Dysfunctional sensory systems, including altered olfactory function, have recently been reported in patients with autism spectrum disorder (ASD). Disturbances in olfactory processing can potentially result from gamma-aminobutyric acid (GABA)ergic synaptic abnormalities. The specific molecular mechanism by which GABAergic transmission affects the olfactory system in ASD remains unclear. Therefore, the present study aimed to evaluate selected components of the GABAergic system in olfactory brain regions and primary olfactory neurons isolated from Shank3-deficient (-/-) mice, which are known for their autism-like behavioral phenotype. Shank3 deficiency led to a significant reduction in GEPHYRIN/GABAAR colocalization in the piriform cortex and in primary neurons isolated from the olfactory bulb, while no change of cell morphology was observed. Gene expression analysis revealed a significant reduction in the mRNA levels of GABA transporter 1 in the olfactory bulb and Collybistin in the frontal cortex of the Shank3-/- mice compared to WT mice. A similar trend of reduction was observed in the expression of Somatostatin in the frontal cortex of Shank3-/- mice. The analysis of the expression of other GABAergic neurotransmission markers did not yield statistically significant results. Overall, it appears that Shank3 deficiency leads to changes in GABAergic synapses in the brain regions that are important for olfactory information processing, which may represent basis for understanding functional impairments in autism.
- MeSH
- GABA metabolismus MeSH
- lidé MeSH
- mikrofilamentové proteiny metabolismus MeSH
- myši MeSH
- neurony metabolismus MeSH
- olfaktoriální kortex * metabolismus MeSH
- poruchy autistického spektra * metabolismus MeSH
- proteiny nervové tkáně genetika metabolismus MeSH
- synapse metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Publikační typ
- abstrakt z konference MeSH
Neuropeptides including oxytocin belong to the group of factors that may play a role in the control of neuronal cell survival, proliferation and differentiation. The aim of the present study was to investigate potential contribution of oxytocin to neuronal differentiation by measuring gene and protein expression of specific neuron and glial markers in the brain. Neonatal and adult oxytocin administration was used to reveal developmental and/or acute effects of oxytocin in Wistar rats. Gene and protein expression of neuron-specific enolase (NSE) in the hippocampus was increased in 21-day and 2-month old rats in response to neonatal oxytocin administration. Neonatal oxytocin treatment induced a significant increase of gene and protein expression of the marker of astrocytes - glial fibrillary acid protein (GFAP). Oxytocin treatment resulted in a decrease of oligodendrocyte marker mRNA - 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNPase) - in 21-day and 2-month old rats, while no change of CD68 mRNA, marker of microglia, was observed. Central oxytocin administration in adult rats induced a significant increase of gene expression of NSE and CNPase. The present study provides the first data revealing the effect of oxytocin on the expression of neuron and glial markers in the brain. It may be suggested that the oxytocin system is involved in the regulation of development of neuronal precursor cells in the brain.
- MeSH
- 2',3'-cyklické nukleotidfosfodiesterasy genetika MeSH
- antigeny diferenciační myelomonocytární genetika MeSH
- CD antigeny genetika MeSH
- hipokampus cytologie účinky léků metabolismus MeSH
- krysa rodu rattus MeSH
- neuroglie cytologie účinky léků metabolismus MeSH
- neurony cytologie účinky léků metabolismus MeSH
- oxytocin farmakologie MeSH
- potkani Wistar MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Although many studies have demonstrated the role of prolactin in the central nervous system, there is a considerable lack of known effects of prolactin on the parameters of neurogenesis and neuronal differentiation. The aim of the present study was to test whether prolactin changes gene expression and protein levels of nestin and microtubule-associated protein 2 (MAP2) in neuroblastoma (SK-N-SH) and glioblastoma (U-87MG) cells. Nestin and MAP2 represent cytoskeletal proteins associated with neuronal differentiation and they contribute to radial growth of the axons, dendrites and glial processes. SK-N-SH and U-87MG cells were exposed to prolactin (10 nM) for 48 h. Total mRNA was extracted. After reverse transcription, qPCR with specific primers for nestin and MAP2 was performed. The levels of proteins were measured by the In-Cell Western assay. Mitochondrial activity test was used to evaluate the viability of cells under the influence of prolactin. Incubation with 10 nM prolactin did not change the viability, either in SK-N-SH or in U-87MG cells. Prolactin significantly increased the gene expression and protein levels of both nestin and MAP2 in SK-N-SH cells, while no significant changes were observed in U-87MG cells. The presented data suggest that prolactin is linked to the regulation of cytoskeletal proteins in the neuronal type of cells and might be important for their differentiation.
- MeSH
- cytoskeletální proteiny biosyntéza genetika MeSH
- glioblastom patologie MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- nádorové proteiny biosyntéza genetika MeSH
- nestin biosyntéza genetika MeSH
- neuroblastom patologie MeSH
- neurogeneze účinky léků MeSH
- neuroglie účinky léků metabolismus MeSH
- neurony účinky léků metabolismus MeSH
- orgánová specificita MeSH
- prolaktin farmakologie MeSH
- proteiny asociované s mikrotubuly biosyntéza genetika MeSH
- regulace genové exprese u nádorů účinky léků MeSH
- rekombinantní proteiny farmakologie MeSH
- upregulace účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
Brain development is determined by neuronal differentiation including changes of cell polarity and asymetric grow- th of neuronal processes. Although, there are many unkown factors contributing to changes of lenght of neuronal cones, mounting experimental and review papers focus on changes of growth conus and role of axonal transport. In particular, mechanisms of actin/microtubule polymerisation and depolymerisation are important. Role of intracellu- lar calcium is also significant. Normal and properly timed changes of lenght of axons and dendrites are dependent on interaction of neurons and glia. Moreover, regeneration of injured axons is dependent on growth factors secre- ted from glial cells. The aim of the present study is characterisation of the most important mechanisms underlying changes of lenght of neurites.
- Klíčová slova
- kofilin,
- MeSH
- aktiny biosyntéza MeSH
- axony fyziologie MeSH
- dendrity fyziologie MeSH
- dyneiny biosyntéza fyziologie MeSH
- lidé MeSH
- mikrotubuly fyziologie MeSH
- morfogeneze MeSH
- mozková kůra růst a vývoj MeSH
- myosiny biosyntéza fyziologie MeSH
- nervový růstový faktor MeSH
- nervový systém - fyziologické jevy MeSH
- neurity MeSH
- neuroglie cytologie MeSH
- neurony fyziologie MeSH
- růst MeSH
- synapse MeSH
- vápník zásobování a distribuce MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
- Publikační typ
- abstrakt z konference MeSH
- Publikační typ
- abstrakt z konference MeSH
Objective of this study was to characterize osmotically-induced insulin secretion in two tumor cell lines. We compared response of freshly isolated rat pancreatic islets and INS-1 and INS-1E tumor cell lines to high glucose, 30 % hypotonic medium and 20 % hypertonic medium. In Ca2+-containing medium glucose induced insulin release in all three cell types. Hypotonicity induced insulin secretion from islets and INS-1 cells but not from INS-1E cells, in which secretion was inhibited despite similar increase in cell volume in both cell types. GdCl3 (100 µmol/l) did not affect insulin response from INS-1E cells to hypotonic challenge. Hypertonic medium inhibited glucose-induced insulin secretion from islets but not from tumor cells. Noradrenaline (1 µmol/l) inhibited glucose-induced but not swelling-induced insulin secretion from INS-1 cells. Surprisingly, perifusion with Ca2+-depleted medium showed distinct secretory response of INS-1E cells to hypotonicity while that of INS-1 cells was partially inhibited. Functioning glucose-induced insulin secretion is not sufficient prerequisite for hypotonicity-induced response in INS- 1E cells suggesting that swelling-induced exocytosis is not essential step in the mechanism mediating glucose-induced insulin secretion. Both cell lines are resistant to inhibitory effect of hyperosmolarity on glucose-induced insulin secretion. Response of INS-1E cells to hypotonicity is inhibited by the presence of Ca2+ in medium.
- Klíčová slova
- Cell volume, Insulin secretion, Pancreatic islets, Tumor cell line, Ca2+ and exocytosis,
- MeSH
- exocytóza MeSH
- financování organizované MeSH
- gadolinium farmakologie MeSH
- glukosa metabolismus MeSH
- hypertonické roztoky MeSH
- hypotonické roztoky MeSH
- inzulin sekrece MeSH
- inzulinom metabolismus sekrece MeSH
- krysa rodu rattus MeSH
- Langerhansovy ostrůvky metabolismus sekrece účinky léků MeSH
- nádorové buněčné linie MeSH
- nádory slinivky břišní metabolismus sekrece MeSH
- noradrenalin metabolismus MeSH
- osmotický tlak MeSH
- potkani Wistar MeSH
- vápník metabolismus nedostatek MeSH
- velikost buňky MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
