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Die Brockenexkursion umfasst die Bergfichtenwälder, Moore und Bergheiden von 850 m ü. NN bis zur Brockenkuppe 1142 m ü. NN. Das Gebiet ist, ausgehend von den Fichtenwäldern und Mooren, relativ artenarm, aber strukturreich. Der Weg zum Brocken führt zunächst zwar durch ausgedehnte Fichtenforsten, dann auch durch die Bergfichtenwälder (Calam agrostio villosae-Piceetum, Sphagno girgensohnii-Piceetum, Betulo carpaticae Piceetum) auf dem Königsberg und am Brocken. Ausgedehnte Moore finden wir zwischen Torfhaus und dem Brocken sowie an den Brockenhängen, mit folgenden Gesellschaften: Eriophoro-Trichophoretum cespitosi, Sphagnetum magellanici, Eriophorum angustifolium-Sphagnum-Gesellschaft, Caricetum nigrae u.a. Von besonderem Interesse, wenn auch nur kleinflächig vorhanden, sind die subalpinen Bergheiden mit der Anemone micrantha-Calluna vulgaris-Gesellschaft auf der Brockenkuppe, die trotz der hohen Besucherzahlen nicht gefährdet ist. Die Kenntnis der Pflanzenarten der Alpen sowie der bedeutendsten Gebirge der übrigen Kontinente kann im 112 Jahre alten Brockengarten vervollständigt werden.
Die Exkursion vermittelt Eindrücke von montanen Grasland-Gesellschaften des Ober- und Unterharzes als Resten einer historischen Kulturlandschaft mit vorwiegend extensiven bis halbintensiven Nutzungseinflüssen. Im Bereich Rotes Bruch - Großer Rappenberg bei Benneckenstein wachsen relativ artenarme Kleinseggensümpfe (Caricion fuscae) und verschiedene Ausprägungen von Sumpfdotterblumen-Wiesen (Calthion palustris). Auf dem Großen Rappenberg gibt es Borstgrasrasen (Violion caninae) und unterschiedlich intensiv bewirtschaftete Goldhafer-Bergwiesen (Polygono-Trisetion). Auf der Jordanshöhe bei St. Andreasberg wachsen verschiedene Ausprägungen der Goldhafer-Bergwiesen mit Übergängen zu Borstgrasrasen. In einem Versuch werden seit 1988 verschiedene Mahdvarianten zur Regeneration und Erhaltung von Bergwiesen auf Dauerflächen verfolgt.
DieNaturschutzgebiete Hainholz und Staufenberg am südwestlichen Harzrand wurden mit den Methoden der Naturwaldforschung untersucht. Beim Hainholz handelt es sich um eines der letzten naturnahen Buchenwaldgebiete in der Gipskarstlandschaft des Zechsteins. Vorherrschend sind trockenheitsertragende bis mäßig frische Kalkbuchenwälder (Carici-Fagetum, Hordelymo-Fagetum ). Der kegelförmige Staufenberg wird durch basenarme Grauwacken- und basenreiche Diabasstandorte geprägt, auf denen von der submontanen bis zur montanen Stufe oligo- bis mesotrophe Buchenwälder (Luzulo-Fagetum, Galio odorati-Fagetum) in allen Expositionen auftreten. Während der Staufenberg seit Ausweisung als Naturwald sich mehr als drei Jahrzehnte weitgehend ungestört entwickeln konnte, wurden im Hainholz 1997 durch einen Sommergewittersturm Buchenbestände z.T. großflächig geworfen. Die Ergebnisse der floristischen und vegetationskundlichen Untersuchungen konzentrieren sich auf drei Aussagen im Zusammenhang mit dem Mosaik-Zyklus-Konzept zur Dynamik von Buchenwäldern: 1. Mit der Einstellung der forstlichen Nutzung gehen die Artenzahlen in Buchenwäldern der Optimalphase zurück. Erst bei großflächigen Störungen wie z.B. Windwurf mit freigelegtem Mineralboden steigen die Artenzahlen wieder an. 2. Fehlende Nutzung begünstigt in der Optimalphase die Buche. Ohne großflächige Störung erfolgt auch die Verjüngung überwiegend durch die schattentolerante Buche. Erst bei großflächiger Störung treten typische Pioniersträucher auf, aber auch dann dominieren auf den basenreichen Böden des Hainholzes die schattentoleranten Baumarten, die bereits vor dem Windwurf in der Verjüngungsschicht des geschlossenen Altbestandes reichlich vorhanden waren. Ein Baumartenwechsel findet nicht statt. 3. Nichtwaldarten verschwinden nach Einstellung der forstlichen Nutzung in Buchenwäldern der Optimalphase. Sie treten nach großflächigen Windwürfen erneut auf, allerdings standörtlich stark unterschiedlich in Raum und Zeit. Insgesamt lassen die Ergebnisse aus dem Hainholz den Schluss zu, dass ohne forstliche Eingriffe auch bei katastrophalen, großflächigen Störungen die Regenerationsprozesse in mitteleuropäischen Buchenwäldern rascher verlaufen als bisher vielfach angenommen wird. Da dies gleichzeitig mit einem Anstieg der Artendiversität verbunden ist, der ohne großflächige Störung ausbleibt, sollte man solchen Katastrophen aus der Sicht des Naturschutzes eher positiv und gelassen gegenüberstehen.
This thesis reports on the results obtained by expression photoactivatable adenylyl cyclase from Beggiatoa spp. (bPAC) in cholinergic neurons from Caenorhabditis elegans (C. elegans) and the characterization of the role of a single neuron, RIS, during locomotion in the adult animal.
Pharmacological activation of adenylyl cyclases through Forskolin is known to induce increased neuronal output in diverse model organisms through a protein kinase A (PKA) dependent mechanism. Nevertheless, pharmacological assays are not spatially restricted, do not allow for precise and acute activation nor to cessation of the signal. Thus, an optogenetic approach for was selected trough the expression of photoactivatable adenylyl cyclase from Beggiatoa spp. (bPAC) in cholinergic neurons of Caenorhabditis elegans (C. elegans). This model organism was chosen due to its transparency, ease of maintenance, fast generation cycles as well as for being an eutelic animal. Further, its genome has been fully sequenced and the connectome of the neuronal network is known, thus allowing for precise analysis of neuronal function. Furthermore, the molecular mechanisms governing neuronal functions are well conserved up to primates. Mainly two optogenetical tools were applied, bPAC and the light gated cation channel channelrhodopsin 2 (ChR2).
Behavioral assays of bPAC photostimulation in cholinergic neurons recapitulated previous work performed with the photoactivatable adenylyl cyclase from Euglena gracilis (EuPACa), in which swimming frequency and speed on solid substrate were increased. Electrophysiological recordings of body wall muscle (BWM) cells by Dr. Jana F. Liewald showed that bPAC photoactivation led to an increase in miniature postsynaptic current (mPSC) rate and, in contrast to ChR2 invoked depolarization, also amplitude. Analysis of mutants deficient in neuropeptidergic signaling (UNC- 31) via electrophysiology performed by Dr. Jana F. Liewald showed that the increase in mPSC amplitude due to bPAC photoactivation requires neuropeptide release. This was confirmed by co-expression of bPAC with the neuropeptide marker NLP-21::Venus and subsequent fluorescence analysis of release, exploiting the fact that released neuropeptides are ultimately degraded by scavenger cells (coelomocytes). These were enriched with NLP-21::Venus after bPAC photostimulation, but no fluorescence could be observed in the UNC-31 mutants.
Additional analysis of the electrophysiological data performed by myself showed no modulation of mPSC kinetics dues to neuropeptidergic release induced by bPAC. Hence, neuropeptide release and action sites were in the cholinergic neurons, the latter including cholinergic motoneurons.
Dr. Szi-chieh Yu provided electron microscopy images of high pressure frozen, bPAC or ChR2 expressing animals. These were tagged by myself for automatic analysis of ultrastructural properties of the cholinergic presynapse, also during photoactivation of both optogenetic tools. Photoactivation of both induced a reduction of synaptic vesicles, with ChR2 showing a more severe effect. In contrast to ChR2, though, bPAC also reduced the amount of dense core vesicles (DCV), the neuropeptide transporters. Additionally, long bPAC photoactivation as well as ChR2 photoactivation led to the appearance of large vesicles (LV), presumably in response to the increased SV fusion rate. bPAC photostimulation also induced an increase in SV size, not observed after ChR2 photostimulation. In UNC-31 mutants, bPAC photostimulation could not lead to the SV size increase, a further argument for the presynaptic effect of the released neuropeptide. Additional analysis of electrophysiology paired with pharmacology, performed by Dr. Jana F. Liewald, showed that mPSC amplitude increase requires the function of the vesicular acetylcholine transporter.
A further effect observed in the ultrastructure of bPAC photostimulated cholinergic presynapses was a shift in the distribution of SV regarding the dense projection. An analysis of cAMP pathway mutants showed that synapsin is required for bPAC induced behavior effects. Synapsin is known to mediate SV tethering to the cytoskeleton. Here, I show evidence for a new role of synapsin in controlling the availability of DCVs for fusion and thus, in neuropeptidergic signaling.
In the second part of my thesis I characterized the function of the GABAergic interneuron RIS in the neuronal network of C. elegans. RIS was shown to induce lethargus, a sleep-like state, during all larval molts, but its function in the adult animal was not yet described. Specific RIS expression of ChR2 achieved by a recombinase based system allowed to acutely depolarize the neuron during locomotion, which led to an acute behavioral stop. Diverse signal transduction pathway mutants were analyzed showing that the phenotype was induced by neuropeptidergic signaling. Through mutagenesis followed by whole genome sequencing data analysis as well as analysis of RIS specific RNA sequencing data further narrowed the signal transduction pathway to mediate the locomotion stop behavior. Since the neuropeptide and, to some extent, the neuron are conserved across nematodes, an argument is outlined in favor of the conservation of this sleep-like state.
In addition, since ChR2 could induce neuropeptidergic signaling from RIS, secretion of vesicles is regulated by variable pathways depending on the neuronal identity. Nevertheless, expression of bPAC in RIS allowed to optogenetically increase the probability of short stops, as observed by expression of a calcium sensor (GCaMP) in RIS and analysis of its intrinsic activity in the adult animal.
In the title compound, C17H18N2O, the central carbon atom with the OH substituent and one of the (E)-benzylideneamino substituents are disordered over two sets of sites with occupancies of 0.851 (4) and 0.149 (4). The relative positions of the two disorder components is equivalent to a rotation of approximately 60° about the C—N single bond. In the crystal, the molecules are held together by O—H...N hydrogen bonds, forming simple C(5) chains along the b-axis direction. In addition, pairs of the chains are further aggregated by weak C—H...π interactions.
Mathematical models of virus dynamics have not previously acknowledged spatial resolution at the intracellular level despite substantial arguments that favor the consideration of intracellular spatial dependence. The replication of the hepatitis C virus (HCV) viral RNA (vRNA) occurs within special replication complexes formed from membranes derived from endoplasmatic reticulum (ER). These regions, termed membranous webs, are generated primarily through specific interactions between nonstructural virus-encoded proteins (NSPs) and host cellular factors. The NSPs are responsible for the replication of the vRNA and their movement is restricted to the ER surface. Therefore, in this study we developed fully spatio-temporal resolved models of the vRNA replication cycle of HCV. Our simulations are performed upon realistic reconstructed cell structures—namely the ER surface and the membranous webs—based on data derived from immunostained cells replicating HCV vRNA. We visualized 3D simulations that reproduced dynamics resulting from interplay of the different components of our models (vRNA, NSPs, and a host factor), and we present an evaluation of the concentrations for the components within different regions of the cell. Thus far, our model is restricted to an internal portion of a hepatocyte and is qualitative more than quantitative. For a quantitative adaption to complete cells, various additional parameters will have to be determined through further in vitro cell biology experiments, which can be stimulated by the results deccribed in the present study.