Photomap of the polythenic chromosomes of Drosophila malerkotliana and in situ mapping of the Hsp83 locus

A photomap of the polythenic chromosomes of D. malerkotliana was constructed to facilitate the identification of the chromosomal arms and their sections, in order to allow the identification of the breaking points of inversions and the location of the bands marked by in situ hybridization. The photomap included the six chromosome arms corresponding to pairs I (chromosome X), II and III, excluding pair IV and Y chromosome, because they were not visualized in the material examined, probably due to their small size. Through the in situ hybridization technique with the use of a biotined probe of a fragment of the D. melanogaster gene, the Hsp83 locus of D. malerkotliana was mapped. The probe hybridized with a frequency of 70% in section 98 of the IIIR chromosome. This is the first mapped gene in the species, and indicates that possibly the IIIR arm of D. malerkotliana corresponds to the IIIL arm of D. melanogaster, where the Hsp83 gene was located.

High-Fat Diet Induces Oxidative Stress and MPK2 and HSP83 Gene Expression inDrosophila melanogaster

The consumption of a high-fat diet (HFD) causes alteration in normal metabolism affecting lifespan of flies; however molecular mechanism associated with this damage in flies is not well known. This study evaluates the effects of ingestion of a diet supplemented with 10% and 20% of coconut oil, which is rich in saturated fatty acids, on oxidative stress and cells stress signaling pathways. After exposure to the diet for seven days, cellular and mitochondrial viability, lipid peroxidation and antioxidant enzymes SOD and CAT activity, and mRNA expression of antioxidant enzymes HSP83 and MPK2 were analyzed. To confirm the damage effect of diet on flies, survival and lifespan were investigated. The results revealed that the HFD augmented the rate of lipid peroxidation and SOD and CAT activity and induced a higher expression of HSP83 and MPK2 mRNA. In parallel, levels of enzymes involved in lipid metabolism (ACSL1 and ACeCS1) were increased. Our data demonstrate that association among metabolic changes, oxidative stress, and protein signalization might be involved in shortening the lifespan of flies fed with a HFD.

Contrasting Histories of Three Gene Regions Associated With In(3L)Payne of Drosophila melanogaster

In the present report, we studied nucleotide variation in three gene regions of Drosophila melanogaster, spanning >5 kb and showing different degrees of association with the cosmopolitan inversion In(3-L)Payne. The analysis of sequence variation in the regions surrounding the breakpoints and the heat shock 83 (Hsp83) gene locus, located close to the distal breakpoint, revealed the absence of shared polymorphisms and the presence of a number of fixed differences between arrangements, indicating absence of genetic exchange. In contrast, for the esterase-6 gene region, located in the center of the inversion, we observed the presence of shared polymorphisms between arrangements suggesting genetic exchange. In the regions close to the breakpoints, the common St arrangement is 10 times more polymorphic than inverted chromosomes. We propose that the lack of recombination between arrangements in these regions coupled with genetic hitchhiking is the best explanation for the low heterozygosity observed in inverted lines. Using the data for the breakpoints, we estimate that this inversion polymorphism is around 0.36 million yr old. Although it is widely accepted that inversions are examples of balanced polymorphisms, none of the current neutrality tests including our Monte Carlo simulations showed significant departure from neutral expectations.

Transcriptional and translational study of the Drosophila subobscura hsp83 gene in normal and heat-shock conditions

In this paper we report a transcriptional and translational study of the hsp83 gene of Drosophila subobscura. This gene is located at the 18C region of the J chromosome. A monoclonal antibody raised against hsp83 was used for the immunological detection of this protein by Western blotting throughout the development of D. subobscura in control and heat-shock conditions. Our results indicate that puff 18C is not only heat-shock inducible but is also expressed during normal development and its level of expression increases at the end of the prepupa period. We detected hsp83 at normal temperatures, in particular developmental stages with the exception of the larval and the beginning of prepupa formation. Hsp83 was induced by heat shock in all stages studied with the exception of 1st instar larvae. We found that temperatures in excess of 26 °C were sufficient to induce hsp83. In addition, at temperatures from 26 to 34 °C, the increase in hsp83 synthesis was accompanied by increased transcription of the hsp83 gene; this positive correlation was not observed at 37 °C.Key words: Drosophila subobscura, hsp83, heat-shock genes, immunoblotting.

Heat shock and developmental regulation of the Drosophila melanogaster hsp83 gene.

In contrast to the hsp70 gene, whose expression is normally at a very low level and increases by more than 2 orders of magnitude during heat shock, the hsp83 gene in Drosophila melanogaster is expressed at high levels during normal development and increases only severalfold in response to heat shock. Developmental expression of the hsp83 gene consists of a high level of tissue-general, basal expression and a very high level of expression in ovaries. We identified regions upstream of the hsp83 gene that were required for its developmental and heat shock-induced expression by assaying beta-galactosidase activity and mRNA levels in transgenic animals containing a series of 5' deletion and insertion mutations of an hsp83-lacZ fusion gene. Deletion of sequences upstream of the overlapping array of a previously defined heat shock consensus (HSC) sequence eliminated both forms of developmental expression of the hsp83 gene. As a result, the hsp83 gene with this deletion mutation was regulated like that of the hsp70 gene. Moreover, an in vivo polymer competition assay revealed that the overlapping HSC sequences of the hsp83 gene and the nonoverlapping HSC sequences of the hsp70 gene had similar affinities for the factor required for heat induction of the two heat shock genes. We discuss the functional similarity of hsp70 and hsp83 heat shock regulation in terms of a revised view of the heat shock-regulatory sequence.

Heat shock and developmental regulation of the Drosophila melanogaster hsp83 gene

In contrast to the hsp70 gene, whose expression is normally at a very low level and increases by more than 2 orders of magnitude during heat shock, the hsp83 gene in Drosophila melanogaster is expressed at high levels during normal development and increases only severalfold in response to heat shock. Developmental expression of the hsp83 gene consists of a high level of tissue-general, basal expression and a very high level of expression in ovaries. We identified regions upstream of the hsp83 gene that were required for its developmental and heat shock-induced expression by assaying beta-galactosidase activity and mRNA levels in transgenic animals containing a series of 5' deletion and insertion mutations of an hsp83-lacZ fusion gene. Deletion of sequences upstream of the overlapping array of a previously defined heat shock consensus (HSC) sequence eliminated both forms of developmental expression of the hsp83 gene. As a result, the hsp83 gene with this deletion mutation was regulated like that of the hsp70 gene. Moreover, an in vivo polymer competition assay revealed that the overlapping HSC sequences of the hsp83 gene and the nonoverlapping HSC sequences of the hsp70 gene had similar affinities for the factor required for heat induction of the two heat shock genes. We discuss the functional similarity of hsp70 and hsp83 heat shock regulation in terms of a revised view of the heat shock-regulatory sequence.