Althaus, J. C. | Search bioRxiv

bioRxiv2026

Protocol for Single-Cell Patch-Proteomics: identify specific proteins associated with electrophysiological properties from individual neurons

Single-cell patch-proteomics is a novel and powerful technique combining electrophysiological and quantitative proteomic characterization from individually sampled neurons. Following whole-cell patch clamp characterization of neuronal excitability, we collect intracellular contents and, using the latest advancements in Liquid Chromatography and Mass Spectrometry, identify and quantify peptides and their corresponding proteins. By cell-matched one-to-one associations between electrophysiological properties and protein abundance, this approach provides a means to elucidate molecular mechanisms of neuronal function on individual neuron basis.

neuroscience↗

bioRxiv2019

RAI1 Regulates Activity-Dependent Nascent Transcription and Synaptic Scaling

Long-lasting forms of synaptic plasticity such as synaptic scaling are critically dependent on transcription. Activity-dependent transcriptional dynamics in neurons, however, have not been fully characterized, because most previous efforts relied on measurement of steady-state mRNAs. Here, we profiled transcriptional dynamics of primary neuronal cultures undergoing network activity shifts using nascent RNA sequencing. We found pervasive transcriptional changes, in which ~45% of expressed genes respond to network activity shifts. Notably, the majority of these genes respond to increases or decreases of network activity uniquely, rather than reciprocally. We further linked the chromatin regulator Retinoic acid induced 1 (RAI1), the Smith-Magenis Syndrome gene, to the specific transcriptional program driven by reduced network activity. Finally, we show that RAI1 is essential for homeostatic synaptic upscaling but not downscaling. These results demonstrate the utility of bona fide transcription profiling to discover mechanisms of activity-dependent chromatin remodeling that underlie normal and pathological synaptic plasticity.

neuroscience↗