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SILAC-based quantitative proteomic analysis of Drosophila gastrula stage embryos mutant for fibroblast growth factor signalling

Quantitative proteomic analyses in combination with genetics provide powerful tools in developmental cell signalling research. Drosophila melanogaster is one of the most widely used genetic models for studying development and disease. Here we combined quantitative proteomics with genetic selection to determine changes in the proteome upon depletion of Heartless (Htl) Fibroblast-Growth Factor (FGF) receptor signalling in Drosophila embryos at the gastrula stage. We present a robust, single generation SILAC (stable isotope labelling with amino acids in cell culture) protocol for labelling proteins in early embryos. For the selection of homozygously mutant embryos at the pre-gastrula stage, we developed an independent genetic marker. Our analyses detected quantitative changes in the global proteome of htl mutant embryos during gastrulation. We identified distinct classes of downregulated and upregulated proteins, and network analyses indicate functionally related groups of proteins in each class. In addition, we identified changes in the abundance of phosphopeptides. In summary, our quantitative proteomic analysis reveals global changes in metabolic, nucleoplasmic, cytoskeletal and transport proteins in htl mutant embryos

Absorption spectrum in the wings of the potassium second resonance doublet broadened by helium

We have measured the reduced absorption coefficients occurring in the wings of the potassium 4S-5P doublet lines at 404.414 nm and at 404.720 nm broadened by helium gas at pressures of several hundred Torr. At the experimental temperature of 900 K, we have detected a shoulder-like broadening feature on the blue wing of the doublet which is relatively flat between 401.8 nm and 402.8 nm and which drops off rapidly for shorter wavelengths, corresponding to absorption from the X doublet Sigma+ state to the C doublet Sigma+ state of the K-He quasimolecule. The accurate measurements of the line profiles in the present work will sharply constrain future calculations of potential energy surfaces and transition dipole moments correlating to the asymptotes He-K(5p), He-K(5s), and He-K(3d).Comment: 2 figure

Genetic structure of Tribolium castaneum populations in mills

We investigated the genetic diversity and differentiation among nine populations of Tribolium castaneum using eight polymorphic loci, including microsatellites and other insertion-deletion polymorphisms (=”indels”). Samples were collected in food processing/storage facilities located in Kansas, Nebraska, California, Louisiana, Florida and Puerto Rico. Standard population genetic analysis was applied, and an assignment test was used to assign individuals to their genetic population. All loci were polymorphic across populations, with the number of alleles per locus-population combination varying from three to fourteen. Among 72 locus-by-population combinations, 31 deviated significantly from Hardy-Weinberg equilibrium, which was associated with a deficiency in heterozygosity. Tribolium castaneum populations show some level of genetic structuring. Genetic differentiation between populations, using FST estimates, was significant, with FST varying from 0.018 to 0.149. AMOVA indicated that 8.32% of the variation in allele frequency resulted from comparisons among populations. Genetic distance was not significantly correlated with geographic distance. Correct assignment to the genetic population was possible in only 56% of all individuals. Together, these results revealed that geographically distinct populations of T. castaneum had low to moderate levels of genetic differentiation that was not correlated with geographic distance, and the genotypic profile of the individuals did not provide enough information for fingerprinting them with their source population. Keywords: Tribolium castaneum, Population genetics, Genetic structure, FST, Genetic fingerprintin