Rat hippocampal neurons in dispersed cell culture

Banker, Gary; Cowan, William

doi:10.1016/0006-8993(77)90594-7

Cited by 1,075 publications

(636 citation statements)

References 23 publications

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“…Cultures were maintained in P-neuronal media (PAA, Pasching, Austria) supplemented with Neuromix (PAA) and 10% horse serum as previously described. 53 After 10 days in vitro (d.i.v), neurons were either transfected with 4D and GFP (gift from F Calegari, Dresden University) 16 constructs and fixed 1 day later. At 14 d.i.v., neurons were exposed for different times to bath application of 40 μM NMDA (Tocris, Bristol, UK).…”

Section: Discussionmentioning

confidence: 99%

Notch signaling in response to excitotoxicity induces neurodegeneration via erroneous cell cycle reentry

et al. 2015

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Neurological disorders such as Alzheimer's disease, stroke and epilepsy are currently marred by the lack of effective treatments to prevent neuronal death. Erroneous cell cycle reentry (CCR) is hypothesized to have a causative role in neurodegeneration. We show that forcing S-phase reentry in cultured hippocampal neurons is sufficient to induce neurodegeneration. We found that kainic-acid treatment in vivo induces erroneous CCR and neuronal death through a Notch-dependent mechanism. Ablating Notch signaling in neurons provides neuroprotection against kainic acid-induced neuronal death. We further show that kainic-acid treatment activates Notch signaling, which increases the bioavailability of CyclinD1 through Akt/GSK3β pathway, leading to aberrant CCR via activation of CyclinD1-Rb-E2F1 axis. In addition, pharmacological blockade of this pathway at critical steps is sufficient to confer resistance to kainic acid-induced neurotoxicity in mice. Taken together, our results demonstrate that excitotoxicity leads to neuronal death in a Notch-dependent manner through erroneous CCR.

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Section: Discussionmentioning

confidence: 99%

Notch signaling in response to excitotoxicity induces neurodegeneration via erroneous cell cycle reentry

et al. 2015

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“…Dissociated hippocampal neurones were prepared as previously described [23,24] from embryonic day 18 hippocampi maintained for 18-28 days. Hippocampal neurons cells were grown at 37 °C in 5% CO 2 in DMEM GlutaMax (Invitrogen, Carlsbad CA, USA) medium supplemented with 10% FBS, 100 µg/mL Pen-Strep, 25 µg/mL zeocin, 5 µg/mL blasticidin and 25 µg/mL hygromycin.…”

Section: Cell Culture Methodsmentioning

confidence: 99%

Tracking of fast moving neuronal vesicles with ageladine A

Bickmeyer¹,

Heisenberg²,

Podbielski³

et al. 2010

Biochemical and Biophysical Research Communications

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This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. Tracking of fast moving neuronal vesicles with ageladine A

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“…Hippocampi were dissected from the brains of 18-or 19-day gestation Wistar rat fetuses (Banker and Cowan, 1977). After dissociation and centrifugation, the tissue was resuspended in serum-free Neurobasal medium (Gibco), enriched with 2% v/v B27 supplement (Brewer et al, 1993), 0.005% gentamicin (Gibco), 0.05% glutamine, and 25 AM glutamate (Sigma).…”

Section: Primary Cell Culturesmentioning

confidence: 99%

Improving lipoplex-mediated gene transfer into C6 glioma cells and primary neurons

Cruz

Simões

Lima

2004

Experimental Neurology

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The development of methodologies for gene transfer into the central nervous system is crucial for gene therapy of neurological disorders. In this study, different cationic liposome formulations were used to transfer DNA into C6 glioma cells and primary hippocampal and cortical neurons by varying the nature of the helper lipid (DOPE, Chol) or a mixture of DOPE and cholesterol (Chol) associated to DOTAP. In addition, the effect of the lipid/DNA (+/À) charge ratio, the association of the ligand transferrin to the lipoplexes, and the stage of differentiation of the primary cells on the levels of transfection activity, transfection efficiency, and duration of gene expression were evaluated. Mechanistic studies were also performed to investigate the route of delivery of the complexes into neurons. Our results indicate that DOTAP:Chol (1:1 mol ratio) was the best formulation to transfer a reporter gene into C6 glioma cells, primary hippocampal neurons, and primary cortical neurons. The use of transferrin-associated lipoplexes resulted in a significant enhancement of transfection activity, as compared to plain lipoplexes, which can be partially attributed to the promotion of their internalization mediated by transferrin. While for hippocampal neurons the levels of luciferase gene expression are very low, for primary cortical neurons the levels of transgene expression are high and relatively stable, although only 4% of the cells has been transfected. The stage of cell differentiation revealed to be critical to the levels of gene expression. Consistent with previous findings on the mechanisms of cell internalization, the experiments with inhibitors of the endocytotic pathway clearly indicate that transferrin-associated lipoplexes are internalized into primary neurons by endocytosis. Promising results were obtained in terms of the levels and duration of gene expression, particularly in cortical neurons when transfected with the Tfassociated lipoplexes, this finding suggesting the usefulness of these lipid-based carriers to deliver genes within the CNS. D 2004 Elsevier Inc. All rights reserved.

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Rat hippocampal neurons in dispersed cell culture

Cited by 1,075 publications

References 23 publications

Notch signaling in response to excitotoxicity induces neurodegeneration via erroneous cell cycle reentry

Notch signaling in response to excitotoxicity induces neurodegeneration via erroneous cell cycle reentry

Tracking of fast moving neuronal vesicles with ageladine A

Improving lipoplex-mediated gene transfer into C6 glioma cells and primary neurons

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