Culturing primary neurons from rat hippocampus and cortex

Sahu, Madhusmita Priyadarshini; Nikkilä, Outi; Lågas, Seija Anneli; Kolehmainen, S.; Ċastrén, Eero

doi:10.1042/ns20180207

Cited by 73 publications

(50 citation statements)

References 22 publications

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“…[23] Second, previous findings report very low levels of glial contamination in these time points. [22,24,25] Finally, it is known that glial proliferation rates are dramatically hindered in the presence of neurons, [26] which corroborates our argument for the use of this assay to quantify neuronal viability. In this way, it is possible to ascertain the biocompatibility of the microreactors in relation to different phases of neuronal differentiation.…”

Section: Neuronal Compatibility and Interactionsupporting

confidence: 83%

“…Importantly, the microreactors were tested with 7 DIV and 14 DIV cultures, since these time points are enriched, respectively, with immature and mature neurons. [ 22 ] The assay used in this work to determine cell viability is based on the quantification of a signal that is proportional to the number of live cells in culture. Thus, it only represents cell viability in non‐proliferative cell culture, being a proxy for cell proliferation in proliferative cultures.…”

Section: Resultsmentioning

confidence: 99%

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Multicompartment Microreactors Prevent Excitotoxic Dysfunctions In Rat Primary Cortical Neurons

et al. 2020

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Excitotoxicity is a cellular phenomenon that comprises the consequences of toxic actions of excitatory neurotransmitters, such as glutamate. This process is usually related to overproduction of reactive oxygen species (ROS) and ammonia (NH4+) toxicity. Platinum nanoparticle (Pt‐NP)‐based microreactors able to degrade hydrogen peroxide (H2O2) and NH4+, are previously described as a novel therapeutical approach against excitotoxicity, conferring protection to neuroblasts. Now, it is demonstrated that these microreactors are compatible with rat primary cortical neurons, show high levels of neuronal membrane interaction, and are able to improve cell survival and neuronal activity when neurons are exposed to H2O2 or NH4+. Additionally, more complex microreactors are assembled, including enzyme‐loaded liposomes containing glutamate dehydrogenase and glutathione reductase, in addition to Pt‐NP. The in vitro activity of these microreactors is characterized and they are compared to the Pt‐NP‐based microreactors in terms of biological activity, concluding that they enhance cell viability similarly or more extensively than the latter. Extracellular electrophysiological recordings demonstrate that these microreactors rescue neuronal functionality lost upon incubation with H2O2 or NH4+. This study provides more evidence for the potential application of these microreactors in a biomedical context with more complex cellular environments.

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Section: Neuronal Compatibility and Interactionsupporting

confidence: 83%

Section: Resultsmentioning

confidence: 99%

Multicompartment Microreactors Prevent Excitotoxic Dysfunctions In Rat Primary Cortical Neurons

et al. 2020

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“…Cerebral cortical cell cultures were prepared from Wistar rat (Harlan Labs) embryos extracted on embryonic day (E)18 ( Sahu et al, 2019 ). The cells were cultured in a serum-free neurobasal medium with supplements (1% penicillin, 1% L-glutamine, 2% B-27) and collected after 7–9 d in vitro (DIV).…”

Section: Methodsmentioning

confidence: 99%

Chondroitinase and Antidepressants Promote Plasticity by Releasing TRKB from Dephosphorylating Control of PTPσ in Parvalbumin Neurons

et al. 2020

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Perineuronal nets (PNNs) are an extracellular matrix structure rich in chondroitin sulfate proteoglycans (CSPGs), which preferentially encase parvalbumin-containing (PV + ) interneurons. PNNs restrict cortical network plasticity but the molecular mechanisms involved are unclear. We found that reactivation of ocular dominance plasticity in the adult visual cortex induced by chondroitinase ABC (chABC)-mediated PNN removal requires intact signaling by the neurotrophin receptor TRKB in PV + neurons. Additionally, we demonstrate that chABC increases TRKB phosphorylation (pTRKB), while PNN component aggrecan attenuates brain-derived neurotrophic factor (BDNF)-induced pTRKB in cortical neurons in culture. We further found that protein tyrosine phosphatase σ (PTPσ, PTPRS), receptor for CSPGs, interacts with TRKB and restricts TRKB phosphorylation. PTPσ deletion increases phosphorylation of TRKB in vitro and in vivo in male and female mice, and juvenile-like plasticity is retained in the visual cortex of adult PTPσ-deficient mice (PTPσ +/− ). The antidepressant drug fluoxetine, which is known to promote TRKB phosphorylation and reopen critical period-like plasticity in the adult brain, disrupts the interaction between TRKB and PTPσ by binding to the transmembrane domain of TRKB. We propose that both chABC and fluoxetine reopen critical period-like plasticity in the adult visual cortex by promoting TRKB signaling in PV + neurons through inhibition of TRKB dephosphorylation by the PTPσ-CSPG complex. SIGNIFICANCE STATEMENT Critical period-like plasticity can be reactivated in the adult visual cortex through disruption of perineuronal nets (PNNs) by chondroitinase treatment, or by chronic antidepressant treatment. We now show that the effects of both chondroitinase and fluoxetine are mediated by the neurotrophin receptor TRKB in parvalbumin-containing (PV + ) interneurons. We found that chondroitinase-induced visual cortical plasticity is dependent on TRKB in PV + neurons. Protein tyrosine phosphatase σ (PTPσ, PTPRS), a receptor for PNNs, interacts with TRKB and inhibits its phosphorylation, and chondroitinase treatment or deletion of PTPσ increases TRKB phosphorylation. Antidepressant fluoxetine disrupts the interaction between TRKB and PTPσ, thereby increasing TRKB phosphorylation. Thus, juvenile-like plasticity induced by both chondroitinase and antidepressant treatment is mediated by TRKB activation in PV + interneurons.

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“…Cortex tissue was harvested from embryonic days 17–18 Wistar rat embryos as described previously 79 . Local authority approved the animal license (County Administrative Board of Southern Finland, ESAVI/10300/04.10.07/2016) to conduct the described procedures.…”

Section: Methodsmentioning

confidence: 99%

Functional characterization of human pluripotent stem cell-derived cortical networks differentiated on laminin-521 substrate: comparison to rat cortical cultures

Hyvärinen

Hyysalo

Kapucu

et al. 2019

Sci Rep

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Human pluripotent stem cell (hPSC)-derived neurons provide exciting opportunities for in vitro modeling of neurological diseases and for advancing drug development and neurotoxicological studies. However, generating electrophysiologically mature neuronal networks from hPSCs has been challenging. Here, we report the differentiation of functionally active hPSC-derived cortical networks on defined laminin-521 substrate. We apply microelectrode array (MEA) measurements to assess network events and compare the activity development of hPSC-derived networks to that of widely used rat embryonic cortical cultures. In both of these networks, activity developed through a similar sequence of stages and time frames; however, the hPSC-derived networks showed unique patterns of bursting activity. The hPSC-derived networks developed synchronous activity, which involved glutamatergic and GABAergic inputs, recapitulating the classical cortical activity also observed in rodent counterparts. Principal component analysis (PCA) based on spike rates, network synchronization and burst features revealed the segregation of hPSC-derived and rat network recordings into different clusters, reflecting the species-specific and maturation state differences between the two networks. Overall, hPSC-derived neural cultures produced with a defined protocol generate cortical type network activity, which validates their applicability as a human-specific model for pharmacological studies and modeling network dysfunctions.

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Culturing primary neurons from rat hippocampus and cortex

Cited by 73 publications

References 22 publications

Multicompartment Microreactors Prevent Excitotoxic Dysfunctions In Rat Primary Cortical Neurons

Multicompartment Microreactors Prevent Excitotoxic Dysfunctions In Rat Primary Cortical Neurons

Chondroitinase and Antidepressants Promote Plasticity by Releasing TRKB from Dephosphorylating Control of PTPσ in Parvalbumin Neurons

Functional characterization of human pluripotent stem cell-derived cortical networks differentiated on laminin-521 substrate: comparison to rat cortical cultures

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