TY - THES A1 - Karakas, Esra T1 - High-resolution studies of epistasis in tomato metabolism T1 - Hochauflösende Studien zur Epistasierung des Tomatenstoffwechsels N2 - The inclusion of exotic germplasm serves as a crucial means to enhance allelic and consequently phenotypic diversity in inbred crop species. Such species have experienced a reduction in diversity due to artificial selection focused on a limited set of traits. The natural biodiversity within ecosystems presents an opportunity to explore various traits influencing plant survival, reproductive fitness and yield potential. In agricultural research, the study of wild species closely related to cultivated plants serves as a means to comprehend the genetic foundations of past domestication events and the polymorphisms essential for future breeding efforts to develop superior varieties. In order to examine the metabolic composition, pinpoint quantitative trait loci (QTL) and facilitate their resolution an extensive large-scale analysis of metabolic QTL (mQTL) was conducted on tomato backcross inbred lines (BILs) derived from a cross between the wild species S. pennellii (5240) incorporated into the background of S. lycopersicum cv. LEA determinate inbred which can be grown in open fields and cv. TOP indeterminate which can be grown in greenhouse conditions. A large number of mQTL associated with primary secondary and lipid metabolism in fruit were identified across the two BIL populations. Epistasis, the interactions between genes at different loci, has been an interest in molecular and quantitative genetics for many decades. The study of epistasis requires the analysis of very large populations with multiple independent genotypes that carry specific genomic regions. In order to understand the genetic basis of tomato fruit metabolism, I extended the work to investigate epistatic interactions of the genomic regions. In addition, two candidate genes were identified through quantitative trait loci underlying fruit-specific sucrose and jasmonic acid derivatives. Finally, in this study, I assessed the genetic framework of fruit metabolic traits with a high level of detail, utilizing the newly created Solanum pennellii (5240) backcrossed introgression lines (n=3000). This investigation resulted in the discovery of promising candidate loci associated with significant fruit quality traits, including those to the abundance of glutamic acid and aspartic acid crucial elements contributing to the development of acidity and flavors. N2 - Die Einbeziehung von exotischem Keimplasma ist ein wichtiges Mittel zur Verbesserung der allelischen und folglich auch der phänotypischen Vielfalt bei Inzuchtpflanzenarten. Bei diesen Arten hat die künstliche Selektion, die sich auf eine begrenzte Anzahl von Merkmalen konzentriert, zu einem Rückgang der Vielfalt geführt. Die natürliche Artenvielfalt in Ökosystemen bietet die Möglichkeit, verschiedene Merkmale zu erforschen, die das Überleben, die Reproduktionsfähigkeit und das Ertragspotenzial von Pflanzen beeinflussen. In der Agrarforschung dient die Untersuchung von Wildarten, die eng mit Kulturpflanzen verwandt sind, als Mittel zum Verständnis der genetischen Grundlagen vergangener Domestizierungsereignisse und der Polymorphismen, die für künftige Züchtungsbemühungen zur Entwicklung besserer Sorten wichtig sind. Um die metabolische Zusammensetzung zu untersuchen, quantitative Merkmalsloci (QTL) zu identifizieren und ihre Auflösung zu erleichtern, wurde eine umfangreiche Analyse metabolischer QTL (mQTL) an Tomaten-Rückkreuzungs-Inzuchtlinien (BILs) durchgeführt, die aus einer Kreuzung zwischen der Wildart S. pennellii (5240), die in den Hintergrund von S. lycopersicum cv. LEA determinate inbred, die im Freiland angebaut werden kann, und cv. TOP indeterminate, die unter Gewächshausbedingungen angebaut werden kann. In den beiden BIL-Populationen wurde eine große Anzahl von mQTL identifiziert, die mit dem primären Sekundär- und Lipidstoffwechsel in der Frucht in Verbindung stehen. Epistase, die Wechselwirkungen zwischen Genen an verschiedenen Loci, ist seit vielen Jahrzehnten ein Thema in der molekularen und quantitativen Genetik. Die Untersuchung der Epistase erfordert die Analyse sehr großer Populationen mit mehreren unabhängigen Genotypen, die bestimmte genomische Regionen tragen. Um die genetischen Grundlagen des Tomatenfruchtstoffwechsels zu verstehen, habe ich die Arbeit erweitert, um epistatische Interaktionen der genomischen Regionen zu untersuchen. Darüber hinaus wurden zwei Kandidatengene identifiziert, die über quantitative Merkmalsloci den fruchttypischen Saccharose- und Jasmonsäurederivaten zugrunde liegen. Schließlich habe ich in dieser Studie das genetische Gerüst der Fruchtstoffwechselmerkmale mit einem hohen Detaillierungsgrad bewertet, wobei ich die neu geschaffenen Solanum pennellii (5240) Rückkreuzungslinien (n=3000) verwendet habe. Diese Untersuchung führte zur Entdeckung vielversprechender Kandidatenloci, die mit bedeutenden Fruchtqualitätsmerkmalen assoziiert sind, einschließlich derjenigen, die mit der Fülle von Glutaminsäure und Asparaginsäure in Verbindung stehen - entscheidende Elemente, die zur Entwicklung von Säure und Aromen beitragen. KW - Epistasis KW - QTL mapping KW - metabolomics KW - backcross inbred line (BIL) KW - Epistase KW - QTL KW - Metabolomik KW - Rückkreuzungsinzuchtlinie (BIL) Y1 - 2024 ER - TY - JOUR A1 - Wendering, Philipp A1 - Nikoloski, Zoran T1 - Genome-scale modeling specifies the metabolic capabilities of Rhizophagus irregularis JF - mSystems N2 - Rhizophagus irregularis is one of the most extensively studied arbuscular mycorrhizal fungi (AMF) that forms symbioses with and improves the performance of many crops. Lack of transformation protocol for R. irregularis renders it challenging to investigate molecular mechanisms that shape the physiology and interactions of this AMF with plants. Here, we used all published genomics, transcriptomics, and metabolomics resources to gain insights into the metabolic functionalities of R. irregularis by reconstructing its high-quality genome-scale metabolic network that considers enzyme constraints. Extensive validation tests with the enzyme-constrained metabolic model demonstrated that it can be used to (i) accurately predict increased growth of R. irregularis on myristate with minimal medium; (ii) integrate enzyme abundances and carbon source concentrations that yield growth predictions with high and significant Spearman correlation (rS = 0.74) to measured hyphal dry weight; and (iii) simulate growth rate increases with tighter association of this AMF with the host plant across three fungal structures. Based on the validated model and system-level analyses that integrate data from transcriptomics studies, we predicted that differences in flux distributions between intraradical mycelium and arbuscles are linked to changes in amino acid and cofactor biosynthesis. Therefore, our results demonstrated that the enzyme-constrained metabolic model can be employed to pinpoint mechanisms driving developmental and physiological responses of R. irregularis to different environmental cues. In conclusion, this model can serve as a template for other AMF and paves the way to identify metabolic engineering strategies to modulate fungal metabolic traits that directly affect plant performance. IMPORTANCE Mounting evidence points to the benefits of the symbiotic interactions between the arbuscular mycorrhiza fungus Rhizophagus irregularis and crops; however, the molecular mechanisms underlying the physiological responses of this fungus to different host plants and environments remain largely unknown. We present a manually curated, enzyme-constrained, genome-scale metabolic model of R. irregularis that can accurately predict experimentally observed phenotypes. We show that this high-quality model provides an entry point into better understanding the metabolic and physiological responses of this fungus to changing environments due to the availability of different nutrients. The model can be used to design metabolic engineering strategies to tailor R. irregularis metabolism toward improving the performance of host plants. KW - Rhizophagus irregularis KW - metabolic modeling Y1 - 2022 U6 - https://doi.org/10.1128/msystems.01216-21 SN - 2379-5077 VL - 7 IS - 1 PB - American Society for Microbiology CY - Washington, DC ER - TY - CHAP A1 - Fayyaz, Susann A1 - Hartmann, Bolette A1 - Hanack, Katja A1 - Michelchen, Sophia A1 - Kreiling, Reinhard T1 - Development of a hematopoietic stem cell (murine system) based system as an alternative for the in vivo T-cell-dependent antibody response (TDAR) assay within the EOGRTS: case-study with Parabens T2 - Toxicology letters Y1 - 2022 U6 - https://doi.org/10.1016/j.toxlet.2022.07.483 SN - 0378-4274 SN - 1879-3169 VL - 368 SP - S175 EP - S176 PB - Elsevier Science CY - Amsterdam [u.a.] ER - TY - THES A1 - Arend, Marius T1 - Comparing genome-scale models of protein-constrained metabolism in heterotrophic and photosynthetic microorganisms N2 - Genome-scale metabolic models are mathematical representations of all known reactions occurring in a cell. Combined with constraints based on physiological measurements, these models have been used to accurately predict metabolic fluxes and effects of perturbations (e.g. knock-outs) and to inform metabolic engineering strategies. Recently, protein-constrained models have been shown to increase predictive potential (especially in overflow metabolism), while alleviating the need for measurement of nutrient uptake rates. The resulting modelling frameworks quantify the upkeep cost of a certain metabolic flux as the minimum amount of enzyme required for catalysis. These improvements are based on the use of in vitro turnover numbers or in vivo apparent catalytic rates of enzymes for model parameterization. In this thesis several tools for the estimation and refinement of these parameters based on in vivo proteomics data of Escherichia coli, Saccharomyces cerevisiae, and Chlamydomonas reinhardtii have been developed and applied. The difference between in vitro and in vivo catalytic rate measures for the three microorganisms was systematically analyzed. The results for the facultatively heterotrophic microalga C. reinhardtii considerably expanded the apparent catalytic rate estimates for photosynthetic organisms. Our general finding pointed at a global reduction of enzyme efficiency in heterotrophy compared to other growth scenarios. Independent of the modelled organism, in vivo estimates were shown to improve accuracy of predictions of protein abundances compared to in vitro values for turnover numbers. To further improve the protein abundance predictions, machine learning models were trained that integrate features derived from protein-constrained modelling and codon usage. Combining the two types of features outperformed single feature models and yielded good prediction results without relying on experimental transcriptomic data. The presented work reports valuable advances in the prediction of enzyme allocation in unseen scenarios using protein constrained metabolic models. It marks the first successful application of this modelling framework in the biotechnological important taxon of green microalgae, substantially increasing our knowledge of the enzyme catalytic landscape of phototrophic microorganisms. N2 - Genomweite Stoffwechselmodelle sind mathematische Darstellungen aller bekannten Reaktionen, die in einer Zelle ablaufen. In Kombination mit Einschränkungen, die auf physiologischen Messungen beruhen, wurden diese Modelle zur genauen Vorhersage von Stoffwechselflüssen und Auswirkungen von Manipulationene (z. B. Knock-outs) sowie zum Entwerfen von Metabolic Engineering Strategien verwendet. In jüngster Zeit hat sich gezeigt, dass proteinlimitierte Modelle, welche die Menge an Proteinen in einer Zelle als Modelbeschränkungen integrieren, ein erweitertes Modellierungspotenzial besitzen (insbesondere beim Überflussstoffwechsel) und gleichzeitig die Messungen der Nährstoffaufnahmerate eines Organismus optional machen. Die resultierenden Modelle quantifizieren die Unterhaltskosten eines bestimmten Stoffwechselflusses als die für die Katalyse erforderliche Mindestmenge an Enzymen. Die beobachtete Verbesserungen in den Voraussagefähigkeiten solcher Modelle werden durch die Parameterisierung mit unterschiedlichen in vitro und in vivo Approximationen der maximalen katalytischen Effizienz (Wechselzahl) aller Enyzme eines Organismus ermöglicht. In dieser Arbeit wurden verschiedene Verfahren zur Schätzung und Verfeinerung dieser Parameter auf der Grundlage von in vivo Proteomikdaten der Organismen Escherichia coli, Saccharomyces cerevisiae und Chlamydomonas reinhardtii entwickelt und angewendet. Der Unterschied zwischen den in vitro und in vivo berechneten katalytischen Raten für die drei Mikroorganismen wurde systematisch analysiert. Die Ergebnisse für die fakultativ heterotrophe Mikroalge C. reinhardtii erweitern die Menge an verfügbaren enzymkatalytischen Parametern für photosynthetische Organismen erheblich. Weiterhin deuten unsere Ergbnisse für C. reinhardtii auf eine globale Verringerung der Enzymeffizienz bei Heterotrophie im Vergleich zu anderen Wachstumsszenarien hin. Unabhängig vom modellierten Organismus konnte gezeigt werden, dass geschätzte in vivo Wechselzahlen die Genauigkeit der Vorhersagen von Proteinmengen im Vergleich zu in vitro Werten verbessern. Um die Vorhersagen von Proteinmengen weiter zu verbessern, wurden Modelle aus dem Bereich des maschinellen Lernens trainiert, die Prediktoren basierend auf der proteinlimitierten Modellierung und der Proteinsequenz integrieren. Die Kombination der beiden Arten von Prediktoren übertraf die Leistung von Modellen mit nur einer Art von Prediktoren und lieferte gute Vorhersageergebnisse, ohne auf experimentelle Transkriptionsdaten angewiesen zu sein. Die vorgestellte Arbeit stellt einen wertvollen Fortschritt bei der Vorhersage der Enzymallokation in unbekannten Szenarien unter Verwendung von proteinlimitierten Stoffwechselmodellen dar. Sie markiert die erste erfolgreiche Anwendung dieses Modellierungsverfahren in dem biotechnologisch wichtigen Taxon der grünen Mikroalgen und erweitert unser Wissen über die enzymkatalytische Landschaft phototropher Mikroorganismen entscheidend. T2 - Vergleich und Analyse genomweiter Modelle des protein-limitierten Metabolismus in heterotrophen und photosynthetischen Microorganismen KW - Metabolic Modeling KW - Systems Biology KW - Computational Biology KW - Proteomics KW - computergestützte Biologie KW - metabolische Modellierung KW - Proteomics KW - Systembiologie Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-651470 ER - TY - JOUR A1 - Sandhage-Hofmann, Alexandra A1 - Angombe, Simon A1 - Kindermann, Liana A1 - Linstädter, Anja A1 - Mörchen, Ramona T1 - Conservation with elephants and agricultural intensification BT - effects on lignin and n-alkanes in soils of sub-Saharan Africa JF - Geoderma : an international journal of soil science N2 - Nature conservation is currently shaping many terrestrial ecosystems in Africa. This is particularly evident in Sub-Saharan Africa (SSA), where conservation is intended to recover wildlife populations, with special focus on elephants. Rising numbers of elephants induce woody biomass losses but increase soil organic carbon (SOC) stocks from decaying wood and dung. We hypothesized that these increases under wildlife conservation in SSA go along with rising contents of plant residues in SOC, traceable by the molecular markers lignin and n-alkanes. In contrast, agricultural intensification would reduce them due to lower C input and faster SOC turnover through tillage. To test this, we analyzed lignin by the CuO oxidation method and n-alkanes by fast pressurized solvent extraction in topsoils (0-10 cm) of Arenosols and corresponding plant samples (trees, grasses and crops). Sampling sites followed conservation gradients with low, medium and high elephant densities and intensification gradients with rangeland and cropland in the woodland savanna of the Namibian Zambezi Region. Patterns of lignin-derived phenols were retained in the soil, whereas n-alkanes showed shifts in chain lengths. n-Alkanes also showed no clear increase or decrease under conservation or intensification, respectively. Differently, lignin-derived phenols showed lower values under intensification than under conservation. Confirming our hypothesis, rising SOC contents with rising elephant densities (from 4.4 at low to 5.7 g kg(-1) SOC at high elephant densities) went along with an increasing accumulation of lignin-derived phenols (24.4-34.8 g kg(-1) VSCOC). This increase is associated with the input of woody debris to the soil, as indicated by V-units and carbon isotopes, modulated by clay and woody biomass. We conclude, that increasing input of woody residues into soil by browsing behaviour of elephants is an important mechanism for controlling SOC supply in the context of wildlife conservation and is traceable with lignin-derived phenols, but not with n-alkanes. KW - lignin-derived phenols KW - n-alkanes KW - soil organic carbon KW - wildlife conservation KW - agricultural intensification Y1 - 2022 U6 - https://doi.org/10.1016/j.geoderma.2022.116009 SN - 0016-7061 SN - 1872-6259 VL - 425 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Zurell, Damaris A1 - König, Christian A1 - Malchow, Anne-Kathleen A1 - Kapitza, Simon A1 - Bocedi, Greta A1 - Travis, Justin M. J. A1 - Fandos, Guillermo T1 - Spatially explicit models for decision-making in animal conservation and restoration JF - Ecography : pattern and diversity in ecology / Nordic Ecologic Society Oikos N2 - Models are useful tools for understanding and predicting ecological patterns and processes. Under ongoing climate and biodiversity change, they can greatly facilitate decision-making in conservation and restoration and help designing adequate management strategies for an uncertain future. Here, we review the use of spatially explicit models for decision support and to identify key gaps in current modelling in conservation and restoration. Of 650 reviewed publications, 217 publications had a clear management application and were included in our quantitative analyses. Overall, modelling studies were biased towards static models (79%), towards the species and population level (80%) and towards conservation (rather than restoration) applications (71%). Correlative niche models were the most widely used model type. Dynamic models as well as the gene-to-individual level and the community-to-ecosystem level were underrepresented, and explicit cost optimisation approaches were only used in 10% of the studies. We present a new model typology for selecting models for animal conservation and restoration, characterising model types according to organisational levels, biological processes of interest and desired management applications. This typology will help to more closely link models to management goals. Additionally, future efforts need to overcome important challenges related to data integration, model integration and decision-making. We conclude with five key recommendations, suggesting that wider usage of spatially explicit models for decision support can be achieved by 1) developing a toolbox with multiple, easier-to-use methods, 2) improving calibration and validation of dynamic modelling approaches and 3) developing best-practise guidelines for applying these models. Further, more robust decision-making can be achieved by 4) combining multiple modelling approaches to assess uncertainty, and 5) placing models at the core of adaptive management. These efforts must be accompanied by long-term funding for modelling and monitoring, and improved communication between research and practise to ensure optimal conservation and restoration outcomes. KW - adaptive management KW - biodiversity conservation KW - cost optimisation KW - ecosystem restoration KW - global change KW - predictive models Y1 - 2021 U6 - https://doi.org/10.1111/ecog.05787 SN - 1600-0587 IS - 4 SP - 1 EP - 16 PB - Wiley-Blackwell CY - Oxford ER - TY - JOUR A1 - Witt, Barbara A1 - Stiboller, Michael A1 - Raschke, Stefanie A1 - Friese, Sharleen A1 - Ebert, Franziska A1 - Schwerdtle, Tanja T1 - Characterizing effects of excess copper levels in a human astrocytic cell line with focus on oxidative stress markers JF - Journal of trace elements in medicine and biology : organ of the Society for Minerals and Trace Elements, GMS N2 - Background: Being an essential trace element, copper is involved in diverse physiological processes. However, excess levels might lead to adverse effects. Disrupted copper homeostasis, particularly in the brain, has been associated with human diseases including the neurodegenerative disorders Wilson and Alzheimer?s disease. In this context, astrocytes play an important role in the regulation of the copper homeostasis in the brain and likely in the prevention against neuronal toxicity, consequently pointing them out as a potential target for the neurotoxicity of copper. Major toxic mechanisms are discussed to be directed against mitochondria probably via oxidative stress. However, the toxic potential and mode of action of copper in astrocytes is poorly understood, so far. Methods: In this study, excess copper levels affecting human astrocytic cell model and their involvement in the neurotoxic mode of action of copper, as well as, effects on the homeostasis of other trace elements (Mn, Fe, Ca and Mg) were investigated. Results: Copper induced substantial cytotoxic effects in the human astrocytic cell line following 48 h incubation (EC30: 250 ?M) and affected mitochondrial function, as observed via reduction of mitochondrial membrane potential and increased ROS production, likely originating from mitochondria. Moreover, cellular GSH metabolism was altered as well. Interestingly, not only cellular copper levels were affected, but also the homeostasis of other elements (Ca, Fe and Mn) were disrupted. Conclusion: One potential toxic mode of action of copper seems to be effects on the mitochondria along with induction of oxidative stress in the human astrocytic cell model. Moreover, excess copper levels seem to interact with the homeostasis of other essential elements such as Ca, Fe and Mn. Disrupted element homeostasis might also contribute to the induction of oxidative stress, likely involved in the onset and progression of neurodegenerative disorders. These insights in the toxic mechanisms will help to develop ideas and approaches for therapeutic strategies against copper-mediated diseases. KW - Copper KW - Astrocytes KW - Toxicity KW - Mitochondria KW - ROS KW - Trace elements Y1 - 2021 U6 - https://doi.org/10.1016/j.jtemb.2021.126711 SN - 1878-3252 VL - 65 PB - Elsevier CY - München ER - TY - GEN A1 - Winkelbeiner, Nicola Lisa A1 - Wandt, Viktoria Klara Veronika A1 - Ebert, Franziska A1 - Lossow, Kristina A1 - Bankoglu, Ezgi E. A1 - Martin, Maximilian A1 - Mangerich, Aswin A1 - Stopper, Helga A1 - Bornhorst, Julia A1 - Kipp, Anna Patricia A1 - Schwerdtle, Tanja T1 - A Multi-Endpoint Approach to Base Excision Repair Incision Activity Augmented by PARylation and DNA Damage Levels in Mice BT - Impact of Sex and Age T2 - Postprints der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe N2 - Investigation of processes that contribute to the maintenance of genomic stability is one crucial factor in the attempt to understand mechanisms that facilitate ageing. The DNA damage response (DDR) and DNA repair mechanisms are crucial to safeguard the integrity of DNA and to prevent accumulation of persistent DNA damage. Among them, base excision repair (BER) plays a decisive role. BER is the major repair pathway for small oxidative base modifications and apurinic/apyrimidinic (AP) sites. We established a highly sensitive non-radioactive assay to measure BER incision activity in murine liver samples. Incision activity can be assessed towards the three DNA lesions 8-oxo-2’-deoxyguanosine (8-oxodG), 5-hydroxy-2’-deoxyuracil (5-OHdU), and an AP site analogue. We applied the established assay to murine livers of adult and old mice of both sexes. Furthermore, poly(ADP-ribosyl)ation (PARylation) was assessed, which is an important determinant in DDR and BER. Additionally, DNA damage levels were measured to examine the overall damage levels. No impact of ageing on the investigated endpoints in liver tissue were found. However, animal sex seems to be a significant impact factor, as evident by sex-dependent alterations in all endpoints investigated. Moreover, our results revealed interrelationships between the investigated endpoints indicative for the synergetic mode of action of the cellular DNA integrity maintaining machinery. T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 1021 KW - maintenance of genomic integrity KW - ageing KW - sex KW - DNA damage KW - base excision repair (incision activity) KW - DNA damage response KW - poly(ADP-ribosyl)ation KW - liver Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-484831 SN - 1866-8372 IS - 1021 ER - TY - JOUR A1 - Ruszkiewicz, Joanna A1 - Endig, Lisa A1 - Güver, Ebru A1 - Bürkle, Alexander A1 - Mangerich, Aswin T1 - Life-cycle-dependent toxicities of mono- and bifunctional alkylating agents in the 3R-compliant model organism C. elegans JF - Cells : open access journal N2 - Caenorhabditis elegans (C. elegans) is gaining recognition and importance as an organismic model for toxicity testing in line with the 3Rs principle (replace, reduce, refine). In this study, we explored the use of C. elegans to examine the toxicities of alkylating sulphur mustard analogues, specifically the monofunctional agent 2-chloroethyl-ethyl sulphide (CEES) and the bifunctional, crosslinking agent mechlorethamine (HN2). We exposed wild-type worms at different life cycle stages (from larvae L1 to adulthood day 10) to CEES or HN2 and scored their viability 24 h later. The susceptibility of C. elegans to CEES and HN2 paralleled that of human cells, with HN2 exhibiting higher toxicity than CEES, reflected in LC50 values in the high µM to low mM range. Importantly, the effects were dependent on the worms’ developmental stage as well as organismic age: the highest susceptibility was observed in L1, whereas the lowest was observed in L4 worms. In adult worms, susceptibility to alkylating agents increased with advanced age, especially to HN2. To examine reproductive effects, L4 worms were exposed to CEES and HN2, and both the offspring and the percentage of unhatched eggs were assessed. Moreover, germline apoptosis was assessed by using ced-1p::GFP (MD701) worms. In contrast to concentrations that elicited low toxicities to L4 worms, CEES and HN2 were highly toxic to germline cells, manifesting as increased germline apoptosis as well as reduced offspring number and percentage of eggs hatched. Again, HN2 exhibited stronger effects than CEES. Compound specificity was also evident in toxicities to dopaminergic neurons–HN2 exposure affected expression of dopamine transporter DAT-1 (strain BY200) at lower concentrations than CEES, suggesting a higher neurotoxic effect. Mechanistically, nicotinamide adenine dinucleotide (NAD+) has been linked to mustard agent toxicities. Therefore, the NAD+-dependent system was investigated in the response to CEES and HN2 treatment. Overall NAD+ levels in worm extracts were revealed to be largely resistant to mustard exposure except for high concentrations, which lowered the NAD+ levels in L4 worms 24 h post-treatment. Interestingly, however, mutant worms lacking components of NAD+-dependent pathways involved in genome maintenance, namely pme-2, parg-2, and sirt-2.1 showed a higher and compound-specific susceptibility, indicating an active role of NAD+ in genotoxic stress response. In conclusion, the present results demonstrate that C. elegans represents an attractive model to study the toxicology of alkylating agents, which supports its use in mechanistic as well as intervention studies with major strength in the possibility to analyze toxicities at different life cycle stages. KW - C. elegans KW - alkylating agents KW - mustards KW - life cycle toxicities KW - neurotoxicity KW - NAD+ Y1 - 2023 U6 - https://doi.org/10.3390/cells12232728 SN - 2073-4409 VL - 12 IS - 23 PB - MDPI CY - Basel ER - TY - JOUR A1 - Jonas, Wenke A1 - Schwerbel, Kristin A1 - Zellner, Lisa A1 - Jähnert, Markus A1 - Gottmann, Pascal A1 - Schürmann, Annette T1 - Alterations of lipid profile in livers with impaired lipophagy JF - International journal of molecular sciences N2 - Non-alcoholic fatty liver disease (NAFLD) is characterized by excessive lipid accumulation in the liver. Various mechanisms such as an increased uptake in fatty acids or de novo synthesis contribute to the development of steatosis and progression to more severe stages. Furthermore, it has been shown that impaired lipophagy, the degradation of lipids by autophagic processes, contributes to NAFLD. Through an unbiased lipidome analysis of mouse livers in a genetic model of impaired lipophagy, we aimed to determine the resulting alterations in the lipidome. Observed changes overlap with those of the human disease. Overall, the entire lipid content and in particular the triacylglycerol concentration increased under conditions of impaired lipophagy. In addition, we detected a reduction in long-chain polyunsaturated fatty acids (PUFAs) and an increased ratio of n-6 PUFAs to n-3 PUFAs, which was due to the depletion of n-3 PUFAs. Although the abundance of major phospholipid classes was reduced, the ratio of phosphatidylcholines to phosphatidylethanolamines was not affected. In conclusion, this study demonstrates that impaired lipophagy contributes to the pathology of NAFLD and is associated with an altered lipid profile. However, the lipid pattern does not appear to be specific for lipophagic alterations, as it resembles mainly that described in relation to fatty liver disease. KW - non-alcoholic fatty liver disease KW - lipophagy KW - lipidomics KW - fatty acid profile KW - long-chain polyunsaturated fatty acids Y1 - 2022 U6 - https://doi.org/10.3390/ijms231911863 SN - 1422-0067 VL - 23 IS - 19 PB - MDPI CY - Basel ER -