Not the outside but the inside matters : functional analysis of Capricious and Tartan during Drosophila tracheal morphogenesis

The tubular structures, which transport essential gases, liquids, or cells from one site to another, are shared among various divergent organisms. These highly organized tubular networks include lung, kidney, vasculature and mammary gland in mammals as well as trachea and salivary gland in Drosophila melanogaster. Many questions regarding the tubular morphogenesis cannot be addressed sufficiently by investigating the mammalian organs because their structures are extremely complex and therefore, systematic analyses of genetic and cellular programs guiding the development is not possible. In contrast, the Drosophila tracheal development provides an excellent model system since many molecular markers and powerful tools for genetic manipulations are available. Two mechanisms were shown to be important for the outgrowth of tracheal cells: the FGF signaling pathway and the interaction between the tracheal cells and the surrounding mesodermal cells. The Drosophila FGF ligand encoded by branchless (bnl) is localized in groups of cells near tracheal metameres. The tracheal cells expressing the FGF receptor breathless (btl) respond to these sources of FGF ligand and extend towards them. However, this FGF signaling pathway is not sufficient for the formation of continuous dorsal trunk, the only muticellular tube in tracheal system. Recently, it was found out that single mesodermal cells called bridge-cells are essential for the formation of continuous dorsal trunk as they direct the outgrowth of dorsal trunk cells towards the correct targets. The results in this PhD thesis demonstrate that a cell adhesion molecule Capricious (Caps), which is specifically localized on the surface of bridge-cells, plays an essential role in guiding the outgrowing dorsal trunk cells towards their correct targets. When caps is lacking, some bridge-cells cannot stretch properly towards the adjacent posterior tracheal metameres and thus fail to interconnect the juxtaposing dorsal trunk cells. Consequently, discontinuous dorsal trunks containing interruptions at several positions are formed. On the other hand, when caps is ectopically expressed in the mesodermal cells through a twi-GAL4 driver, these mesodermal cells acquire a guidance function through ectopic caps and misguide the outgrowing dorsal trunk cells in abnormal directions. As a result, disconnected dorsal trunks are formed. These loss- and gain-of-function studies suggest that Caps presumably establishes the cell-to-cell contact between the bridge-cells and the tracheal cells and thereby mediates directly the guidance function of bridge-cells. The most similar protein known to Caps is another cell adhesion molecule called Tartan (Trn). Interestingly, trn is expressed in the mesodermal cells but not in the bridge-cells. When trn is lacking, the outgrowth of not only the dorsal trunks but also the lateral trunks are disrupted. However, in contrast to the ectopic expression of caps, the misexpression of trn does not affect tracheal development. Whereas Trn requires only its extracellular domain to mediate the matrix function, Caps requires both its extracellular and intracellular domains to function as a guidance molecule in the bridge-cells. These observations suggest that Trn functions differently from Caps during tracheal morphogenesis. Presumably, Trn mediates a matrix function of mesodermal cells, which support the tracheal cells to extend efficiently through the surrounding mesodermal tissue. In order to determine which domains dictate the functional specificity of Caps, two hybrid proteins CapsEdTrnId, which contains the Caps extracellular domain and the Trn intracellular domain, and TrnEdCapsId, which consists of the Trn extracellular domain and the Caps intracellular domain, were constructed. Gain of function and rescue experiments with these hybrid proteins suggest on one hand that the extracellular domains of Caps and Trn are functionally redundant and on the other hand that the intracellular domain dictates the functional specificity of Caps. In order to identify putative interactors of Caps, yeast two-hybrid screening was performed. An in vivo interaction assay in yeast suggests that Ras64B interacts specifically with the Caps intracellular domain. In addition, an in vitro binding assay reveals a direct interaction between an inactive form of Ras64B and the Caps intracellular domain. ras64B, which encodes a small GTPase, is expressed in the mesodermal cells concurrently as caps. Finally, a gain-of-function study with the constitutively active Ras64B suggests that Ras64B presumably functions downstream of Caps. All these results suggest consistently that the small GTPase Ras64B binds specifically to the Caps intracellular domain and may thereby mediate the guidance function of Caps.

Decision Making for Teams of Mobile Robots

In the past years, we could observe a significant amount of new robotic systems in science, industry, and everyday life. To reduce the complexity of these systems, the industry constructs robots that are designated for the execution of a specific task such as vacuum cleaning, autonomous driving, observation, or transportation operations. As a result, such robotic systems need to combine their capabilities to accomplish complex tasks that exceed the abilities of individual robots. However, to achieve emergent cooperative behavior, multi-robot systems require a decision process that copes with the communication challenges of the application domain. This work investigates a distributed multi-robot decision process, which addresses unreliable and transient communication. This process composed by five steps, which we embedded into the ALICA multi-agent coordination language guided by the PROViDE negotiation middleware. The first step encompasses the specification of the decision problem, which is an integral part of the ALICA implementation. In our decision process, we describe multi-robot problems by continuous nonlinear constraint satisfaction problems. The second step addresses the calculation of solution proposals for this problem specification. Here, we propose an efficient solution algorithm that integrates incomplete local search and interval propagation techniques into a satisfiability solver, which forms a satisfiability modulo theories (SMT) solver. In the third decision step, the PROViDE middleware replicates the solution proposals among the robots. This replication process is parameterized with a distribution method, which determines the consistency properties of the proposals. In a fourth step, we investigate the conflict resolution. Therefore, an acceptance method ensures that each robot supports one of the replicated proposals. As we integrated the conflict resolution into the replication process, a sound selection of the distribution and acceptance methods leads to an eventual convergence of the robot proposals. In order to avoid the execution of conflicting proposals, the last step comprises a decision method, which selects a proposal for implementation in case the conflict resolution fails. The evaluation of our work shows that the usage of incomplete solution techniques of the constraint satisfaction solver outperforms the runtime of other state-of-the-art approaches for many typical robotic problems. We further show by experimental setups and practical application in the RoboCup environment that our decision process is suitable for making quick decisions in the presence of packet loss and delay. Moreover, PROViDE requires less memory and bandwidth compared to other state-of-the-art middleware approaches.