ALBERT

Description: The Toll signaling pathway functions in several Drosophila processes, including dorsal-ventral pattern formation and the immune response. Here, we demonstrate that this pathway is required in the epidermis for proper muscle development. Previously, we showed that the zygotic Toll protein is necessary for normal muscle development; in the absence of zygotic Toll, close to 50% of hemisegments have muscle patterning defects consisting of missing, duplicated and misinserted muscle fibers (Halfon, M.S., Hashimoto, C., and Keshishian, H., Dev. Biol. 169, 151-167, 1995). We have now also analyzed the requirements for easter, spatzle, tube, and pelle, all of which function in the Toll-mediated dorsal-ventral patterning pathway. We find that spatzle, tube, and pelle, but not easter, are necessary for muscle development. Mutations in these genes give a phenotype identical to that seen in Toll mutants, suggesting that elements of the same pathway used for Toll signaling in dorsal-ventral development are used during muscle development. By expressing the Toll cDNA under the control of distinct Toll enhancer elements in Toll mutant flies, we have examined the spatial requirements for Toll expression during muscle development. Expression of Toll in a subset of epidermal cells that includes the epidermal muscle attachment cells, but not Toll expression in the musculature, is necessary for proper muscle development. Our results suggest that signals received by the epidermis early during muscle development are an important part of the muscle patterning process.

Description: The Effects of elevated free-stream turbulence (FST) on the natural and periodically excited separation bubbles were studied experimentally, due to the relevance of this flow to low-pressure turbine blades at low Reynolds numbers. A bubble was formed at the leading edge of a flat plate and the FST level was altered by placing a grid across the flow at different locations upstream of the plate. The mixing across the separated shear-layer, forming the free boundary of the bubble, increased due to the elevated FST and due to nominally two-dimensional periodic excitation, both flattening and shortening the bubble. Periodic excitation at frequencies that were at least an order of magnitude lower than those associated with the initial shear-layer instability, were very effective at low FST, because the amplitudes of the excitation frequency and its harmonic were amplified over the bubble. High frequency excitation (F+ 3, based on the length of the baseline low FST bubble) had a major effect close to the separation location, while farther downstream the excited fluctuations rapidly decayed in the reattachment region. Low frequency excitation, that generated waves comparable to the length of the unperturbed bubble (F+ 1) were less effective and their magnitude decayed at a slower rate downstream of reattachment. An increase in the level of the FST reduced the net effect of the periodic excitation on the mixing enhancement and subsequent reattachment process, probably due to a destructive interference between the nominally 2D excitation and the random (in space and time) FST, reducing the spanwise coherence and therefore the effectiveness of the current control strategy. However, even at the reduced effectiveness of 2D periodic excitation at elevated FST, it accelerated the reattachment process and the recovery rate of the reattached boundary layer, enhancing the boundary layer resistance to repeat separation and reducing its momentum loss further downstream.

Description: We have developed a method to target gene expression in the Drosophila embryo to a specific cell without having a promoter that directs expression in that particular cell. Using a digitally enhanced imaging system to identify single cells within the living embryo, we apply a heat shock to each cell individually by using a laser microbeam. A 1- to 2-min laser treatment is sufficient to induce a heat-shock response but is not lethal to the heat-shocked cells. Induction of heat shock was measured in a variety of cell types, including neurons and somatic muscles, by the expression of beta-galactosidase from an hsp26-lacZ reporter construct or by expression of a UAS target gene after induction of hsGAL4. We discuss the applicability of this technique to ectopic gene expression studies, lineage tracing, gene inactivation studies, and studies of cells in vitro. Laser heat shock is a versatile technique that can be adapted for use in a variety of research organisms and is useful for any studies in which it is desirable to express a given gene in only a distinct cell or clone of cells, either transiently or constitutively, at a time point of choice.