Biology subjects

Streit, A.

Publications and source records attributed to Streit, A..

12 recordsLinked to original sources

bioRxiv2026

Spatial Patterning of Otic Vesicles in Human Inner Ear Organoids

During development axial patterning of the otic vesicle sets up the blueprint for the adult inner ear. While human inner ear organoids recapitulate the initial steps of ear formation, whether and how vesicles are properly regionalised remains unknown. Here, we combine re-analysis of published single-cell transcriptomic datasets with 2D and 3D imaging to assess axial patterning in human stem-cell-derived otic vesicles using a new PAX2-reporter line. We find that the transcriptional signatures of individual cells do not conform to clearly defined regional characteristics, with some cells co-expressing conflicting regional markers, while markers that should be co-expressed are not. Some vesicles establish spatially segregated molecular domains. However, patterning is incomplete, stochastic, and restricted to a subset of vesicles. Within the same aggregates, patterned axes lack consistent orientation. Together, our findings point to incomplete segregation of transcriptional programs that are mutually exclusive in vivo and to the absence of morphogen gradients that are crucial for otic vesicle patterning. >One sentence summaryIn human inner ear organoids, otic vesicles show incomplete and disorganized regionalisation pointing to fundamental limits of self-organization, while suggesting new routes to increase cell complexity in vitro.

developmental biology↗

bioRxiv2026

Single-cell, clonal and spatial atlases of cranial placodes illuminate their specification and evolution

The vertebrate head is defined by complex sensory structures derived from cranial placodes. Placodes arise alongside the neural crest at the neural plate border, yet the mechanisms governing their identity, diversification, and evolutionary origins are unclear. We present an integrated single-cell, spatial, and clonal atlas of placode development to resolve the dynamics of their lineage segregation. Combining single-cell RNA-sequencing, spatial transcriptomics, and high-resolution clonal tracing, we show that placodal and neighboring progenitors form a continuous transcriptional landscape with gradual transitions between domains. Domain boundary cells co-express markers of adjacent territories, suggesting transient bipotent states. Consistent with this, clonal analysis reveals sharing of progenitors between neighboring placodes, supporting a model of competitive segregation. Comparisons with amphioxus suggests that vertebrate olfactory placodes emerged from an ancestral neuroectoderm that later partitioned into distinct neural and olfactory domains. Our findings provide a unified framework for understanding the developmental and evolutionary origins of vertebrate sensory organs.

developmental biology↗

bioRxiv2025

Poly(UG)-tailed RNAs are involved in the control of thousands of genes predominantly in the germline in Pristionchus pacificus

In the nematode Caenorhabditis elegans, the terminal transferase RDE-3 adds a poly(UG)-tail to the free 3 end of RNA molecules generated by the action of primary siRNAs or piRNAs. The tailed RNA serves as a template for RNA-dependent RNA polymerases (RdRp) to generate secondary siRNAs, thereby reinforcing the RNAi effect. In this iterative process, progressively shorter tailed RNAs are formed. In C. elegans, injection of poly(UG)-tailed single-stranded RNA (ssRNA) leads to RNAi-mediated gene silencing, thereby bypassing the need for the processing of double-stranded RNA into siRNAs. We wondered if poly(UG)-tailed ssRNAs could also be used for experimental gene knockdown in nematodes where long dsRNA-mediated RNAi does not work reliably, such as the satellite model organism Pristionchus pacificus or parasitic nematodes of the genus Strongyloides. Here we show that injection of poly(UG)-tailed RNA leads to gene knock down in P. pacificus and that the injected RNA, as well as the corresponding endogenous RNA, serve as substrate for the formation of new poly(UG)-tailed RNAs. Different from C. elegans, in P. pacificus, the knockdown effect depends on the redundant activity of the three rde-1 paralogs present in this species. We detected endogenously occurring poly(UG)-tailed RNAs derived from thousands of genes, more than half of which belong to germline-specific co-expression clusters. Mutations in Ppa-rde-3, lead to sterility. In contrast, in Strongyloides spp., we found poly(UG)-tailed RNAs to be much less abundant, if not absent. Our results show that poly(UG)-tailed RNAs are not restricted to C. elegans and suggest that they play an important function in the germ line in P. pacificus.

molecular biology↗

bioRxiv2025

A single cell ATAC-seq atlas uncovers dynamic changes in chromatin accessibility during cell fate specification at the neural plate border

During development, dynamic changes in gene expression and chromatin architecture drive the transition from progenitors to specialised cell types. Here we use single cell ATAC sequencing (scATAC-seq) to investigate changes in chromatin accessibility as neural plate border cells segregate into neural, neural crest and placode cells. We developed a Nextflow pipeline, single cell Advanced Chromatin Exploration (scACE), which integrates scATAC-seq and scRNA-seq data to identify cell state specific accessibility profiles and groups of chromatin regions with coordinated dynamic behaviour, termed accessibility modules (AMs). We find that progenitors are characterised by broadly open chromatin, reflecting their broad potential to generate any ectodermal derivative. As development proceeds, cell-type-specific chromatin signatures are established. Inferring an enhancer-centric gene regulatory network, we predict Foxk2 as new regulator for placode specification and verify this prediction experimentally. Foxk2 target enhancers are open in placodal, but not any other ectodermal cells. This finding suggests that on a regulatory level, cells can use different strategies to control fate choice: differential accessibility of enhancers and broad accessibility controlled by differentially expressed transcription factors.

developmental biology↗

bioRxiv2024

High nutritional conditions influence feeding plasticity in Pristionchus pacificus and render worms non-predatory

Developmental plasticity, the ability of a genotype to produce different phenotypes in response to environmental conditions, has been subject to intense studies in the last four decades. The self-fertilizing nematode Pristionchus pacificus has been developed as a genetic model system for studying developmental plasticity due to its mouth-form polyphenism that results in alternative feeding strategies with a facultative predatory and a non-predatory mouth form. Many studies linked molecular aspects of the regulation of mouth-form polyphenism with investigations of its evolutionary and ecological significance. Also, several environmental factors influencing P. pacificus feeding structure expression were identified including temperature, culture condition and population density. However, the nutritional plasticity of the mouth form has never been properly investigated although polyphenisms are known to be influenced by changes in nutritional conditions. For instance, studies in eusocial insects and scarab beetles have provided significant mechanistic insights into the nutritional regulation of polyphenisms but also other forms of plasticity. Here, we study the influence of nutrition on mouth-form polyphenism in P. pacificus through experiments with monosaccharide and fatty acid supplementation. We show that in particular glucose supplementation renders worms non-predatory. Subsequent transcriptomic and mutant analyses indicate that de novo fatty acid synthesis and peroxisomal beta-oxidation pathways play an important role in the mediation of this plastic response. Finally, the analysis of fitness consequences through fecundity counts suggests that non-predatory animals have an advantage over predatory animals grown in the glucose-supplemented condition. Research highlightsThis study represents the first systematic attempt to investigate the influence of nutrition on mouth-form polyphenism in the genetic model organism Pristionchus pacificus. Through glucose and oleic acid supplementation we show that high nutritional conditions influence feeding plasticity and render worms non-predatory. Mutant analysis indicates a role of de novo fatty acid synthesis and peroxisomal beta-oxidation pathways for these responses.

evolutionary biology↗

bioRxiv2022

Plzf mediates a switch between Fgf signalling regimes in the developing hindbrain

Developing tissues are sequentially patterned by extracellular signals that are turned on and off at specific times. In the zebrafish hindbrain, fibroblast growth factor (Fgf) signalling has different roles at different developmental stages: in the early hindbrain, transient Fgf3 and Fgf8 signalling from rhombomere 4 is required for correct segmentation, whereas later, neuronal Fgf20 expression confines neurogenesis to specific spatial domains within each rhombomere. How the switch between these two signalling regimes is coordinated is not known. We present evidence that the promyelocytic leukaemia zinc finger (Plzf) transcription factor is required for this transition to happen in an orderly fashion. Plzf expression is high in the early anterior hindbrain, then gradually upregulated posteriorly and confined to neural progenitors. In mutants lacking functional Plzf, fgf3 expression fails to be downregulated and persists until a late stage, resulting in excess and more widespread Fgf signalling during neurogenesis. Accordingly, the spatial pattern of neurogenesis is disrupted in plzf mutants. Our results reveal how the distinct stage-specific roles of Fgf signalling are coordinated in the zebrafish hindbrain.

developmental biology↗

bioRxiv2022

A gradient border model for cell fate decisions at the neural plate border

The vertebrate neural plate border is a transient territory located at the edge of the neural plate containing precursors for all ectodermal derivatives: the neural plate; neural crest; placodes; and epidermis. Elegant functional experiments in a range of vertebrate models have provided an in-depth understanding of gene regulatory interactions within the ectoderm. However, these experiments conducted at tissue level raise seemingly contradictory models for fate allocation of individual cells. Here, we carry out single cell RNA sequencing of chick ectoderm from primitive streak to neurulation stage, to explore cell state diversity and heterogeneity. We characterise the dynamics of gene modules containing key factors known to regulate ectodermal cell fates, allowing us to model the order in which these fates are specified. Furthermore, we find that genes previously classified as neural plate border specifiers typically exhibit dynamic expression patterns and are biased towards either placodal or neural crest fates, revealing that the neural plate border should be seen as an anatomical region of the ectoderm and not a discrete transcriptional state. Through co-expression of placodal and neural crest markers, we identify a population of border located unstable progenitors (BLUPs) which gradually reduces in size as fate segregation occurs. Considering our findings, we propose a gradient border model for cell fate choice at the neural plate border, with the probability of cell fate allocation closely tied to the spatiotemporal positioning of cells.

developmental biology↗

bioRxiv2021

Sox8 is sufficient to reprogram ectoderm into ear vesicles and associated neurons

The vertebrate inner ear arises from a pool of progenitors with the potential to contribute to all the sense organs and cranial ganglia in the head. Here we explore the molecular mechanisms that control ear specification from these precursors. Using a multi-omics approach combined with loss-of-function experiments we identify a core transcriptional circuit that imparts ear identity, along with the first genome-wide characterization of non-coding elements that integrate this information. This analysis places the transcription factor Sox8 at the top of the ear determination network. Introducing Sox8 into cranial ectoderm not only converts non-ear cells into ear progenitors, but also activates the cellular programs for ear morphogenesis and neurogenesis. Thus, Sox8 has the unique ability to remodel transcriptional networks in the cranial ectoderm towards ear identity.

developmental biology↗

bioRxiv2021

A gene regulatory network for neural induction

During early vertebrate development, signals from a special region of the embryo, the organizer, can re-direct the fate of non-neural ectoderm cells to form a complete, patterned nervous system. This is called neural induction and has generally been imagined as a single signalling event, causing a switch of fate. Here we undertake a comprehensive analysis, in very fine time-course, of the events following exposure of ectoderm to the organizer. Using transcriptomics and epigenomics we generate a Gene Regulatory Network comprising 175 transcriptional regulators and 5,614 predicted interactions between them, with fine temporal dynamics from initial exposure to the signals to expression of mature neural plate markers. Using in situ hybridization, single-cell RNA-sequencing and reporter assays we show that neural induction by a grafted organizer closely resembles normal neural plate development. The study is accompanied by a comprehensive resource including information about conservation of the predicted enhancers in different vertebrate systems.

developmental biology↗

bioRxiv2021

The chromatin remodelling factor Chd7 protects auditory neurons and sensory hair cells from stress-induced degeneration

Neurons and sensory cells are particularly vulnerable to oxidative stress due to their high oxygen demand during stimulus perception and transmission1-4. The mechanisms that protect them from stress-induced death and degeneration remain elusive. Here we show that embryonic deletion of the chromodomain helicase DNA-binding protein 7 (CHD7) in auditory neurons or hair cells leads to sensorineural hearing loss due to postnatal degeneration of both cell types. Mechanistically, we demonstrate that CHD7 controls the expression of major stress pathway components. In its absence, hair cells are hypersensitive, dying rapidly after brief exposure to stress inducers, suggesting that sound at the onset of hearing triggers their degeneration. In humans, CHD7 haploinsufficiency causes CHARGE syndrome, a disorder affecting multiple organs including the ear5,6. Our findings suggest that CHD7 mutations cause developmentally silent phenotypes that predispose cells to postnatal degeneration due to a failure of protective mechanisms.

developmental biology↗

bioRxiv2019

PRDM1 controls the sequential activation of neural, neural crest and sensory progenitor determinants by regulating histone modification

During early embryogenesis, the ectoderm is rapidly subdivided into neural, neural crest and sensory progenitors. How the onset of lineage-specific determinants and the loss of pluripotency markers are temporally and spatially coordinated in vivo remains an open question. Here we identify a critical role for the transcription factor PRDM1 in the orderly transition from epiblast to defined neural lineages. Like pluripotency factors, PRDM1 is expressed in all epiblast cells prior to gastrulation, but lost as they begin to differentiate. We show that, unlike pluripotency factors, PRDM1 is initially required for the activation of neural, neural crest and sensory progenitor specifiers and for the downregulation of pluripotency-associated genes. In vivo chromatin immunoprecipitation reveals stage-specific binding of PRDM1 to regulatory regions of neural and sensory progenitor genes, PRDM1-dependent recruitment of the histone demethylase Kdm4a to these regions and associated removal of repressive histone marks. Once lineage determinants become expressed, they repress PRDM1, and our data suggest that PRDM1 downregulation is required for cells to maintain their identity. Thus, PRDM1 mediates chromatin modifications that directly control neural and sensory progenitor genes, and its activities switch from an activator at early stages to a repressor once neural fates have been established.

developmental biology↗

bioRxiv2019

FGF signalling regulates enhancer activation during ear progenitor induction

The fibroblast growth factor pathway is essential for inner ear induction in many vertebrates, however how it regulates the chromatin landscape to coordinate the activation of otic genes remains unclear. Here we show that FGF exposure of sensory progenitors leads to rapid deposition of active chromatin marks H3K27ac near hundreds of FGF-responsive, otic-epibranchial progenitor (OEP) genes, while H3K27ac is depleted in the vicinity of non-otic genes. Genomic regions that gain H3K27ac act as cis-regulatory elements controlling OEP gene expression in time and space and define a unique transcription factor signature likely to control their activity. Finally, we provide evidence that in response to FGF signalling the transcription factor dimer AP1 recruits the histone acetyl transferase p300 to OEP enhancers and that de novo acetylation is required for subsequent expression of OEP genes. Thus, during ear induction FGF signalling modifies the chromatin landscape to promote enhancer activation and chromatin accessibility.

developmental biology↗