Toxoplasma gondii und Plasmodium Spezies sind obligat intrazelluläre Parasiten, die Zucker zur Energiehomöostase als auch für die Synthese lebenswichtiger Makromoleküle verwenden. Die hier vorgestellten Daten zeigen, dass der Glukosetransporter von T. gondii, TgGT1, und die homologen Transporter von P. falciparum und P. berghei, PfHT1 und PbHT1, neben Glukose auch Mannose, Fructose und Galactose transportieren. Toxoplasma Tachyzoiten exprimieren neben TgGT1 noch einen weiteren putative Zuckertransporter (TgST2) an der Parasitenoberfläche. Beide Proteine sind nicht essentiell, wie durch ihre individuelle und gleichzeitige Gendeletion belegt wird. Die Deletion von TgGT1 bewirkt einen geringen Wachstumsdefekt. Die Mutante ?tggt1 zeigt keine Glukoseaufnahmeaktivität und folglich eine verminderte glukoseabhängige Motilität. In ?tggt1 Parasiten wird ein verstärkter Glutaminstoffwechsel nachgewiesen, der ausreichend ist dessen Motilität und Replikationsaktivität zu erhalten. Die ?tggt1 Mutante gewährt Einblick in die Anpassungsfähigkeit von T. gondii an unterschiedliche Wirtszellen. Im Gegensatz zu T. gondii benötigen erythrozytäre Plasmodien Glukose und der Transporter PfHT1 wird derzeit als drug-target eingestuft. Hier wird gezeigt, dass das PfHT1-Homolog, PbHT1, essentiell in Blutstadien des Nagerparasiten Plasmodium berghei ist, jedoch auch während des gesamten Lebenszyklus des Parasiten exprimiert wird. Ein PfHT1- und PbHT1-spezifischer Inhibitor (Compound 3361) kann die Entwicklung von P. berghei Leberstadien und Okineten stark hemmen. Um zukünftig PfHT1-Inhibitoren im Hochdurchsatzverfahren zu identifizieren und testen zu können, wurden auf Saccharomyces cerevisiae und P. berghei basierende Expressionssysteme für PfHT1 entwickelt. Abschließend stellt diese Arbeit die Unterschiedlichkeit des zentralen Kohlenstoffwechsels von Toxoplasma und Plasmodium Parasiten durch bisher unbekannter Aspekte heraus. / Toxoplasma gondii and Plasmodium species are obligate intracellular pathogens that utilize host sugars for energy homeostasis and macro molecular synthesis. Here, we report that the T. gondii glucose transporter, TgGT1, and of its homologs of P. falciparum and P. berghei (PfHT1 and PbHT1) transport glucose, mannose, galactose and fructose. Besides TgGT1, Toxoplasma harbours one additional surface localized putative sugar transporter (TgST2). Surprisingly both Proteins are nonessential and only the deletion of TgGT1 inflicts a mild defect in the parasite replication. The ?tggt1 mutant is unable to import glucose and consequently displays an attenuated glucose-dependent motility, which is completely rescued by glutamine. ?tggt1 performs increased glutamine metabolism that is sufficient to sustain motility and replication. The ?tggt1 strain provides a model for further investigating its adaptation to disparate host cells. In contrast to T. gondii, erythrocytic stages of Plasmodium species critically depend on glucose uptake, and the PfHT1 transporter is considered as a drug target against human malaria. Here, we report that PbHT1 (a PfHT1 homolog) is also essential for blood stage development in the rodent malaria parasite P. berghei. PbHT1 is expressed throughout the life cycle. Moreover, a PfHT1- and PbHT1-specific sugar analogue, compound 3361, can inhibit the hepatic development and ookinete formation in P. berghei. These results signify that PbHT1 and exogenous glucose are also required during the ex-erythrocytic stages of P. berghei. To permit a high-throughput screening of selective PfHT1 inhibitors and their subsequent in vivo assessment, we have established a PfHT1-expressing Saccharomyces cerevisiae mutant and generated a PfHT1-dependent ?pbht1 of P. berghei strain. This thesis underscores various previously unknown aspects of sugar metabolism in Toxoplasma and Plasmodium, and unravel their metabolic differences.

In type 1 diabetes (T1D), an autoimmune disease, autoantibodies are preferentially directed against proteins associated with Golgi and post-Golgi secretory vesicles, including insulin secretory granules and synaptic-like microvesicles. Thus, the study of beta-cell autoantigens with yet unknown function may provide novel insight into the secretory machinery of beta-cells and led to the discovery of novel pathways. Islet cell autoantigen of 69 kDa (ICA69) is a T1D autoantigen. It is a cytosolic protein of still unknown function. An impairment in neurotransmitter release upon mutation of its homologue in C. elegans suggests, however, an involvement of ICA69 in neurosecretion. Interestingly, ICA69 contains a BAR domain, present in several proteins involved in intracellular transport. The BAR domain functions as a dimerization motif, provides a general binding interface for different types of GTPases, and is a membrane binding/bending module. Its presence in ICA69 is a further hint supporting the putative involvement of ICA69 in intracellular membrane trafficking. The first part of this thesis was concerned with the characterization of ICA69, and the elucidation of its role in membrane traffic in pancreatic beta-cells. ICA69 was shown to be enriched in the perinuclear region, where also markers of the Golgi region are found. ICA69 was shown to interact with several membrane lipids, preferentially with PI(4)P, enriched on the Golgi complex. During the course of this thesis a combination of biochemical and imaging techniques were applied to investigate the interaction between ICA69 and Rab2, a small GTPase associated with the intermediate compartment and involved in the trafficking between the ER and the Golgi complex. ICA69 was shown to co-immunoprecipitate with Rab2 from INS-1 cells extracts. GST-pull down assays demonstrated that this interaction is GTP-dependent. Furthermore, confocal microscopy indicated that ICA69 and Rab2 extensively colocalize in particulate structures throughout the cytoplasm. Immunocytochemistry and subcellular fractionation experiments suggested that Rab2 recruits ICA69 to membranes. Functional studies indicated that ICA69 over-expression in INS-1 cells has effects that resemble, and in some cases amplify those observed upon Rab2 over-expression. Specifically, it impairs the trafficking between ER and Golgi, measured through the appearance and the conversion of the pro-form of ICA512 in the mature form of the protein. Moreover, it correlates with a redistribution of the beta-COP subunit of the coatomer, participating in the early secretory pathway, between membrane-bound compartments and the cytosol and it reduces stimulated insulin secretion. The data reported in this thesis conclusively point to ICA69 as a novel Rab2 effector, and may therefore contribute to the elucidation the yet poorly understood mechanism of action of Rab2 in the secretory pathway. The second part of the thesis was devoted to the study of an ICA69 paralogue gene, called ICA69-RP. Similarly to ICA69, ICA69-RP mRNA was shown to be primarily present in tissues such as brain and pancreatic islets, showing the expression pattern of a gene preferentially expressed in neuroendocrine cells. Unlike ICA69, however, and similar to other genes associated with the secretory machinery of beta-cells, ICA69-RP appeared to be glucose regulated, as shown by a 1.55 fold increase in mRNA levels upon stimulation of the cells with 25 mM glucose for two hours.Glucose stimulation of beta-cells prompts the activation of post-transcripional mechanisms which quickly up-regulate the expression of secretory granule genes and consequently renew granule stores. The increased expression of ICA69-RP upon glucose stimulation of cells may be part of this process. Unfortunately, all attempts to elucidate the intracellular localization of endogenous ICA69-RP failed, and it was not possible to obtain significant insights about its localization by over-expressing a fusion protein between ICA69-RP and GFP. Unlike other paralogues containing the BAR domain, such as amphiphysin 1 and 2 or Rvs167p and Rvs161p, ICA69 and ICA69-RP were shown not to form heterodimers. Furthermore, ICA69-RP did not show any interaction with Rab2 or Rab1, involved in the anterograde transport between ER and Golgi. Thus, its physiological role remains to be investigated.

Background: Tumor cells are highly dependent on glucose even in the presence of oxygen. This concept called the Warburg effect is a hallmark of cancer and strategies are considered to therapeutically exploit the phenomenon such as ketogenic diets. The success of such strategies is dependent on a profound understanding of tumor cell metabolism. With new techniques it is now possible to thoroughly analyze the metabolic responses to the withdrawal of substrates and their substitution by others. In the present study we used gas chromatography coupled to mass spectrometry (GC-MS) to analyze how glioblastoma brain tumor cells respond metabolically when glucose is withdrawn and substituted by pyruvate. Methods: Glioblastoma brain tumor cells were cultivated in medium with high (25 mM), medium (11 mM) or low (5.5 mM) glucose concentration or with pyruvate (5 mM). After 24 h GC-MS metabolite profiling was performed. Results: The abundances of most metabolites were dependent on the supply of glucose in tendency but not in a linear manner indicating saturation at high glucose. Noteworthy, a high level of sorbitol production and release was observed at high concentrations of glucose and high release of alanine, aspartate and citrate were observed when glucose was substituted by pyruvate. Intermediates of the TCA cycle were present under all nutritional conditions and evidence was found that cells may perform gluconeogenesis from pyruvate. Conclusions: Our experiments reveal a high plasticity of glioblastoma cells to changes in nutritional supply which has to be taken into account in clinical trials in which specific diets are considered for therapy.

In type 1 diabetes (T1D), an autoimmune disease, autoantibodies are preferentially directed against proteins associated with Golgi and post-Golgi secretory vesicles, including insulin secretory granules and synaptic-like microvesicles. Thus, the study of beta-cell autoantigens with yet unknown function may provide novel insight into the secretory machinery of beta-cells and led to the discovery of novel pathways. Islet cell autoantigen of 69 kDa (ICA69) is a T1D autoantigen. It is a cytosolic protein of still unknown function. An impairment in neurotransmitter release upon mutation of its homologue in C. elegans suggests, however, an involvement of ICA69 in neurosecretion. Interestingly, ICA69 contains a BAR domain, present in several proteins involved in intracellular transport. The BAR domain functions as a dimerization motif, provides a general binding interface for different types of GTPases, and is a membrane binding/bending module. Its presence in ICA69 is a further hint supporting the putative involvement of ICA69 in intracellular membrane trafficking. The first part of this thesis was concerned with the characterization of ICA69, and the elucidation of its role in membrane traffic in pancreatic beta-cells. ICA69 was shown to be enriched in the perinuclear region, where also markers of the Golgi region are found. ICA69 was shown to interact with several membrane lipids, preferentially with PI(4)P, enriched on the Golgi complex. During the course of this thesis a combination of biochemical and imaging techniques were applied to investigate the interaction between ICA69 and Rab2, a small GTPase associated with the intermediate compartment and involved in the trafficking between the ER and the Golgi complex. ICA69 was shown to co-immunoprecipitate with Rab2 from INS-1 cells extracts. GST-pull down assays demonstrated that this interaction is GTP-dependent. Furthermore, confocal microscopy indicated that ICA69 and Rab2 extensively colocalize in particulate structures throughout the cytoplasm. Immunocytochemistry and subcellular fractionation experiments suggested that Rab2 recruits ICA69 to membranes. Functional studies indicated that ICA69 over-expression in INS-1 cells has effects that resemble, and in some cases amplify those observed upon Rab2 over-expression. Specifically, it impairs the trafficking between ER and Golgi, measured through the appearance and the conversion of the pro-form of ICA512 in the mature form of the protein. Moreover, it correlates with a redistribution of the beta-COP subunit of the coatomer, participating in the early secretory pathway, between membrane-bound compartments and the cytosol and it reduces stimulated insulin secretion. The data reported in this thesis conclusively point to ICA69 as a novel Rab2 effector, and may therefore contribute to the elucidation the yet poorly understood mechanism of action of Rab2 in the secretory pathway. The second part of the thesis was devoted to the study of an ICA69 paralogue gene, called ICA69-RP. Similarly to ICA69, ICA69-RP mRNA was shown to be primarily present in tissues such as brain and pancreatic islets, showing the expression pattern of a gene preferentially expressed in neuroendocrine cells. Unlike ICA69, however, and similar to other genes associated with the secretory machinery of beta-cells, ICA69-RP appeared to be glucose regulated, as shown by a 1.55 fold increase in mRNA levels upon stimulation of the cells with 25 mM glucose for two hours.Glucose stimulation of beta-cells prompts the activation of post-transcripional mechanisms which quickly up-regulate the expression of secretory granule genes and consequently renew granule stores. The increased expression of ICA69-RP upon glucose stimulation of cells may be part of this process. Unfortunately, all attempts to elucidate the intracellular localization of endogenous ICA69-RP failed, and it was not possible to obtain significant insights about its localization by over-expressing a fusion protein between ICA69-RP and GFP. Unlike other paralogues containing the BAR domain, such as amphiphysin 1 and 2 or Rvs167p and Rvs161p, ICA69 and ICA69-RP were shown not to form heterodimers. Furthermore, ICA69-RP did not show any interaction with Rab2 or Rab1, involved in the anterograde transport between ER and Golgi. Thus, its physiological role remains to be investigated.

Hintergrund: Die Versorgung von Immunzellen mit Energie in Form von ATP ist Grundlage eines funktionstüchtigen Immunsystems. Diese wird durch die mitochondriale OXPHOS oder durch die zytosolische Glykolyse gewährleistet. Sauerstoff und Glukose stellen die Hauptsubstrate dieser Stoffwechselprozesse dar. Fragestellung: Unter pathologischen Bedingungen wie sie in Entzündungsgebieten herrschen, konnte ein relativer Sauerstoffmangel experimentell nachgewiesen werden. Ziel dieser Arbeit war es herauszufinden, in welcher Weise die Funktionen einer definierten Lymphozytenpopulation (CD4+) durch Sauerstoffmangel beeinflusst werden. Methoden: Nach Isolation von CD4+ Zellen aus peripherem Blut gesunder Spender, wurden definierte Zellmengen stimuliert und in einem mit einer Sauerstoffelektrode ausgestatteten Gefäß unter Luftabschluß inkubiert. Zu definierten Zeitpunkten wurden Proben zur ATP-Messung entnommen, sowie Protein- und RNA-Lysate hergestellt. Die Vitalität zu Anfang und zum Ende der Inkubation wurde mittels Propidium-Jodid-Färbung im FACS bestimmt. Aus gesammelten Überständen wurden mittels Multiplex-ELISA die Konzentrationen von IL-1beta, IL-2, IL-6, IL-8, IL-10, TNF-alpha und MCAF gemessen. Als Kontrollen dienten unter Normoxie inkubierte Aliquots der Zellsuspensionen. HIF-1alpha wurde mit Immunoblotting nachgewiesen. Transkriptionsänderungen von SOD1 und HK1 wurden durch SYBR-Green Real-Time-PCR quantifiziert. Ergebnisse: Stimulierte CD4+-Zellen von Normalspendern schütten unter dem Einfluss von Hypoxie vermehrt proinflammatorische und chemotaktisch wirksame Zytokine, sowie zur Differenzierung notwendige antiinflammatorische Zytokine aus. Die Verfügbarkeit von Glukose hat hierauf einen verstärkenden Effekt. Eine hypoxische Umgebung sorgt in Abhängigkeit von der Versorgung mit Glukose für eine Anpassung der zellulären Atmungsrate. Glukose ist für die Aufrechterhaltung eines konstanten ATP-Levels verantwortlich. Die glykolytische Energiegewinnung unter Hypoxie kompensiert den Ausfall der OXPHOS. Hypoxie führt bei stimulierten CD4+-Zellen bei freier Glukoseverfügbarkeit zu einer vermehrten Transkription des Hexokinase1-Gens. Glukosemangel bewirkt dagegen in hypoxischer Umgebung eine Transkriptionssteigerung des SOD1-Gens. / Background: The energy supply of immune cells in form of ATP is the cornerstone of a functional immune system. This supply is realized by either mitochondrial OXPHOS or cytosolic glycolysis. Oxygen and glucose present the main substrates in these metabolic processes. Objective: Relative shortness of oxygen could be determined experimentally under pathological conditions present in inflamed tissues. The aim of this study was to determine the extent of hypoxic influence on the cellular function of CD4+ lymphocytes. Methods: Human CD4+ cells were isolated from peripheral blood of healthy blood donors by MACS sorting. Following a defined protocol cells were stimulated and incubated in a sealed container with a Clark type electrode. Samples were taken for measurements of ATP content. RNA- and Protein lysates were made to quantify the transcription of SOD1 and HK1 by SYBR green RT-PCR and look for the presence of HIF-1alpha by immunoblot analysis respectively. Supernatants were used to measure the expression of IL-1beta, IL-2, IL-6, IL-8, IL-10, TNF-alpha and MCAF using a multiplex ELISA assay. Aliquots of cell supspensions incubated under normoxic conditions served as controls. Results / Conclusion: Under the influence of hypoxia stimulated CD4+ lymphocytes of healthy blood donors express proinflammatory and chemotactically active as well as anti-inflammatory cytokines important for cell differentiation. The availability of glucose leads to an increase of this effect. An hypoxic environment dependant on the availability of glucose leads to an adaptation of cellular respiration. Glucose deficiency provokes an increase in cellular oxygen utilization. The availability of glucose is responsible for a constant intracellular ATP level. This proves that in CD4+ lymphocytes glycolysis is capable of compensating for hypoxically impaired oxidative phosphorylation thus providing enough ATP to enable cellular function. Hypoxia under glucose provision leads to an increase in mRNA expression for HK1, a key enzyme of glycolysis. Lack of glucose under hypoxic conditions results in an increase in mRNA expression for SOD1. Glucose therefore serves in CD4+ cells as an agent of constant energy supply that leads to cell survival and an upkeep of a proinflammatory environment through cytokine expression.

Für die Untersuchung standen 20 tragende Booroola*Merinofleischschaf Kreuzungsgenotypen, meist pluripar, zwischen 2 und 6 Jahren zur Verfügung. Während der Trächtigkeit wurden in wöchentlichem Abstand die Größenveränderungen der Plazentomdurchmesser mittels transkutaner und transrektaler Ultrasonographie erfasst und Blutproben genommen. Post partum erfolgte die detaillierte Auswertung der Plazenta nach Anzahl und Durchmesser der Kotyledonen und des Kotyledonen- und Sekundinengewichts. Aus den gewonnenen Daten wurden Plazentomwachstumskurven im Verlauf der Trächtigkeit erstellt. Die Blutproben wurden hormonanalytisch auf Progesteron, 17ß-Östradiol, und IGF-1, sowie stoffwechselphysiologisch auf die, den Eiweiß- und Energiehaushalt charakterisierenden Substanzen Albumin, Gesamteiweiß, Harnstoff, BHB, Bilirubin und Glukose untersucht. Zudem erfolgte die Bestimmung von Substanzen, die in der Trächtigkeit stark beansprucht werden, wie ASAT, Calcium, Eisen und Cholesterol. Die Auswertung sämtlicher Daten erfolgte in Abhängigkeit von Wurfgröße (WG) und Wurfgewichtsklassen (WGK). Der größte Einfluss von WG und WGK auf die Plazenta zeigt sich bei der Auswertung der morphologischen Aspekte. Dabei sind in erster Linie eine Vergrößerung der Plazentomdurchmesser von uni- zu triparen Tieren sowie von WGK 1 zur WGK 3 festzustellen. In den jeweils höchsten Klassen fällt der Durchmesser wieder. Zudem nimmt in der Regel mit steigender WG und WGK die Anzahl der Plazentome mit kleinen Durchmessern (1-20 mm)ab, die Anzahl der Plazentome mit großen Durchmessern (20-50 mm) dagegen zu. Tripare Tiere und WGK 3 weisen die meisten Plazentome mit den größten Durchmessern (40-50 mm) auf. Bei der Gesamtkontaktfläche zeigt sich ein Anstieg der Fläche von uni- bis zu quadriparen Tieren und der WGK 1 bis WGK 4. Nur WGK 5 weist eine gegenüber WGK 4 verminderte Gesamtkontaktfläche auf. Unabhängig von WG und WGK verkleinern sich die Plazentomdurchmesser und die Gesamtkontaktflächen p.p. gegenüber den Plazentomdurchmessern und Gesamtkontaktflächen a.p. WG und WGK beeinflussen den Hormonhaushalt mäßig. Dabei zeigt sich der größte Einfluss beim Progesteron. Die kleinste WG bzw. WGK präsentiert die niedrigsten Progesteronkonzentrationen. Beim 17ß-Östradiol ist der Verlauf aller Konzentrationskurven im gesamten Trächtigkeitsverlauf und beim IGF-1 ab dem 100. Tag p.c. einheitlich. Durch WG oder WGK werden bei den Booroola*MF Kreuzungsgenotypen keine den Stoffwechsel charakterisierenden Substanzen in einer auffallenden oder für die Tierart untypischen Weise verändert. Die Anzahl Feten bzw. die WGK zeigen im vorliegenden Datenmaterial keinen gravierenden Einfluss auf den maternalen Stoffwechsel während der Gravidität. / The analysis was founded on 20 pregnant Booroola* Merino Mutton crossbreed sheep between 2 and 6 years, most of them pluriparous. During pregnancy the variations of size of the placentomdiameters were drawn up on a weekly basis using transcutaneous and transrectal ultrasonography. Samples of blood were taken likewise in a weekly rhythm. After birth a detailed examination of the placentae followed, considering especially their number and diameter as well as the weight of the cotyledons and the secundinae. The gained data were used to create placentom-growth-diagrams during pregnancy. The samples of blood were hormonally analysed with regard to progesterone, estradiol and IGF-1 as well as to the substances that characterize the protein- and energy metabolism as there are: albumin, protein, urea, BHB, bilirubin, glucose. Moreover substances which are highly required during pregnancy were determined as e.g. ASAT, calcium, iron, cholesterol. The evaluation of all the gained data was made in dependence on the littersize (LS) and total litterweight (TLW). The major influence of the LS and TLW on the placenta is noticed at the morphological aspects. The placentomdiameter increase from uni- to triparous ewes and from TLW 1 to TLW 3. On the other hand the placentomdiameters decrease in the highest groups. With an increase of LS and TLW the amound of placentoms with small diameters (1-20 mm) generally decreased in favour of the placentoms with big diameters (20-50 mm). Triparous ewes and TLW 3 show most of the placentoms with the largest diameters (40-50 mm). The "total contact area" demonstrates an increase from uni-to quadriparous ewes and TLW 1 to TLW 4. Only TLW 5 has a smaller "total contact area" as TLW 4. Independent of LS and TLW of the Booroola* Merino Mutton crossbreed sheep the placentomdiameter and the "total contact area" decrease p.p. in comparison to the placentomdiameter and "total contact area" a.p. The influence of LS and TLW on the endocrinological system is moderate. The highest influence is proved concerning progesterone, the smallest LS and TLW show the lowest blood-progesterone concentration. In contrast to this, the estradiol concentration of both groups seems to be uniform during the whole pregnancy and equally the IGF-1 concentration from the 100 day p.c. None of the metabolism characterising substances seems to be affected by the LS or TLW. The littersize as well as the litterweight do not influence in the gained data the maternal metabolism during pregnancy.

Für die Untersuchung standen 20 tragende Booroola*Merinofleischschaf Kreuzungsgenotypen, meist pluripar, zwischen 2 und 6 Jahren zur Verfügung. Während der Trächtigkeit wurden in wöchentlichem Abstand die Größenveränderungen der Plazentomdurchmesser mittels transkutaner und transrektaler Ultrasonographie erfasst und Blutproben genommen. Post partum erfolgte die detaillierte Auswertung der Plazenta nach Anzahl und Durchmesser der Kotyledonen und des Kotyledonen- und Sekundinengewichts. Aus den gewonnenen Daten wurden Plazentomwachstumskurven im Verlauf der Trächtigkeit erstellt. Die Blutproben wurden hormonanalytisch auf Progesteron, 17ß-Östradiol, und IGF-1, sowie stoffwechselphysiologisch auf die, den Eiweiß- und Energiehaushalt charakterisierenden Substanzen Albumin, Gesamteiweiß, Harnstoff, BHB, Bilirubin und Glukose untersucht. Zudem erfolgte die Bestimmung von Substanzen, die in der Trächtigkeit stark beansprucht werden, wie ASAT, Calcium, Eisen und Cholesterol. Die Auswertung sämtlicher Daten erfolgte in Abhängigkeit von Wurfgröße (WG) und Wurfgewichtsklassen (WGK). Der größte Einfluss von WG und WGK auf die Plazenta zeigt sich bei der Auswertung der morphologischen Aspekte. Dabei sind in erster Linie eine Vergrößerung der Plazentomdurchmesser von uni- zu triparen Tieren sowie von WGK 1 zur WGK 3 festzustellen. In den jeweils höchsten Klassen fällt der Durchmesser wieder. Zudem nimmt in der Regel mit steigender WG und WGK die Anzahl der Plazentome mit kleinen Durchmessern (1-20 mm)ab, die Anzahl der Plazentome mit großen Durchmessern (20-50 mm) dagegen zu. Tripare Tiere und WGK 3 weisen die meisten Plazentome mit den größten Durchmessern (40-50 mm) auf. Bei der Gesamtkontaktfläche zeigt sich ein Anstieg der Fläche von uni- bis zu quadriparen Tieren und der WGK 1 bis WGK 4. Nur WGK 5 weist eine gegenüber WGK 4 verminderte Gesamtkontaktfläche auf. Unabhängig von WG und WGK verkleinern sich die Plazentomdurchmesser und die Gesamtkontaktflächen p.p. gegenüber den Plazentomdurchmessern und Gesamtkontaktflächen a.p. WG und WGK beeinflussen den Hormonhaushalt mäßig. Dabei zeigt sich der größte Einfluss beim Progesteron. Die kleinste WG bzw. WGK präsentiert die niedrigsten Progesteronkonzentrationen. Beim 17ß-Östradiol ist der Verlauf aller Konzentrationskurven im gesamten Trächtigkeitsverlauf und beim IGF-1 ab dem 100. Tag p.c. einheitlich. Durch WG oder WGK werden bei den Booroola*MF Kreuzungsgenotypen keine den Stoffwechsel charakterisierenden Substanzen in einer auffallenden oder für die Tierart untypischen Weise verändert. Die Anzahl Feten bzw. die WGK zeigen im vorliegenden Datenmaterial keinen gravierenden Einfluss auf den maternalen Stoffwechsel während der Gravidität. / The analysis was founded on 20 pregnant Booroola* Merino Mutton crossbreed sheep between 2 and 6 years, most of them pluriparous. During pregnancy the variations of size of the placentomdiameters were drawn up on a weekly basis using transcutaneous and transrectal ultrasonography. Samples of blood were taken likewise in a weekly rhythm. After birth a detailed examination of the placentae followed, considering especially their number and diameter as well as the weight of the cotyledons and the secundinae. The gained data were used to create placentom-growth-diagrams during pregnancy. The samples of blood were hormonally analysed with regard to progesterone, estradiol and IGF-1 as well as to the substances that characterize the protein- and energy metabolism as there are: albumin, protein, urea, BHB, bilirubin, glucose. Moreover substances which are highly required during pregnancy were determined as e.g. ASAT, calcium, iron, cholesterol. The evaluation of all the gained data was made in dependence on the littersize (LS) and total litterweight (TLW). The major influence of the LS and TLW on the placenta is noticed at the morphological aspects. The placentomdiameter increase from uni- to triparous ewes and from TLW 1 to TLW 3. On the other hand the placentomdiameters decrease in the highest groups. With an increase of LS and TLW the amound of placentoms with small diameters (1-20 mm) generally decreased in favour of the placentoms with big diameters (20-50 mm). Triparous ewes and TLW 3 show most of the placentoms with the largest diameters (40-50 mm). The "total contact area" demonstrates an increase from uni-to quadriparous ewes and TLW 1 to TLW 4. Only TLW 5 has a smaller "total contact area" as TLW 4. Independent of LS and TLW of the Booroola* Merino Mutton crossbreed sheep the placentomdiameter and the "total contact area" decrease p.p. in comparison to the placentomdiameter and "total contact area" a.p. The influence of LS and TLW on the endocrinological system is moderate. The highest influence is proved concerning progesterone, the smallest LS and TLW show the lowest blood-progesterone concentration. In contrast to this, the estradiol concentration of both groups seems to be uniform during the whole pregnancy and equally the IGF-1 concentration from the 100 day p.c. None of the metabolism characterising substances seems to be affected by the LS or TLW. The littersize as well as the litterweight do not influence in the gained data the maternal metabolism during pregnancy.

Neurogenesis in the adult hippocampus contributes to learning and memory in the healthy brain but is dysregulated in metabolic and neurodegenerative diseases. The molecular relationships between neural stem cell activity, adult neurogenesis, and global metabolism are largely unknown. Here we applied unbiased systems genetics methods to quantify genetic covariation among adult neurogenesis and metabolic phenotypes in peripheral tissues of a genetically diverse family of rat strains, derived from a cross between the spontaneously hypertensive (SHR/OlaIpcv) strain and Brown Norway (BN-Lx/Cub). The HXB/BXH family is a very well established model to dissect genetic variants that modulate metabolic and cardiovascular diseases and we have accumulated deep phenome and transcriptome data in a FAIR-compliant resource for systematic and integrative analyses. Here we measured rates of precursor cell proliferation, survival of new neurons, and gene expression in the hippocampus of the entire HXB/BXH family, including both parents. These data were combined with published metabolic phenotypes to detect a neurometabolic quantitative trait locus (QTL) for serum glucose and neuronal survival on Chromosome 16: 62.1–66.3 Mb. We subsequently fine-mapped the key phenotype to a locus that includes the Telo2-interacting protein 2 gene (Tti2)—a chaperone that modulates the activity and stability of PIKK kinases. To verify the hypothesis that differences in neurogenesis and glucose levels are caused by a polymorphism in Tti2, we generated a targeted frameshift mutation on the SHR/OlaIpcv background. Heterozygous SHR-Tti2+/- mutants had lower rates of hippocampal neurogenesis and hallmarks of dysglycemia compared to wild-type littermates. Our findings highlight Tti2 as a causal genetic link between glucose metabolism and structural brain plasticity. In humans, more than 800 genomic variants are linked to TTI2 expression, seven of which have associations to protein and blood stem cell factor concentrations, blood pressure and frontotemporal dementia.

No description available.

In den vorliegenden Untersuchungen wurde das Ziel verfolgt die Nutzbarkeit des Milchinhaltsstoffes Laktose als praxistaugliche Managementhilfe zu prüfen. Die Primärdaten stammen aus drei israelischen Hochleistungsherden, über mehrere Laktationen erhoben. Der Parameter Laktosegehalt wurde in der Datenaufbereitung dahingehend geprüft, ob dieser zur Gesundheits- und Leistungsvorhersage ausreicht oder welche zusätzlichen Merkmale für die Verwendung in einem Prognose-Modell von Bedeutung sein könnten. Als leistungs- bzw. gesundheitsrelevante Ereignisse (Events) wurden Brunst, Diarrhoe, Endometritis, Fieber, Infektionen, Klauenerkrankungen, Mastitis, Stress, Stoffwechselstörungen sowie Verletzungen zugeordnet. Die Bewertung der Nützlichkeit einzelner Merkmale für die Prädiktion erfolgte anhand der Erkennungsraten. Zwei- und dreistufige Entscheidungsbäume wurden entwickelt, um diese Events zu identifizieren. Ein einzelnes Merkmal ist oft nicht ausreichend, weshalb verschiedene Kombinationen von Variablen analysiert wurden. Die wichtigste Erkenntnis der vorliegenden Arbeit besteht darin, dass der Abfall der Laktosekonzentration und Laktosemenge immer ein kritisches Ereignis darstellt. Das Hauptziel eines Gesundheitsmonitorings im Milchkuhbestand sollte deshalb darin bestehen, frühzeitig eine Stoffwechselüberlastung "sichtbar" oder "erkennbar" zu machen. Unabhängig davon, welche Erkrankung sich anbahnt, muss das Herdenmanagement darauf hinwirken, die Glukoseversorgungssituation des Einzeltieres zu verbessern. Aus der Analyse für die einzelnen Herden und Laktationen kann grundlegend abgeleitet werden, dass die Ergebnisse der Milchkontrolldaten, die im Zuge der datengestützten Herdenüberwachung erhoben wurden, sich verwenden lassen, um den Leistungs- und Gesundheitsstatus der Kühe im Laktationsverlauf einzuschätzen und zu prognostizieren. Die Verwendung von Informationen zum Laktosegehalt des Gemelks verbesserten in jedem Fall die Erkennungsraten. / The aim of the current studies was to investigate whether the milk ingredient lactose can be used as a practical support management. The primary data comes from three Israeli high-performance herds, collected over several lactations. In the data preparation, the parameter "lactose content" was examined to see whether it is sufficient for a health and performance prediction or whether additional features may be of importance for usage in a forecasting model. Oestrus, diarrhea, endometritis, fever, infections, hoof diseases, mastitis, stress, metabolic disorders, and injuries have been assigned to the performance- and/or health-affecting events. The usefulness of individual features for the prediction was evaluated on the basis of the recognition rates. Thus two- and three-level decision trees have been developed to identify these events. As one single feature is often insufficient, different combinations of variables were analyzed. The most important finding of this study is that the drop in the lactose concentration and lactose quantity always represents a critical event. The main objective of a health monitoring in the dairy herd should therefore be to make a metabolic overload "visible" or "recognisable" at an early stage. Whichever disease begins to take shape, the herd management must work on improving the glucose supply situation of the individual animal. In conclusion from the analysis of the individual herds and lactations it can be inferred that the results of the milk control data collected in the course of the data-based herd monitoring can be used in order to assess and to predict the performance and health status of the cows in the course of lactation. The use of information on the lactose content of the milk improved in any case the recognition rates.