Alterations in the expression of the Atp7a gene in the early postnatal development of the mosaic mutant mice (Atp7amo-ms) – An animal model for Menkes disease

Copper is a trace element that is essential for the normal growth and development of all living organisms. In mammals, the ATP7A Cu-transporting ATPase is a key protein that is required for the maintenance of copper homeostasis. In both humans and mice, the ATP7A protein is coded by the X-linked ATP7A/Atp7a gene. Disturbances in copper metabolism caused by mutations in the ATP7A/Atp7a gene lead to severe metabolic syndromes Menkes disease in humans and the lethal mottled phenotype in mice. Mosaic is one of numerous mottled mutations and may serve as a model for a severe Menkes disease variant. In Menkes patients, mutations in the ATP7A gene often result in a decreased level of the normal ATP7A protein. The aim of this study was to analyse the expression of the Atp7a gene in mosaic mutants in early postnatal development, a critical period for starting copper supplementation therapy in both Menkes patients and mutant mice. Using real-time quantitative RT-PCR, we analysed the expression of the Atp7a gene in the brain, kidney and liver of newborn (P0.5) and suckling (P14) mice. Our results indicate that in mosaic P0.5 mutants, the Atp7a mRNA level is decreased in all analysed organs in comparison with wild-type animals. In two week-old mutants, a significant decrease was observed only in the kidney. In contrast, their hepatic level of Atp7a tended to be higher than in wild-type mice. We speculate that disturbance in the expression of the Atp7a gene and, consequently, change in the copper concentration of the organs, may contribute to the early fatal outcome of mosaic males.     

The presence and polymorphism of genes encoding serine protease autotransporters (spate) among the Escherichia coli isolated in the region of Gdansk from children with diarrhea

The group of 96 strains ofEscherichia coli isolated from children with diarrhea were investigated towards the presence and polymorphism of genes encoding autotransporters that belong to the group of proteins named SPATE (Serine Protease Autotransporters ofEnterobacteriaceae). Based on the results of restriction analysis of the products of PCR reaction 8 different types of genes encoding SPATE were detected. It was found that 39 strains contained one gene of SPATE, 15 strains contained two different genes and 3 different genes were detected in the case of 3 strains. The analysis of combination of presence of genes encoding SPATE let us divide the investigated group of strains into 17 different genotypes. The analysis of polymorphism of genes encoding SPATE seems to be very promising tool for exploring the genetic diversity among pathogenic E. coli.     

Exogenous melatonin expediently modifies proteome of maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.) embryo during seed germination

Melatonin (N-acetyl-5-methoxytryptamine) has a great potential for plant biostimulation. Its role in plant physiology is intensively explored, and its important function in plant stress defence frequently underlined. Melatonin is particularly effective when applied as an additional factor of seed priming. In the presented research, hydroconditioning was chosen experimentally as optimal for maize (Zea mays L.) seeds. The following seed variants were compared: controlled non-treated, hydroprimed with water and hydroprimed with melatonin solutions 50 and 500 μM. To identify modifications in proteome of maize seeds caused by the applied hydroconditioning techniques, protein extracts of germinated seed embryos (24 h, 25 °C) were separated by 2D-PAGE. Next, obtained maps of proteomes were compared (statistically and graphically) using PDQuest software, and characteristic spots of proteins were analysed qualitatively by mass-spectrometric techniques and identified in the Mascot protein databases. Research helped to identify hydropriming-associated proteins, and for the first time those which were expressed only in the presence of melatonin. Study confirmed that suitably selected pre-sowing treatment with melatonin, by embryo proteome modification, effectively prepares plants to adverse environmental conditions. In melatonin treated seeds during the initial state of embryos growth, even under optimal conditions, additional antioxidative, detoxifying, anti-stresses proteins were synthesized. Moreover, the supply of energy from seed storage substances was pretty intensified. The presented results partially explain how melatonin acts in plant stress defence, and why plants with higher melatonin content have exhibited increased capacity for stress tolerance.     

Recognition of interspecific familiar versus unfamiliar odours among bank voles and yellow-necked mice

Two experiments (laboratory and field-laboratory) were designed to determine whether individual bank volesClethrionomys glareolus (Schreber, 1780) and yellow--necked miceApodemus flavicollis (Melchior, 1834) could distinguish heterospecific odour cues from familiar and unfamiliar individuals. In the laboratory experiment each male bank vole was familiarized for 24 h with odour (cotton wool impregnated with urine and faeces) of male yellow-necked mice and yellow-necked mice were familiarized with odour of male bank voles. In the field-laboratory experiment the individual bank voles and yellow-necked mice captured at the same point were considered familiar and transfered to the laboratory. In laboratory, these individuals were tested in a box (for 5 h) affording them the choice between the odours of familiar and unfamiliar heterospecific males. Bank voles discriminated between familiar and unfamiliar yellow--necked mouse odours. Male yellow-necked mice seemed to have a similar ability to recognise odours of familiar bank voles. It is proposed that interactions between these two species occur not only on the species level, but also on the level of individual. This phenomenon (probably asymmetric) can play an important role in spatial orientation, and influence direct contacts between individuals of these species.     

Cytokinin-induced hypocotyl elongation in light-grown<Emphasis Type="Italic"> Arabidopsis</Emphasis> plants with inhibited ethylene action or indole-3-acetic acid transport

Cytokinins inhibit hypocotyl elongation in darkness but have no obvious effect on hypocotyl length in the light. However, we found that cytokinins do promote hypocotyl elongation in the light when ethylene action is blocked. A 50% increase in Arabidopsis thaliana (L.) Heynh. hypocotyl length was observed in response to N⁶-benzyladenine (BA) treatment in the presence of Ag⁺. The level of the ethylene precursor 1-aminocyclopropane-1-carboxylic acid was strongly increased, indicating that ethylene biosynthesis was up-regulated by treatment with cytokinin. Furthermore, the effects of cytokinins on hypocotyl elongation were also tested using a series of mutants in the cascade of the ethylene-signal pathway. In the ethylene-insensitive mutants etr1-3 and ein2-1, cytokinin treatment resulted in hypocotyl lengths comparable to those of wild-type seedlings treated with both Ag⁺ and BA. A similar phenotypical response to cytokinin was observed when auxin transport was blocked by -naphthylphthalamic acid (NPA). Applied cytokinin largely restored cell elongation in the basal and middle parts of the hypocotyls of NPA-treated seedlings and at the same time abolished the NPA-induced decrease in indole-3-acetic acid levels. Our data support the hypothesis that, in the light, cytokinins interact with the ethylene-signalling pathway and conditionally up-regulate ethylene and auxin synthesis.     

Mutation in the Sp1 motif of the bovine leptin gene affects its expression

Leptin is expressed mainly by adipocytes and plays a crucial role in the regulation of energy expenditure, food intake, and adiposity. Using PCR-heteroduplex analysis and sequencing, we investigated a C/G substitution in the promoter region of the bovine leptin gene. Application of the electrophoretic mobility shift assay showed that the C→G transversion decreased the leptin gene promoter binding capacity for nuclear proteins. With real-time PCR and Western blotting, we showed that the leptin expression level was higher in cattle with the CC than with the GG genotype.