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Adult astrocyte isolation

Natália C C A Fernandes, Juliana S Nogueira, Rodrigo A Réssio, Cinthya S Cirqueira, Lidia M Kimura, Karolina R Fernandes, Mariana S Cunha, Renato P Souza, Juliana M Guerra
A widespread epidemic of Zika virus (ZIKV) infection was reported in 2015 in South and Central America, with neurological symptons including meningoencephalitis and Guillain-Barré syndrome in adults, besides an apparent increased incidence of microcephaly in infants born to infected mothers. It is becoming a necessity to have a trustworthy animal model to better understand ZIKV infection. In this study we used newborn white Swiss mice as a model to investigate the ZIKV strain recently isolated in Brazil. ZIKV was inoculated via intracerebral and subcutaneous routes and analysed through gross histopathology and immunohistochemistry...
November 26, 2016: Experimental and Toxicologic Pathology: Official Journal of the Gesellschaft Für Toxikologische Pathologie
Chiara Cervetto, Arianna Venturini, Mario Passalacqua, Diego Guidolin, Susanna Genedani, Kjell Fuxe, Dasiel O Borroto-Esquela, Pietro Cortelli, Amina Woods, Guido Maura, Manuela Marcoli, Luigi F Agnati
Evidence for striatal A2A-D2 heterodimers has led to a new perspective on molecular mechanisms involved in schizophrenia and Parkinson's disease. Despite the increasing recognition of astrocytes' participation in neuropsychiatric disease vulnerability, involvement of striatal astrocytes in A2A and D2 receptor signal transmission has never been explored. Here, we investigated the presence of D2 and A2A receptors in isolated astrocyte processes prepared from adult rat striatum by confocal imaging; the effects of receptor activation were measured on the 4-aminopyridine-evoked release of glutamate from the processes...
November 5, 2016: Journal of Neurochemistry
Yan Wang, Min Yu, Yue Ma, Ruo Ping Wang, Wei Liu, Wei Xia, Ai Li Guan, Cong Hui Xing, Fei Lu, Xiao Ping Ji
Peroxisome proliferator-activated receptors alpha (PPARα) is a therapy target in atherosclerosis and cardiovascular diseases. However, anti-inflammatory effects of PPARα in intracerebral hemorrhage (ICH) remain unknown. We investigated the anti-inflammatory effects of fenofibrate, a ligand of PPARα, in ICH rat model. We found that engagement of fenofibrate increased nissl body and astrocytes, and reduced the neuronal damage, which was observed in paraffin section of ICH rat brain. Fenofibrate also promoted the proliferation of astrocytes that were isolated from adult rat brain...
October 14, 2016: Current Neurovascular Research
F Javier Rubio, Xuan Li, Qing-Rong Liu, Raffaello Cimbro, Bruce T Hope
The study of neuroplasticity and molecular alterations in learned behaviors is switching from the study of whole brain regions to the study of specific sets of sparsely distributed activated neurons called neuronal ensembles that mediate learned associations. Fluorescence Activated Cell Sorting (FACS) has recently been optimized for adult rat brain tissue and allowed isolation of activated neurons using antibodies against the neuronal marker NeuN and Fos protein, a marker of strongly activated neurons. Until now, Fos-expressing neurons and other cell types were isolated from fresh tissue, which entailed long processing days and allowed very limited numbers of brain samples to be assessed after lengthy and complex behavioral procedures...
2016: Journal of Visualized Experiments: JoVE
Hadi Aligholi, Gholamreza Hassanzadeh, Ali Gorji, Hassan Azari
Despite all attempts the problem of regeneration in damaged central nervous system (CNS) has remained challenging due to its cellular complexity and highly organized and sophisticated connections. In this regard, stem cell therapy might serve as a viable therapeutic approach aiming either to support the damaged tissue and hence to reduce the subsequent neurological dysfunctions and impairments or to replace the lost cells and re-establish damaged circuitries. Adult neural stem/progenitor cells (NS/PCs) are one of the outstanding cell sources that can be isolated from the subventricular zone (SVZ) of the lateral ventricles...
2016: Methods in Molecular Biology
Fu-Shi Quan, Jian Chen, Yuan Zhong, Wen-Zhi Ren
The present study evaluated the comparative effect of stereotaxically transplanted immature neuronal or glial cells in brain on motor functional recovery and cytokine expression after cold-induced traumatic brain injury (TBI) in adult rats. A total of 60 rats were divided into four groups (n=15/group): Sham group; TBI only group; TBI plus neuronal cells-transplanted group (NC-G); and TBI plus glial cells-transplanted group (GC-G). Cortical lesions were induced by a touching metal stamp, frozen with liquid nitrogen, to the dura mater over the motor cortex of adult rats...
September 2016: Experimental and Therapeutic Medicine
Marco Magistri, Nathalie Khoury, Emilia Maria Cristina Mazza, Dmitry Velmeshev, Jae K Lee, Silvio Bicciato, Pantelis Tsoulfas, Mohammad Ali Faghihi
Astrocytes are a morphologically and functionally heterogeneous population of cells that play critical roles in neurodevelopment and in the regulation of central nervous system homeostasis. Studies of human astrocytes have been hampered by the lack of specific molecular markers and by the difficulties associated with purifying and culturing astrocytes from adult human brains. Human neural progenitor cells (NPCs) with self-renewal and multipotent properties represent an appealing model system to gain insight into the developmental genetics and function of human astrocytes, but a comprehensive molecular characterization that confirms the validity of this cellular system is still missing...
November 2016: European Journal of Neuroscience
Tal Iram, Dorit Trudler, David Kain, Sivan Kanner, Ronit Galron, Robert Vassar, Ari Barzilai, Pablo Blinder, Zvi Fishelson, Dan Frenkel
In Alzheimer's disease (AD), astrocytes undergo morphological changes ranging from atrophy to hypertrophy, but the effect of such changes at the functional level is still largely unknown. Here, we aimed to investigate whether alterations in astrocyte activity in AD are transient and depend on their microenvironment, or whether they are irreversible. We established and characterized a new protocol for the isolation of adult astrocytes and discovered that astrocytes isolated from old 5xFAD mice have higher GFAP expression than astrocytes derived from WT mice, as observed in vivo...
August 17, 2016: Neurobiology of Disease
Linda V Blomster, Dorte Strøbaek, Charlotte Hougaard, Jessica Klein, Lars H Pinborg, Jens D Mikkelsen, Palle Christophersen
The KCa 3.1 channel (KCNN4) is an important modulator of microglia responses in rodents, but no information exists on functional expression on microglia from human adults. We isolated and cultured microglia (max 1% astrocytes, no neurons or oligodendrocytes) from neocortex surgically removed from epilepsy patients and employed electrophysiological whole-cell measurements and selective pharmacological tools to elucidate functional expression of KCa 3.1. The channel expression was demonstrated as a significant increase in the voltage-independent current by NS309, a KCa 3...
July 29, 2016: Glia
Jana Seele, Roland Nau, Chittappen K Prajeeth, Martin Stangel, Peter Valentin-Weigand, Maren Seitz
Streptococcus (S.) suis infections are the most common cause of meningitis in pigs. Moreover, S. suis is a zoonotic pathogen, which can lead to meningitis in humans, mainly in adults. We assume that glial cells may play a crucial role in host-pathogen interactions during S. suis infection of the central nervous system. Glial cells are considered to possess important functions during inflammation and injury of the brain in bacterial meningitis. In the present study, we established primary astrocyte-microglial cell co-cultures to investigate interactions of S...
2016: Pathogens
M S Sandhu, H H Ross, K Z Lee, B K Ormerod, P J Reier, D D Fuller
Following spinal cord injury (SCI), intraspinal transplantation of neural progenitor cells (NPCs) harvested from the forebrain sub-ventricular zone (SVZ) can improve locomotor outcomes. Cervical SCI often results in respiratory-related impairments, and here we used an established model cervical SCI (C2 hemisection, C2Hx) to confirm the feasibility of mid-cervical transplantation of SVZ-derived NPCs and the hypothesis that that this procedure would improve spontaneous respiratory motor recovery. NPCs were isolated from the SVZ of enhanced green fluorescent protein (GFP) expressing neonatal rats, and then intraspinally delivered immediately caudal to an acute C2Hx lesion in adult non-GFP rats...
June 11, 2016: Experimental Neurology
Claire Lépinoux-Chambaud, Kristell Barreau, Joël Eyer
UNLABELLED: Targeting neural stem cells (NSCs) in the adult brain represents a promising approach for developing new regenerative strategies, because these cells can proliferate, self-renew, and differentiate into new neurons, astrocytes, and oligodendrocytes. Previous work showed that the NFL-TBS.40-63 peptide, corresponding to the sequence of a tubulin-binding site on neurofilaments, can target glioblastoma cells, where it disrupts their microtubules and inhibits their proliferation...
July 2016: Stem Cells Translational Medicine
Muriel D Mardones, Gabriela A Andaur, Manuel Varas-Godoy, Jenny F Henriquez, Felipe Salech, María Isabel Behrens, Andrés Couve, Nibaldo C Inestrosa, Lorena Varela-Nallar
BACKGROUND: In the adult hippocampus new neurons are continuously generated from neural stem cells (NSCs) present at the subgranular zone of the dentate gyrus. This process is controlled by Wnt signaling, which plays a complex role in regulating multiple steps of neurogenesis including maintenance, proliferation and differentiation of progenitor cells and the development of newborn neurons. Differential effects of Wnt signaling during progression of neurogenesis could be mediated by cell-type specific expression of Wnt receptors...
2016: Molecular Brain
H E Marei, A Althani, S Rezk, A Farag, S Lashen, N Afifi, A Abd-Elmaksoud, R Pallini, P Casalbore, C Cenciarelli, T Caceci
STUDY DESIGN: Adult human olfactory bulb neural stem cells (OBNSCs) were isolated from human patients undergoing craniotomy for tumor resection. They were genetically engineered to overexpresses green fluorescent protein (GFP) to help trace them following engraftment. Spinal cord injury (SCI) was induced in rats using standard laminectomy protocol, and GFP-OBNSC were engrafted into rat model of SCI at day 7 post injury. Three rat groups were used: (i) Control group, (ii) Sham group (injected with cerebrospinal fluid) and treated group (engrafted with OBNSCs)...
February 16, 2016: Spinal Cord
Audrey Giocanti-Auregan, Ophélie Vacca, Romain Bénard, Sijia Cao, Lourdes Siqueiros, Cecilia Montañez, Michel Paques, José-Alain Sahel, Florian Sennlaub, Xavier Guillonneau, Alvaro Rendon, Ramin Tadayoni
Understanding retinal vascular development is crucial because many retinal vascular diseases such as diabetic retinopathy (in adults) or retinopathy of prematurity (in children) are among the leading causes of blindness. Given the localization of the protein Dp71 around the retinal vessels in adult mice and its role in maintaining retinal homeostasis, the aim of this study was to determine if Dp71 was involved in astrocyte and vascular development regulation. An experimental study in mouse retinas was conducted...
May 2016: Glia
Nicolas Vedrenne, Vincent Sarrazy, Serge Battu, Nelly Bordeau, Laurence Richard, Fabrice Billet, Valérie Coronas, Alexis Desmoulière
Astrocytes encompass a heterogeneous cell population. Using sedimentation field flow fractionation method, different, almost pure, astrocyte subpopulations were isolated. Cells were collected from cortex of newborn rats and sorted by sedimentation field flow fractionation to obtain different fractions which were subjected to protein analysis and characterized by immunocytofluorescence. The behavior of the cells was analyzed <i>in vitro</i>, under culture conditions used for neural stem cells. These culture conditions were also applied to cells derived from an adult cortical tissue following traumatic brain injury...
December 9, 2015: Rejuvenation Research
Seyed Mojtaba Hosseini, Mohammad Farahmandnia, Sepehr Kazemi, Benafshe Shakibajahromi, Fatemeh Sabet Sarvestani, Zahra Khodabande
BACKGROUND: Nowadays, stroke leads to a significant part of the adult mortality and morbidity and also it could result in some neurological deficits in the patients' lives. Cell therapy has opened a new approach to treat the brain ischemia and reduce its terrible effects on the patients' lives. There are several articles which show that the cell therapy could be beneficial for treating brain stroke. In this study, we have planned to present a new cell therapy method for stroke by administration of Mesenchymal stem cells and differentiated neural stem cells without astrocytes...
November 2015: International Journal of Stem Cells
Raffaella Spina, Dillon M Voss, Laura Asnaghi, Andrew Sloan, Eli E Bar
Glioblastoma multiforme (GBM) are the most common primary malignant brain tumor in adults, with a median survival of about one year. This poor prognosis is attributed primarily to therapeutic resistance and tumor recurrence after surgical removal, with the root cause suggested to be found in glioblastoma stem cells (GSCs). Using glial fibrillary acidic protein (GFAP) as a reporter of astrocytic differentiation, we isolated multiple clones from three independent GSC lines which express GFAP in a remarkably stable fashion...
January 5, 2016: Oncotarget
Sophie Péron, Benedikt Berninger
Cells with neural stem cell (NSC)-like properties can be isolated from the cortex of adult brains following injury, but their origins and function are unclear. Now in Cell Stem Cell, Faiz et al. (2015) show that subventricular-zone-derived NSCs home to injured cortical area following stroke, where they generate reactive astrocytes.
November 5, 2015: Cell Stem Cell
Maryam Faiz, Nadia Sachewsky, Sergio Gascón, K W Annie Bang, Cindi M Morshead, Andras Nagy
Reactive astrocytes (RAs) have been reported to convert to multipotent neural stem cells (NSCs) capable of neurosphere (NS) formation and multilineage differentiation in vitro. Using genetic tagging, we determined that subventricular zone (SVZ) NSCs give rise to NSs derived from the stroke-injured cortex. We demonstrate that these cells can be isolated from the cortex in two different models of stroke and from different stroke-lesioned cortical regions. Interestingly, SVZ NSCs give rise to a subpopulation of RAs in the cortex that contribute to astrogliosis and scar formation...
November 5, 2015: Cell Stem Cell
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