Sirois, C. L. | Search bioRxiv

bioRxiv2025

A Semi-Automated MEA Spike sorting (SAMS) method for high throughput assessment of cultured neurons

Neurons derived from human pluripotent stem cells (hPSCs) are valuable models for studying brain development and developing therapies for brain disorders. Evaluating human-derived neurons requires assessing their electrical activity, which can be achieved using multi-electrode arrays (MEAs) for extracellular recordings. Because each electrode channel generally detects activity from multiple neurons, resolving the activity of single neurons requires a process called spike sorting. However, currently available spike sorting methods are not optimized for the analysis of hPSC-derived neurons, and require complex workflows and time-consuming manual intervention. Here, we introduce a Semi-Automated MEA Spike sorting software (SAMS) designed specifically for low-density MEA recordings of cultured neurons. SAMS outperforms commercially available automated spike sorting algorithms in terms of accuracy and greatly reduces computational and human processing time. By providing an accessible, efficient, and integrated platform for spike sorting, SAMS enhances the resolution and utility of MEA in disease modeling and drug development using human-derived neurons. HighlightsO_LISAMS is designed and optimized for high throughput analysis of hPSC-derived neurons. C_LIO_LISAMS is more efficient and accurate compared to recommended spike-sorting software. C_LIO_LISAMS resolves phenotypic differences previously not observed without spike sorting. C_LIO_LISAMS is an open-source software. C_LI

neuroscience↗

bioRxiv2020

Abundance and localization of human UBE3A protein isoforms

Loss of UBE3A expression, a gene regulated by genomic imprinting, causes Angelman Syndrome (AS), a rare neurodevelopmental disorder. The UBE3A gene encodes an E3 ubiquitin ligase with three known protein isoforms in humans. Studies in mouse suggest that the human isoforms may have differences in localization and neuronal function. A recent case study reported mild AS phenotypes in individuals lacking one specific isoform. Here we have used CRISPR/Cas9 to generate isogenic human embryonic stem cells (hESCs) that lack the individual protein isoforms. We demonstrate that isoform 1 accounts for the majority of UBE3A protein in hESCs and neurons. We also show that UBE3A predominantly localizes to the cytoplasm in both wild type and isoform-null cells. Finally, we show that neurons lacking isoform 1 display a less severe electrophysiological AS phenotype.

genetics↗