably also involved. © 2019 International Society for Advancement of Cytometry.

• MeSH
• buněčné jádro genetika   metabolismus   MeSH
• Chiroptera MeSH
• chromozomy genetika   MeSH
• cytogenetické vyšetření MeSH
• délka genomu MeSH
• fragmentace DNA MeSH
• gonády cytologie   MeSH
• lidé MeSH
• ploidie MeSH
• pohlavní chromozomy genetika   MeSH
• průtoková cytometrie MeSH
• štěnice genetika   metabolismus   MeSH
• zárodečné buňky chemie   metabolismus   MeSH
• zastoupení bazí genetika   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH

We performed the genetic analysis of Rasopathy syndromes in patients from Central European by direct sequencing followed by next generation sequencing of genes associated with Rasopathies. All 51 patients harboured the typical features of Rasopathy syndromes. Thirty-five mutations were identified in the examined patients (22 in PTPN11, two in SOS1, one in RIT1, one in SHOC2, two in HRAS, three in BRAF, two in MAP2K1 and two in the NF1 gene). Two of them (p.Gly392Glu in the BRAF gene and p.Gln164Lys in the MAP2K1 gene) were novel with a potentially pathogenic effect on the structure of these proteins. Statistically significant differences in the presence of pulmonary stenosis (63.64% vs. 23.81%, P = 0.013897) and cryptorchidism (76.47% vs. 30%, P = 0.040224) were identified as the result of comparison of the prevalence of phenotypic features in patients with the phenotype of Noonan syndrome and mutation in the PTPN11 gene, with the prevalence of the same features in patients without PTPN11 mutation. Cryptorchidism as a statistically significant feature in our patients with PTPN11 mutation was not reported as significant in other European countries (Germany, Italy and Greece). The majority of mutations were clustered in exons 3 (45.45%), 8 (22.73%), and 13 (22.73%) of the PTPN11 gene.

• MeSH
• běloši genetika   MeSH
• dítě MeSH
• dospělí MeSH
• ektodermální dysplazie genetika   MeSH
• exony MeSH
• faciální stigmatizace MeSH
• fenotyp MeSH
• intracelulární signální peptidy a proteiny genetika   MeSH
• kojenec MeSH
• kryptorchismus genetika   MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mutační analýza DNA * MeSH
• neprospívání genetika   MeSH
• Noonanové syndrom genetika   MeSH
• předškolní dítě MeSH
• protein SOS1 genetika   MeSH
• ras proteiny genetika   MeSH
• stenóza pulmonální chlopně genetika   MeSH
• tyrosinfosfatasa nereceptorového typu 11 genetika   MeSH
• vrozené srdeční vady genetika   MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• kojenec MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Spatial working memory or short-term place memory is impaired in schizophrenia. The efficiency of antipsychotic drugs, particularly of typical antipsychotics, on cognitive deficit in schizophrenia remains disputable. Inhibition of serotonin (5-HT) 2A/2C receptors is important for cognitive improvement in schizophrenic patients treated with antipsychotics. The aim of the present work was to establish the effect of the 5-HT2A/2C receptor antagonist ritanserin (2.5 or 5 mg/kg), the dopamine D2 antagonist haloperidol (0.1 or 1 mg/kg), and the atypical antipsychotic risperidone (0.1 mg/kg or 1 mg/kg), which is an antagonist of both 5-HT2A/2C and D2 receptors, on cognitive deficit induced by subchronic administration of dizocilpine (MK-801, 0.1 mg/kg). We used the active allothetic place avoidance (AAPA) task, requiring the rat to differentiate between relevant and irrelevant stimuli, in a way similar to disruption of information processing disturbed in schizophrenic patients. Our results show that treatment with 5-HT2A/2C receptor antagonists, regardless of their effect on D2 receptors, blocked the cognitive impairment produced by MK-801. Haloperidol did not sufficiently reduce the deficit in AAPA induced by MK-801. Interestingly, administration of risperidone and haloperidol alone, but not ritanserin, impaired the AAPA performance in intact rats. Ritanserin and risperidone actually improve cognition independently of their effect on locomotor activity in an animal model of schizophrenia-like behavior. This finding is in accordance with the assumption that some antipsychotics are primarily effective against cognitive dysfunction in schizophrenia.

• MeSH
• časové faktory MeSH
• chování zvířat účinky léků   MeSH
• dizocilpinmaleát farmakologie   MeSH
• financování organizované MeSH
• haloperidol farmakologie   MeSH
• krysa rodu rattus MeSH
• potkani Wistar MeSH
• risperidon farmakologie   MeSH
• ritanserin farmakologie   MeSH
• učení vyhýbat se účinky léků   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH

Juvenile myelomonocytic leukemia (JMML) is an aggressive myeloproliferative disorder of early childhood characterized by mutations activating RAS signaling. Established clinical and genetic markers fail to fully recapitulate the clinical and biological heterogeneity of this disease. Here we report DNA methylome analysis and mutation profiling of 167 JMML samples. We identify three JMML subgroups with unique molecular and clinical characteristics. The high methylation group (HM) is characterized by somatic PTPN11 mutations and poor clinical outcome. The low methylation group is enriched for somatic NRAS and CBL mutations, as well as for Noonan patients, and has a good prognosis. The intermediate methylation group (IM) shows enrichment for monosomy 7 and somatic KRAS mutations. Hypermethylation is associated with repressed chromatin, genes regulated by RAS signaling, frequent co-occurrence of RAS pathway mutations and upregulation of DNMT1 and DNMT3B, suggesting a link between activation of the DNA methylation machinery and mutational patterns in JMML.

• MeSH
• biopsie MeSH
• chromatin genetika   metabolismus   MeSH
• dítě MeSH
• DNA-(cytosin-5-)methyltransferasa metabolismus   MeSH
• DNA-(cytosin-5)-methyltransferasa 1 metabolismus   MeSH
• epigenomika MeSH
• juvenilní myelomonocytární leukemie genetika   mortalita   patologie   terapie   MeSH
• kojenec MeSH
• lidé MeSH
• metylace DNA * MeSH
• mutace MeSH
• mutační analýza DNA MeSH
• Noonanové syndrom genetika   patologie   MeSH
• předškolní dítě MeSH
• prognóza MeSH
• prospektivní studie MeSH
• protinádorové látky terapeutické užití   MeSH
• protoonkogenní proteiny c-cbl MeSH
• protoonkogenní proteiny p21(ras) genetika   metabolismus   MeSH
• regulace genové exprese u leukemie MeSH
• signální transdukce genetika   MeSH
• transplantace hematopoetických kmenových buněk MeSH
• tyrosinfosfatasa nereceptorového typu 11 genetika   metabolismus   MeSH
• upregulace MeSH
• Check Tag
• dítě MeSH
• kojenec MeSH
• lidé MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• klinické zkoušky MeSH
• multicentrická studie MeSH
• pozorovací studie MeSH
• práce podpořená grantem MeSH