Monitoring Astrocyte Reactivity and Proliferation &lt;em&gt;in Vitro&lt;/em&gt; Under Ischemic-Like Conditions

Ferrer-Acosta, Yancy; Gonzalez-Vega, Maxine N.; Rivera-Aponte, David E.; Martinez-Jimenez, Solianne M.; Martins, Ana

doi:10.3791/55108

Cited by 9 publications

(7 citation statements)

References 38 publications

(11 reference statements)

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“…Note that we used the culture medium with a low serum concentration (only 0,4%) which is much less than usually used for glial cultures (10%) [ 67 – 69 ]. A neuron/glia ratio decreased after two weeks of culture of hippocampal cells with high density in the conditions of low serum concentration, indicating proliferation of glial cells [ 39 ].…”

Section: Discussionmentioning

confidence: 99%

Theta rhythm-like bidirectional cycling dynamics of living neuronal networks in vitro

et al. 2018

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The phenomena of synchronization, rhythmogenesis and coherence observed in brain networks are believed to be a dynamic substrate for cognitive functions such as learning and memory. However, researchers are still debating whether the rhythmic activity emerges from the network morphology that developed during neurogenesis or as a result of neuronal dynamics achieved under certain conditions. In the present study, we observed self-organized spiking activity that converged to long, complex and rhythmically repeated superbursts in neural networks formed by mature hippocampal cultures with a high cellular density. The superburst lasted for tens of seconds and consisted of hundreds of short (50–100 ms) small bursts with a high spiking rate of 139.0 ± 78.6 Hz that is associated with high-frequency oscillations in the hippocampus. In turn, the bursting frequency represents a theta rhythm (11.2 ± 1.5 Hz). The distribution of spikes within the bursts was non-random, representing a set of well-defined spatio-temporal base patterns or motifs. The long superburst was classified into two types. Each type was associated with a unique direction of spike propagation and, hence, was encoded by a binary sequence with random switching between the two “functional” states. The precisely structured bidirectional rhythmic activity that developed in self-organizing cultured networks was quite similar to the activity observed in the in vivo experiments.

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Section: Discussionmentioning

confidence: 99%

Theta rhythm-like bidirectional cycling dynamics of living neuronal networks in vitro

et al. 2018

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“…As described previously (Ferrer-Acosta et al, 2017), oxygenglucose deprivation/reoxygenation (OGD/R) is a classic in vitro model of I/R injury. Briefly, astrocytes were washed three times with glucose-free DMEM and cultured in the same medium in a hypoxia chamber with a mixture of 95% N 2 and 5% CO 2 for 12 h. Then, the astrocytes were cultured in normal DMEM medium and re-oxygenated under normoxic conditions (95% air, 5% CO 2 ) for 6 h.…”

Section: Oxygen-glucose Deprivation/reoxygenation (Ogd/r) In Vitro Modelmentioning

confidence: 99%

Vinpocetine Protects Against Cerebral Ischemia-Reperfusion Injury by Targeting Astrocytic Connexin43 via the PI3K/AKT Signaling Pathway

et al. 2020

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Vinpocetine (Vinp) is known for its neuroprotective properties. However, the protective mechanism of Vinp against cerebral ischemia/reperfusion (I/R) injury should be further explored. This study was designed to investigate the neuroprotective effects of Vinp against oxygen-glucose deprivation/reoxygenation (OGD/R) injury in vitro and cerebral I/R injury in vivo and explore whether this mechanism would involve enhancement of astrocytic connexin 43 (Cx43) expression via the phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) pathway. In vitro, we detected astrocytic viability and extracellular nitric oxide by an assay kit, intracellular reactive oxygen species by a DCFH-DA probe, inflammation and apoptosis-related protein expression by immunofluorescence staining, and the astrocytic apoptosis rate by flow cytometry. In vivo, we measured the cerebral infarction volume, superoxide dismutase activity, malondialdehyde content, and the expression of inflammation and apoptosis-related proteins. The results indicated that Vinp ameliorated the detrimental outcome of I/R injury. Vinp attenuated astrocytic injury induced by OGD/R and reduced cerebral infarction volume and cerebral edema in rats with cerebral I/R injury. Moreover, Vinp reduced oxidative stress, inflammation, and apoptosis induced by cerebral I/R injury in brain tissues. Meanwhile, Vinp increased p-Cx43 and p-AKT expression, and the p-Cx43/Cx43 and p-AKT/AKT ratio, which was decreased by cerebral I/R injury. Coadministration of PI3K inhibitors LY294002 and BKM120 blunted the effects of Vinp. This study suggests that Vinp protects against cerebral I/R injury via Cx43 phosphorylation by activating the PI3K/AKT pathway.

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“…The protocol of primary astrocyte culture was similar to that described previously [21]. Cortices from 1-4 days old rat pups were used for astrocyte isolation.…”

Section: Methodsmentioning

confidence: 99%

The Role of S100B/RAGE-Enhanced ADAM17 Activation in Endothelial Glycocalyx Shedding After Traumatic Brain Injury

Zou

et al. 2021

Preprint

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Background: Traumatic brain injury (TBI) remains one of the main causes for disability and death worldwide. While the primary mechanical injury cannot be avoided, the prevention of secondary injury is the focus of TBI research. Present study aimed to elucidate the effects and mechanisms of S100B and its receptor RAGE on mediating secondary injury after TBI. Methods: This study established TBI animal model by fluid percussion injury in rats, cell model by stretch-injured in astrocytes, and endothelial injury model with conditioned medium stimulation. Pharmacological intervention was applied to interfere the activities of S100B/RAGE/ADAM17 signaling pathway, respectively. The expressions or contents of S100B, RAGE, syndecan-1 and ADAM17 in brain and serum, as well as in cultured cells and medium, were detected by western blot. The distribution of relative molecules was observed with immunofluorescence. Results: We found that TBI could activate the release of S100B, mostly from astrocytes, and S100B and RAGE could mutually regulate their expression and activation. Most importantly, present study revealed an obvious increase of syndecan-1 in rat serum or in endothelial cultured medium after injury, and a significant decrease in tissue and in cultured endothelial cells, indicating TBI-induced shedding of endothelial glycocalyx. The data further proved that the activation of S100B/RAGE signaling could promote the shedding of endothelial glycocalyx by enhancing the expression, translocation and activity of ADAM17, an important sheddase, in endothelial cells. The damage of endothelial glycocalyx consequently aggravated blood brain barrier (BBB) dysfunction and systemic vascular hyper-permeability, overall resulting in secondary brain and lung injury. Conclusions: TBI triggers the activation of S100B/RAGE signal pathway. The regulation S100B/RAGE on ADAM17 expression, translocation and activation further promotes the shedding of endothelial glycocalyx, aggravates the dysfunction of BBB, and increases the vascular permeability, leading to secondary brain and lung injury. Present study may open a new corridor for the more in-depth understanding of the molecular processes responsible for cerebral and systemic vascular barrier impairment and secondary injury after TBI.

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Monitoring Astrocyte Reactivity and Proliferation <em>in Vitro</em> Under Ischemic-Like Conditions

Cited by 9 publications

References 38 publications

Theta rhythm-like bidirectional cycling dynamics of living neuronal networks in vitro

Theta rhythm-like bidirectional cycling dynamics of living neuronal networks in vitro

Vinpocetine Protects Against Cerebral Ischemia-Reperfusion Injury by Targeting Astrocytic Connexin43 via the PI3K/AKT Signaling Pathway

The Role of S100B/RAGE-Enhanced ADAM17 Activation in Endothelial Glycocalyx Shedding After Traumatic Brain Injury

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