bioRxiv Science⌕ Search

bioRxiv · 10.1101/2024.08.26.609746

Transcriptomically-measured gene expression predicts physiological variation across single neurons in humans and mice

Abstract

Single-cell transcriptomics measures the molecular landscape of individual neurons with unprecedented efficiency and scale. This insight has the potential to advance our understanding of the molecular basis of neuronal function, and to identify druggable targets for disease treatments. However, transcriptomics also suffers from greater measurement noise than traditional techniques (e.g., RT-PCR), which raises questions about its ability to offer insight into function at true single-cell resolution. We tested if transcriptomic data could yield insight into function of individual neurons in human and mouse neocortex by analyzing two datasets collected via Patch-Seq, a powerful technique for obtaining transcriptomic and physiology data from the same neuron. We found that computational models trained on single-cell transcriptomic data robustly predicted physiology of individual neurons. Critically, models trained on single cells outperformed those trained on cell type averages when predicting single-cell physiology. Thus, the standard approach of denoising single-cell transcriptomic data by averaging on cell types sacrifices functionally-relevant information. Our analysis also revealed novel relationships between gene expression and physiology, including a potential molecular substrate of human- mouse cross-species differences in the speed of single-neuron computation. Broadly, our findings highlight the promise of Patch-Seq for generating new insight into the molecular basis of neuronal function.

Explore related subjects

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Kendrick, R. M., Linderman, S., Owen, S. F.. 2024-08-26. Transcriptomically-measured gene expression predicts physiological variation across single neurons in humans and mice. https://doi.org/10.1101/2024.08.26.609746

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

A Mechanistic Account of Dedifferentiation in the Aging Visual System

To uncover the mechanisms of age-related dedifferentiation, we simulated neural degradation in a computational model of the ventral visual stream. Severing connections or adding noise, whether in inferotemporal cortex (IT) or upstream, reduced IT differentiation, indicating that dedifferentiation need not originate locally. Degrading peripheral V1, which preferentially drove place-selective IT neurons, selectively reduced place differentiation, a candidate mechanism for the stronger place dedifferentiation reported in aging.

neuroscience↗

Reusing pipettes in Patch-seq: a foundation for automated multimodal profiling

Patch-seq is a technique for single-cell multimodal characterization. It combines whole-cell electrophysiology, single-cell transcriptomics, and morphological reconstruction in single neurons. Patch-seq morphologies are a critical bridge among cell types, genes, physiology, and large-scale connectivity platforms such as EM connectomes and single-cell-resolution projectomes. However, patch-seq experimental complexity and operator dependence limit throughput. Automation could enable scalable Patch-seq but requires procedural adaptations, including detergent-based pipette cleaning, robotic nucleus extraction and collection. The impact of these automations on Patch-seq quality has not been systematically evaluated. Here, we show that each of these modifications preserves or improves key Patch-seq outcomes. Automated nucleus extraction performs comparably to, or better than, expert manual operation, preserving key RNA and morphological metrics. Detergent-cleaned, reused pipettes supported a viable sealing success rate across multiple cleaning cycles and preserved electrophysiological quality and transcriptomic yield without introducing excess contamination. Sonication effectively allows collection of nuclear material without damaging RNA and supports subsequent pipette cleaning and reuse. Enhanced automated sealing and break-in procedures increased throughput regardless of brain region, cell type or species. Together, these results validate the core procedural adaptations required for automated Patch-seq and establish a foundation for high-throughput, standardized multimodal single-cell characterization.

neuroscience↗

Development of direction selectivity in the Xenopus tadpole optic tectum and midbrain tegmentum

Detecting motion direction is a fundamental function performed by direction selective (DS) neurons. DS neurons in the optic tectum (superior colliculus in mammals) enable the tracking of moving objects. DS neurons of the accessory optic system support reflexive eye / head movements that stabilize gaze during self-motion. By developmental stage 48 (10 days postfertilization), Xenopus tadpoles display visually guided behaviors that require both types of DS neurons. Our previous work has shown that these two regions are not only functionally distinct but also follow different developmental programs. Here we characterized and compared how direction selectivity develops in both visual processing centers. Using in vivo electrophysiological recordings during moving bar visual stimulation, we measured DS synaptic inputs and spiking outputs in both regions across three key developmental stages of the retinotectal projection. Tegmental neurons were direction selective ab initio, and both input and output tuning remained stable across development. Tectal neurons were observed to receive DS inputs early, and their input selectivity also remained stable across development. However, their spiking output grew substantially more DS at the later developmental stage. This was due at least in part to a naturally occurring decrease in intrinsic excitability which preferentially dampened overall spike rates, but especially in response to the non-preferred direction of motion, thereby increasing direction selectivity. These results reveal distinct developmental trajectories in parallel visual pathways. Tegmental direction selectivity is established early and stays stable, whereas tectal output tuning sharpens during development through a decrease in intrinsic excitability.

neuroscience↗