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Cholinergic effects mediated by M2 muscarinic receptor in human Schwann-like cells induced from adipose mesenchymal stem cells
No full textSchwann cells (SCs) have an important role in peripheral nerve regeneration but there are several restrictions hindering their clinical application. Adipose derived stem cells (ASCs) feature good properties for cell therapies. When exposed to specific growth factors in vitro, they can acquire a SC-like phenotype (dASCs), expressing key SCs markers. Our group has demonstrated that M2 muscarinic receptor in rat dASCs causes a reversible arrest of cell proliferation, increasing SCs myelinating phenotype. Human dASCs, as rat dASCs, express muscarinic receptors. In the present work we evaluate if M2 muscarinic receptor activation may contribute to human dASCs proliferation, migration and phenotype. M2 selective activation by selective agonist Arecaidine Propargyl Ester (APE) causes a decreased cell proliferation, modulating the expression of genes involved in the proliferation/differentiation (i.e. c-jun and egr2) and several neurotrophic factors. Moreover, M2 selective activation is able to decrease cell migration. Although further analyses are needed to best characterise the role of M2 receptor, these results are the first evidence that its selective activation may have effects also on human dASCs proliferation and migration. This may improve our knowledge of these promising therapeutic cells for potential use in nerve regeneration
Muscarinic receptor in Schwann-like adipose derived stem cells: implications in nerve regeneration.
No full textSchwann Cells (SCs) play a central role in the physiology and in the response of the axon injury. The capacity of SCs to proliferate, to secrete growth factors, to modulate immune response, to migrate and to re-myelinate regenerating nerves have been reported (Jessen et al, 2016). However, SCs present limited clinical application, such as the difficulty in collection and culture and the slow rate of in vitro expansion.
Some papers describe that Adipose-derived stem cells (ASCs) have the ability to differentiate towards SCs phenotype (Schwann-like, dASC) following exposure to suitable culture media (Kingham et al, 2007). dASC, like SCs, express functional receptors for different neurotrasmitters, including muscarinic receptor subtypes (M1-M4) that regulate some physiological events.
In the present work, we have characterised the effects mediated by muscarinic receptors on proliferation and neurotrophic factors (NFs) expression and production
Cholinergic system and neuroinflammation: Implication in multiple sclerosis.
No full textMultiple sclerosis (MS) is an inflammatory and neurodegenerative disease of the central nervous system (CNS) characterized by leucocytes infiltration, demyelination, axonal degeneration and neuronal death. Although the etiology of MS is still unknown, inflammation and autoimmunity are considered to be key players of the disease. The severe alterations affecting the nervous system contribute to the motor and cognitive disabilities and are in large part dependent on severe inflammatory processes active in both central nervous system and immune system. Acetylcholine (ACh) appears to be involved in the modulation of central and peripheral inflammation. Immune cells as well as astrocytes and microglia respond to ACh stimuli by activation of cholinergic receptors. Muscarinic and nicotinic receptors differently contribute to the modulation of immunological and inflammatory processes stimulating pro- and anti-inflammatory cytokines respectively. The role played by ACh in MS is not yet fully understood, although some results point to its involvement in different neurological disorders such as Alzheimer's disease and schizophrenia. In the present review we summarize the evidence indicating the correlation between nervous system dysfunction in MS, with inflammation and cholinergic system alterations. Experiments performed in EAE models and analyses on biological fluids from MS patients such as blood, serum and cerebrospinal fluid suggest that cholinergic alterations may contribute to the dysregulated inflammatory processes of MS. Many current therapeutic approaches in MS are based on anti-inflammatory drugs. We also discuss how the use of cholinesterase inhibitors or ACh mimetics may represent a new interesting therapeutic approach in MS
Muscarinic cholinergic receptors in dorsal root ganglia of chick embryo: a radioligand binding and immunocytochemical study.
No full textThe presence and micro-anatomical localization of muscarinic cholinergic receptors were assessed in dorsal root ganglia of chick embryo during development using radioligand binding and immunocytochemical techniques, respectively. The non-selective muscarinic cholinergic receptor radioligand [3H]quinuclidinyl benzilate was specifically bound to sections of chick dorsal root ganglia with a dissociation constant value (Kd) of 0.75 +/- 0.02 nM and a maximum density of binding sites (Bmax) of 7.2 +/- 0.5 fmol/mg tissue. [3H]Quinuclidinyl benzilate binding was partially sensitive to pirenzepine displacement. This suggests that muscarinic cholinergic receptors expressed by dorsal root ganglia of chick embryo at least in part belong to the M1 muscarinic receptor subtype. Immunocytochemical analysis confirmed the presence of muscarinic receptors in the ganglia. These findings suggest that neurons of dorsal root ganglia, which are known to express cholinergic markers such as choline acetyltransferase, acetylcholinesterase and high affinity choline uptake, are also cholinoceptive
Muscarinic receptor activation modulates neurotrophic factors production in rat Schwann-like cells derived from adipose mesenchymal stem cells.
No full textMuscarinic receptor activation modulates neurotrophic factors production in rat Schwann-like cells derived from adipose mesenchymal stem cells
Piovesana R1, Faroni A2, Soligo M3, Manni L3, Reid AJ2 & Tata AM1
1Dept. Biol and Biotech. C. Darwin, University of Rome “Sapienza”, Rome, Italy; 2Blond McIndoe Lab, Faculty of Biology, Medicine and Health, University of Manchester, Manchester, UK; 3Institute of Translational Pharmacology-CNR, Rome, Italy
Peripheral nerve injury is commonly caused by direct mechanical trauma. Regeneration depends on the ability of Schwann cells (SCs) to create a favourable environment, by producing neurotrophic factors. Although SCs are effective in promoting nerve regeneration, they are not a convenient source of transplantable cells to improve outcomes after injury. Mesenchymal Stem Cells derived from adipose tissue (ASCs) seem to be a promising alternative source considering their ability to differentiate towards SC phenotype (Schwann-like).
SCs express different receptors for neurotransmitters. In particular cholinergic stimulation of M2 muscarinic receptor decreases SCs proliferation whilst upregulating myelination. Previously, we demonstrated that Schwann-like cells express muscarinic receptors; in particular the M2 receptor activation resulted in decreased proliferation and reduced migration.
In present work, we have characterised the effects mediated by muscarinic receptors on neurotrophic factors (NFs) expression and production. The selective activation of M2 receptors by arecaidine propargyl ester (APE) caused a significant decrease of the transcript levels for NFs (NGF, BDNF and GDNF), while the non-selective agonist muscarine did not influence NFs mRNA expression. By custom made Elisa Assay, we analysed the production of two different NGF forms, precursor (proNGF) and mature NGF (mNGF). APE treatment induced a decreased release of both NGF forms, whereas muscarine treatment stimulated an increased release of mNGF. Western blot analysis indicated that both agonists caused a significant decrease in the expression of the proNGF isoform at 25 kDa, which is likely involved in the modulation of apoptotic processes.
The data obtained suggest a relevant role of muscarinic receptors in the modulation of NFs production in Schwann-like cells. In particular the ability of both muscarinic agonists to negatively modulate the proNGF isoform, thereby suggesting a neuroprotective role of muscarinic receptors towards regenerating axons
Muscarinic receptor activation modulates proliferation and neurotrophic factors production in rat Schwann-like cells: implications in nerve regeneration.
No full textMuscarinic receptor activation modulates proliferation and neurotrophic factors production in rat Schwann-like cells: implications in nerve regeneration.
Theme B: Neural Excitability, Synapses, and Glia: Cellular Mechanisms
*Roberta Piovesana1,2, Alessandro Faroni2, Marzia Soligo3, Alessandro Matera1, Luigi Manni3, Valerio Magnaghi4, Adam J Reid2 & Ada Maria Tata1
1Dept. Biol and Biotech. C. Darwin, University of Rome “Sapienza”, Rome, Italy;
2Blond McIndoe Lab, Faculty of Biology, Medicine and Health, University of Manchester, Manchester, UK;
3Institute of Translational Pharmacology-CNR, Rome, Italy;
4Dept.of Pharmacol and Biomol Sciences, Univ of Milan, Milan, Italy
Introduction: Peripheral nerve injury is caused by direct mechanical trauma or surgical resection secondary to tumor excision. Patients show sensory and motor loss and chronic pain. The peripheral nervous system has an intrinsic regeneration capability; nevertheless, full functional recovery is limited. Schwann Cells (SCs) play a pivotal role in the response of the axon injury, thus, SCs are an attractive therapeutic target. In fact SCs enhance nerve regeneration, producing neurotrophic factors (NFs). Unfortunately, SCs have some clinical limitations, such as the difficulty in collection and culture and a slow proliferation rate. The ideal transplantable cells should be easily accessible, proliferate rapidly in culture and successfully integrate into host tissue.
Methods: Adipose-derived stem cells (ASCs) can be differentiated in SC phenotype (Schwann-like, dASCs) following exposure to a growth factor cocktail. Gene expression was analysed by Real Time PCR; 25 kDa proNGF isoform was detected by Western Blotting analysis. By custom made Elisa Assay, the production of two different NGF forms, precursor (proNGF) and mature NGF (mNGF), have been detected. The cell migration has been analysed by wound healing assay.
Results: SCs express receptors for different neurotransmitters. Rat SCs express different muscarinic receptor subtypes. In particular, the most expressed is M2 receptor. Its activation causes a negative effect on SC proliferation, upregulating transcription factors involved in the promyelinating phase (e.g., Sox10 and Krox20) and downregulating proteins involved in the maintenance of the proliferative state (c-jun, Notch-1).
dASCs, like SCs, express functional receptors for different neurotrasmitters, including all muscarinic receptor subtypes. In present work, we characterized the effects mediated by M2 receptors in rat dASCs. As for SCs, M2 receptor activation caused a reversible decrease of cell proliferation and the inhibition of cell migration without affecting cell survival. After 24hs of M2 agonist treatment (Arecaidine propargyl ester, APE), it has been observed a significant decrease of NF expression (i.e. NGF, BDNF and GDNF) and an increase of P0 transcript level. APE treatment induced a decreased release of both NGF forms, whereas the non-selective agonist muscarine stimulated an increased release of mNGF. Both agonists caused a significant decreased expression of the 25 kDa proNGF isoform, which is involved in the modulation of apoptotic processes. Furthermore, the selective activation of M2 receptors enhanced a pronounced spindle shaped morphology in dASCs similarly to native SCs.
Conclusions: Our data demonstrate that M2 receptor activity inhibits dASCs proliferation and migration and it could improve their differentiation. Conversely muscarinic receptors activation positively modulates NFs production. These data suggest that cholinergic agonists may differently contribute to the dASC functions during nerve regeneration
Moderate Static Magnetic Field (6 mT)-induced lipid rafts rearrangement increases silver NPs uptake in Human Lymphocytes
No full textNovel Therapeutic Delivery of Nanocurcumin in Central Nervous System Related Disorders
Full text linkNutraceuticals represent complementary or alternative beneficial products to the expensive and high-tech therapeutic tools in modern medicine. Nowadays, their medical or health benefits in preventing or treating different types of diseases is widely accepted, due to fewer side effects than synthetic drugs, improved bioavailability and long half-life. Among herbal and natural compounds, curcumin is a very attractive herbal supplement considering its multipurpose propertiesThe potential effects of curcumin on glia cells and its therapeutic and protective properties in central nervous system (CNS)-related disorders is relevant. However, curcumin is unstable and easily degraded or metabolized into other forms posing limits to its clinical development. This is particularly important in brain pathologies determined blood brain barrier (BBB) obstacle. To enhance the stability and bioavailability of curcumin, many studies focused on the design and development of curcumin nanodelivery systems (nanoparticles, micelles, dendrimers, and diverse nanocarriers)These nanoconstructs can increase curcumin stability, solubility, in vivo uptake, bioactivity and safetyRecently, several studies have reported on a curcumin exosome-based delivery system, showing great therapeutical potential. The present work aims to review the current available data in improving bioactivity of curcumin in treatment or prevention of neurological disorders
NGS e nuove tecnologie per il riposizionamento di farmaci e la terapia di precisione
No full textLe tcnologie "Next Generation Sequencing" (NGS) e le nuov tecnologie OMICS stanno permettendo la stratificazione di pazienti rispetto al genotipo non solo riguardante il difetto primario di malattie genetiche o acquisite, ma anche SNPs modificatori che potrebbero giocare un ruolo chiave nella risposta a terapie personalizzate in medicina di precisione. Inoltre, le metodologie basate sulla biopsia liquida potrebbero permettere un'analisi del DNA circolante e dell'RNA circolante (inclusi i microRNA) in grado insieme di conferire un quadro globale delle modificazioni dell'espressione genica in pazienti. Questo tipo di analisi globale potrebbe permettere di identificare da un lato nuovi bio-marcatori, e dall'altro nuove molecole potenzialmente bersaglio per lo sviluppo di nuove forme di terapia personalizzata. Se queste ricerche vengono combinate con risultati derivanti da trattamenti clinici, si potrebbe rendere sempre più possibile un approccio personalizzato alla terapia. Questo potrebbe giocare un ruolo molto importante anche nel riposizionamento di farmaci e nella loro valorizzazione
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