An In Vitro Trauma Model to Study Rodent and Human Astrocyte Reactivity

Author(s):  
Ina-Beate Wanner
Keyword(s):  
2018 ◽  
Vol 8 (1) ◽  
Author(s):  
Robert P. Weinberg ◽  
Vera V. Koledova ◽  
Kirsten Schneider ◽  
T. G. Sambandan ◽  
Adlai Grayson ◽  
...  
Keyword(s):  

2018 ◽  
Vol 137 (4) ◽  
pp. 350-358 ◽  
Author(s):  
Keita Kitamura ◽  
Ryo Ito ◽  
Kenta Umehara ◽  
Hanae Morio ◽  
Kosuke Saito ◽  
...  

2020 ◽  
Author(s):  
Elsa Papadimitriou ◽  
Paraskevi N. Koutsoudaki ◽  
Timokratis Karamitros ◽  
Dimitra Karagkouni ◽  
Dafni Chroni-Tzartou ◽  
...  

AbstractmiR-124 plays a major regulatory role in neurogenesis and neuronal differentiation during brain development through control of its multiple non-neuronal targets and has therefore been employed in direct reprogramming protocols supplementary to neurogenic TFs, and other miRNAs to enhance neurogenic conversion. However, its capacity to instruct neurogenic conversion of astrocytes and its independent mechanism of direct reprogramming action have been poorly investigated. Aim of the study was to investigate whether miR-124 is a master-regulatory reprogramming agent, potent to drive direct reprogramming of astrocytes to induced-neurons (iNs) on its own and to elucidate its mechanism of reprogramming action. To this end we overexpressed miR-124 either alone or in combination with the small neurogenic compound ISX9 both in vitro and in in vivo in a mouse mechanical cortical trauma model and analyzed their mechanism of reprogramming action. Our data indicate that miR-124 and ISX9 exhibit both unique and convergent molecular contributions in the reprogramming process to iNs. miR-124 is a potent driver of the astrocytic reprogramming switch of astrocytes towards an immature neuronal fate by repressing genes regulating astrocytic function, among which we identified the RNA-binding protein Zfp36l1 as a novel miR-124 direct target. We also provide evidence that ISX9 greatly improves both miR-124-induced reprogramming efficiency and functional maturation of iNs. Importantly, miR-124 either alone or along with ISX9 is potent to guide direct neuronal reprogramming of reactive astrocytes to iNs of cortical identity in vivo, a novel finding confirming the robust direct reprogramming action of the two molecules in activated astrocytes in vivo.


2020 ◽  
Vol 15 (2) ◽  
pp. 105-109 ◽  
Author(s):  
Burak Yulug ◽  
Mehmet Ozansoy ◽  
Merve Alokten ◽  
Muzaffer B.C. Ozansoy ◽  
Seyda Cankaya ◽  
...  

Background: Antibiotic therapies targeting multiple regenerative mechanisms have the potential for neuroprotective effects, but the diversity of experimental strategies and analyses of non-standardised therapeutic trials are challenging. In this respect, there are no cases of successful clinical application of such candidate molecules when it comes to human patients. Methods: After 24 hours of culturing, three different minocycline (Sigma-Aldrich, M9511, Germany) concentrations (1 μM, 10 μM and 100 μM) were added to the primary cortical neurons 15 minutes before laser axotomy procedure in order to observe protective effect of minocycline in these dosages. Results: Here, we have shown that minocycline exerted a significant neuroprotective effect at 1 and 100μM doses. Beyond confirming the neuroprotective effect of minocycline in a more standardised and advanced in-vitro trauma model, our findings could have important implications for future studies that concentrate on the translational block between animal and human studies. Conclusion: Such sophisticated approaches might also help to conquer the influence of humanmade variabilities in critical experimental injury models. To the best of our knowledge, this is the first study showing that minocycline increases in-vitro neuronal cell survival after laser-axotomy.


Neuroscience ◽  
2014 ◽  
Vol 274 ◽  
pp. 1-10 ◽  
Author(s):  
C. Augustine ◽  
G. Cepinskas ◽  
D.D. Fraser

Molecules ◽  
2020 ◽  
Vol 25 (22) ◽  
pp. 5403
Author(s):  
Emanuela Colombo ◽  
Rosaria Pascente ◽  
Daniela Triolo ◽  
Claudia Bassani ◽  
Anthea De Angelis ◽  
...  

Astrocytes greatly participate to inflammatory and neurotoxic reactions occurring in neurodegenerative diseases and are valuable pharmacological targets to support neuroprotection. Here we used human astrocytes generated from reprogrammed fibroblasts as a cellular model to study the effect of the compound Laquinimod and its active metabolite de-Laquinimod on astrocyte functions and the astrocyte–neuron interaction. We show that human iAstrocytes expressed the receptor for the inflammatory mediator IL1 and responded to it via nuclear translocation of NFκB, an event that did not occur if cells were treated with Laquinimod, indicating a direct anti-inflammatory activity of the drug on the human astrocyte. Similarly, while exposure to IL1 downregulated glial glutamate transporters GLAST and GLT1, treatment with Laquinimod supported maintenance of physiological levels of these proteins despite the inflammatory milieu. Laquinimod also induced nuclear translocation of the aryl hydrocarbon receptor (AHR), suggesting that drug action was mediated by activation of the AHR pathway. However, the drug was effective despite AHR inhibition via CH223191, indicating that AHR signaling in the astrocyte is dispensable for drug responses. Finally, in vitro experiments with rat spinal neurons showed that laquinimod did not exert neuroprotection directly on the neuron but dampened astrocyte-induced neurodegeneration. Our findings indicate that fibroblast-derived human astrocytes represent a suitable model to study astrocyte–neuron crosstalk and demonstrate indirect, partial neuroprotective efficacy for laquinimod.


2016 ◽  
Vol 7 ◽  
Author(s):  
Yuli Cao ◽  
Mårten Risling ◽  
Elisabeth Malm ◽  
Anders Sondén ◽  
Magnus Frödin Bolling ◽  
...  

2012 ◽  
Vol 2012 ◽  
pp. 1-6 ◽  
Author(s):  
P. Mommsen ◽  
T. Barkhausen ◽  
C. Zeckey ◽  
H. Andruszkow ◽  
C. Krettek ◽  
...  

Splenic immune function is known to be depressed following hemorrhage. The present study investigates the effects of femoral shaft fracture, isolated or in combination with hemorrhage, on early stage cytokine production capacity of splenocytes and observes the role of IL-6 under these conditions. Male IL-6 knockout (IL-6−/−) and wild-type mice (WT) were randomly divided into three groups: sham (S), isolated femoral fracture (Fx), and femoral fracture + volume controlled hemorrhage (TH-Fx) ( per group). Animals were sacrificed four hours after induction of hemorrhage and fracture. Cytokine release (TNF-α, IL-6, and IL-10) of isolated and LPS-stimulated splenocytes was determined by cytometric bead array. Femoral fracture with or without hemorrhage caused a suppression of in vitro cytokine production capacity of splenocytes at an early posttraumatic stage in WT and IL-6−/−. In the absence of IL-6, the profile of splenic cytokine secretion is significantly altered, identifying this cytokine as a potential therapeutic target to modulate the posttraumatic immune response.


PeerJ ◽  
2017 ◽  
Vol 5 ◽  
pp. e2829 ◽  
Author(s):  
V. Bleu Knight ◽  
Elba E. Serrano

Biomaterial scaffolds have the potential to enhance neuronal development and regeneration. Understanding the genetic responses of astrocytes and neurons to biomaterials could facilitate the development of synthetic environments that enable the specification of neural tissue organization with engineered scaffolds. In this study, we used high throughput transcriptomic and imaging methods to determine the impact of a hydrogel, PuraMatrix™, on human glial cellsin vitro. Parallel studies were undertaken with cells grown in a monolayer environment on tissue culture polystyrene. When the Normal Human Astrocyte (NHA) cell line is grown in a hydrogel matrix environment, the glial cells adopt a structural organization that resembles that of neuronal-glial cocultures, where neurons form clusters that are distinct from the surrounding glia. Statistical analysis of next generation RNA sequencing data uncovered a set of genes that are differentially expressed in the monolayer and matrix hydrogel environments. Functional analysis demonstrated that hydrogel-upregulated genes can be grouped into three broad categories: neuronal differentiation and/or neural plasticity, response to neural insult, and sensory perception. Our results demonstrate that hydrogel biomaterials have the potential to transform human glial cell identity, and may have applications in the repair of damaged brain tissue.


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