@phdthesis{12809, abstract = {Understanding the mechanisms of learning and memory formation has always been one of the main goals in neuroscience. Already Pavlov (1927) in his early days has used his classic conditioning experiments to study the neural mechanisms governing behavioral adaptation. What was not known back then was that the part of the brain that is largely responsible for this type of associative learning is the cerebellum. Since then, plenty of theories on cerebellar learning have emerged. Despite their differences, one thing they all have in common is that learning relies on synaptic and intrinsic plasticity. The goal of my PhD project was to unravel the molecular mechanisms underlying synaptic plasticity in two synapses that have been shown to be implicated in motor learning, in an effort to understand how learning and memory formation are processed in the cerebellum. One of the earliest and most well-known cerebellar theories postulates that motor learning largely depends on long-term depression at the parallel fiber-Purkinje cell (PC-PC) synapse. However, the discovery of other types of plasticity in the cerebellar circuitry, like long-term potentiation (LTP) at the PC-PC synapse, potentiation of molecular layer interneurons (MLIs), and plasticity transfer from the cortex to the cerebellar/ vestibular nuclei has increased the popularity of the idea that multiple sites of plasticity might be involved in learning. Still a lot remains unknown about the molecular mechanisms responsible for these types of plasticity and whether they occur during physiological learning. In the first part of this thesis we have analyzed the variation and nanodistribution of voltagegated calcium channels (VGCCs) and α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid type glutamate receptors (AMPARs) on the parallel fiber-Purkinje cell synapse after vestibuloocular reflex phase reversal adaptation, a behavior that has been suggested to rely on PF-PC LTP. We have found that on the last day of adaptation there is no learning trace in form of VGCCs nor AMPARs variation at the PF-PC synapse, but instead a decrease in the number of PF-PC synapses. These data seem to support the view that learning is only stored in the cerebellar cortex in an initial learning phase, being transferred later to the vestibular nuclei. Next, we have studied the role of MLIs in motor learning using a relatively simple and well characterized behavioral paradigm – horizontal optokinetic reflex (HOKR) adaptation. We have found behavior-induced MLI potentiation in form of release probability increase that could be explained by the increase of VGCCs at the presynaptic side. Our results strengthen the idea of distributed cerebellar plasticity contributing to learning and provide a novel mechanism for release probability increase. }, author = {Alcarva, Catarina}, issn = {2663-337X}, pages = {115}, publisher = {Institute of Science and Technology Austria}, title = {{Plasticity in the cerebellum: What molecular mechanisms are behind physiological learning}}, doi = {10.15479/at:ista:12809}, year = {2023}, } @phdthesis{14280, abstract = {Cell division in Escherichia coli is performed by the divisome, a multi-protein complex composed of more than 30 proteins. The divisome spans from the cytoplasm through the inner membrane to the cell wall and the outer membrane. Divisome assembly is initiated by a cytoskeletal structure, the so-called Z-ring, which localizes at the center of the E. coli cell and determines the position of the future cell septum. The Z-ring is composed of the highly conserved bacterial tubulin homologue FtsZ, which forms treadmilling filaments. These filaments are recruited to the inner membrane by FtsA, a highly conserved bacterial actin homologue. FtsA interacts with other proteins in the periplasm and thus connects the cytoplasmic and periplasmic components of the divisome. A previous model postulated that FtsA regulates maturation of the divisome by switching from an oligomeric, inactive state to a monomeric and active state. This model was based mostly on in vivo studies, as a biochemical characterization of FtsA has been hampered by difficulties in purifying the protein. Here, we studied FtsA using an in vitro reconstitution approach and aimed to answer two questions: (i) How are dynamics from cytoplasmic, treadmilling FtsZ filaments coupled to proteins acting in the periplasmic space and (ii) How does FtsA regulate the maturation of the divisome? We found that the cytoplasmic peptides of the transmembrane proteins FtsN and FtsQ interact directly with FtsA and can follow the spatiotemporal signal of FtsA/Z filaments. When we investigated the underlying mechanism by imaging single molecules of FtsNcyto, we found the peptide to interact transiently with FtsA. An in depth analysis of the single molecule trajectories helped to postulate a model where PG synthases follow the dynamics of FtsZ by a diffusion and capture mechanism. Following up on these findings we were interested in how the self-interaction of FtsA changes when it encounters FtsNcyto and if we can confirm the proposed oligomer-monomer switch. For this, we compared the behavior of the previously identified, hyperactive mutant FtsA R286W with wildtype FtsA. The mutant outperforms WT in mirroring and transmitting the spatiotemporal signal of treadmilling FtsZ filaments. Surprisingly however, we found that this was not due to a difference in the self-interaction strength of the two variants, but a difference in their membrane residence time. Furthermore, in contrast to our expectations, upon binding of FtsNcyto the measured self-interaction of FtsA actually increased. We propose that FtsNcyto induces a rearrangement of the oligomeric architecture of FtsA. In further consequence this change leads to more persistent FtsZ filaments which results in a defined signalling zone, allowing formation of the mature divisome. The observed difference between FtsA WT and R286W is due to the vastly different membrane turnover of the proteins. R286W cycles 5-10x faster compared to WT which allows to sample FtsZ filaments at faster frequencies. These findings can explain the observed differences in toxicity for overexpression of FtsA WT and R286W and help to understand how FtsA regulates divisome maturation.}, author = {Radler, Philipp}, isbn = {978-3-99078-033-6}, issn = {2663-337X}, keywords = {Cell Division, Reconstitution, FtsZ, FtsA, Divisome, E.coli}, pages = {156}, publisher = {Institute of Science and Technology Austria}, title = {{Spatiotemporal signaling during assembly of the bacterial divisome}}, doi = {10.15479/at:ista:14280}, year = {2023}, } @phdthesis{12781, abstract = {Most energy in humans is produced in form of ATP by the mitochondrial respiratory chain consisting of several protein assemblies embedded into lipid membrane (complexes I-V). Complex I is the first and the largest enzyme of the respiratory chain which is essential for energy production. It couples the transfer of two electrons from NADH to ubiquinone with proton translocation across bacterial or inner mitochondrial membrane. The coupling mechanism between electron transfer and proton translocation is one of the biggest enigma in bioenergetics and structural biology. Even though the enzyme has been studied for decades, only recent technological advances in cryo-EM allowed its extensive structural investigation. Complex I from E.coli appears to be of special importance because it is a perfect model system with a rich mutant library, however the structure of the entire complex was unknown. In this thesis I have resolved structures of the minimal complex I version from E. coli in different states including reduced, inhibited, under reaction turnover and several others. Extensive structural analyses of these structures and comparison to structures from other species allowed to derive general features of conformational dynamics and propose a universal coupling mechanism. The mechanism is straightforward, robust and consistent with decades of experimental data available for complex I from different species. Cyanobacterial NDH (cyanobacterial complex I) is a part of broad complex I superfamily and was studied as well in this thesis. It plays an important role in cyclic electron transfer (CET), during which electrons are cycled within PSI through ferredoxin and plastoquinone to generate proton gradient without NADPH production. Here, I solved structure of NDH and revealed additional state, which was not observed before. The novel “resting” state allowed to propose the mechanism of CET regulation. Moreover, conformational dynamics of NDH resembles one in complex I which suggest more broad universality of the proposed coupling mechanism. In summary, results presented here helped to interpret decades of experimental data for complex I and contributed to fundamental mechanistic understanding of protein function. }, author = {Kravchuk, Vladyslav}, isbn = {978-3-99078-029-9}, issn = {2663-337X}, pages = {127}, publisher = {Institute of Science and Technology Austria}, title = {{Structural and mechanistic study of bacterial complex I and its cyanobacterial ortholog}}, doi = {10.15479/at:ista:12781}, year = {2023}, } @phdthesis{12891, abstract = {The tight spatiotemporal coordination of signaling activity determining embryo patterning and the physical processes driving embryo morphogenesis renders embryonic development robust, such that key developmental processes can unfold relatively normally even outside of the full embryonic context. For instance, embryonic stem cell cultures can recapitulate the hallmarks of gastrulation, i.e. break symmetry leading to germ layer formation and morphogenesis, in a very reduced environment. This leads to questions on specific contributions of embryo-specific features, such as the presence of extraembryonic tissues, which are inherently involved in gastrulation in the full embryonic context. To address this, we established zebrafish embryonic explants without the extraembryonic yolk cell, an important player as a signaling source and for morphogenesis during gastrulation, as a model of ex vivo development. We found that dorsal-marginal determinants are required and sufficient in these explants to form and pattern all three germ layers. However, formation of tissues, which require the highest Nodal-signaling levels, is variable, demonstrating a contribution of extraembryonic tissues for reaching peak Nodal signaling levels. Blastoderm explants also undergo gastrulation-like axis elongation. We found that this elongation movement shows hallmarks of oriented mesendoderm cell intercalations typically associated with dorsal tissues in the intact embryo. These are disrupted by uniform upregulation of BMP signaling activity and concomitant explant ventralization, suggesting that tight spatial control of BMP signaling is a prerequisite for explant morphogenesis. This control is achieved by Nodal signaling, which is critical for effectively downregulating BMP signaling in the mesendoderm, highlighting that Nodal signaling is not only directly required for mesendoderm cell fate specification and morphogenesis, but also by maintaining low levels of BMP signaling at the dorsal side. Collectively, we provide insights into the capacity and organization of signaling and morphogenetic domains to recapitulate features of zebrafish gastrulation outside of the full embryonic context.}, author = {Schauer, Alexandra}, issn = {2663-337X}, pages = {190}, publisher = {Institute of Science and Technology Austria}, title = {{Mesendoderm formation in zebrafish gastrulation: The role of extraembryonic tissues}}, doi = {10.15479/at:ista:12891}, year = {2023}, } @phdthesis{14697, abstract = {During my Ph.D. research, I managed a series of projects, each focused on the mechanisms underlying cell migration. My work involved an in-depth examination of the complex strategies employed by neutrophils, with a specific focus on their ability to synchronize spatial-temporal cues and optimize their gradient perception. However, it is essential to acknowledge that not all projects yielded successful results, as some ideas were discontinued and are archived for future reference within this thesis. My main project investigated how neutrophils decode spatial cues for precise navigation. Human neutrophils showcased distinct movement patterns based on source type – linear or point-like. By combining single-cell tracking in 3D environments with proxy dyes, this project linked cell behaviors to gradient changes, revealing a stronger response to semi-exponential gradients from point sources. In addition, neutrophils exhibited oscillating migration speeds, using speed minima to adjust trajectories toward sources. Experiencing continuous concentration changes, they accelerated over time and employed a "Run and Fumble" strategy, alternating between consistent runs and strategic "tumbles" for efficient navigation. The project extended to the possibility of cells amplifying perceived gradients by enclosing their immediate surroundings, pushing attractants forward for enrichment while depleting it at the cell rear. Microfluidic devices were employed, and various experimental parameters configurations were optimized. Although significant differences in migratory efficacy were detected across pore sizes and device heights, quantifying gradient manipulation effects proved challenging. The "Laser-Assisted Protein Adsorption by Photobleaching" (LAPAP) project was promising, as it allowed the printing of gradients. Initially successful with dendritic cells, we aimed to adapt it for neutrophils. Through extensive experimentation with multiple parameters, we attempted to trigger responses from neutrophils. Despite these efforts and collaboration, the project failed due to practical challenges and limitations. Facing a lack of neutrophil-like cells at IST, we initially established the SCF-HoxB8 primary murine cell line. Despite their existence, their migratory behavior was largely unexplored due to potential limitations. Through differentiation protocol refinements we enhanced their migratory capabilities, though their capacity still lagged behind human neutrophils. Despite this, the improved migration potential of these cells pointed toward their utility for in vitro murine neutrophil migration studies.}, author = {Stopp, Julian A}, isbn = {978-3-99078-038-1}, issn = {2663-337X}, pages = {226}, publisher = {Institute of Science and Technology Austria}, title = {{Neutrophils on the hunt : Migratory strategies employed by neutrophils to fulfill their effector function}}, doi = {10.15479/at:ista:14697}, year = {2023}, } @phdthesis{13081, abstract = {During development, tissues undergo changes in size and shape to form functional organs. Distinct cellular processes such as cell division and cell rearrangements underlie tissue morphogenesis. Yet how the distinct processes are controlled and coordinated, and how they contribute to morphogenesis is poorly understood. In our study, we addressed these questions using the developing mouse neural tube. This epithelial organ transforms from a flat epithelial sheet to an epithelial tube while increasing in size and undergoing morpho-gen-mediated patterning. The extent and mechanism of neural progenitor rearrangement within the developing mouse neuroepithelium is unknown. To investigate this, we per-formed high resolution lineage tracing analysis to quantify the extent of epithelial rear-rangement at different stages of neural tube development. We quantitatively described the relationship between apical cell size with cell cycle dependent interkinetic nuclear migra-tions (IKNM) and performed high cellular resolution live imaging of the neuroepithelium to study the dynamics of junctional remodeling. Furthermore, developed a vertex model of the neuroepithelium to investigate the quantitative contribution of cell proliferation, cell differentiation and mechanical properties to the epithelial rearrangement dynamics and validated the model predictions through functional experiments. Our analysis revealed that at early developmental stages, the apical cell area kinetics driven by IKNM induce high lev-els of cell rearrangements in a regime of high junctional tension and contractility. After E9.5, there is a sharp decline in the extent of cell rearrangements, suggesting that the epi-thelium transitions from a fluid-like to a solid-like state. We found that this transition is regulated by the growth rate of the tissue, rather than by changes in cell-cell adhesion and contractile forces. Overall, our study provides a quantitative description of the relationship between tissue growth, cell cycle dynamics, epithelia rearrangements and the emergent tissue material properties, and novel insights on how epithelial cell dynamics influences tissue morphogenesis.}, author = {Bocanegra, Laura}, issn = {2663-337X}, pages = {93}, publisher = {Institute of Science and Technology Austria}, title = {{Epithelial dynamics during mouse neural tube development}}, doi = {10.15479/at:ista:13081}, year = {2023}, } @phdthesis{14323, abstract = {Morphogens are signaling molecules that are known for their prominent role in pattern formation within developing tissues. In addition to patterning, morphogens also control tissue growth. However, the underlying mechanisms are poorly understood. We studied the role of morphogens in regulating tissue growth in the developing vertebrate neural tube. In this system, opposing morphogen gradients of Shh and BMP establish the dorsoventral pattern of neural progenitor domains. Perturbations in these morphogen pathways result in alterations in tissue growth and cell cycle progression, however, it has been unclear what cellular process is affected. To address this, we analysed the rates of cell proliferation and cell death in mouse mutants in which signaling is perturbed, as well as in chick neural plate explants exposed to defined concentrations of signaling activators or inhibitors. Our results indicated that the rate of cell proliferation was not altered in these assays. By contrast, both the Shh and BMP signaling pathways had profound effects on neural progenitor survival. Our results indicate that these pathways synergise to promote cell survival within neural progenitors. Consistent with this, we found that progenitors within the intermediate region of the neural tube, where the combined levels of Shh and BMP are the lowest, are most prone to cell death when signaling activity is inhibited. In addition, we found that downregulation of Shh results in increased apoptosis within the roof plate, which is the dorsal source of BMP ligand production. This revealed a cross-interaction between the Shh and BMP morphogen signaling pathways that may be relevant for understanding how gradients scale in neural tubes with different overall sizes. We further studied the mechanism acting downstream of Shh in cell survival regulation using genetic and genomic approaches. We propose that Shh transcriptionally regulates a non-canonical apoptotic pathway. Altogether, our study points to a novel role of opposing morphogen gradients in tissue size regulation and provides new insights into complex interactions between Shh and BMP signaling gradients in the neural tube.}, author = {Kuzmicz-Kowalska, Katarzyna}, issn = {2663-337X}, pages = {151}, publisher = {Institute of Science and Technology Austria}, title = {{Regulation of neural progenitor survival by Shh and BMP in the developing spinal cord}}, doi = {10.15479/at:ista:14323}, year = {2023}, } @phdthesis{14422, abstract = {Animals exhibit a remarkable ability to learn and remember new behaviors, skills, and associations throughout their lifetime. These capabilities are made possible thanks to a variety of changes in the brain throughout adulthood, regrouped under the term "plasticity". Some cells in the brain —neurons— and specifically changes in the connections between neurons, the synapses, were shown to be crucial for the formation, selection, and consolidation of memories from past experiences. These ongoing changes of synapses across time are called synaptic plasticity. Understanding how a myriad of biochemical processes operating at individual synapses can somehow work in concert to give rise to meaningful changes in behavior is a fascinating problem and an active area of research. However, the experimental search for the precise plasticity mechanisms at play in the brain is daunting, as it is difficult to control and observe synapses during learning. Theoretical approaches have thus been the default method to probe the plasticity-behavior connection. Such studies attempt to extract unifying principles across synapses and model all observed synaptic changes using plasticity rules: equations that govern the evolution of synaptic strengths across time in neuronal network models. These rules can use many relevant quantities to determine the magnitude of synaptic changes, such as the precise timings of pre- and postsynaptic action potentials, the recent neuronal activity levels, the state of neighboring synapses, etc. However, analytical studies rely heavily on human intuition and are forced to make simplifying assumptions about plasticity rules. In this thesis, we aim to assist and augment human intuition in this search for plasticity rules. We explore whether a numerical approach could automatically discover the plasticity rules that elicit desired behaviors in large networks of interconnected neurons. This approach is dubbed meta-learning synaptic plasticity: learning plasticity rules which themselves will make neuronal networks learn how to solve a desired task. We first write all the potential plasticity mechanisms to consider using a single expression with adjustable parameters. We then optimize these plasticity parameters using evolutionary strategies or Bayesian inference on tasks known to involve synaptic plasticity, such as familiarity detection and network stabilization. We show that these automated approaches are powerful tools, able to complement established analytical methods. By comprehensively screening plasticity rules at all synapse types in realistic, spiking neuronal network models, we discover entire sets of degenerate plausible plasticity rules that reliably elicit memory-related behaviors. Our approaches allow for more robust experimental predictions, by abstracting out the idiosyncrasies of individual plasticity rules, and provide fresh insights on synaptic plasticity in spiking network models. }, author = {Confavreux, Basile J}, issn = {2663-337X}, pages = {148}, publisher = {Institute of Science and Technology Austria}, title = {{Synapseek: Meta-learning synaptic plasticity rules}}, doi = {10.15479/at:ista:14422}, year = {2023}, } @phdthesis{14510, abstract = {Clathrin-mediated endocytosis (CME) is vital for the regulation of plant growth and development by controlling plasma membrane protein composition and cargo uptake. CME relies on the precise recruitment control of protein regulators for vesicle maturation and release. During the early stages of endocytosis, an area of flat membrane is remodelled by proteins to create a spherical vesicle against intracellular forces. After the Clathrin-coated vesicle (CCV) is fully formed, scission machinery releases it from the plasma membrane, and cargo proceeds for recycling or degradation through early endosomes / Trans Golgi network. Protein machineries that mediate membrane bending and vesicle release in plants are unknown. However, studies show, that plant endocytosis is actin independent, thus indicating that plants utilize a unique mechanism to mediate membrane bending against highturgor pressure compared to other model systems. First, by using biochemical and advanced live microscopy approaches we investigate the TPLATE complex, a plant-specific endocytosis protein complex. We found that TPLATE is peripherally associated with clathrin-coated vesicles and localises at the rim of endocytosis events. Next, our study of plant Dynamin-related protein 1C (DRP1C), which was hypothesised previously to play a role in vesicle release, shows the recruitment of the protein already at the early stages of endocytosis. Moreover, DRP1C assembles into organised ring-like structures and is able to induce membrane deformation and tubulation, suggesting its role also in membrane bending during early CME. Based on the data from mammalian and yeast systems, plant DynaminRelated Proteins 2 and SH3P2 protein are strong candidates to be part of the plant vesicle scission machinery; however, their precise role in plant CME has not been yet elucidated. Here, we characterised DRP2s and SH3P2 roles in CME by combining high-resolution imaging of endocytic events in vivo and protein characterisation. Although DRP2s and SH3P2 arrive together during late CME and physically interact, genetic analysis using ∆sh3p1,2,3 mutant and complementation with non-DRP2-interacting SH3P2 variants suggest that SH3P2 does not directly recruit DRP2s to the site of endocytosis. Summarising our research, these observations provide new important insights into the mechanism of plant CME and show that, despite plants posses many homologues of mammalian and yeast CME components, they do not necessarily act in the same manner. }, author = {Gnyliukh, Nataliia}, isbn = {978-3-99078-037-4}, issn = {2663-337X}, keywords = {Clathrin-Mediated Endocytosis, vesicle scission, Dynamin-Related Protein 2, SH3P2, TPLATE complex, Total internal reflection fluorescence microscopy, Arabidopsis thaliana}, pages = {180}, publisher = {Institute of Science and Technology Austria}, title = {{Mechanism of clathrin-coated vesicle formation during endocytosis in plants}}, doi = {10.15479/at:ista:14510}, year = {2023}, } @phdthesis{12470, abstract = {The brain is an exceptionally sophisticated organ consisting of billions of cells and trillions of connections that orchestrate our cognition and behavior. To decode its complex connectivity, it is pivotal to disentangle its intricate architecture spanning from cm-sized circuits down to tens of nm-small synapses. To achieve this goal, I developed CATS – Comprehensive Analysis of nervous Tissue across Scales, a versatile toolbox for obtaining a holistic view of nervous tissue context with (superresolution) fluorescence microscopy. CATS combines comprehensive labeling of the extracellular space, that is compatible with chemical fixation, with information on molecular markers, superresolved data acquisition and machine-learning based data analysis for segmentation and synapse identification. I used CATS to analyze key features of nervous tissue connectivity, ranging from whole tissue architecture, neuronal in- and output-fields, down to synapse morphology. Focusing on the hippocampal circuitry, I quantified synaptic transmission properties of mossy fiber boutons and analyzed the connectivity pattern of dentate gyrus granule cells with CA3 pyramidal neurons. This shows that CATS is a viable tool to study hallmarks of neuronal connectivity with light microscopy.}, author = {Michalska, Julia M}, isbn = {978-3-99078-026-8}, issn = {2663-337X}, pages = {201}, publisher = {Institute of Science and Technology Austria}, title = {{A versatile toolbox for the comprehensive analysis of nervous tissue organization with light microscopy}}, doi = {10.15479/at:ista:12470}, year = {2023}, } @phdthesis{14622, abstract = {This Ph.D. thesis presents a detailed investigation into Variational Quantum Algorithms (VQAs), a promising class of quantum algorithms that are well suited for near-term quantum computation due to their moderate hardware requirements and resilience to noise. Our primary focus lies on two particular types of VQAs: the Quantum Approximate Optimization Algorithm (QAOA), used for solving binary optimization problems, and the Variational Quantum Eigensolver (VQE), utilized for finding ground states of quantum many-body systems. In the first part of the thesis, we examine the issue of effective parameter initialization for the QAOA. The work demonstrates that random initialization of the QAOA often leads to convergence in local minima with sub-optimal performance. To mitigate this issue, we propose an initialization of QAOA parameters based on the Trotterized Quantum Annealing (TQA). We show that TQA initialization leads to the same performance as the best of an exponentially scaling number of random initializations. The second study introduces Transition States (TS), stationary points with a single direction of descent, as a tool for systematically exploring the QAOA optimization landscape. This leads us to propose a novel greedy parameter initialization strategy that guarantees for the energy to decrease with increasing number of circuit layers. In the third section, we extend the QAOA to qudit systems, which are higher-dimensional generalizations of qubits. This chapter provides theoretical insights and practical strategies for leveraging the increased computational power of qudits in the context of quantum optimization algorithms and suggests a quantum circuit for implementing the algorithm on an ion trap quantum computer. Finally, we propose an algorithm to avoid “barren plateaus”, regions in parameter space with vanishing gradients that obstruct efficient parameter optimization. This novel approach relies on defining a notion of weak barren plateaus based on the entropies of local reduced density matrices and showcases how these can be efficiently quantified using shadow tomography. To illustrate the approach we employ the strategy in the VQE and show that it allows to successfully avoid barren plateaus in the initialization and throughout the optimization. Taken together, this thesis greatly enhances our understanding of parameter initialization and optimization in VQAs, expands the scope of QAOA to higher-dimensional quantum systems, and presents a method to address the challenge of barren plateaus using the VQE. These insights are instrumental in advancing the field of near-term quantum computation.}, author = {Sack, Stefan}, issn = {2663-337X}, pages = {142}, publisher = {Institute of Science and Technology Austria}, title = {{Improving variational quantum algorithms: Innovative initialization techniques and extensions to qudit systems}}, doi = {10.15479/at:ista:14622}, year = {2023}, } @phdthesis{11128, abstract = {Although we often see studies focusing on simple or even discrete traits in studies of colouration, the variation of “appearance” phenotypes found in nature is often more complex, continuous and high-dimensional. Therefore, we developed automated methods suitable for large datasets of genomes and images, striving to account for their complex nature, while minimising human bias. We used these methods on a dataset of more than 20, 000 plant SNP genomes and corresponding fower images from a hybrid zone of two subspecies of Antirrhinum majus with distinctly coloured fowers to improve our understanding of the genetic nature of the fower colour in our study system. Firstly, we use the advantage of large numbers of genotyped plants to estimate the haplotypes in the main fower colour regulating region. We study colour- and geography-related characteristics of the estimated haplotypes and how they connect to their relatedness. We show discrepancies from the expected fower colour distributions given the genotype and identify particular haplotypes leading to unexpected phenotypes. We also confrm a signifcant defcit of the double recessive recombinant and quite surprisingly, we show that haplotypes of the most frequent parental type are much less variable than others. Secondly, we introduce our pipeline capable of processing tens of thousands of full fower images without human interaction and summarising each image into a set of informative scores. We show the compatibility of these machine-measured fower colour scores with the previously used manual scores and study impact of external efect on the resulting scores. Finally, we use the machine-measured fower colour scores to ft and examine a phenotype cline across the hybrid zone in Planoles using full fower images as opposed to discrete, manual scores and compare it with the genotypic cline.}, author = {Matejovicova, Lenka}, isbn = {978-3-99078-016-9}, issn = {2663-337X}, pages = {112}, publisher = {Institute of Science and Technology Austria}, title = {{Genetic basis of flower colour as a model for adaptive evolution}}, doi = {10.15479/at:ista:11128}, year = {2022}, } @phdthesis{11473, abstract = {The polaron model is a basic model of quantum field theory describing a single particle interacting with a bosonic field. It arises in many physical contexts. We are mostly concerned with models applicable in the context of an impurity atom in a Bose-Einstein condensate as well as the problem of electrons moving in polar crystals. The model has a simple structure in which the interaction of the particle with the field is given by a term linear in the field’s creation and annihilation operators. In this work, we investigate the properties of this model by providing rigorous estimates on various energies relevant to the problem. The estimates are obtained, for the most part, by suitable operator techniques which constitute the principal mathematical substance of the thesis. The first application of these techniques is to derive the polaron model rigorously from first principles, i.e., from a full microscopic quantum-mechanical many-body problem involving an impurity in an otherwise homogeneous system. We accomplish this for the N + 1 Bose gas in the mean-field regime by showing that a suitable polaron-type Hamiltonian arises at weak interactions as a low-energy effective theory for this problem. In the second part, we investigate rigorously the ground state of the model at fixed momentum and for large values of the coupling constant. Qualitatively, the system is expected to display a transition from the quasi-particle behavior at small momenta, where the dispersion relation is parabolic and the particle moves through the medium dragging along a cloud of phonons, to the radiative behavior at larger momenta where the polaron decelerates and emits free phonons. At the same time, in the strong coupling regime, the bosonic field is expected to behave purely classically. Accordingly, the effective mass of the polaron at strong coupling is conjectured to be asymptotically equal to the one obtained from the semiclassical counterpart of the problem, first studied by Landau and Pekar in the 1940s. For polaron models with regularized form factors and phonon dispersion relations of superfluid type, i.e., bounded below by a linear function of the wavenumbers for all phonon momenta as in the interacting Bose gas, we prove that for a large window of momenta below the radiation threshold, the energy-momentum relation at strong coupling is indeed essentially a parabola with semi-latus rectum equal to the Landau–Pekar effective mass, as expected. For the Fröhlich polaron describing electrons in polar crystals where the dispersion relation is of the optical type and the form factor is formally UV–singular due to the nature of the point charge-dipole interaction, we are able to give the corresponding upper bound. In contrast to the regular case, this requires the inclusion of the quantum fluctuations of the phonon field, which makes the problem considerably more difficult. The results are supplemented by studies on the absolute ground-state energy at strong coupling, a proof of the divergence of the effective mass with the coupling constant for a wide class of polaron models, as well as the discussion of the apparent UV singularity of the Fröhlich model and the application of the techniques used for its removal for the energy estimates. }, author = {Mysliwy, Krzysztof}, issn = {2663-337X}, pages = {138}, publisher = {Institute of Science and Technology Austria}, title = {{Polarons in Bose gases and polar crystals: Some rigorous energy estimates}}, doi = {10.15479/at:ista:11473}, year = {2022}, } @phdthesis{11626, abstract = {Plant growth and development is well known to be both, flexible and dynamic. The high capacity for post-embryonic organ formation and tissue regeneration requires tightly regulated intercellular communication and coordinated tissue polarization. One of the most important drivers for patterning and polarity in plant development is the phytohormone auxin. Auxin has the unique characteristic to establish polarized channels for its own active directional cell to cell transport. This fascinating phenomenon is called auxin canalization. Those auxin transport channels are characterized by the expression and polar, subcellular localization of PIN auxin efflux carriers. PIN proteins have the ability to dynamically change their localization and auxin itself can affect this by interfering with trafficking. Most of the underlying molecular mechanisms of canalization still remain enigmatic. What is known so far is that canonical auxin signaling is indispensable but also other non-canonical signaling components are thought to play a role. In order to shed light into the mysteries auf auxin canalization this study revisits the branches of auxin signaling in detail. Further a new auxin analogue, PISA, is developed which triggers auxin-like responses but does not directly activate canonical transcriptional auxin signaling. We revisit the direct auxin effect on PIN trafficking where we found that, contradictory to previous observations, auxin is very specifically promoting endocytosis of PIN2 but has no overall effect on endocytosis. Further, we evaluate which cellular processes related to PIN subcellular dynamics are involved in the establishment of auxin conducting channels and the formation of vascular tissue. We are re-evaluating the function of AUXIN BINDING PROTEIN 1 (ABP1) and provide a comprehensive picture about its developmental phneotypes and involvement in auxin signaling and canalization. Lastly, we are focusing on the crosstalk between the hormone strigolactone (SL) and auxin and found that SL is interfering with essentially all processes involved in auxin canalization in a non-transcriptional manner. Lastly we identify a new way of SL perception and signaling which is emanating from mitochondria, is independent of canonical SL signaling and is modulating primary root growth.}, author = {Gallei, Michelle C}, isbn = {978-3-99078-019-0}, issn = {2663-337X}, pages = {248}, publisher = {Institute of Science and Technology Austria}, title = {{Auxin and strigolactone non-canonical signaling regulating development in Arabidopsis thaliana}}, doi = {10.15479/at:ista:11626}, year = {2022}, } @phdthesis{11777, abstract = {In this dissertation we study coboundary expansion of simplicial complex with a view of giving geometric applications. Our main novel tool is an equivariant version of Gromov's celebrated Topological Overlap Theorem. The equivariant topological overlap theorem leads to various geometric applications including a quantitative non-embeddability result for sufficiently thick buildings (which partially resolves a conjecture of Tancer and Vorwerk) and an improved lower bound on the pair-crossing number of (bounded degree) expander graphs. Additionally, we will give new proofs for several known lower bounds for geometric problems such as the number of Tverberg partitions or the crossing number of complete bipartite graphs. For the aforementioned applications one is naturally lead to study expansion properties of joins of simplicial complexes. In the presence of a special certificate for expansion (as it is the case, e.g., for spherical buildings), the join of two expanders is an expander. On the flip-side, we report quite some evidence that coboundary expansion exhibits very non-product-like behaviour under taking joins. For instance, we exhibit infinite families of graphs $(G_n)_{n\in \mathbb{N}}$ and $(H_n)_{n\in\mathbb{N}}$ whose join $G_n*H_n$ has expansion of lower order than the product of the expansion constant of the graphs. Moreover, we show an upper bound of $(d+1)/2^d$ on the normalized coboundary expansion constants for the complete multipartite complex $[n]^{*(d+1)}$ (under a mild divisibility condition on $n$). Via the probabilistic method the latter result extends to an upper bound of $(d+1)/2^d+\varepsilon$ on the coboundary expansion constant of the spherical building associated with $\mathrm{PGL}_{d+2}(\mathbb{F}_q)$ for any $\varepsilon>0$ and sufficiently large $q=q(\varepsilon)$. This disproves a conjecture of Lubotzky, Meshulam and Mozes -- in a rather strong sense. By improving on existing lower bounds we make further progress towards closing the gap between the known lower and upper bounds on the coboundary expansion constants of $[n]^{*(d+1)}$. The best improvements we achieve using computer-aided proofs and flag algebras. The exact value even for the complete $3$-partite $2$-dimensional complex $[n]^{*3}$ remains unknown but we are happy to conjecture a precise value for every $n$. %Moreover, we show that a previously shown lower bound on the expansion constant of the spherical building associated with $\mathrm{PGL}_{2}(\mathbb{F}_q)$ is not tight. In a loosely structured, last chapter of this thesis we collect further smaller observations related to expansion. We point out a link between discrete Morse theory and a technique for showing coboundary expansion, elaborate a bit on the hardness of computing coboundary expansion constants, propose a new criterion for coboundary expansion (in a very dense setting) and give one way of making the folklore result that expansion of links is a necessary condition for a simplicial complex to be an expander precise.}, author = {Wild, Pascal}, isbn = {978-3-99078-021-3}, issn = {2663-337X}, pages = {170}, publisher = {Institute of Science and Technology Austria}, title = {{High-dimensional expansion and crossing numbers of simplicial complexes}}, doi = {10.15479/at:ista:11777}, year = {2022}, } @phdthesis{11945, abstract = {G protein-coupled receptors (GPCRs) respond to specific ligands and regulate multiple processes ranging from cell growth and immune responses to neuronal signal transmission. However, ligands for many GPCRs remain unknown, suffer from off-target effects or have poor bioavailability. Additional challenges exist to dissect cell-type specific responses when the same GPCR is expressed on several cell types within the body. Here, we overcome these limitations by engineering DREADD-based GPCR chimeras that selectively bind their agonist clozapine-N-oxide (CNO) and mimic a GPCR-of-interest in a desired cell type. We validated our approach with β2-adrenergic receptor (β2AR/ADRB2) and show that our chimeric DREADD-β2AR triggers comparable responses on second messenger and kinase activity, post-translational modifications, and protein-protein interactions. Since β2AR is also enriched in microglia, which can drive inflammation in the central nervous system, we expressed chimeric DREADD-β2AR in primary microglia and successfully recapitulate β2AR-mediated filopodia formation through CNO stimulation. To dissect the role of selected GPCRs during microglial inflammation, we additionally generated DREADD-based chimeras for microglia-enriched GPR65 and GPR109A/HCAR2. In a microglia cell line, DREADD-β2AR and DREADD-GPR65 both modulated the inflammatory response with a similar profile as endogenously expressed β2AR, while DREADD-GPR109A showed no impact. Our DREADD-based approach provides the means to obtain mechanistic and functional insights into GPCR signaling on a cell-type specific level.}, author = {Schulz, Rouven}, issn = {2663-337X}, pages = {133}, publisher = {Institute of Science and Technology Austria}, title = {{Chimeric G protein-coupled receptors mimic distinct signaling pathways and modulate microglia function}}, doi = {10.15479/at:ista:11945}, year = {2022}, } @phdthesis{12358, abstract = {The complex yarn structure of knitted and woven fabrics gives rise to both a mechanical and visual complexity. The small-scale interactions of yarns colliding with and pulling on each other result in drastically different large-scale stretching and bending behavior, introducing anisotropy, curling, and more. While simulating cloth as individual yarns can reproduce this complexity and match the quality of real fabric, it may be too computationally expensive for large fabrics. On the other hand, continuum-based approaches do not need to discretize the cloth at a stitch-level, but it is non-trivial to find a material model that would replicate the large-scale behavior of yarn fabrics, and they discard the intricate visual detail. In this thesis, we discuss three methods to try and bridge the gap between small-scale and large-scale yarn mechanics using numerical homogenization: fitting a continuum model to periodic yarn simulations, adding mechanics-aware yarn detail onto thin-shell simulations, and quantitatively fitting yarn parameters to physical measurements of real fabric. To start, we present a method for animating yarn-level cloth effects using a thin-shell solver. We first use a large number of periodic yarn-level simulations to build a model of the potential energy density of the cloth, and then use it to compute forces in a thin-shell simulator. The resulting simulations faithfully reproduce expected effects like the stiffening of woven fabrics and the highly deformable nature and anisotropy of knitted fabrics at a fraction of the cost of full yarn-level simulation. While our thin-shell simulations are able to capture large-scale yarn mechanics, they lack the rich visual detail of yarn-level simulations. Therefore, we propose a method to animate yarn-level cloth geometry on top of an underlying deforming mesh in a mechanics-aware fashion in real time. Using triangle strains to interpolate precomputed yarn geometry, we are able to reproduce effects such as knit loops tightening under stretching at negligible cost. Finally, we introduce a methodology for inverse-modeling of yarn-level mechanics of cloth, based on the mechanical response of fabrics in the real world. We compile a database from physical tests of several knitted fabrics used in the textile industry spanning diverse physical properties like stiffness, nonlinearity, and anisotropy. We then develop a system for approximating these mechanical responses with yarn-level cloth simulation, using homogenized shell models to speed up computation and adding some small-but-necessary extensions to yarn-level models used in computer graphics. }, author = {Sperl, Georg}, isbn = {978-3-99078-020-6}, issn = {2663-337X}, pages = {138}, publisher = {Institute of Science and Technology Austria}, title = {{Homogenizing yarn simulations: Large-scale mechanics, small-scale detail, and quantitative fitting}}, doi = {10.15479/at:ista:12103}, year = {2022}, } @phdthesis{12368, abstract = {Metazoan development relies on the formation and remodeling of cell-cell contacts. The binding of adhesion receptors and remodeling of the actomyosin cell cortex at cell-cell interaction sites have been implicated in cell-cell contact formation. Yet, how these two processes functionally interact to drive cell-cell contact expansion and strengthening remains unclear. Here, we study how primary germ layer progenitor cells from zebrafish bind to supported lipid bilayers (SLB) functionalized with E-cadherin ectodomains as an assay system for monitoring cell-cell contact formation at high spatiotemporal resolution. We show that cell-cell contact formation represents a two-tiered process: E-cadherinmediated downregulation of the small GTPase RhoA at the forming contact leads to both depletion of Myosin-2 and decrease of F-actin. This is followed by centrifugal actin network flows at the contact triggered by a sharp gradient of Myosin-2 at the rim of the contact zone, with Myosin-2 displaying higher cortical localization outside than inside of the contact. These centrifugal cortical actin flows, in turn, not only further dilute the actin network at the contact disc, but also lead to an accumulation of both F-actin and Ecadherin at the contact rim. Eventually, this combination of actomyosin downregulation and flows at the contact contribute to the characteristic molecular organization implicated in contact formation and maintenance: depletion of cortical actomyosin at the contact disc, driving contact expansion by lowering interfacial tension at the contact, and accumulation of both E-cadherin and F-actin at the contact rim, mechanically linking the contractile cortices of the adhering cells. Thus, using a biomimetic assay, we exemplify how adhesion signaling and cell mechanics function together to modulate the spatial organization of cell-cell contacts.}, author = {Arslan, Feyza N}, isbn = {978-3-99078-025-1 }, issn = {2663-337X}, pages = {113}, publisher = {Institute of Science and Technology Austria}, title = {{Remodeling of E-cadherin-mediated contacts via cortical flows}}, doi = {10.15479/at:ista:12153}, year = {2022}, } @phdthesis{12390, abstract = {The scope of this thesis is to study quantum systems exhibiting a continuous symmetry that is broken on the level of the corresponding effective theory. In particular we are going to investigate translation-invariant Bose gases in the mean field limit, effectively described by the Hartree functional, and the Fröhlich Polaron in the regime of strong coupling, effectively described by the Pekar functional. The latter is a model describing the interaction between a charged particle and the optical modes of a polar crystal. Regarding the former, we assume in addition that the particles in the gas are unconfined, and typically we will consider particles that are subject to an attractive interaction. In both cases the ground state energy of the Hamiltonian is not a proper eigenvalue due to the underlying translation-invariance, while on the contrary there exists a whole invariant orbit of minimizers for the corresponding effective functionals. Both, the absence of proper eigenstates and the broken symmetry of the effective theory, make the study significantly more involved and it is the content of this thesis to develop a frameworks which allows for a systematic way to circumvent these issues. It is a well-established result that the ground state energy of Bose gases in the mean field limit, as well as the ground state energy of the Fröhlich Polaron in the regime of strong coupling, is to leading order given by the minimal energy of the corresponding effective theory. As part of this thesis we identify the sub-leading term in the expansion of the ground state energy, which can be interpreted as the quantum correction to the classical energy, since the effective theories under consideration can be seen as classical counterparts. We are further going to establish an asymptotic expression for the energy-momentum relation of the Fröhlich Polaron in the strong coupling limit. In the regime of suitably small momenta, this asymptotic expression agrees with the energy-momentum relation of a free particle having an effectively increased mass, and we find that this effectively increased mass agrees with the conjectured value in the physics literature. In addition we will discuss two unrelated papers written by the author during his stay at ISTA in the appendix. The first one concerns the realization of anyons, which are quasi-particles acquiring a non-trivial phase under the exchange of two particles, as molecular impurities. The second one provides a classification of those vector fields defined on a given manifold that can be written as the gradient of a given functional with respect to a suitable metric, provided that some mild smoothness assumptions hold. This classification is subsequently used to identify those quantum Markov semigroups that can be written as a gradient flow of the relative entropy. }, author = {Brooks, Morris}, issn = {2663-337X}, pages = {196}, publisher = {Institute of Science and Technology Austria}, title = {{Translation-invariant quantum systems with effectively broken symmetry}}, doi = {10.15479/at:ista:12390}, year = {2022}, } @phdthesis{10759, abstract = {In this Thesis, I study composite quantum impurities with variational techniques, both inspired by machine learning as well as fully analytic. I supplement this with exploration of other applications of machine learning, in particular artificial neural networks, in many-body physics. In Chapters 3 and 4, I study quasiparticle systems with variational approach. I derive a Hamiltonian describing the angulon quasiparticle in the presence of a magnetic field. I apply analytic variational treatment to this Hamiltonian. Then, I introduce a variational approach for non-additive systems, based on artificial neural networks. I exemplify this approach on the example of the polaron quasiparticle (Fröhlich Hamiltonian). In Chapter 5, I continue using artificial neural networks, albeit in a different setting. I apply artificial neural networks to detect phases from snapshots of two types physical systems. Namely, I study Monte Carlo snapshots of multilayer classical spin models as well as molecular dynamics maps of colloidal systems. The main type of networks that I use here are convolutional neural networks, known for their applicability to image data.}, author = {Rzadkowski, Wojciech}, issn = {2663-337X}, pages = {120}, publisher = {Institute of Science and Technology Austria}, title = {{Analytic and machine learning approaches to composite quantum impurities}}, doi = {10.15479/at:ista:10759}, year = {2022}, }