Survival and maturation of cerebral neurons on poly(l-lysine) surfaces in the absence of serum

Yavin, Ziva; Yavin, Ephraïm

doi:10.1016/0012-1606(80)90176-1

Cited by 88 publications

(33 citation statements)

References 17 publications

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“…Mouse Cortical Neurons. The procedure of preparation of primary cultures of mouse cortical neurons is based on techniques described in detail by Yavin and Yavin (1980) and Yu et al (1984). The cultures are highly enriched in well differentiated GABAergic cortical neurons.…”

Section: E In Vitro Models To Study Regulation Of Gaba a Receptor Exmentioning

confidence: 99%

Regulation of GABA_AReceptor Subunit Expression by Pharmacological Agents

2010

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Section: E In Vitro Models To Study Regulation Of Gaba a Receptor Exmentioning

confidence: 99%

Regulation of GABA_AReceptor Subunit Expression by Pharmacological Agents

2010

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“…Cortical neurons-Primary cultures of cortical neurons were prepared from the cerebral hemispheres of 16 day gestation rat embryos using a modification of the methods of Yavin and Yavin (1980). All dissections were performed in dissecting medium (Leibovitz's L-15 with glutamine).…”

Section: Cell Culturementioning

confidence: 99%

Isolation of mitochondria with high respiratory control from primary cultures of neurons and astrocytes using nitrogen cavitation

Kristián

Hopkins

McKenna

et al. 2006

Journal of Neuroscience Methods

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To study neurons or glia-specific mitochondria one needs to isolate these organelles from primary neuronal or astrocytic cell culture. This work provides novel method for isolation of functional and morphologically intact mitochondria from neurons and astrocytes in cell cultures. In the first step, mitochondria are released from cells by disruption of cell membranes using a nitrogen cavitation technique. This technique is based on rapid decompression of a cell suspension from within a pressure vessel. Mitochondria released from cell bodies are then separated from the rest of cell homogenate by Percoll gradient centrifugation. This is a relatively rapid technique that yields to very well coupled mitochondria that exhibited functional and morphological characteristics comparable to mitochondria isolated from brain tissue using common techniques. This technique thus will allow examination of mitochondria that are exclusively cell specific in origin.

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“…Cortical cultures enriched in GABAergic neurons (Ͼ95%) were grown in the presence of valine (25 g/ml), pyruvate (2 mM), and ␣-ketoglutarate (5 mM) and in the absence of glutamine. Hippocampal cultures enriched in glutamatergic neurons (Ͼ87%) were maintained in medium supplemented with 2 mM glutamine as described previously (76). Cultures were incubated at 37°C in a humidified atmosphere containing 5% CO 2 .…”

Section: Generation and Characterization Of Ib␣-sr Bigenic Micementioning

confidence: 99%

NF-κB/Rel Regulates Inhibitory and Excitatory Neuronal Function and Synaptic Plasticity

O’Mahony

Raber

Montaño

et al. 2006

Molecular and Cellular Biology

100

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Changes in synaptic plasticity required for memory formation are dynamically regulated through opposing excitatory and inhibitory neurotransmissions. To explore the potential contribution of NF-B/Rel to these processes, we generated transgenic mice conditionally expressing a potent NF-B/Rel inhibitor termed IB␣ superrepressor (IB␣-SR). Using the prion promoter-enhancer, IB␣-SR is robustly expressed in inhibitory GABAergic interneurons and, at lower levels, in excitatory neurons but not in glia. This neuronal pattern of IB␣-SR expression leads to decreased expression of glutamate decarboxylase 65 (GAD65), the enzyme required for synthesis of the major inhibitory neurotransmitter, ␥-aminobutyric acid (GABA) in GABAergic interneurons. IB␣-SR expression also results in diminished basal GluR1 levels and impaired synaptic strength (input/output function), both of which are fully restored following activity-based task learning. Consistent with diminished GAD65-derived inhibitory tone and enhanced excitatory firing, IB␣-SR ؉ mice exhibit increased late-phase long-term potentiation, hyperactivity, seizures, increased exploratory activity, and enhanced spatial learning and memory. IB␣-SR ؉ neurons also express higher levels of the activity-regulated, cytoskeleton-associated (Arc) protein, consistent with neuronal hyperexcitability. These findings suggest that NF-B/Rel transcription factors act as pivotal regulators of activity-dependent inhibitory and excitatory neuronal function regulating synaptic plasticity and memory.Stimulus-coupled changes in synaptic plasticity are required for the storage, retrieval, and removal of acquired information collectively referred to as memory formation (28,32,39). Such changes are facilitated by both modifications of existing synaptic effectors and the de novo synthesis of new gene products regulated by various transcriptional regulators. These processes are tightly controlled by the coordinated action of both excitatory and inhibitory neurotransmitters derived from glutamatergic neurons and GABAergic (where GABA is ␥-aminobutyric acid) interneurons, respectively (47, 54). While the vast majority of studies to date have focused on the cyclic AMPresponsive transcription factor (CREB) regulating excitatory neuron function (7, 32-34, 62, 72), more recently, other transcription factors, including members of the NF-B/Rel family of transcription factors, have been implicated in experiencebased synaptic adaptations (38,45,49,55). However, our understanding of their precise role in regulating synaptic plasticity remains rudimentary at best.Although NF-B/Rel factors were originally implicated as central regulators of the immune and inflammatory responses, both basal expression and stimulus-coupled induction of NF-B/Rel factors occur in neurons and glial cells (23,30,31,45,48,55). Activation of NF-B/Rel proceeds through the sitespecific phosphorylation, polyubiquitylation, and proteasomemediated degradation of the major NF-B/Rel inhibitor protein, IB␣ (41). The newly liberated NF-B/Rel complex ra...

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Survival and maturation of cerebral neurons on poly(l-lysine) surfaces in the absence of serum

Cited by 88 publications

References 17 publications

Regulation of GABA_AReceptor Subunit Expression by Pharmacological Agents

Regulation of GABA_AReceptor Subunit Expression by Pharmacological Agents

Isolation of mitochondria with high respiratory control from primary cultures of neurons and astrocytes using nitrogen cavitation

NF-κB/Rel Regulates Inhibitory and Excitatory Neuronal Function and Synaptic Plasticity

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Survival and maturation of cerebral neurons on poly(l-lysine) surfaces in the absence of serum

Cited by 88 publications

References 17 publications

Regulation of GABAAReceptor Subunit Expression by Pharmacological Agents

Regulation of GABAAReceptor Subunit Expression by Pharmacological Agents

Isolation of mitochondria with high respiratory control from primary cultures of neurons and astrocytes using nitrogen cavitation

NF-κB/Rel Regulates Inhibitory and Excitatory Neuronal Function and Synaptic Plasticity

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Regulation of GABA_AReceptor Subunit Expression by Pharmacological Agents

Regulation of GABA_AReceptor Subunit Expression by Pharmacological Agents