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PubMed1994

RFLP analysis of chromosomal fragments in genetic mosaic strains of Bombyx mori.

Mottled striped (pSm), a genetic mosaic strain for larval body marking of the silkworm, Bombyx mori, contains a small chromosomal fragment generated by breakage near the end of the 2nd chromosome. This fragment carries the striped marking (pS) gene and part of the chorion gene clusters. To determine the structural features of this fragmented chromosome, we studied the organization of the chorion genes on the fragment using restriction fragment length polymorphism (RFLP) analysis. Two of three classes of chorion gene family probes detected RFLPs in the two phenotypes, pSm (p/p/pS), and p (p/p), which had lost the fragment, segregated among the siblings of a mottled striped strain. Although hybridization patterns were basically identical between them, one or two additional bands were always observed associating with the mosaic phenotype (pSm). This suggests that the additional bands correspond to extra copies of chorion genes on the chromosomal fragment, which have a different structure from ones on the intrinsic 2nd chromosomes. Such heterogeneity of chorion genes may have been maintained since the beginning of mosaic induction, due to the absence of recombinational events between the two chromosomes. We are unable to detect any RFLPs by hybridization with the early class of chorion genes, implying that chromosomal breakage might have occurred between the two chorion clusters, Chl-2 and Ch3, which are located approximately 4 cM apart from each other. Based on RFLP analyses for two independent mosaic strains (788 and 872), we postulate a common chromosomal origin with independent breakpoints and construct structural models for the two kinds of chromosomal fragments.

Animals↗
PubMed1995

RNA metabolism in situ at the 93D heat shock locus in polytene nuclei of Drosophila melanogaster after various treatments.

Quantitative in situ hybridization to RNA on polytene chromosome spreads, using the 93D exon-, intron- and repeat-specific 35S-labeled antisense RNA probes, revealed treatment- (heat shock, benzamide, colchicine, heat shock followed by benzamide and heat shock in the presence of colchicine) specific differences in the metabolism (synthesis and/or accumulation at the puff site) of the various hsr-omega transcripts, namely hsr-omega-nuclear (omega-n), omega-pre-cytoplasmic (omega-pre-c) and omega-cytoplasmic (omega-c). While heat shock increased the levels of all the three transcripts at the 93D puff site in a coordinated manner, benzamide led to a significant increase in the levels of hsr-omega-n and pre-c; on the other hand, colchicine caused increased levels of the omega-n and omega-c RNA species at 93D. The results also suggested splicing of hsr-omega-pre-c RNA at the site of synthesis with the spliced-out 'free' intron (hsr-omega-fi) accumulating at the puff site. The rate of splicing and/or turnover of the hsr-omega-fi varied in a treatment-specific manner. Although a combined treatment to salivary glands with heat shock and benzamide or colchicine is known to inhibit puffing and [3H]uridine incorporation at 93D, the two treatments resulted in a treatment-specific increase in the in situ levels of different hsr-omega transcripts at the 93D site, suggesting a reduced turnover of specific transcripts from the site under these conditions. We suggest that the different 93D transcripts have roles in turnover and/or transport of RNA in nucleus as well as some role in cytoplasmic translation.

Animals↗
PubMed1997

A framework physical map of Drosophila virilis based on P1 clones: applications in genome evolution.

The analysis of patterns of genome evolution may help to evaluate the evolutionary forces that shape the composition and organization of the genome. Comparisons between the physical maps of divergent species can be used to identify conserved blocks of closely linked genes whose synteny is possibly under selective constraint. We have used in situ hybridization to determine the genomic position of 732 randomly selected clones from a bacteriophage P1 library of Drosophila virilis. The resulting map includes at least one clone in each of 69% of the subdivisions into which the D. virilis polytene chromosomes are divided. A subset of these clones was used to carry out a comparative physical analysis of chromosome 2 from D. virilis and from Drosophila montana. A number of discrepancies with the classical scenario of chromosome evolution were noted. The D. virilis P1 clones were also used to determine the physical relations between ten genes that are located in the X chromosome of Drosophila melanogaster between the markers crn (2F1) and omb (4C5-6). In this region, which is approximately 2 Mb in length, there have been at least six breakpoints since the divergence of the species, and six of the genes are found at widely scattered locations in the D. virilis X chromosome. However, a block of four functionally unrelated genes, including white, roughest, Notch, and dunce, seems to be conserved between the two species.

Animals↗
PubMed1999

Histochemical localization of reverse transcriptase in polytene chromosomes of chironomids.

The localization of a reverse transcriptase-related protein in salivary gland polytene chromosomes was investigated by immunohistochemistry in two species of Chironomus. The antibodies used were raised against a recombinant protein containing phylogenetically conserved motifs of reverse transcriptases and derived from an abundant non-LTR element previously identified in Chironomus. Immunoreactive protein was found in some telomeres, in a centromeric region, in a few interstitial bands and in Balbiani ring 3. The telomeric signal was probably dependent on transcription and increased dramatically when telomeric heat shock puffs were induced. A correlation with transcription was also seen in Balbiani ring 3, the immunobinding of which disappeared after inhibition of transcription with actinomycin D.

Animals↗
PubMed1994

Induction of chromosome aberrations and micronuclei in pulmonary alveolar macrophages of rats following inhalation of mosquito coil smoke.

The genotoxic potential of inhalation of mosquito coil (MC) smoke was evaluated by using metaphase chromosome aberration and micronucleus assays in pulmonary alveolar macrophages (PAMs) of rats following short-term as well as long-term whole body intermittent exposure. For short-term exposure, the animals were exposed for 15 min/h, 8 h/day to smoke collected for 1, 5 or 10 min, and they were killed 16 or 24 h after the final exposure. For long-term exposure, they were exposed for 15 min/h, 8 h/day, 7 days/week to smoke collected for 10 min and then they were killed 24 h after the final exposure. Each time before exposure, fresh smoke was collected by burning a mosquito coil. Pulmonary lavage was collected, and conventional flame-drying preparation was done for metaphase chromosome analysis and micronuclei (MN) were analyzed from smear preparations. Significantly higher frequencies of chromosome aberrations, including as well as excluding gaps, and micronucleated PAMs in smoke-exposed animals, compared to controls, indicated genotoxic capacity of MC smoke. The increases significantly correlated with the "concentration" of the gas. Mitotic indices also showed a significant and concentration-dependent increase. The frequencies of chromosome aberrations and MN following 7-day exposure were very similar to those for 1-day exposure. This was probably due to the transient nature of PAMs. A post-exposure gap of 24 h, compared to the 16-h gap, yielded a higher incidence of both mitoses and chromosome aberrations.

Animals↗
PubMed2006

Mapping Drosophila genomic aberration breakpoints with comparative genome hybridization on microarrays.

Chromosomal aberrations are genetic "reagents" that are commonly used in Drosophila research. Stocks containing chromosomes carrying large deletions of DNA (deficiency stocks, designated Df) as well as stocks carrying an extra copy of a chromosomal region (duplication stocks, designated Dp) are essential for a variety of genetic analyses. The extent of what is deleted or duplicated has typically been determined cytologically by salivary gland polytene chromosome squashes, which identify the edges of the aberration (so-called breakpoints) of each Df or Dp at low resolution. The margin of error for this technique can be quite high, however, because it is dependent on the quality of the squash and the experience of the scientist interpreting the data. Comparative genome hybridization on microarrays provides a precise molecular method to identify which regions of the genome are deleted or duplicated in these stocks by examining a change in chromosomal ploidy across the whole genome. Furthermore, this technique allows genetic data obtained with these strains to be placed in a molecular genomic context.

Animals↗
PubMed1996

The transmembrane tyrosine phosphatase DLAR controls motor axon guidance in Drosophila.

DLAR is a receptor-like, transmembrane protein-tyrosine phosphatase in Drosophila that is expressed almost exclusively by developing neurons. Analysis of Dlar loss-of-function mutations shows that DLAR plays a key role during motoneuron growth cone guidance. Segmental nerve b (SNb) motor axons normally exit the common motor pathway, enter the ventral target region, and then synapse on specific ventral muscles. In Dlar mutant embryos, SNb axons bypass their normal target region and instead continue to extend along the common pathway. SNd motor axons also make pathfinding errors, while SNa and SNc axons appear normal. Thus, DLAR controls the ability of certain motor axons to navigate specific choices points in the developing Drosophila nervous system.

Amino Acid Sequence↗
PubMed2000

dtrap-1 encodes a novel member of the heat shock super family of proteins and is expressed in derivatives of all three germ layers during Drosophila embryogenesis.

Heat shock proteins (Hsps) comprise a highly conserved superfamily of proteins that are required for stress tolerance in living cells. At physiological conditions, these proteins act as chaperones during protein folding and protein assembly processes (Parsell and Lindquist, 1993. Annu. Rev. Genet. 27, 427-496). Members of the subfamily of Hsp90 proteins were shown to be additionally involved in the presentation and structural modification of components of diverse cellular signal transduction pathways including steroid hormone reception and regulatory kinase activities (Pratt, 1993. J. Biol. Chem. 268, 21455-21458); Cutforth et al., 1994. Cell 77, 1027-1036; van der Straten et al., 1997. EMBO J. 16, 1961-1997; Hunter and Poon, 1997. Trends Cell Biol. 7, 157-161). We have identified a Drosophila gene, called dtrap-1, which encodes a Hsp of a novel subfamily that is related to the Hsp90 family of proteins. During oogenesis dtrap-1 is expressed in nurse cells and its transcripts accumulate in the oocyte. The maternal transcripts remain in the egg but rapidly degrade during early embryogenesis, except in the posterior pole region. Zygotic expression is initiated after the onset of gastrulation showing dynamic patterns of transcripts in the developing mid- and hindgut as well as a subset of mesoderm derivatives.

Animals↗
PubMed1995

Linkage analysis of the gene encoding precursor protein of diapause hormone and pheromone biosynthesis-activating neuropeptide in the silkmoth, Bombyx mori.

We have determined the map position of the gene encoding a common precursor protein for diapause hormone and pheromone biosynthesis-activating neuropeptide (the DH-PBAN gene, Dh) in the silkmoth, Bombyx mori. First we compared the structure of introns in the DH-PBAN gene by the polymerase chain reaction, and found that the Dh locus carried three alleles, DhA1, DhA2 and DhB. The DhA1 and DhA2 alleles contained a fourth intron consisting of 740 bp, whereas DhB had a longer fourth intron of 770 bp. DhA1 and DhA2 contained a fifth intron consisting of 940 bp, whereas the fifth intron in DhB was much longer and consisted of 1700 bp. DhA1 was distinguished from DhA2 by an RFLP in the fifth intron after digestion with Rsa I. Linkage analyses using these polymorphisms showed that Dh was linked to the bp gene on chromosome 11, and independent of markers on chromosomes 1, 2, 3, 4, 5, 6, 7 and 13. To determine the map position, we obtained F1 hybrids between the n501 strain (K DhA1) and the w30 strain (+K DhB), and backcrossed the F1 hybrid to females of the w30 strain. From the segregation of K and Dh in 864 individuals in the next generation, the recombination value was calculated as 25.5% between K and Dh. Similarly we obtained backcross progeny between the No. 744 strain (Bu DhA1) and the w30 strain (+Bu DhB), and calculated the recombination value between Bu and Dh as 30.4% from 487 progeny.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2006

An independent non-linear latitudinal cline for the sn-glycerol-3-phosphate (alpha- Gpdh ) polymorphism of Drosophila melanogaster from eastern Australia.

Latitudinal variation of the polymorphic sn-glycerol-3-phosphate (alpha-Gpdh) locus in Drosophila melanogaster has been characterized on several continents; however, apparent clinal patterns are potentially confounded by linkage with an inversion, close associations with other genetic markers that vary clinally, and a tandem alpha-Gpdh pseudogene. Here we compare clinal patterns in alpha-Gpdh with those of other linked markers by testing field flies from eastern Australian locations collected in two separate years. The alpha-Gpdh variation exhibited a consistent non-linear cline reflecting an increase in the alpha-GpdhF allele at extreme latitudes. This pattern was not influenced by the In(2L)t inversion wherein this locus is located, nor was it influenced by the presence of the alpha-Gpdh pseudogene, whose presence was ubiquitous and highly variable among populations. The alpha-Gpdh pattern was also independent of a cline in allozyme frequencies at the alcohol dehydrogenase (Adh) locus, and two length polymorphisms in the Adh gene. These results suggest clinal selection at the alpha-Gpdh locus that is partially or wholly unrelated to linear climatic gradients along the eastern coast of Australia.

Alleles↗
PubMed2002

In situ hybridisation analysis of the X-linked genes in the species of the virilis group of Drosophila.

When estimating the level of DNA sequence variation within and between populations or when planning QTL analysis, it is essential to know the location of the genes under study. In the present work, five X chromosomal genes, earlier localised in Drosophila virilis and D. littoralis, were mapped by in situ hybridisation on the larval polytene chromosomes of four other virilis group species, D. a. americana, D. flavomontana, D. lacicola and D. montana. Conjugation of X chromosomes of the most interesting species pairs was studied in interspecific hybrids. Three of the marker genes were used as RFLP markers to examine the occurrence of recombination in D. flavontontana and D. montana hybrid females. The gene arrangement of all species studied, appeared to be different at the proximal end of the X chromosome, which prevented normal conjugation along the most part of the X chromosome. The data illustrating the locations of five X chromosomal marker genes are presented for D. a. americana, D. flavomontana, D. lacicola and D. montana.

Animals↗
PubMed2000

Karyotypic analysis of the cotton boll weevil, Anthonomus grandis Boheman.

The diploid chromosome number of the cotton boll weevil, Anthonomus grandis Boheman, is 44. Both C- and N-banding techniques of mitotic cells demonstrated constitutive heterochromatin in the p arm of the eight largest chromosomes, the p arm of the X chromosome, and the centromeric region of autosomal groups A-D. Neither the y nor the group E autosomes appeared to contain constitutive heterochromatin. Supernumerary chromosomes were not found in the boll weevil. Restriction endonuclease banding of primary spermatocytes revealed a rod-shaped Xy tetrad in which the X and y were terminally associated. The p arm of the large, submetacentric X was C-band positive. While two of the autosomal tetrads were typically ring-shaped in primary spermatocytes, the remaining 19 autosomal tetrads were rod-shaped.

Animals↗