Common and distinct roles of DFos and DJun during Drosophila development.

The Drosophila homolog of c-Jun regulates epithelial cell shape changes during the process of dorsal closure in mid-embryogenesis. Here, mutations in the DFos gene are described. In dorsal closure, DFos cooperates with DJun by regulating the expression of dpp; Dpp acts as a relay signal that triggers cell shape changes and DFos expression in neighboring cells. In addition to the joint requirement of DFos and DJun during dorsal closure, DFos functions independently of DJun during early stages of embryogenesis. These findings demonstrate common and distinct roles of DFos and DJun during embryogenesis and suggest a conserved link between AP-1 (activating protein-1) and TGF-beta (transforming growth factor-beta) signaling during epithelial cell shape changes.

Drosophila Jun kinase regulates expression of decapentaplegic via the ETS-domain protein Aop and the AP-1 transcription factor DJun during dorsal closure.

During Drosophila embryogenesis, ectodermal cells of the lateral epithelium stretch in a coordinated fashion to internalize the amnioserosa cells and close the embryo dorsally. This process, dorsal closure, requires two signaling pathways: the Drosophila Jun-amino-terminal kinase (DJNK) pathway and the Dpp pathway. We have identified mutations in DJun and show that DJNK controls dorsal closure by activating DJun and inactivating the ETS repressor Aop/Yan by phosphorylation. DJun and Aop regulate dpp expression in the most dorsal row of cells. Secreted Dpp then instructs more ventrally located cells to stretch. Our results provide a causal link between the DJNK and Dpp pathways during dorsal closure. Interestingly, in vertebrates, transforming growth factor-beta and c-Jun regulate collagenase gene expression during wound healing, a process that also involves the closing of an epithelial sheath.

The maxillary palp of Drosophila: ultrastructure and physiology depends on the lozenge gene.

The ultrastructure and physiology of the maxillary palp of Drosophila melanogaster have been studied in wild-type and lozenge mutants. Olfactory physiology in the maxillary palp is shown to depend upon the lozenge(lz) gene. Reduced response amplitudes were recorded for all odorants tested, and the physiological defect was shown to map to the lz locus. The structure of the maxillary palp sensilla is described by scanning electron microscopy (SEM) at high magnification, initially in the wild-type. A linear arrangement of pores, connected by furrows, was found in one class of sensilla, the basiconic sensilla. In the lz3 mutant, morphological alterations in the basiconic sensilla and duplications of sensilla are documented by SEM. The correlation of structural abnormalities in the lz sensilla and physiological abnormalities in odorant response are consistent with an olfactory role for the basiconic sensilla of the maxillary palp.

The Drosophila antenna: ultrastructural and physiological studies in wild-type and lozenge mutants.

The physiology and ultrastructure of the antenna in Drosophila melanogaster have been examined in wild-type and lozenge mutants. Scanning electron microscopy (SEM) of sensilla on the antennal surface has revealed that in the wild-type the basiconic sensilla contain linear arrays of pores connected by longitudinal furrows and transected by shorter furrows. Sensilla trichodea also are shown to have pores, as revealed by examining transverse sections by transmission electron microscopy (TEM); these data directly address a longstanding controversy. Coeloconic sensilla, previously described as "pit sensilla" and as "grooved" sensilla, are shown to rise directly from the antennal surface, as opposed to lying below the antennal surface in pits; the previously observed grooves correspond to the junctions between bundled, finger-like projections. This description of coeloconic sensilla is supported by analysis of lz mutants, in which the projections of coeloconic sensilla splay apart. Coeloconic sensilla are also shown to undergo duplication on the lz3 antenna. Physiological recordings from the antenna show that responses to all odorants tested are severely decreased in lz mutants. Measurements made from different parts of the antenna show similar defects. Evidence is provided that both the physiological and ultrastructural defects map to the lz locus.

The Drosophila Jun-N-terminal kinase is required for cell morphogenesis but not for DJun-dependent cell fate specification in the eye.

We cloned and characterized the Drosophila homolog of mammalian Jun-N-terminal kinases (DJNK). We show that DJNK is encoded by basket (bsk). Like hemipterous (hep), which encodes the Drosophila JNK kinase, bsk is required in the embryo for dorsal closure, a process involving coordinate cell shape changes of ectodermal cells. Dorsal closure can also be blocked by dominant negative Drosophila cdc42, which has been shown to act upstream of JNKK in vertebrates. Therefore it appears that the JNK pathway is conserved and that it is involved in controlling cell morphogenesis in Drosophila. Although DJNK efficiently phosphorylates DJun in vitro, bsk function is not required for the specification of cell fate in the developing eye, a process that requires MAP kinase and DJun function.

Olfactory physiology in the Drosophila maxillary palp requires the visual system gene rdgB.

We describe the kinetics of odorant response in the maxillary palp of Drosophila, and show that the rate of recovery from odorant stimulation is affected by mutation of the rdgB (retinal degeneration B) gene. We use immunocytochemistry to confirm that the rdgB gene product is expressed in the maxillary palp. rdgB has recently been shown to encode a protein with Ca(2+)-binding sites and sequence similarity to rat brain phosphatidylinositol transfer protein; it is located near the rhabdomeric membranes in photoreceptor cells, where it has been suggested to play a role in membrane transport. The delay in recovery kinetics that we observe in olfactory tissue may reflect a defect in membrane restoration at the conclusion of the olfactory transduction cascade. The use of common molecules in the physiology of two olfactory organs, and in both visual and olfactory physiology, is discussed.

Characterization of calcium uptake in chick retinal pigment epithelium.

45Ca uptake was studied in isolated chick retinal pigment epithelial cells. 45Ca was accumulated by a saturable, temperature-dependent system with a KM of 400 microM and a Vmax of 0.13 mumoles2mg protein/min, which depends on the external sodium concentrations. The transport system was present early during embryonic development. RPE cells of three breeds of chicks with different degrees of pigmentation accumulated calcium proportionally to the melanin content of the cells, suggesting that pigment granules participate in the storage and regulation of intracellular calcium.

A glutamate dehydrogenase-based method for the assay of L-glutamic acid: formation of pyridine nucleotide fluorescent derivatives.

A method for the quantitation of L-glutamic acid in the picomole range was developed by finding conditions which allowed the production of NADH by the action of the L-glutamate dehydrogenase (EC 1.4.1.3) and its subsequent transformation to a highly fluorescent derivative. The method measures linearly glutamate from 250 pmol to 5 nmol. For its simplicity and low cost it is ideally suited to the assay of a large number of samples within a single working day. Its application to the determination of regional glutamate levels in the rat brain, as well as to the measurement of ornithine aminotransferase (EC 2.6.1.13) activity from several tissues is described. The results are similar to those obtained by different methodologies in several laboratories, but the present method offers additional advantages.