Climbing on the cage lid, a regular component of locomotor activity in the mouse.

The aim of the study was to obtain base values of climbing behaviour in mice maintained under standardized conditions in Makrolon-cages. Therefore three adult male mice each of the inbred strains BALB/cJ and C57BL/6J were kept separately and two C57BL/6J females as a group in Makrolon-cages type III. In addition, the same BALB/c mice were later kept in a cage with an eightfold floor area. Behavioural observations were carried out by video technique using a light-sensitive camera and a time-lapse recorder. Locomotor activity on the cage floor and climbing on the top of the cage were measured over a period of 48 h for each animal. The duration of locomotion on the ground ranged from 24-65 min/day, climbing between 49-122 (males) and 159 min/day (females) respectively. Climbing showed a more pronounced daily periodicity than locomotion, especially in the case of the BALB/cJ strain, where the average duration of climbing was about 28 min/h during the first hour after light off. In the mouse, climbing is obviously a regular component of activity, which deserves not only attention in the discussion concerning the needs of laboratory animals, but also in measurements of locomotor activity.     

Differential expression of two cadherins in Xenopus laevis

Using a cadherin fraction from Xenopus tissue culture cells as an immunogen, two monoclonal antibodies were obtained that allowed the characterization of two distinct cadherins in the Xenopus embryo. The two cadherins differ in molecular weight, in their time of appearance during development and in their spatial pattern of expression. One of the antigens was identified as E-cadherin. It appears in the embryonic ectoderm during gastrulation when epidermal differentiation commences and it disappears from the neural plate area upon neural induction. The second antigen could not be allocated to any of the known cadherin subtypes and was termed U-cadherin. It is present in the egg and becomes deposited in newly formed inner cell membranes during cleavage, the outer apical membranes of the embryo remaining devoid of the cadherin throughout development. U-cadherin is found on membranes of all cells up to the late neurula stages. A conspicuous polarized expression of the antigen on the membranes of individual inner cells suggests its participation in the segregation of cell layers and organ anlagen. These findings are discussed in the context of current hypotheses on the role of cadherins in establishing the spatial structure of the embryo.     

Co-localization of ecdysteroid receptors and c-fos-like protein in the brain of Manduca sexta larvae

Summary The presence of c-fos, a marker for cell activation, was investigated in cerebral neurons actively expressing ecdysteroid receptors during larval-pupal development in the tobacco hornworm, Manduca sexta. Colocalization was accomplished by ecdysteroid autoradiography using the tritiated high affinity 20-hydroxyecdysone agonist ponasterone A and immunocytochemistry with an antibody to a peptide sequence which is highly conserved in both human and murine c-fos. Immunoreactivity to a c-fos-like protein(s) was present in nuclei of many neurons of all the developmental stages examined. However, with the exception of the optic lobe, cells expressing nuclear ecdysteroid receptors were more immunoreactive than non-ecdysteroid-binding neurons. These data suggest that ecdysteroid-induced gene activation and translation may involve c-fos expression. Offprint requests to: H.-J. Bidmon     

Are there seasonal variations of trace element concentrations (Cd,Pb, Zn) in wood of Fagus trees in Germany?

Concentrations of Cd, Pb and Zn were determined in stem wood of beech trees (Fagus sylvatica L.) from 3 sites in northern Germany. Distinct radial distribution patterns of the elements were observed in the xylem. Concentrations of Cd and Pb increased from the youngest, outermost annual rings towards the center of the stem. With Zn intermediate concentrations were observed in the sapwood and higher levels at the center of the stem.Temporal and spatial stability of such distribution patterns in the trunks was investigated. Wood samples taken from the same individual tree in different months of the year were analysed. Marked seasonal variations of mineral concentrations were observed. Also the shape of the distribution patterns of the elements varied with the season. Such variations were larger than those observed with samples taken simultaneously from different sides of the trunk. Furthermore, Pb concentrations in the stem showed variations with height above ground.The results indicate, that radial distribution patterns of Cd, Pb and Zn in xylem rings of beech are not stable. Biomonitoring trace element pollution levels by analysis of beech wood is, thus, questionable. To obtain a reliable historical record of pollution from tree rings, the distribution patterns should be stable over a long period of time. This basic requirement of the dendroanalytical method does not hold for the examined beech. Still, with other tree species and under more favourable conditions the dendroanalytical biomonitoring method may prove valuable.Presented as symposium paper at the V International Congress of Ecology, Yokohama, August 23–30, 1990.     

Siberian species of the riihimakiensis-group in the genusChironomus (Diptera,Chironomidae). 1. Karyotypes and morphology

Karyotypes and morphology of three species with close relations toC. riihimakiensis Wülker 1971 are described from material of Tuva and Altai in Siberia. Only one of them could be reared to the male adult and was namedC. tuvanicus n.sp.     

A specific protein, p92, detected in flat revertants derived from NIH/3T3 transformed by human activated c-Ha-ras oncogene

Total proteins from a mouse embryo fibroblast cell line NIH/3T3, NIH/3T3 cells transformed by human activated c-Ha-ras (EJ-ras) oncogene (EJ-NIH/3T3), and the two flat revertant cell lines, R1 and R2, were analyzed by two-dimensional gel electrophoresis (IEF and NEPHGE). Several hundred polypeptides were resolved as seen by silver staining. Common alterations in four polypeptide spots were observed in the revertants when compared with NIH/3T3 and EJ-NIH/3T3 cells. In these alterations, a new polypeptide spot p92-5.7 (designated by molecular weight x 10(-3) and pI) was detected only in the revertants and not in NIH/3T3 and EJ-NIH/3T3 cells. Furthermore, the expression level of p92-5.7 seemed to be associated with the flat morphology and the reduced tumorigenicity of the revertants. Polypeptide p92-5.7 was also not detected in the total proteins extracted from BALB/3T3 cells, NIH Swiss mouse primary embryo fibroblasts, NRK (normal rat kidney) cells, and L6 (rat myoblast). Subcellular fractionation of total protein from R1 cells revealed that the p92-5.7 was present in the cytosol. Western blot analysis using an anti-gelsolin antibody demonstrated that the p92-5.7 might be a variant form of gelsolin which is thought to be an actin regulatory protein or a gelsolin-like polypeptide. These results may suggest that the expression of p92-5.7 detected only in the revertants is associated, at least in part, with the reversion. This may be the first demonstration of specific protein expression in the flat revertants.     

Activation of multiple signal transduction pathways by endothelin in cultured human vascular smooth muscle cells

Cellular responses to the vasoconstrictor peptide, endothelin, have been investigated in quiescent cultured human vascular smooth muscle cells (hVSMC). Endothelin caused intracellular alkalinization and activation of the protein synthetic enzyme S6-kinase, but such responses were not associated with any mitogenic effects of endothelin on hVSMC. In myo-[3H]inositol-prelabelled hVSMC endothelin elicited a rapid increase in inositol bis- and tris-phosphates and concomitant hydrolysis of polyphosphoinositol lipids. In [3H]arachidonate-prelabelled hVSMC endothelin promoted production of diacylglycerol, the early kinetics of which parallelled polyphosphoinositol lipid hydrolysis. Such phospholipase C activation by endothelin was sustained in hVSMC with accumulation of inositol polyphosphates being markedly protracted and the decay of diacylglycerol slow. Endothelin promoted extracellular release of [3H]arachidonate-labelled material from hVSMC which derived via deacylation of both phosphatidylinositol and phosphatidylcholine. This process was inhibited by phospholipase A2 and lipoxygenase inhibitors, but insensitive to phospholipase C and cyclooxygenase inhibitors. Endothelin-induced activation of phospholipase C and phospholipase A2 signal transduction pathways (EC50 approximately 5-8 nM for both) in hVSMC apparently proceed in an independent parallel manner rather than a sequential one.     

Gene expression during tuber development in potato plants

Potato tubers are modified stems that have differentiated into storage organs. Factors such as day-length, nitrogen supply, and levels of the phytohormones cytokinin and gibberellic acid, are known to control tuberization. Morphological changes during tuber initiation are accompanied by the accumulation of a characteristic set of proteins, thought to be involved in N-storage (i.e. patatin) or defense against microbial or insect attack (i.e. proteinase inhibitor II). Additionally, deposition of large amounts of starch occurs during tuber formation, which is paralleled by an increase in sucrose synthase and other enzymes involved in starch biosynthesis (i.e. ADP-glucose pyrophosphorylase, starch synthases, and branching enzyme). Potential controlling mechanisms for genes expressed during tuberization are discussed.