[Morphoecological Aspects of Preimaginal Stages of Development of the Parasitic Wasp Minotetrastichus frontalis (Nees, 1834) (Insecta, Hymenoptera, Eulophidae)].

This work is devoted to studying the morphology and biology of preimaginal stages of development of the parasitic wasp Minotetrastichus frontalis. The morphological characteristics of eggs and larvae, the features of interaction of the parasitic wasp with the host (Phyllonorycter issikii), and other biological characteristics were determined. For the species identification of the studied specimens of insects, we performed molecular genetic analysis using a nuclear 28S RNA gene fragment as a molecular marker. The result showed a strong genetic polymorphism of the populations of the studied species for the selected marker.

Mutagenesis testing using the LacZ reporter activity of the reparation gene mus209 in Drosophila melanogaster.

We studied a set of Drosophila melanogaster strains that could be potentially suitable for testing a variety of mutagenic factors. Their genomes contained insertions of the enhancer trap P{lacW} in which the activity of the LacZ reporter is under the control of the reparation genes' regulatory region. We demonstrated that the beta-galactosidase reporter, which is encoded by insertion of P{lacW} element in the gene mus209, is induced by irradiation in the cells of the salivary glands and wing imaginal discs. Despite the fact that the reporting coloration is not associated with the dose of radiation treatment, we found that the induction threshold of the reporter is different for these tissues. Thus, coloration in salivary glands is detectable after the dose of 200 rad and above, whereas the imaginal discs get colored with 500 rad and above. Thereby, multiple thresholds for induction of the reporter in the various tissues allow approximating the received dose.

Raman spectroscopy for DNA quantification in cell nucleus.

Here we demonstrate the feasibility of a novel approach to quantify DNA in cell nuclei. This approach is based on spectroscopy analysis of Raman light scattering, and avoids the problem of nonstoichiometric binding of dyes to DNA, as it directly measures the signal from DNA. Quantitative analysis of nuclear DNA contribution to Raman spectrum could be reliably performed using intensity of a phosphate mode at 1096 cm(-1) . When compared to the known DNA standards from cells of different animals, our results matched those values at error of 10%. We therefore suggest that this approach will be useful to expand the list of DNA standards, to properly adjust the duration of hydrolysis in Feulgen staining, to assay the applicability of fuchsines for DNA quantification, as well as to measure DNA content in cells with complex hydrolysis patterns, when Feulgen densitometry is inappropriate.

[hrs Gene and Borders of Compartments of Imaginal Wing Disc in Drosophila melanogaster].

Hepatocyte growth factor receptor tyrosine kinase substrate (Hrs) is an endosome protein involved in the sorting and transport of receptor tyrosine kinases and other proteins (which are absorbed by the cell during endocytosis) from early endosomes to lysosomes. Since receptor tyrosine kinases are important components of different cellular signaling systems, Hrs protein defects can result in the appearance of a number of developmental anomalies. In particular, it was demonstrated that ectopic Hrs expression in the wing kidney results in the disappearance of one or several rows of marginal wing setae in imago (which indicates hrs gene involvement in the development of the disc D/V border). We previously confirmed Hrs involvement in the developmental process of imaginal wing disc D/V border and demonstrated a change in the apterous gene expression pattern during ectopic hrs expression. The present report is devoted to the clarification and detailing of the Hrs effect on D/V, as well as on A/P borders of the imaginal wing disc compartments in Drosophila melanogaster.

[COMPARATIVE STUDY OF Hrs AND OTHER ENDOSOMAL MARKERS CELLULAR LOCALIZATION IN DROSOPHILA MELANOGASTER SPERMATOGENESIS BY GFP-CHIMERICAL PROTEIN APPROACH].

Acrosome is a special organelle in spermatozoids necessary for fertilizing oocyte and originates, according to various theories, either from Golgi apparatus, or from endosomes and lysosomes. One of the proteins, found at mammalian acrosome, is Hgs, a homologue of Drosophila melanogaster Hrs (Hepatocyte growth factor regulated tyrosine kinase substrate), a known marker of multivesicular bodies (MVBs). However, although Drosophila acrosome was extensively studied, it is yet unknown whether Hrs localizes at acrosome similar to Hgs and, more generally, whether the spectrum of acrosomal proteins in Drosophila is the same as in mammals. Hrs (hepatocyte growth factor regulated tyrosine kinase substrate) is the multidomain vesicular protein participating in the endosome-lysosome protein sorting. We demonstrated that two protein variants of the Drosophila Hrs are expressed in testes: a longer isoform B, and a shorter isoform A, which lacks VHS and FYVE domains that are necessary for anchoring Hrs in endosomes. We found that Hrs isoform B is concentrated at fusoma of spermatocytes in contrast to mammalian Hrs. This localization requires the C-terminus of the protein, starting from the aminoacid residue 383. In situ hybridization of hrs RNA probe showed that the gene is expressed early in spermatogenesis consistently with Hrs localization in early fusoma. Additionally, we demonstrated that Hrs is dispensable for cytokinesis. Finally, it was found that although Drosophila Hrs does not localize at acrosome, the other endosomal markers--Rab4, Rab7, and Rab11--are detected at the organelle.

A GFP trap study uncovers the functions of Gilgamesh protein kinase in Drosophila melanogaster spermatogenesis.

The function of the gene gilgamesh (89B9-12) encoding a casein kinase in Drosophila spermatogenesis was studied. The chimeric Gilgamesh-GFP protein in spermatocytes is cortically located. In the polar and apolar spermatocytes, it concentrates at the terminal ends of the fusome, the organelle that passes through the system of ring canals of the spermatocyte cyst. At the stage of spermatid elongation, the protein associates with the nucleus. A spot of the highest Gilgamesh-GFP concentration in the nucleus co-localizes with gamma-tubulin in the basal body. At later stages, Gilgamesh is localized to the individualization complex (IC), leaving the nuclei somewhat before the IC investment cones, as detected by actin binding. The sterile mutation due to the gilgamesh gene leads to the phenotype of scattered nuclei and altered structure of actin cones in the individualizing spermatid cyst. Ultrastructural evidence confirmed defective spermatid individualization due to the mutation. The phylogenetic origin of the protein, and the connection between vesicular trafficking and spermatid individualization, are discussed.

[Genetics of the cell cycle: adaptive modification of the cycB2g mutation expression in dividing cells of Drosophila melanogaster].

The effect of mutation CycB2g on mitosis in neural ganglia and imaginal disks was studied in third-instar larvae of Drosophila melanogaster. Chromosome condensation and segregation were shown to be impaired in dividing cells of mutant larvae. During the three-year period of maintenance of the mutation in heterozygote, frequencies of some defects decreased via cellular adaptive modification.