Helix neuronal ensembles with controlled cell type composition and placement develop functional polysynaptic circuits on Micro-Electrode Arrays

Massobrio, Paolo; Tedesco, Mariateresa; Giachello, Carlo Natale Giuseppe; Ghirardi, Mirella; Fiumara, Ferdinando; Martinoia, Sergio

doi:10.1016/j.neulet.2009.10.019

Cited by 13 publications

(17 citation statements)

References 16 publications

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“…On average 29.1% of helSynKD neurons maintained their firing after PTZ washout versus a 3.7% of control neurons (data not shown). In addition, Helix neurons rarely showed spontaneous firing when cultured on MEAs, as previously reported (Massobrio et al, 2009;Massobrio et al, 2013). In our experiments, 24 h after cell plating on MEAs, and before the first PTZ treatment, no spontaneous activity was recorded.…”

Section: Epileptic-like Activity Induced By Ptz In Syn-silenced C1 Pasupporting

confidence: 82%

“…Furthermore, it has been extensively studied concerning its electrophysiological (Chalazonitis and Takeuchi, 1968;Schulze et al, 1975;Altrup et al, 1991), morphological (Steffens, 1980), functional Ghirardi et al, 2004;Fiumara et al, 2005;Fiumara et al, 2007;Massobrio et al, 2009;Giachello et al, 2010;Massobrio et al, 2013) and epileptological…”

Section: Discussionmentioning

confidence: 99%

“…When Helix neurons are coupled 1:1 to microelectrodes, the MEA system allows an easy assessment of synaptic functionality and cross-correlograms (CC) can be used to characterize and classify the functional synapses developed between paired Helix neurons, as previously reported by Massobrio et al (2009;. Functional synapses were classified as chemical or electrical depending on the delay between the pre-and post-synaptic cell responses.…”

Section: Ptz Modulation Of Synaptogenesis In Syn-silenced C1 Paired Wmentioning

confidence: 99%

“…They are identifiable, form monosynaptic connections in vitro (Fiumara et al, 2001;Fiumara et al, 2005), and allow a variety of experimental manipulations that are not feasible with mammalian neurons, such as single cell-electrode coupling on microelectrode arrays (MEAs) (Massobrio et al, 2009;Massobrio et al, 2013) and intranuclear DNA injection (Brenes et al, 2015a;Brenes et al, 2015b). The convenient genetic organization (one single Syn gene) of this organism allows antisense RNA (asRNA)-dependent constitutive Syn depletion without cellular compensatory mechanisms.…”

Section: Introductionmentioning

confidence: 99%

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Subconvulsant doses of pentylenetetrazol uncover the epileptic phenotype of cultured synapsin-deficient Helix serotonergic neurons in the absence of excitatory and inhibitory inputs

Brenes

Carabelli²,

Gosso³

et al. 2016

Epilepsy Research

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Synapsins are a family of presynaptic proteins related to several processes of synaptic functioning. A variety of reports have linked mutations in synapsin genes with the development of epilepsy. Among the proposed mechanisms, a main one is based on the synapsin-mediated imbalance towards network hyperexcitability due to differential effects on neurotransmitter release in GABAergic and glutamatergic synapses. Along this line, a non-synaptic effect of synapsin depletion increasing neuronal excitability has recently been described in Helix neurons. To further investigate this issue, we examined the effect of synapsin knock-down on the development of pentylenetetrazol (PTZ)-induced epileptic-like activity using single neurons or isolated monosynaptic circuits reconstructed on microelectrode arrays (MEAs). Compared to control neurons, synapsin-silenced neurons showed a lower threshold for the development of epileptic-like activity and prolonged periods of activity, together with the occurrence of spontaneous firing after recurrent PTZ-induced epileptic-like activity. These findings highlight the crucial role of synapsin on neuronal excitability regulation in the absence of inhibitory or excitatory inputs. KeywordsSynapsin; pentylenetetrazol (PTZ); invertebrate neurons; convulsants; Helix snail. Highlights• Synapsin-silenced cells lacking synaptic inputs exhibit greater PTZ susceptibility.• PTZ-induced epileptic-like behaviors were increased in synapsin-silenced cells.• Recurrent epileptic-like activity impairs control neuron synaptogenesis.• Recurrent epileptic-like activity is associated with spontaneous spiking activity.

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Section: Epileptic-like Activity Induced By Ptz In Syn-silenced C1 Pasupporting

confidence: 82%

Section: Discussionmentioning

confidence: 99%

Section: Ptz Modulation Of Synaptogenesis In Syn-silenced C1 Paired Wmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Subconvulsant doses of pentylenetetrazol uncover the epileptic phenotype of cultured synapsin-deficient Helix serotonergic neurons in the absence of excitatory and inhibitory inputs

Brenes

Carabelli²,

Gosso³

et al. 2016

Epilepsy Research

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“…When paired in culture, these neurons reliably form excitatory synaptic connections, and display several forms of activity-dependent synaptic enhancement, including PTP [44]. Both synaptically isolated and chemically interconnected B2 neurons are usually silent in culture [47]. Soon after the application of PTZ, we observed in these cells a peculiar neural activity that can be divided into three phases (Fig.…”

Section: Resultsmentioning

confidence: 79%

Pentylenetetrazol-Induced Epileptiform Activity Affects Basal Synaptic Transmission and Short-Term Plasticity in Monosynaptic Connections

et al. 2013

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Epileptic activity is generally induced in experimental models by local application of epileptogenic drugs, including pentylenetetrazol (PTZ), widely used on both vertebrate and invertebrate neurons. Despite the high prevalence of this neurological disorder and the extensive research on it, the cellular and molecular mechanisms underlying epileptogenesis still remain unclear. In this work, we examined PTZ-induced neuronal changes in Helix monosynaptic circuits formed in vitro, as a simpler experimental model to investigate the effects of epileptiform activity on both basal release and post-tetanic potentiation (PTP), a form of short-term plasticity. We observed a significant enhancement of basal synaptic strength, with kinetics resembling those of previously described use-dependent forms of plasticity, determined by changes in estimated quantal parameters, such as the readily releasable pool and the release probability. Moreover, these neurons exhibited a strong reduction in PTP expression and in its decay time constant, suggesting an impairment in the dynamic reorganization of synaptic vesicle pools following prolonged stimulation of synaptic transmission. In order to explain this imbalance, we determined whether epileptic activity is related to the phosphorylation level of synapsin, which is known to modulate synaptic plasticity. Using western blot and immunocytochemical staining we found a PTZ-dependent increase in synapsin phosphorylation at both PKA/CaMKI/IV and MAPK/Erk sites, both of which are important for modulating synaptic plasticity. Taken together, our findings suggest that prolonged epileptiform activity leads to an increase in the synapsin phosphorylation status, thereby contributing to an alteration of synaptic strength in both basal condition and tetanus-induced potentiation.

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Insects Neural Model: Potential Alternate to Mammals for Electrophysiological Studies

Gaburro

Nahavandi

Bhatti

2017

Series in BioEngineering

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Helix neuronal ensembles with controlled cell type composition and placement develop functional polysynaptic circuits on Micro-Electrode Arrays

Cited by 13 publications

References 16 publications

Subconvulsant doses of pentylenetetrazol uncover the epileptic phenotype of cultured synapsin-deficient Helix serotonergic neurons in the absence of excitatory and inhibitory inputs

Subconvulsant doses of pentylenetetrazol uncover the epileptic phenotype of cultured synapsin-deficient Helix serotonergic neurons in the absence of excitatory and inhibitory inputs

Pentylenetetrazol-Induced Epileptiform Activity Affects Basal Synaptic Transmission and Short-Term Plasticity in Monosynaptic Connections

Insects Neural Model: Potential Alternate to Mammals for Electrophysiological Studies

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