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Subventricular zone neural progenitors reverse TNF-alpha effects in cortical neurons.

Morini R, Ghirardini E, Butti E, Verderio C, Martino G, Matteoli M - Stem Cell Res Ther (2015)

Bottom Line: In this study, we demonstrate that aNPCs from the subventricular zone reverse the effects induced by the cytokine.Moreover, we show that the effect of aNPCs on cortical neurons is mediated by cannabinoid CB1 receptor activation.These data suggest that the role of aNPCs in preventing excitatory neurotransmission potentiation induced by TNFα on cortical neurons may have important implications for pathologies characterized by an inflammatory component affecting cortical neurons such as Alzheimer's disease.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Biotechnology and Traslational Medicine, University of Milano, via Vanvitelli 32, 20129, Milan, Italy. raffaella.morini@humanitasresearch.it.

ABSTRACT

Introduction: Tumor necrosis factor alpha (TNFα) plays a physiological role in controlling synaptic transmission and plasticity in the healthy central nervous system by modulating glutamate receptor trafficking to the plasma membrane. TNFα expression is also rapidly induced in response to tissue injury and infection. By promoting the insertion of Ca(2+) permeable-AMPA receptors into the neuronal plasma membrane, this cytokine may cause excessive Ca(2+) influx into neurons, thus enhancing neuronal death.

Methods: Primary cultures of cortical neurons were obtained from E18 foetal mice and incubated for 24 h with adult neural stem cells (aNPCs) either stimulated with lipopolysaccharide (LPS(+)aNPCs) or not (aNPCs). Cultures were treated with TNFα (100 ng/ml), and electrophysiological recordings were performed in different conditions to evaluate the effect of the cytokine on neuronal transmission.

Results: In this study, we demonstrate that aNPCs from the subventricular zone reverse the effects induced by the cytokine. Moreover, we show that the effect of aNPCs on cortical neurons is mediated by cannabinoid CB1 receptor activation.

Conclusion: These data suggest that the role of aNPCs in preventing excitatory neurotransmission potentiation induced by TNFα on cortical neurons may have important implications for pathologies characterized by an inflammatory component affecting cortical neurons such as Alzheimer's disease.

No MeSH data available.


Related in: MedlinePlus

aNPCs treated with LPS prevent the increase in mEPSC frequency produced by TNFα in cortical neurons. a Representative fluorescence microscopy images of cortical neurons incubated for 24 h with aNPCs either lipopolysaccharide-stimulated (LPS+aNPCs) or not (aNPCs) (green cells). The right panel shows higher magnification (scale bar: 100 μm; 50 μm). Only cortical neurons surrounded by aNPCs were selected to record (white arrows). b Details of (a) show GFP-positive membranous structures of small and medium size resembling round membrane vesicles. c Representative traces of voltage-clamp recordings of mEPSCs from cultured cortical neurons in control conditions or from neurons treated with LPS+aNPCs/aNPCs, TNFα, or LPS+aNPCs/aNPCs with TNFα (100 ng/ml 30 min) (calibration bars: 20 pA, 200 ms). d Group data of average mEPSC amplitude and frequency of control (n = 15), LPS+aNPCs-treated (n = 12), aNPCs-untreated (n = 8), TNFα-treated (n = 13), LPS+aNPC and TNFα-treated (n = 10), and aNPC-untreated and TNFα-treated (n = 8) cells. The increase in mEPSC frequency due to TNFα is completely reverted by LPS+aNPCs (P < 0.05). The increase in mEPSC amplitude due to TNFα is completely reverted by aNPC treatment (P < 0.05). A clear, though not significant, tendency toward reduction is detected in mEPSC amplitude or frequency in neuronal cultures exposed to LPS+aNPCs or aNPCs, respectively. All data are expressed as mean ± standard error of the mean. Statistical test: one-way analysis of variance, Dunn’s multiple comparisons test. aNPC adult neural stem cell, GFP green fluorescent protein, LPS lipopolysaccharide, LPS+aNPC lipopolysaccharide-stimulated adult neural stem cell, mEPSC miniature excitatory post-synaptic current, TNFα tumor necrosis factor alpha
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Fig2: aNPCs treated with LPS prevent the increase in mEPSC frequency produced by TNFα in cortical neurons. a Representative fluorescence microscopy images of cortical neurons incubated for 24 h with aNPCs either lipopolysaccharide-stimulated (LPS+aNPCs) or not (aNPCs) (green cells). The right panel shows higher magnification (scale bar: 100 μm; 50 μm). Only cortical neurons surrounded by aNPCs were selected to record (white arrows). b Details of (a) show GFP-positive membranous structures of small and medium size resembling round membrane vesicles. c Representative traces of voltage-clamp recordings of mEPSCs from cultured cortical neurons in control conditions or from neurons treated with LPS+aNPCs/aNPCs, TNFα, or LPS+aNPCs/aNPCs with TNFα (100 ng/ml 30 min) (calibration bars: 20 pA, 200 ms). d Group data of average mEPSC amplitude and frequency of control (n = 15), LPS+aNPCs-treated (n = 12), aNPCs-untreated (n = 8), TNFα-treated (n = 13), LPS+aNPC and TNFα-treated (n = 10), and aNPC-untreated and TNFα-treated (n = 8) cells. The increase in mEPSC frequency due to TNFα is completely reverted by LPS+aNPCs (P < 0.05). The increase in mEPSC amplitude due to TNFα is completely reverted by aNPC treatment (P < 0.05). A clear, though not significant, tendency toward reduction is detected in mEPSC amplitude or frequency in neuronal cultures exposed to LPS+aNPCs or aNPCs, respectively. All data are expressed as mean ± standard error of the mean. Statistical test: one-way analysis of variance, Dunn’s multiple comparisons test. aNPC adult neural stem cell, GFP green fluorescent protein, LPS lipopolysaccharide, LPS+aNPC lipopolysaccharide-stimulated adult neural stem cell, mEPSC miniature excitatory post-synaptic current, TNFα tumor necrosis factor alpha

Mentions: Both injury and neurodegenerative diseases increase the amount of TNFα in the brain and this contributes to neuronal death. Recent studies demonstrated that SVZ-derived aNPCs protect striatal neurons from glutamate-mediated toxicity in pathological conditions associated with inflammation and excitotoxicity, such as epilepsy and ischemic stroke [24]. To assess whether the neuroprotective effects of aNPCs extend to different neuronal types, we exposed neurons to TNFα before or after incubating them for 24 h with LPS+aNPCs in order to mimic the effects of different danger (or recognition) signals triggering the inflammatory response. aNPCs were infected in order to express GFP [29], and only neurons surrounded by aNPCs were selected to record (Fig. 2a). Notably, membranous structures of small and medium size, including round membrane vesicles, were detected on the cell surface of aNPCs (Fig. 2b left panel), as recently described [33]. These vesicles were released by aNPCs (Fig. 2b middle panel) and adhered to GFP-negative neuronal cells (Fig. 2b right panel).Fig. 2


Subventricular zone neural progenitors reverse TNF-alpha effects in cortical neurons.

Morini R, Ghirardini E, Butti E, Verderio C, Martino G, Matteoli M - Stem Cell Res Ther (2015)

aNPCs treated with LPS prevent the increase in mEPSC frequency produced by TNFα in cortical neurons. a Representative fluorescence microscopy images of cortical neurons incubated for 24 h with aNPCs either lipopolysaccharide-stimulated (LPS+aNPCs) or not (aNPCs) (green cells). The right panel shows higher magnification (scale bar: 100 μm; 50 μm). Only cortical neurons surrounded by aNPCs were selected to record (white arrows). b Details of (a) show GFP-positive membranous structures of small and medium size resembling round membrane vesicles. c Representative traces of voltage-clamp recordings of mEPSCs from cultured cortical neurons in control conditions or from neurons treated with LPS+aNPCs/aNPCs, TNFα, or LPS+aNPCs/aNPCs with TNFα (100 ng/ml 30 min) (calibration bars: 20 pA, 200 ms). d Group data of average mEPSC amplitude and frequency of control (n = 15), LPS+aNPCs-treated (n = 12), aNPCs-untreated (n = 8), TNFα-treated (n = 13), LPS+aNPC and TNFα-treated (n = 10), and aNPC-untreated and TNFα-treated (n = 8) cells. The increase in mEPSC frequency due to TNFα is completely reverted by LPS+aNPCs (P < 0.05). The increase in mEPSC amplitude due to TNFα is completely reverted by aNPC treatment (P < 0.05). A clear, though not significant, tendency toward reduction is detected in mEPSC amplitude or frequency in neuronal cultures exposed to LPS+aNPCs or aNPCs, respectively. All data are expressed as mean ± standard error of the mean. Statistical test: one-way analysis of variance, Dunn’s multiple comparisons test. aNPC adult neural stem cell, GFP green fluorescent protein, LPS lipopolysaccharide, LPS+aNPC lipopolysaccharide-stimulated adult neural stem cell, mEPSC miniature excitatory post-synaptic current, TNFα tumor necrosis factor alpha
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Related In: Results  -  Collection

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Fig2: aNPCs treated with LPS prevent the increase in mEPSC frequency produced by TNFα in cortical neurons. a Representative fluorescence microscopy images of cortical neurons incubated for 24 h with aNPCs either lipopolysaccharide-stimulated (LPS+aNPCs) or not (aNPCs) (green cells). The right panel shows higher magnification (scale bar: 100 μm; 50 μm). Only cortical neurons surrounded by aNPCs were selected to record (white arrows). b Details of (a) show GFP-positive membranous structures of small and medium size resembling round membrane vesicles. c Representative traces of voltage-clamp recordings of mEPSCs from cultured cortical neurons in control conditions or from neurons treated with LPS+aNPCs/aNPCs, TNFα, or LPS+aNPCs/aNPCs with TNFα (100 ng/ml 30 min) (calibration bars: 20 pA, 200 ms). d Group data of average mEPSC amplitude and frequency of control (n = 15), LPS+aNPCs-treated (n = 12), aNPCs-untreated (n = 8), TNFα-treated (n = 13), LPS+aNPC and TNFα-treated (n = 10), and aNPC-untreated and TNFα-treated (n = 8) cells. The increase in mEPSC frequency due to TNFα is completely reverted by LPS+aNPCs (P < 0.05). The increase in mEPSC amplitude due to TNFα is completely reverted by aNPC treatment (P < 0.05). A clear, though not significant, tendency toward reduction is detected in mEPSC amplitude or frequency in neuronal cultures exposed to LPS+aNPCs or aNPCs, respectively. All data are expressed as mean ± standard error of the mean. Statistical test: one-way analysis of variance, Dunn’s multiple comparisons test. aNPC adult neural stem cell, GFP green fluorescent protein, LPS lipopolysaccharide, LPS+aNPC lipopolysaccharide-stimulated adult neural stem cell, mEPSC miniature excitatory post-synaptic current, TNFα tumor necrosis factor alpha
Mentions: Both injury and neurodegenerative diseases increase the amount of TNFα in the brain and this contributes to neuronal death. Recent studies demonstrated that SVZ-derived aNPCs protect striatal neurons from glutamate-mediated toxicity in pathological conditions associated with inflammation and excitotoxicity, such as epilepsy and ischemic stroke [24]. To assess whether the neuroprotective effects of aNPCs extend to different neuronal types, we exposed neurons to TNFα before or after incubating them for 24 h with LPS+aNPCs in order to mimic the effects of different danger (or recognition) signals triggering the inflammatory response. aNPCs were infected in order to express GFP [29], and only neurons surrounded by aNPCs were selected to record (Fig. 2a). Notably, membranous structures of small and medium size, including round membrane vesicles, were detected on the cell surface of aNPCs (Fig. 2b left panel), as recently described [33]. These vesicles were released by aNPCs (Fig. 2b middle panel) and adhered to GFP-negative neuronal cells (Fig. 2b right panel).Fig. 2

Bottom Line: In this study, we demonstrate that aNPCs from the subventricular zone reverse the effects induced by the cytokine.Moreover, we show that the effect of aNPCs on cortical neurons is mediated by cannabinoid CB1 receptor activation.These data suggest that the role of aNPCs in preventing excitatory neurotransmission potentiation induced by TNFα on cortical neurons may have important implications for pathologies characterized by an inflammatory component affecting cortical neurons such as Alzheimer's disease.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Biotechnology and Traslational Medicine, University of Milano, via Vanvitelli 32, 20129, Milan, Italy. raffaella.morini@humanitasresearch.it.

ABSTRACT

Introduction: Tumor necrosis factor alpha (TNFα) plays a physiological role in controlling synaptic transmission and plasticity in the healthy central nervous system by modulating glutamate receptor trafficking to the plasma membrane. TNFα expression is also rapidly induced in response to tissue injury and infection. By promoting the insertion of Ca(2+) permeable-AMPA receptors into the neuronal plasma membrane, this cytokine may cause excessive Ca(2+) influx into neurons, thus enhancing neuronal death.

Methods: Primary cultures of cortical neurons were obtained from E18 foetal mice and incubated for 24 h with adult neural stem cells (aNPCs) either stimulated with lipopolysaccharide (LPS(+)aNPCs) or not (aNPCs). Cultures were treated with TNFα (100 ng/ml), and electrophysiological recordings were performed in different conditions to evaluate the effect of the cytokine on neuronal transmission.

Results: In this study, we demonstrate that aNPCs from the subventricular zone reverse the effects induced by the cytokine. Moreover, we show that the effect of aNPCs on cortical neurons is mediated by cannabinoid CB1 receptor activation.

Conclusion: These data suggest that the role of aNPCs in preventing excitatory neurotransmission potentiation induced by TNFα on cortical neurons may have important implications for pathologies characterized by an inflammatory component affecting cortical neurons such as Alzheimer's disease.

No MeSH data available.


Related in: MedlinePlus