Cytoskeletons of retinal pigment epithelial cells: Interspecies differences of expression patterns indicate independence of cell function from the specific complement of cytoskeletal proteins

Summary In vertebrate tissue development a given cell differentiation pathway is usually associated with a pattern of expression of a specific set of cytoskeletal proteins, including different intermediate filament (IF) and junctional proteins, which is identical in diverse species. The retinal pigment epithelium (RPE) is a layer of polar cells that have very similar morphological features and practically identical functions in different vertebrate species. However, in biochemical and immunolocalization studies of the cytoskeletal proteins of these cells we have noted remarkable interspecies differences. While chicken RPE cells contain only IFs of the vimentin type and do not possess desmosomes and desmosomal proteins RPE cells of diverse amphibian (Rana ridibunda, Xenopus laevis) and mammalian (rat, guinea pig, rabbit, cow, human) species express cytokeratins 8 and 18 either as their sole IF proteins, or together with vimentin IFs as in guinea pig and a certain subpopulation of bovine RPE cells. Plakoglobin, a plaque protein common to desmosomes and the zonula adhaerens exists in RPE cells of all species, whereas desmoplakin and desmoglein have been identified only in RPE desmosomes of frogs and cows, including bovine RPE cell cultures in which cytokeratins have disappeared and vimentin IFs are the only IFs present. These challenging findings show that neither cytokeratin IFs nor desmosomes are necessary for the establishment and function of a polar epithelial cell layer and that the same basic cellular architecture can be achieved by different programs of expression of cytoskeletal proteins. The differences in the composition of the RPE cytoskeleton further indicate that, at least in this tissue, a specific program of expression of IF and desmosomal proteins is not related to the functions of the RPE cell, which are very similar in the various species.     

Zur Morphologie und Systematik desTrebouxia-Phycobionten im Thallus vonUsnea longissima (Lecanorales)

In the lichen genusUsnea different species ofTrebouxia-phycobionts as well as different haustorial types are known. The isolated and cultivated phycobiont ofUsnea longissima Ach. was studied by light- and electron microscopy and resembles in cytomorphological details the type ofTrebouxia impressa Ahmad. In addition to simple wall-to-wall contacts between the symbiotic components also intraparietal (=intrawall-)haustoria could be observed as the normal interaction type.

Frau Prof. Dr.Elisabeth Tschermak-Woess zu ihrem 70. Geburtstag gewidmet.     

Phenetic relationships inClerodendrum (Verbenaceae) and some phylogenetic considerations

Cluster analyses by different methods and a minimum spanning tree were used to study phenetic relationships in the genusChlerodendrum. 129 species were scored for 52 morphological characters corresponding to 119 character states. The phenetic results suggest a classification into 7 distinct groups, which may be grouped into two subgenera. This classification is supported by the iridoid distribution as well as by some phylogenetic considerations.     

Chemical signalling and monopolization of nectar resources by territorial Pachymelus limbatus (Hymenoptera Anthophoridae) male bees in Madagascar

Observations on territoriality and the associated odour-marking and foraging behaviours in male Pachymelus limbarus Saussure, a Malagasy endemic anthophorid bee, are reported. Data indicated a monandrous mating system where virgin females are attracted, probably by pheromonal cues, to male-patrolled territories associated with flowering, nectariferous Plectran- rhussensu lato species (Lamiaceae). One male observed in detail applied odour marks every 30–80 minutes on twigs around a food-resource area that contained c. 850 flowers. This marking behaviour always preceded territory residence. The male spent c. 80% of his time patrolling. Meanwhile, any intruding flying insect was inspected or chased, which caused conspecific males, as well as often females, to leave the site. The resident male fed frequently on flowers within his territory (c. 3,600 flower-visits on fine days), except immediately following odour-marking bouts. His flower-visiting was often released by interactions with intruding insects entering the territory. The observations provide the first example of male territoriality based on resource defence polygyny in the bee family Anthophoridae. The importance of resource distribution in the evolution of male territoriality in bees is discussed.     

The muskrat in biomedical research

Muskrats are aquatic rodents of moderate size which are plentiful throughout North America, but are not used commonly in the laboratory. Recently, we tested the feasibility of muskrats as experimental models and have found them to be acquired and cared for easily in conventional laboratory animal facilities. Some of their natural characteristics and diseases are described. The husbandry techniques that we used are presented and form a base for the preparation of future guidelines for the maintenance and use of feral animals in research. The results of some initial experiments testing the muskrat's utility for investigations of cardiorespiratory control mechanisms also are presented. Our data show that even anesthetized muskrats possess brisk and dramatic cardiovascular and respiratory reflexes. Our findings that their brains possess the cytoarchitectural and myeloarchitectural features comparable to other mammals, combined with their relative uniformity in size, has allowed us to locate specific neuronal loci stereotaxically. We suggest that the muskrat be considered as an experimental animal model for studies of the neural control of cardiorespiratory systems.     

Surface characteristics of Pseudomonas cepacia

Two major surface characteristics of Pseudomonas cepacia were examined in this study: reactivity with lectins and hydrophobicity. The results indicated that the surfaces of P. cepacia strains are heterogeneous with regard to the distribution of lectin receptors. Only lima bean agglutinin was found to strongly agglutinate all strains. The strains were also heterogeneous with regard to hydrophobicity as determined by adhesion to hexadecane. The degree of hydrophobicity, however, was not significantly altered when selected strains were mixed with either fibronectin or bovine serum albumin. In addition, the strains exhibited no apparent affinities for buccal epithelial cells and gave no evidence for an ability to haemagglutinate human red cells.     

A selection cartridge for rapid detection and analysis of spontaneous mutations including insertions of transposable elements in Enterobacteriaceae

Summary We present a method that allows positive selection and rapid analysis of mutations in Enterobacteriaceae. Mutations are detected in a 2630 bp selection cartridge inserted in two different bacterial mutlicopy plasmid vectors. Spontaneous mutations in Escherichia coli, Enterobacter cloacae and Citrobacter freundii include insertions, deletions and point mutations. The small size of the target sequence facilitates rapid analysis of DNA rearrangements by cleavage with restriction enzymes and of any type of mutation by DNA sequence analysis. While in E. coli insertions of the mobile elements IS1, IS2 and IS5 were readily found, insertions of putative new transposable elements were detected in Enterobacter cloacae. The selection cartridge can thus serve as a tool for studying the spectrum of insertion mutations in Enterobacteriaceae and probably other Gramnegative bacteria, and the dependency of this spectrum on physiological and environmental factors and the host's genetic background can be investigated.     

MPTP in mice: treatment, distribution and possible source of contamination

Mice were treated daily with [3H]MPTP (30 mg/kg, 1 uCi, s.c.) for 1, 3, and 10 days to determine the fate and localization of tritiated compounds. An untreated mouse was housed either in the same cage ("cage-mate control") or in an adjacent cage separated by mesh-wire ("near-neighbor control"). The radioactivity measured in blood, brain, liver, and remaining body of [3H]MPTP-treated mice was dependent on the total dose of the drug the animals received and did not vary with the type of tissue analyzed. Significant amounts of radioactivity were found in the tissues of the "cage-mate control" mice, but not of the "near-neighbor control" mice. The route of transmission appears to be through the urine, as the urine of [3H]MPTP-treated mice was highly radioactive after the drug injection. Only traces of radioactivity were found in their feces and there was no increase in the background radiation in the environment of the cages, indicating that the tritiated compounds were not exhaled. Proper disposal of urinary products of MPTP-treated animals is therefore necessary to reduce the risk of possible drug contamination in humans.     

Refined structure of human carbonic anhydrase II at 2.0 Å resolution

The structure of human erythrocytic carbonic anhydrase II has been refined by constrained and restrained structure–factor least-squares refinement at 2.0 Å resolution. The conventional crystallographic R value is 17.3%. Of 167 solvent molecules associated with the protein, four are buried and stabilize secondary structure elements. The zinc ion is ligated to three histidyl residues and one water molecule in a nearly tetrahedral geometry. In addition to the zinc-bound water, seven more water molecules are identified in the active site. Assuming that Glu-106 is deprotonated at pH 8.5, some of the hydrogen bond donor–acceptor relations in the active site can be assigned and are described here in detail. The Oγ1 atom of Thr-199 donates its proton to the Oε1 atom of Glu-106 and can function as a hydrogen bond acceptor only in additional hydrogen bonds.     

Weighting functions and parameter resolvability for oxygenation data subject to error in the independent variable.

Parameter resolvability and bias has been investigated for weighted nonlinear regression of data where the independent variable is subject to instrumental uncertainty. The specific example of cooperative oxygenation of hemoglobin was studied, where fractional saturation is determined spectrophotometrically and the oxygen activity is measured with a Clark polarographic electrode. For this system the instrumental uncertainty in the oxygen electrode was measured directly and the influence of the uncertainties on resolution of oxygen binding parameters was determined by Monte Carlo simulations. Four weighting functions were tested for their ability to minimize parameter uncertainty and bias: (1) uniform weighting; (2) "propagated weighting" whereby uncertainties in the independent variable are propagated into and added to uncertainties of the dependent variable; (3) Hill plot transform, or "end weighting"; and (4) maximum likelihood analysis, where deviations between fitting function and data are minimized as weighted horizontal and vertical distance vectors. Results of the Monte Carlo simulations favor the use of either uniform weighting, propagated weighting, or maximum likelihood weighting methods. Use of the Hill transform as a weighting function produced poorer parameter resolvability and inaccurate representation of the data in general. Bias error was negligible for all weighting functions.