@phdthesis{Post2014, author = {Post, Antonia}, title = {Snap25 heterozygous knockout mice as a potential model for attention deficit/hyperactivity disorder (ADHD)}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-122899}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2014}, abstract = {SNAP25 (Synaptosomal-Associated Protein of 25 kDa; part of the SNARE complex) is involved in the docking and fusion of synaptic vesicles in presynaptic neurons necessary for the regulation of neurotransmitter release, as well as in axonal growth and synaptic plasticity. In humans, different single nucleotide polymorphisms of SNAP25 have repeatedly been associated with attention deficit/hyperactivity disorder (ADHD). Thus, in this study heterozygous Snap25 knockout mice were investigated as a model of ADHD. Heterozygous (+/-) Snap25 knockout mice as well as their wild-type (+/+) littermates were reared under control conditions or underwent a Maternal Separation (MS) procedure. Starting at the age of 2 months, mice were tested for locomotor activity in a repeated long-term Open Field (OF) task, for attention deficits and impulsive behavior in the 5 Choice Serial Reaction Time Task (5CSRTT), for anxiety-like behavior in the Light-Dark Box (LDB) and for depression-like behavior in the Porsolt Forced Swim Test (FST). The brains of these mice were subsequently tested for the expression of several ADHD related genes in a quantitative Real-Time PCR (qRT-PCR) study. Another group of female mice (+/+; +/-) underwent a one hour OF test after oral administration of 45 mg/kg Methylphenidate (MPH) or placebo. To find an optimized dosage for this MPH challenge, a pilot study was performed. Wild-type C57BL/6 mice were tested in a long-term OF with several dosages of MPH both intraperitoneally (i.p.) and orally. The brains of these animals were afterwards investigated for neurotransmitter concentrations. In this pilot study the dosages of MPH that were similarly behaviorally effective without causing symptoms of overdosing were 7.5-15 mg/kg intraperitoneally and 30-60 mg/kg orally. However, even though it was possible to find intraperitoneal and oral doses that correlate behaviorally, the neurochemistry was mostly different. In the study on Snap25-deficient mice, unstressed controls showed a hyperactive phenotype in the second of two long-term OF sessions (60 min) spaced three weeks apart. Considering all groups, there was a significant interaction of stress and genotype in the second session, with animals subjected to MS being overall hyperactive with no genotype differences. In the training phase of the 5CSRTT only effects of stress were found, with MS animals finding and consuming fewer rewards. In the single test trial, several genotype effects became apparent, with tendencies for the number of correct nose pokes and the number of rewards eaten, and a significant effect for the number of rewards eaten directly after the correct response. In all of these variables +/- mice performed worse than their wild-type littermates. In the LDB +/- mice entered the lit compartment of the arena earlier than the controls, thus showing attenuated anxiety-like behavior. Regarding depressive-like behavior in the FST, male +/- mice spent significantly less time struggling than male +/+ mice. In the gene expression study, +/- mice had lower expression levels of Maoa and Comt, and higher expression levels of Nos1 than wild-types. Finally, the locomotor activity response to MPH was exaggerated in +/- mice as compared to controls. Heterozygous Snap25 knockout mice show some of the behavioral characteristics of ADHD, as for example a mild hyperactivity in a familiar environment, difficulties in the correct execution of a given task and even some behavior that can be interpreted as delay aversion. Additionally, expression levels of three ADHD related genes were changed in these animals. Although the exaggerated locomotor activity response to MPH is not to be expected of an ADHD model, the difference in the response between +/+ and +/- mice nonetheless implicates a potential dysfunction of the brain dopaminergic system.}, subject = {Aufmerksamkeitsdefizit-Syndrom}, language = {en} } @phdthesis{Proft2014, author = {Proft, Florian Lukas Patrick}, title = {Molekulare Wirkmechanismen des Antidepressivums Venlafaxin - genetische Untersuchungen in Maus und Mensch}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-109201}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2014}, abstract = {Depressive Erkrankungen verursachen sowohl das pers{\"o}nliche Leid der erkrankten Individuen als auch volkswirtschaftlichen Schaden durch krankheitsbedingten Arbeitsausfall und Belastung der Gesundheitsversorgungssysteme. Therapeutische Konzepte wie die Anwendung pharmakotherapeutischer Intervention sind in unterschiedlichem Maß von Erfolg gekr{\"o}nt. Zahlreiche somatische Faktoren wurden mit der {\"A}tiologie depressiver St{\"o}rungen in Verbindung gebracht. Die prim{\"a}r verfolgten pharmakologischen Ans{\"a}tze basieren nach wie vor auf Erkenntnissen aus der Mitte des vergangenen Jahrhunderts. In erster Linie setzt die Pharmakotherapie Substanzen ein, die die Wiederaufnahme monoaminerger Neurotransmitter (Serotonin, Noradrenalin, zum Teil auch Dopamin) aus dem synaptischen Spalt inhibieren und nach einer allerdings meist mehrw{\"o}chigen, regelm{\"a}ßigen Einnahme des Pr{\"a}parates zu einem R{\"u}ckgang der depressiven Symptomatik f{\"u}hren. Andererseits kann jedoch bei zahlreichen Erkrankten auch nach fortgesetzter Therapie mit verschiedenen Behandlungsans{\"a}tzen keine Remission verzeichnet werden und es stellt sich die Frage nach der Ursache dieser Diskrepanz. Im Fokus der vorliegenden Arbeit stand der als Antidepressivum eingesetzte selektive Serotonin- / Noradrenalin-Wiederaufnahme-Inhibitor Venlafaxin. Durch Blockade des pr{\"a}synaptischen Serotonin- und Noradrenalin-Transporters f{\"u}hrt Venlafaxin initial zu einer intensivierten Neurotransmission. Die Zielstrukturen von Venlafaxin sind der pr{\"a}synaptische Serotonin- und der Noradrenalin-Transporter, wobei aufgrund unterschiedlicher Affinit{\"a}t eine geringe Dosis beziehungsweise Konzentration als rein serotonerg betrachtet wird und bei einer hohen Dosis beziehungsweise Konzentration sowohl die Wiederaufnahme von Serotonin als auch Noradrenalin inhibiert wird. Es wurden in dieser Arbeit zwei Ziele verfolgt. Im ersten Teil wurde mittels Gen-expressionsuntersuchungen nach potentiellen Effektoren von Venlafaxin gesucht, um prinzipielle Mechanismen der antidepressiven Wirkung zu identifizieren und auf ihrer Basis die Entwicklung spezifischerer Intervention zu erm{\"o}glichen. Der zweite Teil beinhaltet eine pharmakogenetische Untersuchung am Menschen. Ziel war zu evaluieren, inwieweit die Expressionsaktivit{\"a}t von SLC6A2 und SLC6A4 und damit die pr{\"a}synaptische Transportkapazit{\"a}t in Kombination mit der Serumkonzentration aktiver Substanz als Pr{\"a}diktor des therapeutischen Effektes dienen kann. Die Kenntnis dieser Zusammenh{\"a}nge w{\"u}rde bei Vorliegen eines bestimmten Genotyps eine gezieltere Titration der individuell ben{\"o}tigten Konzentration erm{\"o}glichen und k{\"o}nnte die Effektivit{\"a}t der Therapie steigern. F{\"u}r die Genexpressionsuntersuchungen erhielten DBA/2-M{\"a}use {\"u}ber einen Zeitraum von 30 Tagen Venlafaxin in verschiedenen Dosierungen {\"u}ber das Trinkwasser. Anschließend wurden die Hippokampi der Tiere mittels genomweiter Microarray-Analyse hypothesenfrei auf zwischen den Dosisgruppen differentiell exprimierte Gene hin untersucht. Der Hippokampus wird als zentrales Element der Steuerung, Ausbildung und Ver{\"a}nderung von Verhaltensmustern gesehen. Signifikant differentiell exprimierte Gene, die in vorherigen Studien mit depressiver Erkrankung beziehungsweise einem Effekt psychiatrischer Medikation assoziiert worden waren, wurden mittels qRT-PCR-Analyse validiert. Im Anschluss an die Analyse im Tier wurden als differentiell exprimiert best{\"a}tigte Gene per qRT-PCR analog in humanen Leukozyten untersucht. Die Blutproben waren in einem klinisch-naturalistischen Design w{\"a}hrend der ersten und der f{\"u}nften Woche einer Venlafaxin-Pharmakotherapie von Patienten der Klinik f{\"u}r Psychiatrie, Psychosomatik und Psychotherapie des Universit{\"a}tsklinikums W{\"u}rzburg gewonnen worden, das heißt vor und nach potentiellem Eintreten der antidepressiven Wirkung. Trotz der unterschiedlichen Herkunft der analysierten Gewebe k{\"o}nnten auf diesem Weg Hinweise auf Vorg{\"a}nge im menschlichen Gehirn gefunden werden, wie in vergleichenden post mortem Untersuchungen zwischen peripherem und zentralem humanem Material erkannt worden war. Die in der Tierstudie identifizierten Gene kodieren f{\"u}r Transkriptionsfaktoren sowie Proteine die als Teil von second messenger-Kaskaden bekannt sind. Von statistischer Signifikanz erwies sich in der Analyse der humanen Leukozyten die Expressionsreduktion der mRNA der Transkriptionsfaktor-Untereinheit Fos. Befunde zu einer Funktion von Fos, die eine Interpretation im Bezug auf den antidepressiven Effekt von Venlafaxin erm{\"o}glichen, liegen lediglich aus Tierstudien vor. Fos-ko im Hippo-kampus von M{\"a}usen wurde mit reduziertem Angstverhalten und h{\"o}herer Exzitabilit{\"a}t von hippokampalen Neuronen assoziiert. Auch wurde eine Assoziation mit Vorg{\"a}ngen bei synaptischer Plastizit{\"a}t und damit potentiell bei Lernvorg{\"a}ngen gefunden. Auf der anderen Seite wurde depressions-{\"a}hnliches Verhalten bei Ratten mit niedriger hippokampaler Fos-Expression und dessen erfolgreiche pharmakologische "Therapie" mit einer Induktion der Fos-Expression assoziiert. Es scheinen also bereits zwischen nicht-menschlichen Spezies ausgepr{\"a}gte Unterschiede der Rolle von Fos beziehungsweise Fos zu bestehen. Aufgrund der unterschiedlichen Spezies und Gewebe in den hier durchgef{\"u}hrten Untersuchungen sowie den uneinheitlichen Befunden bez{\"u}glich der Rolle von Fos beziehungsweise Fos in vorangegangenen Studien kann abschließend lediglich konstatiert werden, dass Fos vermutlich an der Entstehung depressionsbeg{\"u}nstigender Physiologie beteiligt ist und auch, dass eine antidepressive Pharmakotherapie mit Venlafaxin ihre Wirkung vermutlich unter Beteiligung von Fos entfaltet. Die Entwicklung innovativer Antidepressiva die unter Umgehung der monoaminergen Transmissionssysteme durch gezielte Reduktion der Fos-Abundanz das therapeutische Ziel erreichen lassen, k{\"o}nnte auf Basis der vorliegenden Studie angedacht werden, scheint allerdings aufgrund der ubiquit{\"a}ren Mediatorent{\"a}tigkeit des Proteins und insbesondere aufgrund seiner nicht endg{\"u}ltig definierten Rolle bei der Entstehung von Krebs nicht praktikabel. Zuk{\"u}nftige Untersuchungen sollten daher auf andere im Microarray differentiell exprimiert gefundene Gene fokussieren. In die Untersuchung der Expressionsaktivit{\"a}t der f{\"u}r die prim{\"a}ren Zielstrukturen von Venlafaxin (Serotonin- beziehungsweise Noradrenalin-Transporter) kodierenden Gene (SLC6A4 beziehungsweise SLC6A2) und der Serumkonzentration an aktiver Substanz nach Venlafaxin-Applikation im Hinblick auf deren Pr{\"a}diktivit{\"a}t des therapeutischen Effektes, wurden in einem klinisch-naturalistischen Design Patienten der Klinik f{\"u}r Psychiatrie, Psychosomatik und Psychotherapie des Universit{\"a}tsklinikums W{\"u}rzburg eingeschlossen. Genotypisiert wurden f{\"u}r SLC6A2 der SNP rs28386840 und f{\"u}r SLC6A4 der Polymorphismus 5-HTTLPR. Die Genotypen wurden jeweils in niedrig- und hoch-exprimierend unterteilt und damit auf die ph{\"a}notypische Transportkapazit{\"a}t der pr{\"a}synaptischen Membran Bezug genommen. Der therapeutische Erfolg wurde anhand der CGI-I-Skala evaluiert und f{\"u}r die Analysen in "gutes Ansprechen" und "schlechtes Ansprechen" dichotomisiert. Der SLC6A2-Polymorphismus zeigte sich als nicht mit dem therapeutischen Effekt assoziiert. Der hochexprimierende SLC6A4-Genotyp wurde signifikant mit einem schlechteren Ansprechen assoziiert. Dies war in den nach Serumkonzentration aktiver Substanz stratifizierten Unterkollektiven insbesondere in dem Bereich zwischen 200 und 400 ng / ml zu erkennen, wohingegen unter- und oberhalb dieses Bereiches keine Assoziation zu finden war. Aus diesen Resultaten kann gefolgert werden, dass sich aus der Genotypisierung von rs28386840 keine therapeutischen Instruktionen ableiten lassen. Bei Kenntnis des 5-HTTLPR-Genotyps k{\"o}nnte f{\"u}r den klinischen Alltag die Empfehlung ergehen, falls Venlafaxin als sSNRI bei Patienten mit hochexprimierendem Genotyp eingesetzt werden soll, eine Serumsummenkonzentration jenseits des durch die AGNP empfohlenen Bereiches (100 - 400 ng / ml) anzustreben. Da hier jedoch lediglich eine Stichprobe von 56 Patienten untersucht und insbesondere, da zahlreiche potentielle Kofaktoren des therapeutischen Effektes nicht in die Analyse einbezogen werden konnten, ist die Assoziation vor Anwendung in der Therapiesteuerung anhand umfassenderer prospektiver kontrollierter Studien zu validieren.}, subject = {Wirkmechanismus}, language = {de} } @phdthesis{Wurster2014, author = {Wurster, Sebastian}, title = {Die Bedeutung von LIN9 f{\"u}r die Regulation der Genexpression, die genomische Stabilit{\"a}t und die Tumorsuppression}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-114967}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2014}, abstract = {Pocket proteins and E2F transcription factors regulate the expression of cell cycle associated genes and play a central role in the coordination of cell division, differentiation, and apoptosis. Disorders of these pathways contribute to the development of various human tumor entities. Despite intensive research in the field of cell cycle regulation many details are not yet understood. The LIN complex (LINC / DREAM) is a recently discovered human multiprotein complex, which dynamically interacts with pocket proteins and E2F transcription factors. An essential component of the LIN complex is the LIN9 protein. In order to obtain a better insight into the function of this protein in cell cycle regulation and tumorigenesis, a conditional Lin9 knockout mouse model was established in our laboratory. The primary objective of this study was the phenotypic characterization of embryonic fibroblasts (MEFs) from these mice. Shortly after inactivation of Lin9 cell proliferation was massively impaired. Multiple types of mitotic defects such as structural abnormalities of the spindle apparatus, aberrant nuclei, failed nuclear segregation and cytokinesis failure have been observed in Lin9-depleted cells leading to a dramatic increase in polyploid and aneuploid cells. Ultimately these serious aberrations result in premature cellular senescence. If the senescence of Lin9-deficient cells is overcome by the Large T antigen the cells can adhere to the loss of Lin9, but show severe genomic instability and grow anchorage-independently in soft-agar as a sign of oncogenic transformation. In the second part of the thesis the gene expression of Lin9-deficient cells was assessed by quantitative real time PCR analyses to determine, whether the mitotic abnormalities are caused by transcriptional defects. Here a significant reduction of mitotic gene expression was observed in Lin9-depleted cells. Additionally chromatin immunoprecipitation experiments were performed to clarify the underlying molecular mechanisms. Compared to control cells epigenetic alterations at the promoters of mitotic target genes with regard to activating histone modifications were found in Lin9-deficient MEFs. In the last section of this study, the effects of Lin9 heterozygosity were analyzed. Lin9 heterozygous MEFs showed normal proliferation, although expression of different mitotic genes was slightly reduced. It appeared, however, that the mitotic spindle checkpoint of Lin9 heterozygous MEFs is weakened and thus over several cell generations an increase in polyploid cells was observed. Soft-agar assays showed that Lin9 heterozygosity contributes to oncogenic transformation. Taken together, these results document a crucial role of LIN9 in the regulation of cell cycle-associated gene expression. LIN9 is an essential factor for cell proliferation on one hand, while at the same time it functions as a tumor suppressor.}, subject = {Zellzyklus}, language = {de} } @phdthesis{Hondke2014, author = {Hondke, Sylvia}, title = {Elucidation of WISP3 function in human mesenchymal stem cells and chondrocytes}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-109641}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2014}, abstract = {WISP3 is a member of the CCN family which comprises six members found in the 1990's: Cysteine-rich,angiogenic inducer 61 (CYR61, CCN1), Connective tissue growth factor (CTGF, CCN2), Nephroblastoma overexpressed (NOV, CNN3) and the Wnt1 inducible signalling pathway protein 1-3 (WISP1-3, CCN4-6).They are involved in the adhesion, migration, mitogenesis, chemotaxis, proliferation, cell survival, angiogenesis, tumorigenesis, and wound healing by the interaction with different integrins and heparan sulfate proteoglycans. Until now the only member correlated to the musculoskeletal autosomal disease Progressive Pseudorheumatoid Dysplasia (PPD) is WISP3. PPD is characterised by normal embryonic development followed by cartilage degradation over time starting around the age of three to eight years. Animal studies in mice exhibited no differences between knock out or overexpression compared to wild type litter mates, thus were not able to reproduce the symptoms observed in PPD patients. Studies in vitro and in vivo revealed a role for WISP3 in antagonising BMP, IGF and Wnt signalling pathways. Since most of the knowledge of WISP3 was gained in epithelial cells, cancer cells or chondrocyte cell lines, we investigated the roll of WISP3 in primary human mesenchymal stem cells (hMSCs) as well as primary chondrocytes. WISP3 knock down was efficiently established with three short hairpin RNAs in both cell types, displaying a change of morphology followed by a reduction in cell number. Simultaneous treatment with recombinant WISP3 was not enough to rescue the observed phenotype nor increase the endogenous expression of WISP3. We concluded that WISP3 acts as an essential survival factor, where the loss resulted in the passing of cell cycle control points followed by apoptosis. Nevertheless, Annexin V-Cy3 staining and detection of active caspases by Western blot and immunofluorescence staining detected no clear evidence for apoptosis. Furthermore, the gene expression of the death receptors TRAILR1 and TRAILR2,important for the extrinsic activation of apoptosis, remained unchanged during WISP3 mRNA reduction. Autophagy as cause of cell death was also excluded, given that the autophagy marker LC3 A/B demonstrated to be uncleaved in WISP3-deficient hMSCs. To reveal correlated signalling pathways to WISP3 a whole genome expression analyses of WISP3-deficient hMSCs compared to a control (scramble) was performed. Microarray analyses exhibited differentially regulated genes involved in cell cycle control, adhesion, cytoskeleton and cell death. Cell death observed by WISP3 knock down in hMSCs and chondrocytes might be explained by the induction of necroptosis through the BMP/TAK1/RIPK1 signalling axis. Loss of WISP3 allows BMP to bind its receptor activating the Smad 2/3/4 complex which in turn can activate TAK1 as previously demonstrated in epithelial cells. TAK1 is able to block caspase-dependent apoptosis thereby triggering the assembly of the necrosome resulting in cell death by necroptosis. Together with its role in cell cycle control and extracellular matrix adhesion, as demonstrated in human mammary epithelial cells, the data supports the role of WISP3 as tumor suppressor and survival factor in cells of the musculoskeletal system as well as epithelial cells.}, subject = {Knorpelzelle}, language = {en} } @phdthesis{Zhang2014, author = {Zhang, Yi}, title = {Regulation of Agrobacterial Oncogene Expression in Host Plants}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-102578}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2014}, abstract = {Virulent Agrobacterium tumefaciens strains transfer and integrate a DNA region of the tumor-inducing (Ti) plasmid, the T-DNA, into the plant genome and thereby cause crown gall disease. The most essential genes required for crown gall development are the T-DNA-encoded oncogenes, IaaH (indole-3-acetamide hydrolase), IaaM (tryptophan monooxygenase) for auxin, and Ipt (isopentenyl transferase) for cytokinin biosynthesis. When these oncogenes are expressed in the host cell, the levels of auxin and cytokinin increase and cause cell proliferation. The aim of this study was to unravel the molecular mechanisms, which regulate expression of the agrobacterial oncogenes in plant cells. Transcripts of the three oncogenes were expressed in Arabidopsis thaliana crown galls induced by A. tumefaciens strain C58 and the intergenic regions (IGRs) between their coding sequences (CDS) were proven to have promoter activity in plant cells. These promoters possess eukaryotic sequence structures and contain cis-regulatory elements for the binding of plant transcription factors. The high-throughput protoplast transactivation (PTA) system was used and identified the Arabidopsis thaliana transcription factors WRKY18, WRKY40, WRKY60 and ARF5 to activate the Ipt oncogene promoter. No transcription factor promoted the activity of the IaaH and IaaM promoters, despite the fact that the sequences contained binding elements for type B ARR transcription factors. Likewise, the treatment of Arabidopsis mesophyll protoplasts with cytokinin (trans-zeatin) and auxin (1-NAA) exerted no positive effect on IaaH and IaaM promoter activity. In contrast, the Ipt promoter strongly responded to a treatment with auxin and only modestly to cytokinin. The three Arabidopsis WRKYs play a role in crown gall development as the wrky mutants developed smaller crown galls than wild-type plants. The WRKY40 and WRKY60 genes responded very quickly to pathogen infection, two and four hours post infection, respectively. Transcription of the WRKY18 gene was induced upon buffer infiltration, which implicates a response to wounding. The three WRKY proteins interacted with ARF5 and with each other in the plant nucleus, but only WRKY40 together with ARF5 increased activation of the Ipt promoter. Moreover, ARF5 activated the Ipt promoter in an auxin-dependent manner. The severe developmental phenotype of the arf5 mutant prevented studies on crown gall development, nevertheless, the reduced crown gall growth on the transport inhibitor response 1 (TIR1) tir1 mutant, lacking the auxin sensor, suggested that auxin signaling is required for optimal crown gall development. In conclusion, A. tumefaciens recruits the pathogen defense related WRKY40 pathway to activate Ipt expression in T-DNA-transformed plant cells. IaaH and IaaM gene expression seems not to be controlled by transcriptional activators, but the increasing auxin levels are signaled via ARF5. The auxin-depended activation of ARF5 boosts expression of the Ipt gene in combination with WRKY40 to increase cytokinin levels and induce crown gall development.}, subject = {Agrobacterium tumefaciens}, language = {en} } @phdthesis{Merker2014, author = {Merker, S{\"o}ren}, title = {Genome-wide screenings in attention-deficit/hyperactivity disorder (ADHD): investigation of novel candidate genes SLC2A3 and LPHN3}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-100129}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2014}, abstract = {Attention-deficit/hyperactivity disorder (ADHD) is a highly prevalent childhood-onset neurodevelopmental disorder that involves a substantial risk of persisting into adolescence and adulthood. A number of genome-wide screening studies in ADHD have been conducted in recent years, giving rise to the discovery of several variants at distinct chromosomal loci, thus emphasising the genetically complex and polygenic nature of this disorder. Accordingly, promising novel candidate genes have emerged, such as the gene encoding the glucose transporter isoform 3 (SLC2A3) and the gene encoding the latrophilin isoform 3 (LPHN3). In this thesis, both genes were investigated in form of two separated projects. The first focused on SLC2A3 polymorphisms associated with ADHD and their potential physiological impact. For this purpose, gene expression analyses in peripheral cell models were performed as well as functional EEG measurements in humans. The second project concerned the murine gene Lphn3 including the goal of developing a mouse line containing a genetically modified Lphn3 with conditional knockout potential. In this respect, a specific DNA vector was applied to target the Lphn3 gene locus in murine embryonic stem (ES) cells as a prerequisite for the generation of appropriate chimeric mice. The results of the first project showed that SLC2A3 duplication carriers displayed increased SLC2A3 mRNA expression in peripheral blood cells and significantly altered event-related potentials (ERPs) during tests of cognitive response control and working memory, possibly involving changes in prefrontal brain activity and memory processing. Interestingly, ADHD patients with the rs12842 T-allele, located within and tagging the SLC2A3 gene, also exhibited remarkable effects during these EEG measurements. However, such effects reflected a reversed pattern to the aforementioned SLC2A3 duplication carriers with ADHD, thus indicative of an opposed molecular mechanism. Besides, it emerged that the impact of the aforementioned SLC2A3 variants on different EEG parameters was generally much more pronounced in the group of ADHD patients than the healthy control group, implying a considerable interaction effect. Concerning the second project, preliminary results were gathered including the successful targeting of Lphn3 in murine ES cells as well as the production of highly chimeric, phenotypically unremarkable and mostly fertile mouse chimeras. While germline transmission of the modified Lphn3 allele has not yet occurred, there are still several newborn chimeric mice that will be tested in the near future. In conclusion, the findings suggest that SLC2A3 variants associated with ADHD are accompanied by transcriptional and functional changes in humans. Future research will help to elucidate the molecular network and neurobiological basis involved in these effects and apparently contributing to the complex clinical picture of ADHD. Moreover, given the increasing number of publications concerning latrophilins in recent years and the multitude of research opportunities provided by a conditional knockout of Lphn3 in mice, the establishment of a respective mouse line, which currently is in progress, constitutes a promising approach for the investigation of this gene and its role in ADHD.}, subject = {Genexpression}, language = {en} }