Regulation of Drosophila neural development by a putative secreted protein

The Drosophila strawberry (sty) locus was isolated by P-element insertion mutagenesis in a screen for mutations affecting eye development. Analysis of the mutant phenotype and the putative expression pattern of the sty gene suggested that it has multiple functions. Mutations in the sty gene lead to irregular spacing of ommatidia, an increase in the number of photoreceptor cells, as well as abnormal axonal projections to the lamina and disrupted structure of the optic lobes in the adult fly. The sty mutation also causes abnormal head involution, a change in a number of sensilla in the antennomaxillary complex in the embryonic stage and abnormal morphogenesis of the maxillary palp and wings in later stages. We examined the presumptive expression of the sty gene during development by histochemical staining for lacZ expression from enhancer trap elements inserted within the sty gene. During embryogenesis, expression of lacZ showed a segmental pattern in the ectoderm and in the nervous system. In the eye imaginal discs, lacZ was expressed in photoreceptor cells beginning a few rows posterior to the morphogenetic furrow. The lacZ was also expressed in the wing disc. In the adult, lacZ was expressed in the retina and lamina. We cloned the sty gene by P-element tagging and found that it encodes a putative secreted protein containing a cysteine-rich region similar to the epidermal growth factor (EGF) repeat. On the basis of the loss of functional phenotype, the expression pattern and the predicted structure of its product, we propose that sty encodes a diffusible protein acting as a signal involved in lateral inhibition within the developing nervous system and also as a factor involved either directly or indirectly in axonal guidance and optic lobe development.     

Proteomics identified nuclear N-myc downstream-regulated gene 1 as a prognostic tissue biomarker candidate in renal cell carcinoma

The aim of this study was to identify proteins with aberrant expression in clear cell renal cell carcinoma (ccRCC), and elucidate their clinical utilities. The protein expression profiles of primary ccRCC tumor tissues and neighboring non-tumor tissues were obtained from 9 patients by two-dimensional difference gel electrophoresis and mass spectrometry. Comparative analysis of 3771 protein spots led to the identification of 73 proteins that were expressed at aberrant levels in tumor tissues compared with non-tumor tissues. Among these 73 proteins, we further focused on N-myc downstream-regulated gene 1 protein (NDRG1). NDRG1 expression is regulated by members of myc family as well as by p53, HIF1A, and SGK1. The biological and clinical significance of NDRG1 is controversial for various malignancies and no detailed studies on NDRG1 have been reported in ccRCC until our study. For the 82 newly enrolled ccRCC patients, immunohistochemical analysis revealed a significant association between nuclear NDRG1 and favorable prognosis (p < 0.05). Multivariate analysis demonstrated the role of NDRG1 as an independent factor of progression-free survival (p = 0.01). Subsequent in vitro gene suppression assay demonstrated that NDRG1 silencing significantly enhanced cell proliferation and invasion of RCC cells. The cytotoxic effects of NDRG1 up-regulation induced by an iron chelator were also confirmed. These findings suggest that nuclear NDRG1 has tumor suppressive effects, and the NDRG1 expression may have clinical values in ccRCC. Nuclear NDRG1 may provide additional insights on molecular backgrounds of ccRCC progression, and contribute to the development of novel therapeutic strategy.     

Multiple receptor proteins for sweet taste inDrosophila discriminated by papain treatment

Summary Treatment of the labellar chemosensory setae of the fruit fly,Drosophila melanogaster, with 0.1 % papain for 3 min induced a complete elimination of the taste nerve response to fructose (Fig. 1). Responses to other sugars examined were not affected (Table 1). Responses to 20 mM LiCl and 0.1 M NaCl also remained unchanged by the treatment. The experiment on the time-dependency of the papain treatment showed a clear difference in the proteasesensitivity between the response to fructose and to glucose and sucrose (Fig. 2). The treatment with 0.005% trypsin for 3 min produced the same results. The response to fructose which was eliminated with the papain treatment, was restored after 3 hrs. These findings reveal the presence of a specific receptor site for fructose and its protease-sensitive nature and suggest the involvement of multiple receptor proteins in the sugar receptor ofDrosophila.We thank Dr. A. Shiraishi for valuable suggestions for electrophysiological recordings. This work was supported in part by a Grant-in Aid for Scientific Research from the Ministry of Education, Science and Culture of Japan.     

Genetic dimorphism in the taste sensitivity to trehalose inDrosophila melanogaster

Summary A quantitative behavioral assay was developed for the measurement of taste responses to sugars inDrosophila. The amount of the intake of a sugar solution was measured colorimetrically after homogenization of flies which had consumed sugar solutions mixed with a food-dye. A two-choice method was utilized to determine the taste sensitivity to sugars. Two kinds of sugar solutions were marked with either blue or red food-dye and placed alternately in the wells of a micro test plate. Flies were allowed to choose between the two sugar solutions. By classifying and counting the coloured flies, the relative taste sensitivity could be determined. Employing these methods, a genetic dimorphism in the taste sensitivity to trehalose was found among some laboratory strains ofDrosophila melanogaster. No difference in the taste sensitivity to glucose, fructose and sucrose was found between the trehalose high-sensitivity (T-1) and the low-sensitivity (Oregon-R) strains. Trehalose concentration equivalent to 2 mmol/1 sucrose, in terms of stimulating activity, was 57 mmol/1 inOregon-R and was 10 mmol/1 inT-1. Genetic analysis showed that theTre gene, whose locus is closely linked tocx (13.6) on theX chromosome, is responsible for the difference in the taste sensitivity to trehalose.     

Isolation of Festuclavine and Three New Indole Alkaloids,Roquefortine A,B and C from the Cultures of Penicillium roqueforti

The structure and absolute configuration of a metabolite isolated from Aspergillus silvaticus was studied by an X-ray diffraction method. Crystals were orthorhombic, with space group P2₁2₁2₁, and a=9.894 (5), b = 6.803 (4), c=22.519 (6) Å, and Z=4. The structure was solved by a direct method and refined by a block-diagonal least-square method to R=0.07 for 1143 non-zero reflections. The metabolite was identified as naphthalic anhydride obtained from atrovenetin. The absolute configuration determined by the Bijvoet method is consistent with that shown chemically. The molecule has two intramolecular hydrogen bonds.