Biology subjects

Wutz, A.

Publications and source records attributed to Wutz, A..

10 recordsLinked to original sources

bioRxiv2026

Brain-body timing gates conscious access across sensory systems

Conscious awareness fluctuates with the brains internal state at the moment of perception, yet how these states are temporally coordinated remains unclear. Although such internal states are reflected in ongoing neural activity, their temporal structure may arise from intrinsic interactions between the brain and the body. To test this hypothesis, we simultaneously recorded neural, cardiac, and respiratory activity while participants detected auditory, tactile, or visual stimuli presented at perceptual threshold. Across sensory modalities, stimulus detection was preceded by transient bursts of alpha-band (8-13 Hz) activity localized to lateralized prefrontal and premotor regions. Crucially, these neural events were aligned with specific phases of the cardiac and respiratory cycles, revealing coordinated brain-body states at the time of perception. This cross-system coupling predicted whether sensory stimuli reached conscious awareness. Together, these findings show that brain-body synchrony acts as a temporal gate for conscious access across sensory systems.

neuroscience↗

bioRxiv2023

The scramblases VMP1 and TMEM41b are required for primitive endoderm specification by targeting WNT signaling

The ER resident proteins VMP1 and TMEM41b share a conserved DedA domain, which confers lipid scramblase activity. Loss of either gene results in embryonic lethality in mice and defects in autophagy and lipid droplet metabolism. We set out to investigate their role in pluripotency and specification. For this purpose, we generated Vmp1 and Tmem41b mutations in mouse embryonic stem cells (ESCs). We observed that ESCs carrying mutations in Vmp1 and Tmem41b show robust self-renewal and an unperturbed pluripotent expression profile but accumulate LC3-positive autophagosomes and lipid droplets consistent with defects in autophagy and lipid metabolism. ESCs carrying combined mutations in Vmp1 and Tmem41b can differentiate into a wide range of embryonic cell types. However, differentiation into primitive endoderm-like cells in culture is impaired, and the establishment of extra- embryonic endoderm stem (XEN) cells is delayed. Mechanistically, we show the deregulation of genes that are associated with WNT signaling. This is further confirmed by cell surface proteome profiling, which identified a significant reduction of the WNT receptor FZD2 at the plasma membrane in Vmp1 and Tmem41b double mutant ESCs. Importantly, we show that transgenic expression of Fzd2 rescues XEN differentiation. Our findings identify the role of the lipid scramblases VMP1 and TMEM41b in WNT signaling during extra-embryonic endoderm development and characterize their distinct and overlapping functions.

developmental biology↗

bioRxiv2023

Comprehensive comparison of female germ cell development in vitro and in vivo identifies epigenetic gene regulation crucial for oocyte development and embryonic competence

Germ cells are the origin of new individuals. Hence, specifying germ cell identity is crucial for reproduction. The recent establishment of in vitro culture systems for generating oocytes from mouse pluripotent stem cells provides a basis for progress in studies of oogenesis and reproductive technology. However, currently the developmental competence of in vitro generated oocytes is low compared to in vivo grown oocytes. The causes underlying poor oocyte quality remain to be determined. By reconstituting germ cell development in culture from different developmental starting points within gametogenesis, we show that the differentiation of primordial germ cells (PGCs) and primordial germ cell-like cells (PGCLCs) to growing oocytes (GROs), as well as the subsequent growth of follicles are critical culture steps for specifying competence of fully-grown oocytes (FGOs) for preimplantation development. A systematic comparison of transcriptomes of single oocytes having undergone different in vitro culture trajectories identifies genes normally upregulated during oocyte growth to be susceptible for mis-regulation during in vitro oogenesis. Many of such genes have been described as targets of Polycomb repressive complexes (PRCs). Deregulation of Polycomb repression therefore likely perturbs the accumulation of cytoplasmic factors and/or setting of chromatin states in FGOs that are required for embryonic development after fertilization. Conversely, in vitro derived oocytes often displayed failure of zygotic genome activation (ZGA) and abnormal acquisition of 5-hydroxymethylcytosine (5hmC) on maternal chromosomes after activation. In addition, subcellular delocalization of pyruvate dehydrogenase (PDH) and of STELLA were observed suggesting new molecular markers for defective oocyte development. Our study identifies epigenetic regulation at an early stage of oogenesis as crucial for developmental competence and suggests specific in vitro culture steps as targets for improving oocyte quality. HighlightsO_LISingle cell transcriptomics and functional assessment of oocyte development from pluripotent stem cells in culture in a stage-specific manner provides a comprehensive resource for comparisons to oogenesis in vivo. C_LIO_LICulture steps for growth and differentiation of reconstituted follicles are critical for defining embryonic competence of in vitro generated oocytes. C_LIO_LIZygotic genome activation failure and epigenetic impairment are hallmarks of in vitro-generated oocytes that fail to develop after activation or fertilization. C_LIO_LIComputational analysis of gene expression changes and chromatin modification patterns identifies specific gene sets that indicate that Polycomb mediated repression is vulnerable during in vitro folliculogenesis. C_LI

developmental biology↗

bioRxiv2022

XIST ribonucleoproteins promote female sex-biased autoimmunity

Autoimmune diseases disproportionately affect females more than males. The XX sex chromosome complement is strongly associated with susceptibility to autoimmunity. Xist long noncoding RNA (lncRNA) is expressed only in females to randomly inactivate one of the two X chromosomes to achieve gene dosage compensation. Here, we show that the Xist ribonucleoprotein (RNP) complex, comprised of numerous autoantigenic components, is an important driver of sex-biased autoimmunity. Inducible transgenic expression of a non-silencing form of Xist in male mice introduced Xist RNP complexes and sufficed to produce autoantibodies. Male SJL/J mice expressing transgenic Xist developed more severe multiorgan pathology in pristane-induced model of lupus than wild-type males. Xist expression in males reprogrammed T and B cell population and chromatin states to more resemble wild type females. Human patients with autoimmune diseases displayed significant autoantibodies to multiple components of XIST RNP. Thus, a sex-specific lncRNA scaffolds ubiquitous RNP components to drive sex-biased immunity. HIGHLIGHTSO_LITransgenic mouse models inducibly express Xist in male animals. C_LIO_LIXist expression in males induce autoantibodies and autoimmune pathology. C_LIO_LIXist in males reprograms T and B cell populations to female-like patterns. C_LIO_LIAutoantibodies to Xist RNP characterize female-biased autoimmune diseases. C_LI

molecular biology↗

bioRxiv2021

Efficient prestimulus network integration of FFA biases face perception during binocular rivalry

Ongoing fluctuations in neural excitability and connectivity influence whether or not a stimulus is seen. Do they also influence which stimulus is seen? We recorded magnetoencepahlography data while participants viewed face or house stimuli, either one at a time or under bi-stable conditions induced through binocular rivalry. Multivariate pattern analysis revealed common neural substrates for rivalrous vs. non-rivalrous stimuli with an additional delay of ~36ms for the bi-stable stimulus and poststimulus signals were source-localized to the fusiform face area (FFA). Prior to stimulus onset followed by a face-vs. house-report, FFA showed stronger connectivity to primary visual cortex and to the rest of the cortex in the alpha frequency range (8-13 Hz) but there were no differences in local oscillatory alpha power. The pre-stimulus connectivity metrics predicted the accuracy of post-stimulus decoding and the delay associated with rivalry disambiguation suggesting that perceptual content is shaped by ongoing neural network states.

neuroscience↗

bioRxiv2020

Haploid mouse germ cell precursors from embryonic stem cells reveal Xist activation from a single X chromosome

Mammalian haploid cells have applications for genetic screening and substituting gametic genomes. Here we characterize a culture system for obtaining haploid primordial germ cell-like cells (PCGLCs) from haploid mouse embryonic stem cells (ESCs). We find that a haploid genome is maintained in PGCLCs with a high frequency indicating a substantially lower rate of diploidization than somatic cells. Characterization of the differentiating haploid ESCs reveals that Xist is activated from the single X chromosome. This observation suggests that X chromosome inactivation is initiated in haploid cells consistent with a model where autosomal blocking factors set a threshold for X-linked activators. The germline segregates from the epiblast and differs from somatic lineages in gene expression and epigenetic mechanisms. The ability of primordial germ cells for repressing Xist might contribute to the maintenance of a haploid genome.

developmental biology↗

bioRxiv2020

Polyploidy of semi-cloned embryos generated from parthenogenetic haploid embryonic stem cells

In mammals, the fusion of two gametes, an oocyte and a spermatozoon, during fertilization forms a totipotent zygote. There has been no reported case of natural parthenogenesis, in which embryos develop from unfertilized oocytes. The genome and epigenetic information of haploid gametes are crucial for the proper development of embryos. Haploid embryonic stem cells (haESCs) are unique stem cells established from uniparental blastocysts and possess only one set of chromosomes. Previous studies have shown that sperm or oocyte genome can be replaced by haESCs with or without manipulation of genomic imprinting for generation of mice. Recently, these remarkable semi-cloning methods have been applied for screening of key factors of mouse embryonic development. While haESCs have been applied as substitute of gametic genome, the fundamental mechanism how haESCs contribute to the genome of totipotent embryos is unclear. Here, we show the generation of fertile semi-cloned mice by injection of parthenogenetic haESCs (phaESCs) into oocytes after deletions of two differentially methylated regions (DMRs), the IG-DMR and H19-DMR. For characterizing the genome of semi-cloned embryos further we establish ESC lines from semi-cloned blastocysts. We report that polyploid karyotypes are observed frequently in semi-cloned ESCs (scESCs). Our results confirm that mitotically arrested phaESCs provide high efficiency for semi-cloning when the IG-DMR and H19-DMR are deleted. In addition, we highlight the occurrence of polyploidy that needs to be considered for further improvement for development of semi-cloned embryos derived by haESC injection.

developmental biology↗

bioRxiv2020

Tmed2 regulates Smoothened trafficking and Hedgehog signalling

Hedgehog (HH) signalling plays a key role in embryonic pattering and stem cell differentiation. Compounds that selectively bind Smoothened (SMO) can induce cell death in mouse embryonic stem cells (ESCs). Here we perform a genetic screen in haploid ESCs and discover that SMO inhibits a cell death pathway that resembles dissociation induced death of human ESCs and Anoikis. In mouse ESCs, SMO acts through a G-protein coupled mechanism that is independent of GLI activation. Our screen also identifies the Golgi proteins Tmed2 and Tmed10. We show that TMED2 binds SMO and controls its abundance at the plasma membrane. In neural differentiation and neural tube pattering Tmed2 acts as a repressor of HH signalling strength. We demonstrate that the interaction between SMO and TMED2 is regulated by HH signalling suggesting SMO release form the ER-Golgi is critical for controlling G-protein and GLI mediated functions of mammalian HH signalling.

cell biology↗

bioRxiv2018

Structural basis of sterol recognition by human hedgehog receptor PTCH1

Hedgehog signaling is central in embryonic development and tissue regeneration. Disruption of the pathway is linked to genetic diseases and cancer. Binding of the secreted ligand, Sonic hedgehog (ShhN) to its receptor Patched (PTCH1) activates the signaling pathway. Here, we describe a 3.4 [A] cryo-EM structure of the human PTCH1 bound to ShhNC24II, a modified hedgehog ligand mimicking its palmitoylated form. The membrane-embedded part of PTCH1 is surrounded by ten sterol molecules at the inner and outer lipid bilayer portion of the protein. The annular sterols interact at multiple sites with both the sterol sensing domain, SSD, and the SSD-like domain, SSDL, which are located on opposite sides of PTCH1. The structure reveals a possible route for sterol translocation across the lipid bilayer by PTCH1 and homologous transporters.

biochemistry↗

bioRxiv2018

Pre-stimulus feedback connectivity biases the content of visual experiences

Ongoing, spontaneous fluctuations in neural excitability and in network-wide activity patterns before stimulus onset have been proposed to underlie variability in near-threshold stimulus detection paradigms, i.e. whether an object is perceived or not. Here, we investigated the impact of pre-stimulus neural fluctuations on the content of perception, i.e. whether one or another object is perceived. We recorded neural activity with magnetoencephalography before and while participants briefly viewed a bi-stable image, the Rubin face/vase illusion, and required them to report their perceived interpretation on each trial. Using multi-variate pattern analysis, we showed robust decoding of the perceptual report during the post-stimulus period. Applying source localization to the classifier weights suggested early recruitment of V1 and ~160 ms recruitment of category-sensitive FFA. These post-stimulus effects were driven by stronger oscillatory power in the gamma frequency band for face vs vase reports. In pre-stimulus intervals, we found no differences in oscillatory power between face vs. vase reports in V1 nor in FFA, indicating similar levels of neural excitability. Despite this, we found stronger connectivity between V1 and FFA prior to face reports for low-frequency oscillations. Specifically, the strength of pre-stimulus feedback connectivity (i.e. Granger causality) from FFA to V1 predicted not only the category of the upcoming percept, but also the strength of post-stimulus neural activity associated with the percept. Our work identifies pre-stimulus network states that shape future processing in category-sensitive brain regions and in this way determine the content of visual experiences.

neuroscience↗