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Habibey, R.

Publications and source records attributed to Habibey, R..

4 recordsLinked to original sources

Crosstalk between the Methyl-Cytosine Dioxygenase TET3 and the Methyl-CpG-binding protein MECP2 Controls Neuronal Maturation

Active DNA demethylation depends on Ten-Eleven-Translocation (TET) enzymes, which oxidize 5-methylcytosine (mC) to 5-hydroxymethylcytosine (hmC) and further derivatives. Mutations in TET3, encoding the predominant neuronal isoform, lead to Beck-Fahrner syndrome, a neurodevelopmental disorder. Using human iPSC-derived neurons, we show that TET3 is dispensable for neuronal specification but critical for subsequent maturation. Differentiating TET3-deficient neurons exhibit delayed transcriptional and proteomic transitions, altered synaptic signatures, and impaired network activity, indicating delayed functional maturation. Mechanistically, we identified an interaction between TET3 and the mC/hmC-binding protein MECP2, pathogenic variants of which cause Rett syndrome. MECP2 negatively regulates TET3 activity, as demonstrated in functional assays and by inverse hmC patterns in MECP2- and TET3-deficient neurons. Despite this, MECP2- and TET3-deficient neurons exhibit highly similar phenotypes later in differentiation. Our findings uncover a functional interplay between TET3 and MECP2 that coordinates DNA methylation and chromatin dynamics during neuronal maturation, suggesting a shared pathogenic mechanism in Beck-Fahrner and Rett syndromes.

neuroscience↗

Empirical validation of ephaptic coupling in printed human neural circuits

Ephaptic coupling is a phenomenon describing the influence of endogenous electric fields on neuronal activity1,2. Although ephaptic coupling is deemed to contribute to computations in the brain1,3,4, the olfactory system5, the retina6,7, and to cardiac conduction8,9, and being associated with diseases like epilepsy10,11 and arrhythmia12,13, it is still poorly understood since it is notoriously difficult to investigate in vivo and in vitro. In vitro electrophysiology allows accessible and flexible experimentation, but circuits form randomly, leading to a lack of precision and reproducibility. Here, we present a method for reproducibly constructing human neuronal networks in vitro with single-cell precision. We constructed neuronal circuits from the bottom up by bringing their axons into close contact. This enabled us to measure the effects of ephaptic coupling and validate theoretical predictions, such as reduced action potential velocity, increased activity synchronization14-16 and reduced stimulation threshold17. Our precise measurements of electrophysiological activity support the importance of ephaptic coupling in neuronal circuit function. Printed neuronal circuits allow detailed in vitro studies of neuronal interactions and may serve as a platform for disease modeling related to synaptic, ephaptic, or myelination processes.

bioengineering↗

Microengineered 2D and 3D modular neuronal networks represent structure-function relationship

Brain function is substantially linked to the highly organized structure of neuronal networks. Emerging three-dimensional (3D) neuronal cell culture technologies attempt to mimic the complexity of brain circuits as in vitro microphysiological systems. Nevertheless, structures of in vitro assembled neuronal circuits often varies between samples and changes over time that makes it challenging to reliably record network functional output and link it to the network structure. Hence, engineering neuronal structures with pre- defined geometry and reproducible functional features are essential to model in vivo neuronal circuits in a robust way. Here, we engineered thin microchannel devices to assemble 2D and 3D modular networks. Microchannel devices were coupled with multi-electrode array (MEA) electrophysiology system to enable long-term electrophysiology recordings from microengineered circuits. Each network was composed of 64 micromodules which were connected through micron size channels to their adjacent modules. Microstructures physically confined neurons to the recording electrodes that considerably enhanced the electrophysiology readout efficiency. In addition, microstructures preserved modular network structure over weeks. Modular circuits within microfluidic devices showed consistent spatial patterns of activity over weeks, which was missing in the randomly formed circuits. Number of physical connections per module was shown to be influencing the measured activity and functional connectivity parameters, that represents the impact of network structure on its functional output. We show that microengineered 3D modular networks with a profound activity and higher number of functional connections recapitulate key functional features of developing cortex. Structurally and functionally stable 2D and 3D network mimic the modular architecture of brain circuits and offers a robust and reproducible in vitro microphysiolopgical system to serve basic and translational neuroscience research.

bioengineering↗

Tracking long-term functional connectivity maps in human stem-cell-derived neuronal networks by holographic-optogenetic stimulation

Neuronal networks derived from human induced pluripotent stem cells (hiPSCs) have been exploited widely for modelling neuronal circuits, neurological diseases and drug screening. As these networks require extended culturing periods to functionally mature in vitro, most studies are based on immature networks. To obtain insights on long-term functional features of human networks, we improved a long-term glia-co-culture culturing protocol directly on multi-electrode arrays (MEA), facilitating long-term assessment of electrical features at weekly intervals. We applied optogenetic stimulation to induce neuronal activity, which resulted in accelerated neuronal responses during network development. Using holographic stimulation with single-cell-resolution, propagating evoked activities of 400 individually stimulated neurons per MEA were traceable, and precise network functional connectivity motifs were revealed. Our integrated holographic optogenetic stimulation platform on MEAs facilitates studying long-term functional dynamics of human neuronal networks in vitro. This is an important step towards establishing hiPSC-derived neurons as profound functional testbeds for basic and biomedical research. HighlightsO_LIIntegrated platform allowed long-term optogenetic experiments on hiPSC-derived networks. C_LIO_LIFull-field optogenetic stimulation boosted hiPSC-derived neuronal network activity. C_LIO_LISingle-neuron resolution holographic stimulation evoked local responses in the network. C_LIO_LIHolographic stimulation of each neuron revealed its functional connectivity patterns. C_LIO_LISubsequent holographic stimulation of more than 400 neurons revealed the whole network connectivity map. C_LI

neuroscience↗