gcm and pointed synergistically control glial transcription of the Drosophila gene loco.

In Drosophila lateral glial cell development is initiated by the transcription factor encoded by glial cells missing. glial cells missing activates downstream transcription factors such as repo and pointed which subsequently control terminal glial differentiation. The gene loco has been identified as a potential target gene of pointed and is involved in terminal glial differentiation. It encodes an RGS domain protein expressed specifically by the lateral glial cells in the developing embryonic CNS. Here we analyzed the loco promoter and the control of the glial-specific transcription pattern. Using promoter-reporter gene fusions we identified a 1.9 kb promoter element capable of directing the almost complete loco gene expression pattern. Sequence analysis suggested the presence of gcm and pointed DNA binding sites. Following in vitro mutagenesis of these sites we demonstrated their relevance in vivo. The expression of loco is initially dependent on gcm. During subsequent stages of embryonic development gcm and pointed appear to activate loco transcription synergistically. In addition, at least two other factors appear to repress loco expression in the ectoderm and in the CNS midline cells.

A primary cation transport by a V-type ATPase of low specificity

The enzyme involved in outward K+ transport in insect epithelia belongs to the family of V-ATPases. Evidence has been reported relating the generation of the K+ gradient to a primary electrogenic proton transport via a distinct electrophoretic nH+/K+ antiport. The subject of this paper is the transport of K+ at a thread hair sensillum of the cockroach in situ. We recorded changes in the voltage and resistance of the ion-transporting membrane and of shifts in pH caused by inhibition of energy metabolism and by putative inhibitors of a proton/cation exchanger. The results are supplemented by previous determinations of the K+ activities in the same preparation. 1. In cockroach hair sensilla, the ion transport generates a membrane voltage of 105 mV. We found that the transport rendered the positive output compartment alkaline with respect to the cytoplasm by 1.0 pH unit compared with the pH at equilibrium distribution, and we infer that proton transport cannot be the process that energizes the generation of the K+ gradient. 2. The ion transport created an electrochemical potential difference for protons, DeltaetaH, of approximately 4.5 kJ mol-1, while the potential difference for K+, DeltaetaK, amounted to approximately 11 kJ mol-1. Both potential differences are directed to the cytosol. It follows from DeltaetaK/DeltaetaH that an antiport would have to be electrophoretic to drive K+ by DeltaetaH and it should, therefore, contribute to the membrane conductance. Amiloride and harmaline did not significantly change the pH in the adjacent spaces and did not affect the voltage or the resistance of the transporting membrane. Previous determinations of the impedance have shown that the ATP-independent conductance of this membrane is small, supporting the conclusion that it lacks an electrophoretic antiport. From these results, we deduce that K+ transport in cockroach sensilla is not secondary to a proton transport and an electrochemical proton gradient. The phenomena observed match the performance of a primary, electrogenic, cation-translocating ATPase of the type deduced from analyses of the short-circuit current at the midgut epithelium of lepidopteran larvae. The validity of the H+ transport/antiport hypothesis is discussed.