Determinants of spontaneous activity in networks of cultured hippocampus

Cohen, Eyal; Ivenshitz, Miriam; Amor-Baroukh, Veronique; Greenberger, Varda; Segal, Menahem

doi:10.1016/j.brainres.2008.06.022

Cited by 86 publications

(100 citation statements)

References 18 publications

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“…Above all, we describe a serum -free culturing method for long -term neuronal cultures with preserved functionality and connectivity, reminiscent of that seen in the intact tissue. The three culturing protocols that we compared were: I-FBS based medium (traditionally used in our lab 26 and by other research groups 17,27,28 ; II-Neurobasal/B27 medium as described in Beaudoin 7 ; III-our serum -free ACM medium. The exact composition for all the three growing media is reported in the Methods section and the conditions are referred to as FBS, B27 and ACM, respectively.…”

Section: Introduction *Manuscript (With Page Numbers) Click Here To Dmentioning

confidence: 99%

An improved method for growing neurons: Comparison with standard protocols

Pozzi

Ban

Iseppon

et al. 2017

Journal of Neuroscience Methods

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Highlights An improved protocol for primary hippocampal cell cultures is proposed.  The method relies on serum-free astrocytes conditioned medium (ACM).  The ACM method is extensively compared with other two commonly used protocols.  ACM improved morphology and function of both short-and long-term cultures. AbstractBackground: Since different culturing parameters -such as media composition or cell density -lead to different experimental results, it is important to define the protocol used for neuronal cultures. The vital role of astrocytes in maintaining homeostasis of neurons -both in vivo and in vitro -is well established: the majority of improved culturing conditions for primary dissociated neuronal cultures rely on astrocytes.New Method: Our culturing protocol is based on a novel serum-free preparation of astrocyteconditioned medium (ACM). We compared the proposed ACM culturing method with other two commonly used methods: Neurobasal/B27-and FBS-based media. We performed morphometric characterization by immunocytochemistry and functional analysis by calcium imaging for all three culture methods at 1,7,14 and 60 days in vitro (DIV).Results: ACM-based cultures gave the best results for all tested criteria, i.e. growth cone's size and shape, neuronal outgrowth and branching, network activity and synchronization, maturation and long-term survival. The differences were more pronounced when compared with FBS-based medium. Neurobasal/B27 cultures were comparable to ACM for young cultures (DIV1), but not for culturing times longer than DIV7. Comparison with Existing Method(s):ACM-based cultures showed more robust neuronal outgrowth at DIV1. At DIV7 and 60, the activity of neuronal network grown in ACM had a more vigorous spontaneous electrical activity and a higher degree of synchronization. Conclusions:We propose our ACM-based culture protocol as an improved and more suitable method for both short-and long-term neuronal cultures.

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Section: Introduction *Manuscript (With Page Numbers) Click Here To Dmentioning

confidence: 99%

An improved method for growing neurons: Comparison with standard protocols

Pozzi

Ban

Iseppon

et al. 2017

Journal of Neuroscience Methods

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“…Yet, while the forms of synchrony observed in vitro differ in many respects from those associated with low arousal levels in the intact brain, their underlying biophysical mechanisms share important similarities. Both in vivo [10,15,17,21,44] and in vitro [27,33,34,37,[45][46][47] experiments, as well as modeling studies [21,39,40,48,49], indicate that these forms of synchrony are not imposed by some external circuitry, global inhibition or pacemaker cells, but probably arise from the interplay of spontaneous synaptic activity, nonlinear neuronal recruitment cascades, refractoriness and network wide synaptic depression (summarized in [17]), effectively giving rise to a default activity mode, as it has been referred to [35,36] (see also [39]). Furthermore, and in full concordance with their activities in vivo [44,[50][51][52][53][54][55][56][57][58][59][60][61], cholinergic and adrenergic agonists suppress network synchrony in cell culture and slice preparations, shifting spontaneous activity away from this 'default' mode towards desynchronized, tonic firing modes [35,36,[62][63][64][65].…”

Section: Introductionmentioning

confidence: 99%

Adaptation to prolonged neuromodulation in cortical cultures: an invariable return to network synchrony

2014

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Background: Prolonged neuromodulatory regimes, such as those critically involved in promoting arousal and suppressing sleep-associated synchronous activity patterns, might be expected to trigger adaptation processes and, consequently, a decline in neuromodulator-driven effects. This possibility, however, has rarely been addressed. Results: Using networks of cultured cortical neurons, acetylcholine microinjections and a novel closed-loop 'synchrony-clamp' system, we found that acetylcholine pulses strongly suppressed network synchrony. Over the course of many hours, however, synchrony invariably reemerged, even when feedback was used to compensate for declining cholinergic efficacy. Network synchrony also reemerged following its initial suppression by noradrenaline, but this did not occlude the suppression of synchrony or its gradual reemergence following subsequent cholinergic input. Importantly, cholinergic efficacy could be restored and preserved over extended time scales by periodically withdrawing cholinergic input. Conclusions: These findings indicate that the capacity of neuromodulators to suppress network synchrony is constrained by slow-acting, reactive processes. A multiplicity of neuromodulators and ultimately neuromodulator withdrawal periods might thus be necessary to cope with an inevitable reemergence of network synchrony.

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“…Although existing 3D anatomical connections are lost during the preparation of primary neurons (e.g. extracted from mouse embryos), bust, synchronized network activity 14,18 . Neuronal electrical activity can be visualized by means of voltage or calcium sensors, both of which are available as synthetic dyes or genetically encoded fluorescent proteins [30][31][32][33] .…”

Section: Models For Studying Neuronal Connectivitymentioning

confidence: 99%

Image Informatics Strategies for Deciphering Neuronal Network Connectivity

Detrez

Verstraelen

Gebuis

et al. 2016

Focus on Bio-Image Informatics

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Brain function relies on an intricate network of highly dynamic neuronal connections that rewires dramatically under the impulse of various external cues and pathological conditions. Among the neuronal structures that show morphological plasticity are neurites, synapses, dendritic spines and even nuclei. This structural remodelling is directly connected with functional changes such as intercellular communication and the associated calcium-bursting behaviour. In vitro cultured neuronal networks are valuable models for studying these morpho-functional changes. Owing to the automation and standardisation of both image acquisition and image analysis, it has become possible to extract statistically relevant readout from such networks. Here, we focus on the current state-of-the-art in image informatics that enables quantitative microscopic interrogation of neuronal networks. We describe the major correlates of neuronal connectivity and present workflows for analysing them. Finally, we provide an outlook on the challenges that remain to be addressed, and discuss how imaging algorithms can be extended beyond in vitro imaging studies.2

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Determinants of spontaneous activity in networks of cultured hippocampus

Cited by 86 publications

References 18 publications

An improved method for growing neurons: Comparison with standard protocols

An improved method for growing neurons: Comparison with standard protocols

Adaptation to prolonged neuromodulation in cortical cultures: an invariable return to network synchrony

Image Informatics Strategies for Deciphering Neuronal Network Connectivity

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