Biology subjects

Otor, Y.

Publications and source records attributed to Otor, Y..

2 recordsLinked to original sources

bioRxiv2024

Electrical Coordinated Reset Stimulation Induces Network Desynchronization in an in Vivo Model of Status Epilepticus

Epilepsy, a neurological disorder characterized by recurrent seizures, profoundly impacts individuals worldwide. Various electrical stimulation protocols have been investigated to mitigate epileptic seizures, among which Coordinated Reset (CR) stimulation may have potential for inducing long-lasting neural desynchronization. This study explores the acute effects of CR stimulation on synchronization dynamics during Status Epilepticus (SE) in an in vivo animal model. An electrographically sustained seizure-state was induced via 4-aminopyridine (4AP) administration to CA3. Custom-designed electrode probes were implanted to facilitate simultaneous recording and electrical stimulation. Analytical univariate and bivariate features were constructed from the LFP time-series recording. Feature metrics focused on spike synchronization metrics and continuous signal analysis of amplitude, spectral power and phase synchronization across electrode pairs and frequency bands. Significance of modulation was assessed through permutation testing of the observed differences between the CR-stimulated group (N=5) compared to the control (no stimulation) group (N=3) during SE. Results showed overall decrease in amplitude and power univariate features, and a significant modulation of bivariate synchronization and connectivity measures across the spectrum between the CR stimulation and control group. Our findings underscore the potential effectiveness of CR stimulation in attenuating excessive neural synchronization, paving the way for further exploration of CR stimulation as a viable intervention for network desynchronization of epileptiform activity and subsequently treatment of seizures.

neuroscience↗

bioRxiv2023

A novel polycistronic method tailored for engineering split GECIs

We assessed the feasibility of using stop-codons as means to obtain polycistronic expression in eukaryotic cells. We show robust bicistronic expression of different open reading frames (ORFs), when these are cloned in-sequence and simply separated by stop codons (in-or out-of-frame), in heterologous expression systems and primary neurons. We further find this method to support polycistronic expression of three stop-codon-separated ORFs in vivo, which guided us to develop a technicolor Genetically-Encoded Functional Rainbow Indicators (GEFRIs) for monitoring cellular morphology and neuronal firing, concomitantly. These findings guided us to develop a new technique we denote SPLIT--Stop-codon mediated Polycistronic Induction in HeTerologous expression systems-- for rapid and easy development of fragmented proteins by the sole use of stop codons. We validated the SPLIT method by generating several new split-GFP variants, then engineer a palette of functional split-GCaMP6s variants and, lastly, generate a split ca2+-probe localized at ER and mitochondria junctions, denoted split-MEGIC. With the use of the probe, we show presence and activity of mito-ER contact sites within individual dendritic spines. Split-MEGIC can thereby be imaged by two-photon excitation in vivo in mice brains and, by standard confocal microscope in transgenic zebrafish larvae. Together, we explore non-canonical translation mechanisms and show these to be highly pervasive in various cell types in vitro and in vivo. We harness translation re-initiation to express multiple ORFs, to engineer rainbow indicators and to swiftly produce functional split-proteins and probes.

neuroscience↗