arXiv subjects

Thomas Gregor

Publications and source records attributed to Thomas Gregor.

At least 19 recordsLinked to original sources

arXiv2025

An integrated quantitative single-objective light-sheet microscope for subcellular dynamics in embryos and cultured multicellular systems

Quantitative imaging of subcellular processes in living embryos, stem-cell systems, and organoid models requires microscopy platforms that combine high spatial resolution, fast volumetric acquisition, long-term stability, and minimal phototoxicity. Single-objective light-sheet approaches based on oblique plane microscopy (OPM) are well suited for live imaging in standard sample geometries, but most existing implementations lack the optical calibration, timing precision, and end-to-end integration required for reproducible quantitative measurements. Here we present a fully integrated and quantitatively characterized OPM platform engineered for dynamic studies of transcription and nuclear organization in embryos, embryonic stem cells, and three-dimensional culture systems. The system combines high numerical aperture remote refocusing with tilt-invariant light-sheet scanning and hardware-timed synchronization of laser excitation, galvo scanning, and camera readout. We provide a comprehensive characterization of the optical performance, including point spread function, sampling geometry, usable field of view, and system stability, establishing a well-defined framework for quantitative volumetric imaging. To support high-throughput operation, we developed a unified acquisition and reconstruction pipeline that enables real time volumetric imaging at hardware-limited rates while preserving deterministic timing and reproducible geometry. Using this platform, we demonstrate quantitative three-dimensional imaging of MS2-labeled transcription sites in living Drosophila embryos, cultured mouse embryonic stem cells, and mESC-derived gastruloids, enabling extraction of transcriptional intensity traces across diverse biological contexts. This work establishes OPM as a robust and quantitatively calibrated single-objective light-sheet platform for transcription imaging in complex living systems.

q-bio.QM↗

arXiv2025

The Physics of Sustainability: Material and Power Constraints for the Long Term

Much of today's sustainability discourse emphasizes efficiency, clean technologies, and smart systems, but largely underestimates fundamental physical constraints relating to energy-matter interactions. These constraints stem from the fact that Earth is a materially closed yet energetically open system, driven by the sustained but low power-density flux of solar radiation. This Perspective reframes sustainability within these axiomatic limits, integrating relevant timescales and orders of magnitude. We argue that fossil-fueled industrial metabolism is inherently incompatible with long-term viability, while post-fossil systems are surface-, materials-, and power-intensive. Long-term sustainability must therefore be defined not only by how much energy or material is used, but also by how it is used: favoring organic, carbon-based chemistry with limited reliance on purified metals, operating at low power density, and maintaining low throughput rates. Achieving this requires radical technological shifts toward life-compatible systems and biogeochemical circular processes, and, likely as a consequence, a paradigm change toward degrowth to a steady-state. These two shifts are mutually reinforcing and together provide the necessary foundation for any viable future.

physics.soc-ph↗

arXiv2025

Invariant non-equilibrium dynamics of transcriptional regulation optimize information flow

Eukaryotic gene regulation is based on stochastic yet controlled promoter switching, during which genes transition between transcriptionally active and inactive states. Despite the molecular complexity of this process, recent studies reveal a surprising invariance of the "switching correlation time" ($T_C$), which characterizes promoter activity fluctuations, across gene expression levels in diverse genes and organisms. A biophysically plausible explanation for this invariance remains missing. Here, we show that this invariance imposes stringent constraints on minimal yet plausible models of transcriptional regulation, requiring at least four system states and non-equilibrium dynamics that break detailed balance. Using Bayesian inference on Drosophila gap gene expression data, we demonstrate that such models (i) accurately reproduce the observed $T_C$-invariance; (ii) remain robust to parameter perturbations; and (iii) maximize information transmission from transcription factor concentration to gene expression. These findings suggest that eukaryotic gene regulation has evolved to balance precision with reaction rate and energy dissipation constraints, favoring non-equilibrium architectures for optimal information transmission.

q-bio.MN↗

arXiv2025

Bridging spatial and temporal scales of developmental gene regulation

The development of multicellular organisms relies on the precise coordination of molecular events across multiple spatial and temporal scales. Understanding how information flows from molecular interactions to cellular processes and tissue organization during development is crucial for explaining the remarkable reproducibility of complex organisms. This review explores how chromatin-encoded information is transduced from localized transcriptional events to global gene expression patterns, highlighting the challenge of bridging these scales. We discuss recent experimental findings and theoretical frameworks, emphasizing polymer physics as a tool for describing the relationship between chromatin structure and dynamics across scales. By integrating these perspectives, we aim to clarify how gene regulation is coordinated across levels of biological organization and suggest strategies for future experimental approaches.

physics.bio-ph↗

arXiv2024

Transcription factor clusters as information transfer agents

Deciphering how genes interpret information from the concentration of transcription factors (TFs) within the cell nucleus remains a fundamental question in gene regulation. Recent advancements have unveiled the heterogeneous distribution of TF molecules in the nucleus, posing challenges to the precise decoding of concentration signals. To explore this phenomenon, we employ high-resolution single-cell imaging of a fluorescently tagged TF protein, Bicoid, in living fly embryos. We show that accumulation of Bicoid in submicron clusters preserves the spatial information of the maternal Bicoid gradient, and that cluster intensity, size, and frequency offer remarkably precise spatial cues. We further discover that various known gene targets of Bicoid activation colocalize with clusters and that for the target gene Hunchback, this colocalization is dependent on its enhancer binding affinity. Modeling information transfer through these clusters suggests that clustering offers a more rapid sensing mechanism for global nuclear concentrations than freely diffusing TF molecules detected by simple enhancers.

physics.bio-ph↗

arXiv2023

Scale invariance in early embryonic development

The body plan of the fruit fly is determined by the expression of just a handful of genes. We show that the spatial patterns of expression for several of these genes scale precisely with the size of the embryo. Concretely, discrete positional markers such as the peaks in striped patterns have absolute positions along the anterior-posterior axis that are proportional to embryo length, with better than 1% accuracy. Further, the information (in bits) that graded patterns of expression provide about position can be decomposed into information about fractional or scaled position and information about absolute position or embryo length; all of the available information is about scaled position, again with ~1% accuracy. These observations suggest that the underlying genetic network exhibits scale invariance in a deeper mathematical sense. Taking this mathematical statement seriously requires that the network dynamics have a zero mode, which connects to many other observations on this system.

q-bio.MN↗

arXiv2023

Finding the last bits of positional information

In a developing embryo, information about the position of cells is encoded in the concentrations of "morphogen" molecules. In the fruit fly, the local concentrations of just a handful of proteins encoded by the gap genes are sufficient to specify position with a precision comparable to the spacing between cells along the anterior--posterior axis. This matches the precision of downstream events such as the striped patterns of expression in the pair-rule genes, but is not quite sufficient to define unique identities for individual cells. We demonstrate theoretically that this information gap can be bridged if positional errors are spatially correlated, with relatively long correlation lengths. We then show experimentally that these correlations are present, with the required strength, in the fluctuating positions of the pair-rule stripes, and this can be traced back to the gap genes. Taking account of these correlations, the available information matches the information needed for unique cellular specification, within error bars of ~2%. These observation support a precisionist view of information flow through the underlying genetic networks, in which accurate signals are available from the start and preserved as they are transformed into the final spatial patterns.

q-bio.MN↗

arXiv2023

Functional analysis of a gene locus in response to non-canonical combinations of transcription factors

Transcription factor combinations determine gene locus activity and thereby cell identity. However, the precise link between concentrations of such activating transcription factors and target-gene activity is ambiguous. Here we investigate this link for the gap gene dependent activation of the even-skipped (eve) locus in the Drosophila embryo. We simultaneously measure the spatiotemporal gap gene concentrations in hemizygous and homozygous gap mutants, and link these to eve activity. Although changes in expression extend well beyond the genetically manipulated gene, nearly all expression alternations approximate the canonical combinations of activating levels in wild-type, sometimes necessitating pattern shifts. Expression levels that diverge from the wild-type repertoire still drive locus activation. Specific stripes in the homozygous mutants show partial penetrance, justifying their renown variable phenotypes. However, all eve stripes appear at highly reproducible positions, even though a broader span of gap gene expression levels activates eve. Our results suggest a correction capacity of the gap gene network and set constraints on the activity of multi-enhancer gene loci.

q-bio.MN↗

arXiv2023

Gene activity fully predicts transcriptional bursting dynamics

Transcription commonly occurs in bursts, with alternating productive (ON) and quiescent (OFF) periods, governing mRNA production rates. Yet, how transcription is regulated through bursting dynamics remains unresolved. Here, we conduct real-time measurements of endogenous transcriptional bursting with single-mRNA sensitivity. Leveraging the diverse transcriptional activities in early fly embryos, we uncover stringent relationships between bursting parameters. Specifically, we find that the durations of ON and OFF periods are linked. Regardless of the developmental stage or body-axis position, gene activity levels predict individual alleles' average ON and OFF periods. Lowly transcribing alleles predominantly modulate OFF periods (burst frequency), while highly transcribing alleles primarily tune ON periods (burst size). These relationships persist even under perturbations of cis-regulatory elements or trans-factors and account for bursting dynamics measured in other species. Our results suggest a novel mechanistic constraint governing bursting dynamics rather than a modular control of distinct parameters by distinct regulatory processes.

q-bio.MN↗

arXiv2023

Precise and scalable self-organization in mammalian pseudo-embryos

Gene expression is inherently noisy, posing a challenge to understanding how precise and reproducible patterns of gene expression emerge in mammals. We investigate this phenomenon using gastruloids, an in vitro model for early mammalian development. Our study reveals intrinsic reproducibility in the self-organization of gastruloids, encompassing growth dynamics and gene expression patterns. We observe a remarkable degree of control over gene expression along the main body axis, with pattern boundaries positioned at single-cell precision. Furthermore, as gastruloids grow, both their physical proportions and gene expression patterns scale proportionally with system size. Notably, these properties emerge spontaneously in self-organizing cell aggregates, distinct from many in vivo systems constrained by fixed boundary conditions. Our findings shed light on the intricacies of developmental precision, reproducibility, and size scaling within a mammalian system, suggesting that these phenomena might constitute fundamental features of multicellularity.

q-bio.CB↗

arXiv2023

Deriving a genetic regulatory network from an optimization principle

Many biological systems approach physical limits to their performance, motivating the idea that their behavior and underlying mechanisms could be determined by such optimality. Nevertheless, optimization as a predictive principle has only been applied in very simplified setups. Here, in contrast, we explore a mechanistically-detailed class of models for the gap gene network of the Drosophila embryo, and determine its 50+ parameters by optimizing the information that gene expression levels convey about nuclear positions, subject to physical constraints on the number of available molecules. Optimal networks recapitulate the architecture and spatial gene expression profiles of the real organism. Our framework makes precise the many tradeoffs involved in maximizing functional performance, and allows us to explore alternative networks to address the questions of necessity vs contingency. Multiple solutions to the optimization problem may be realized in closely related organisms.

physics.bio-ph↗

arXiv2022

Structured foraging of soil predators unveils functional responses to bacterial defenses

Predators and their foraging strategies often determine ecosystem structure and function. Yet, the role of protozoan predators in microbial soil ecosystems remains elusive despite the importance of these ecosystems to global biogeochemical cycles. In particular, amoebae -- the most abundant soil protozoan predators of bacteria -- remineralize soil nutrients and shape the bacterial community. However, their foraging strategies and their role as microbial ecosystem engineers remain unknown. Here we present a multi-scale approach, connecting microscopic single-cell analysis and macroscopic whole ecosystem dynamics, to expose a phylogenetically widespread foraging strategy, in which an amoeba population spontaneously partitions between cells with fast, polarized movement and cells with slow, unpolarized movement. Such differentiated motion gives rise to efficient colony expansion and consumption of the bacterial substrate. From these insights we construct a theoretical model that predicts how disturbances to amoeba growth rate and movement disrupt their predation efficiency. These disturbances correspond to distinct classes of bacterial defenses, which allows us to experimentally validate our predictions. All considered, our characterization of amoeba foraging identifies amoeba mobility, and not amoeba growth, as the core determinant of predation efficiency and a key target for bacterial defense systems.

q-bio.PE↗

arXiv2021

Eukaryotic gene regulation at equilibrium, or non?

Models of transcriptional regulation that assume equilibrium binding of transcription factors have been very successful at predicting gene expression from sequence in bacteria. However, analogous equilibrium models do not perform as well in eukaryotes, most likely due to the largely out-of-equilibrium nature of eukaryotic regulatory processes. These processes come with unavoidable energy expenditure at the molecular level to support precise, reliable, or fast gene expression responses that could correspond to evolutionarily optimized regulatory strategies. Unfortunately, the space of possible non-equilibrium mechanisms is vast and predominantly uninteresting. The key question is therefore how this space can be navigated efficiently, to focus on mechanisms and models that are biologically relevant. In this review, we advocate for the normative role of theory - theory that prescribes rather than just describes - in providing such a focus. Theory should expand its remit beyond inferring models from data (by fitting), towards identifying non-equilibrium gene regulatory schemes (by optimizing) that may have been evolutionarily selected due to their favorable functional characteristics which outperform regulation at equilibrium. This approach can help us navigate the expanding complexity of regulatory architectures, and refocus the questions about the observed biological mechanisms from how they work towards why they have evolved in the first place. We illustrate our reasoning by toy examples for which we provide simulation code.

q-bio.MN↗

arXiv2020

Transcription-dependent spatial organization of a gene locus

There is growing appreciation that gene function is connected to the dynamic structure of the chromosome. Here we explore the interplay between three-dimensional structure and transcriptional activity at the single cell level. We show that inactive loci are spatially more compact than active ones, and that within active loci the enhancer driving transcription is closest to the promoter. On the other hand, even this shortest distance is too long to support direct physical contact between the enhancer-promoter pair when the locus is transcriptionally active. Artificial manipulation of genomic separations between enhancers and the promoter produces changes in physical distance and transcriptional activity, recapitulating the correlation seen in wild-type embryos, but disruption of topological domain boundaries has no effect. Our results suggest a complex interdependence between transcription and the spatial organization of cis-regulatory elements.

q-bio.MN↗

arXiv2020

Trading bits in the readout from a genetic network

In genetic networks, information of relevance to the organism is represented by the concentrations of transcription factor molecules. In order to extract this information the cell must effectively "measure"' these concentrations, but there are physical limits to the precision of these measurements. We explore this trading between bits of precision in measuring concentration and bits of relevant information that can be extracted, using the gap gene network in the early fly embryo as an example. We argue that cells in the embryo can extract all the available information about their position, but only if the concentration measurements approach the physical limits to information capacity. These limits necessitate the observed proliferation of enhancer elements with sensitivities to combinations of transcription factors, but fine tuning of the parameters of these multiple enhancers is not required.

q-bio.MN↗

arXiv2019

Action at a distance in transcriptional regulation

There is increasing evidence that protein binding to specific sites along DNA can activate the reading out of genetic information without coming into direct physical contact with the gene. There also is evidence that these distant but interacting sites are embedded in a liquid droplet of proteins which condenses out of the surrounding solution. We argue that droplet-mediated interactions can account for crucial features of gene regulation only if the droplet is poised at a non-generic point in its phase diagram. We explore a minimal model that embodies this idea, show that this model has a natural mechanism for self-tuning, and suggest direct experimental tests.

q-bio.SC↗

arXiv2018

Precise spatial scaling in the early fly embryo

The early fly embryo offers a relatively pure version of the problem of spatial scaling in biological pattern formation. Within three hours, a "blueprint" for the final segmented body plan of the animal is visible in striped patterns of gene expression. We measure the positions of these stripes in an ensemble of 100+ embryos from a laboratory strain of Drosophila melanogaster, under controlled conditions. These embryos vary in length by only 4% (rms), yet stripes are positioned with 1% accuracy; precision and scaling of the pattern are intertwined. We can see directly the variation of absolute stripe positions with length, and the precision is so high as to exclude alternatives, such as combinations of unscaled signals from the two ends of the embryo.

q-bio.MN↗

arXiv2017

Diverse spatial expression patterns emerge from common transcription bursting kinetics

In early development, regulation of transcription results in precisely positioned and highly reproducible expression patterns that specify cellular identities. How transcription, a fundamentally noisy molecular process, is regulated to achieve reliable embryonic patterning remains unclear. In particular, it is unknown how gene-specific regulation mechanisms affect kinetic rates of transcription, and whether there are common, global features that govern these rates across a genetic network. Here, we measure nascent transcriptional activity in the gap gene network of early Drosophila embryos and characterize the variability in absolute activity levels across expression boundaries. We demonstrate that boundary formation follows a common transcriptional principle: a single control parameter determines the distribution of transcriptional activity, regardless of gene identity, boundary position, or enhancer-promoter architecture. By employing a minimalist model of transcription, we infer kinetic rates of transcriptional bursting for these patterning genes; we find that the key regulatory parameter is the fraction of time a gene is in an actively transcribing state, while the rate of Pol II loading appears globally conserved. These results point to a universal simplicity underlying the apparently complex transcriptional processes responsible for early embryonic patterning and indicate a path to general rules in transcriptional regulation.

q-bio.SC↗