G-protein coupled receptors (GPCRs) exist in an equilibrium of multiple conformational states, including different active states, which depend on the nature of the bound ligand. In consequence, different conformational states can initiate specific signal transduction pathways. The study identified compound 7e, which acts as a potent 5-hydroxytryptamine type 6 receptor (5-HT6R) neutral antagonist at Gs and does not impact neurite growth (process controlled by Cdk5). MD simulations highlighted receptor conformational changes for 7e and inverse agonist PZ-1444. In cell-based assays, neutral antagonists of the 5-HT6R (7e and CPPQ), but not inverse agonists (SB-258585, intepirdine, PZ-1444), displayed glioprotective properties against 6-hydroxydopamine-induced and doxorubicin-induced cytotoxicity. These suggest that targeting the activated conformational state of the 5-HT6R with neutral antagonists implicates the protecting properties of astrocytes. Additionally, 7e prevented scopolamine-induced learning deficits in the novel object recognition test in rats. We propose 7e as a probe for further understanding of the functional outcomes of different states of the 5-HT6R.
- MeSH
- agonisté serotoninových receptorů farmakologie MeSH
- antagonisté serotoninu chemická syntéza farmakologie MeSH
- astrocyty účinky léků MeSH
- imidazoly chemická syntéza farmakologie MeSH
- krysa rodu rattus MeSH
- lidé MeSH
- molekulární konformace MeSH
- neurity účinky léků MeSH
- neuroglie účinky léků MeSH
- neuroprotektivní látky chemická syntéza farmakologie MeSH
- poruchy učení chemicky indukované prevence a kontrola MeSH
- potkani Sprague-Dawley MeSH
- pyridiny chemická syntéza farmakologie MeSH
- receptory serotoninové účinky léků MeSH
- receptory spřažené s G-proteiny účinky léků MeSH
- vztahy mezi strukturou a aktivitou MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- lidé MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
In the field of tissue engineering, much research has been devoted to the surface topography of conductive materials. However, less work has been carried out on how the electrical stimulation of such materials influences nerve regeneration. Here, we investigated the effect of electrical stimulation on randomly- and uniaxially-aligned polypyrrole-coated cellulose acetate butyrate (PPy/CAB) nanofibers. First, SEM revealed that the conducting PPy coverage resulted in dramatic changes to the nanofiber morphology. In turn, these changes led to an increase in the sample wettability. Fourier transform spectroscopy (FTIR) and X-ray photoelectron spectroscopy (XPS) confirmed the presence of a PPy layer. Second, human neuroblastoma cells (SH-SY5Y) were seeded on the PPy/CAB nanofibers and stimulated by 100 mV mm-1 at 1 Hz pulses in vitro. We demonstrated that either with or without this electrical stimulation both nanofiber alignment and PPy coverage had a strong influence on cell morphology and attachment. Moreover, fluorescence microscopy revealed that the cells stimulated on PPy/CAB had longer neurite outgrowth. Collectively, our results shed light on the combined effect of scaffold morphology and external stimulation on neuronal cell behavior.
- MeSH
- buněčná adheze účinky léků MeSH
- celulosa analogy a deriváty farmakologie toxicita MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- nanovlákna chemie toxicita MeSH
- neurity účinky léků MeSH
- neuronální růst účinky léků MeSH
- polymery farmakologie toxicita MeSH
- proliferace buněk účinky léků MeSH
- pyrroly farmakologie toxicita MeSH
- smáčivost MeSH
- viabilita buněk účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
4,5-Dihydroxyanthraquinone-2-carboxylic acid (Rhein) has been shown to have various physiological and pharmacological properties including anticancer activity and modulatory effects on bioenergetics. In this study, we explored the impact of rhein on protein profiling of undifferentiated (UC) and differentiated (DC) SH-SY5Y cells. Besides that, the cellular morphology and expression of differentiation markers were investigated to determine the effect of rhein on retinoic acidinduced neuronal cell differentiation. Using two-dimensional gel electrophoresis and matrix-assisted laser desorption/ ionization-time-of-flight mass spectrometry we evaluated the changes in the proteome of both UC and DC SH-SY5Y cells after 24 h treatment with rhein. Validation of selected differentially expressed proteins and the assessment of neuronal differentiation markers were performed by western blotting. Proteomic analysis revealed significant changes in the abundance of 15 proteins linked to specific cellular processes such as cytoskeleton structure and regulation, mitochondrial function, energy metabolism, protein synthesis and neuronal plasticity. We also observed that the addition of rhein to the cultured cells during differentiation resulted in a significantly reduced neurite outgrowth and decreased expression of neuronal markers. These results indicate that rhein may strongly interfere with the differentiation process of SH-SY5Y neuroblastoma cells and is capable of inducing marked proteomic changes in these cells.
- MeSH
- anthrachinony farmakologie MeSH
- buněčná diferenciace účinky léků MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- nervové kmenové buňky účinky léků MeSH
- neurity účinky léků patologie MeSH
- neuroblastom farmakoterapie genetika patologie MeSH
- neuronální růst účinky léků MeSH
- neurony účinky léků MeSH
- proteomika * MeSH
- regulace genové exprese u nádorů účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Chondroitin sulfate (CS) proteoglycans in perineuronal nets (PNNs) from the central nervous system (CNS) are involved in the control of plasticity and memory. Removing PNNs reactivates plasticity and restores memory in models of Alzheimer's disease and ageing. Their actions depend on the glycosaminoglycan (GAG) chains of CS proteoglycans, which are mainly sulfated in the 4 (C4S) or 6 (C6S) positions. While C4S is inhibitory, C6S is more permissive to axon growth, regeneration and plasticity. C6S decreases during critical period closure. We asked whether there is a late change in CS-GAG sulfation associated with memory loss in aged rats. Immunohistochemistry revealed a progressive increase in C4S and decrease in C6S from 3 to 18 months. GAGs extracted from brain PNNs showed a large reduction in C6S at 12 and 18 months, increasing the C4S/C6S ratio. There was no significant change in mRNA levels of the chondroitin sulfotransferases. PNN GAGs were more inhibitory to axon growth than those from the diffuse extracellular matrix. The 18-month PNN GAGs were more inhibitory than 3-month PNN GAGs. We suggest that the change in PNN GAG sulfation in aged brains renders the PNNs more inhibitory, which lead to a decrease in plasticity and adversely affect memory.
- MeSH
- buněčné extrakty MeSH
- krysa rodu rattus MeSH
- messenger RNA MeSH
- mozek metabolismus MeSH
- nervová síť fyziologie MeSH
- neurity účinky léků MeSH
- poruchy paměti etiologie MeSH
- proteoglykany metabolismus MeSH
- regulace genové exprese fyziologie MeSH
- stárnutí fyziologie MeSH
- sulfotransferasy genetika metabolismus MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- MeSH
- antitumorózní látky MeSH
- cisplatina farmakologie MeSH
- karboplatina farmakologie MeSH
- krysa rodu rattus MeSH
- neurity růst a vývoj účinky léků MeSH
- spinální ganglia růst a vývoj účinky léků MeSH
- techniky in vitro MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
- Publikační typ
- srovnávací studie MeSH
