Culture of Rodent Cortical, Hippocampal, and Striatal Neurons

Facci, Laura; Skaper, Stephen D.

doi:10.1007/978-1-4939-7571-6_3

Cited by 12 publications

(14 citation statements)

References 16 publications

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“…Primary cultured hippocampal neurons are frequently harvested from hippocampi of newborn (<24-h-old) rodents, owing to their localization and the amount of available material (23,24), and these cells have been used to establish experimental models to study the function of neurons in vitro . Therefore, in the present study, the hippocampal neurons from fetal rats were selected as a model to study I/R injury.…”

Section: Discussionmentioning

confidence: 99%

Overexpression of COX6B1 protects against I/R‑induced neuronal injury in rat hippocampal neurons

Shan

Xiao

et al. 2019

Mol Med Report

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Cerebrovascular disease (CVD) is one of the leading causes of mortality worldwide. The role of cytochrome c oxidase subunit 6B1 (COX6B1) in the central nervous system remains unclear. The present study aimed to analyze the role of COX6B1 in rat hippocampal neurons extracted from fetal rats. The subcellular localization of the neuron-specific marker microtubule-associated protein 2 was detected by immunofluorescence assay. Cell viability was assessed using a cell counting kit, and the levels of apoptosis and cytosolic Ca 2+ were analyzed by flow cytometry. The expression levels of the molecular factors downstream to COX6B1 were determined using reverse transcription-quantitative polymerase chain reaction and western blotting. Reoxygenation following oxygen-glucose deprivation (OGD) decreased cell viability and the expression levels of COX6B1 in a time-dependent manner, and 60 min of reoxygenation was identified as the optimal time period for establishing an ischemia/reperfusion (I/R) model. Overexpression of COX6B1 was demonstrated to reverse the viability of hippocampal neurons following I/R treatment. Specifically, COX6B1 overexpression decreased the cytosolic concentration of Ca 2+ and suppressed neuronal apoptosis, which were increased following I/R treatment. Furthermore, overexpression of COX6B1 increased the protein expression levels of apoptosis regulator BCL-2 and mitochondrial cytochrome c (cyt c), and decreased the protein expression levels of apoptosis regulator BCL2-associated X and cytosolic cyt c in I/R model cells. Collectively, the present study results suggested that COX6B1 overexpression may reverse I/R-induced neuronal damage by increasing the viability of neurons, by decreasing the cytosolic levels of Ca 2+ and by suppressing apoptosis. These results may facilitate the development of novel strategies for the prevention and treatment of CVD.

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Section: Discussionmentioning

confidence: 99%

Overexpression of COX6B1 protects against I/R‑induced neuronal injury in rat hippocampal neurons

Shan

Xiao

et al. 2019

Mol Med Report

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“…For mouse cell culture, a protocol was established based on earlier methods [ 36 ]. Newborn pups between P 0–P 1 were sacrificed by cephalic-segregation, cortices were taken immediately and placed into ice-cold HBSS-/-.…”

Section: Methodsmentioning

confidence: 99%

Acid Sphingomyelinase Impacts Canonical Transient Receptor Potential Channels 6 (TRPC6) Activity in Primary Neuronal Systems

Zeitler

Schumacher

Monti

et al. 2020

Cells

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The acid sphingomyelinase (ASM)/ceramide system exhibits a crucial role in the pathology of major depressive disorder (MDD). ASM hydrolyzes the abundant membrane lipid sphingomyelin to ceramide that regulates the clustering of membrane proteins via microdomain and lipid raft organization. Several commonly used antidepressants, such as fluoxetine, rely on the functional inhibition of ASM in terms of their antidepressive pharmacological effects. Transient receptor potential canonical 6 (TRPC6) ion channels are located in the plasma membrane of neurons and serve as receptors for hyperforin, a phytochemical constituent of the antidepressive herbal remedy St. John’s wort. TRPC6 channels are involved in the regulation of neuronal plasticity, which likely contributes to their antidepressant effect. In this work, we investigated the impact of reduced ASM activity on the TRPC6 function in neurons. A lipidomic analysis of cortical brain tissue of ASM deficient mice revealed a decrease in ceramide/sphingomyelin molar ratio and an increase in sphingosine. In neurons with ASM deletion, hyperforin-mediated Ca2+-influx via TRPC6 was decreased. Consequently, downstream activation of nuclear phospho-cAMP response element-binding protein (pCREB) was changed, a transcriptional factor involved in neuronal plasticity. Our study underlines the importance of balanced ASM activity, as well as sphingolipidome composition for optimal TRPC6 function. A better understanding of the interaction of the ASM/ceramide and TRPC6 systems could help to draw conclusions about the pathology of MDD.

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“…Primary mouse neuronal cultures were prepared according to an established protocol [8] with some modifications. Briefly, newborn α7nAChR flox /flox mice (P0-P1) brains were collected and freed of meninges.…”

Section: Primary Chrna7 Flox/flox Mice Culturesmentioning

confidence: 99%

Aβ1-16 controls synaptic vesicle pools at excitatory synapses via cholinergic modulation of synapsin phosphorylation

Anni

Weiß

Guhathakurta

et al. 2021

Cell. Mol. Life Sci.

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Amyloid beta (Aβ) is linked to the pathology of Alzheimer’s disease (AD). At physiological concentrations, Aβ was proposed to enhance neuroplasticity and memory formation by increasing the neurotransmitter release from presynapse. However, the exact mechanisms underlying this presynaptic effect as well as specific contribution of endogenously occurring Aβ isoforms remain unclear. Here, we demonstrate that Aβ1-42 and Aβ1-16, but not Aβ17-42, increased size of the recycling pool of synaptic vesicles (SV). This presynaptic effect was driven by enhancement of endogenous cholinergic signalling via α7 nicotinic acetylcholine receptors, which led to activation of calcineurin, dephosphorylation of synapsin 1 and consequently resulted in reorganization of functional pools of SV increasing their availability for sustained neurotransmission. Our results identify synapsin 1 as a molecular target of Aβ and reveal an effect of physiological concentrations of Aβ on cholinergic modulation of glutamatergic neurotransmission. These findings provide new mechanistic insights in cholinergic dysfunction observed in AD.

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Culture of Rodent Cortical, Hippocampal, and Striatal Neurons

Cited by 12 publications

References 16 publications

Overexpression of COX6B1 protects against I/R‑induced neuronal injury in rat hippocampal neurons

Overexpression of COX6B1 protects against I/R‑induced neuronal injury in rat hippocampal neurons

Acid Sphingomyelinase Impacts Canonical Transient Receptor Potential Channels 6 (TRPC6) Activity in Primary Neuronal Systems

Aβ1-16 controls synaptic vesicle pools at excitatory synapses via cholinergic modulation of synapsin phosphorylation

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