During the development of multicellular organisms, organs grow to well-defined shapes and sizes. The proper size and patterning of tissues are ensured by signaling molecules as e.g. morphogens. Secreted from a restricted source, morphogens spread into the adjacent target tissue where they form a graded concentration profile. Upon binding of the morphogens to receptors on the cell surfaces, the morphogenetic signal is transduced inside the cell via the phosphorylation of transcription factors, which subsequently regulate the expression of different target genes. Thus, cell fates are determined by the local concentration of such morphogens. In this work, we investigate three key aspects of morphogenetic signaling processes in growing tissues. First, we study the mechanics of tissue growth via cell division and cell death. We examine the rearrangements of cells on large scales and times by developing a continuum theory which describes the growing tissue as an active complex fluid. In our description we include anisotropic stresses generated by oriented cell division, and we show that average cellular trajectories exhibit anisotropic scaling behaviors. Our description is used to study experimentally measured shape changes of the developing wing disk of the fruit fly Drosophila melanogaster. Second, we focus on the spreading of morphogens in growing tissues. We show that the flow field of cell movements due to oriented cell division and cell death causes a drift term in the morphogen transport equation, which captures the stretching and dilution of the concentration profile. Comparing our theoretical results to recent experiments in the Drosophila wing disk, we find that the transport of the morphogen Dpp is mainly intracellular. We moreover show that the decay length of the Dpp gradient increases during development as a result of changing degradation rate and diffusion coefficient, whereas the drift of molecules due to growth is negligible. Thus growth does not affect the decay length of the gradient, but the decay length of the gradient might affect the tissue growth rate as discussed in this work. Finally, we develop a microscopic theoretical description of the intracellular transduction machinery of morphogenetic signals within an individual cell. Our description captures the kinetics of the trafficking of proteins between different cellular compartments in response to receptor-bound signaling molecules. Analyzing experimental data at the Drosophila neuromuscular junction, we show that the morphogenetic signaling is modulated by synaptic signaling via neuronal action potentials.

Early events during axolotl limb regeneration include an immune response and the formation of a wound epithelium. These events are linked to a clearance of damaged tissue prior to blastema formation and regeneration of the missing structures. Here, we report the resorption of calcified skeletal tissue as an active, cell-driven, and highly regulated event. This process, carried out by osteoclasts, is essential for a successful integration of the newly formed skeleton. Indeed, the extent of resorption is directly correlated with the integration efficiency, and treatment with zoledronic acid resulted in osteoclast function inhibition and failed tissue integration. Moreover, we identified the wound epithelium as a regulator of skeletal resorption, likely releasing signals involved in recruitment/differentiation of osteoclasts. Finally, we reported a correlation between resorption and blastema formation, particularly, a coordination of resorption with cartilage condensation. In sum, our results identify resorption as a major event upon amputation, playing a critical role in the overall process of skeletal regeneration.

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Wir fanden konservierte molekulare Signaturen der Regulation durch Δ7-DA und Ascarosid bei Dauer- und infektiösen Larven. Danach wurde die hohe Konservierung durch unsere Analyse in Dauer- und Postdauer-Stadium zwischen den zwei nah verwandten freilebenden Arten C. elegans und C. briggsae identifiziert. Das heißt, dass die relative Veränderung auf mRNA- oder Protein- Ebene zwischen zwei Arten stark korreliert ist. Aber die relative Veränderung innerhalb derselben Art zeigt keine hochgradige Korrelation zwischen mRNA- und Protein-Ebene. Unsere Ergebnisse zeigen in C. elegans Dauerlarven die signifikante Reduzierung der RNA-Mengen in 20 Stoffwechselwegen. Im Gegensatz dazu speicherten Dauerlarven reichlich RNA-Mengen in GO Termen wie Ribosome und Aminoacyl-tRNA biosynthesis. Auf Protein-Ebene sind die Stoffwechselwege von Proteinsynthese und Proteinverarbeitung im endoplasmatischen Retikulum in Dauerlarven herunterreguliert und GO Terme wie Lysosome sind hochreguliert. Durch die Zeitreihenanalyse der Proteom-Remodellierung der molekularen Signaturen beim Austritt aus dem Dauer-Stadium fand wir, dass GO Terme wie metal ion binding signifikant herunterreguliert sind und der Proteinabbau hochreguliert ist. Unsere Ergebnisse vom pSILAC Experiment deuten an, dass die Proteine für Energieerzeugung und Chaperone/Proteinfaltung beim Daueraustritt schnell verbraucht sind und wieder hergestellt werden. Zum Schluss haben wir als Erste den popomR-Assay in C. elegans etabliert und ein Screening der vermeintlichen Proteinbindestellen auf poly-A-RNA durchgeführt, um in der Zukunft die konservierten Mechanismen der post-transkriptionellen Regulation durch RBPs im Dauer-Stadium zu analysieren. / We found the conservation of molecular signatures by regulating with Δ7-DA and Ascarosid in dauer larvae and infective larvae. Then by our comparative analysis, the high degree of conservation between two closely related free-living species C. elegans and C. briggsae was identified in dauer and post-dauer stages. This means that the relative changes are strongly correlated on the mRNA or the protein level between two species. But the relative changes in the same species don’t show any strong correlation between the mRNA and the protein levels. Our results showed a significantly reduced amount of RNA in 20 metabolic pathways in C. elegans dauer larvae. In contrast, dauer larvae stored a large amount of RNA in GO terms such as ribosome and aminoacyl-tRNA biosynthesis. On the protein level, the metabolic pathways of protein synthesis and protein processing in endoplasmic reticulum were downregulated in dauer larvae and the term of lysosome was up-regulated. Due to time course analysis for proteome remodeling of molecular signatures during exit process from dauer stage, we found that GO terms such as metal ion binding were significantly downregulated during dauer exit and at the same time the protein degradation was up-regulated. Our results of pSILAC experiment suggest that the proteins for energy generation and chaperone/protein folding are quickly spent and rebuilded during dauer exit. Finally, we were the first to establish the popomR assay in C. elegans and performed a screening of the putative protein binding sites on poly-A RNA to analyze the conserved mechanisms of post-transcriptional regulation by RBPs in dauer larvae in the future.

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