Oxidized phospatidylcholine but not native phosphatidylcholine inhibits inducible nitric oxide synthase in RAW 264.7 macrophages

This study was designed to compare the effects of oxidized 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine (PAPC) and native PAPC on the inducible nitric oxide synthase (iNOS) in the macrophage cell line RAW 264.7. Macrophages stimulated by bacterial lipopolysaccharide (1 microg/ml) were incubated with increasing amounts of native or oxidized PAPC (oxPAPC, 10-20 microg/ml). Cells incubated with oxPAPC showed a dose-dependent inhibition of inducible nitric oxide synthesis, as well as reduced iNOS protein expression and mRNA levels. Additionally, chromatin immunoprecipitation assay revealed that oxPAPC reduced the interaction of the active NF-kappaB subunit p65 with the iNOS promoter region when compared to native PAPC.     

Orientation of birds in total darkness

Magnetic compass orientation of migratory birds is known to be light dependent, and radical-pair processes have been identified as the underlying mechanism. Here we report for the first time results of tests with European robins, Erithacus rubecula, in total darkness and, as a control, under 565 nm green light. Under green light, the robins oriented in their normal migratory direction, with southerly headings in autumn and northerly headings in spring. By contrast, in darkness they significantly preferred westerly directions in spring as well as autumn. This failure to show the normal seasonal change characterizes the orientation in total darkness as a "fixed direction" response. Tests in magnetic fields with the vertical or the horizontal component inverted showed that the preferred direction depended on the magnetic field but did not involve the avian inclination compass. A high-frequency field of 1.315 MHz did not affect the behavior, whereas local anesthesia of the upper beak resulted in disorientation. The behavior in darkness is thus fundamentally different from normal compass orientation and relies on another source of magnetic information: It does not involve the radical-pair mechanism but rather originates in the iron-containing receptors in the upper beak.     

Photosynthetic acclimation to light gradients in plant stands comes out of shade

Dense plant populations or canopies exhibit a strong enrichment in far-red wavelengths which leads to unequal excitation of the two photosystems. In the long-term plants acclimate to changes in light quality by adjusting photosystem stoichiometry and antenna structure, a mechanism called here long-term response (LTR). Using an artificial light system it is possible to mimic such naturally occurring gradients in light quality under controlled laboratory conditions. By this means we recently demonstrated that the LTR is crucial for plant fitness and survival of Arabidopsis. We could also demonstrate that the chlorophyll fluorescence parameter Fs/Fm is a genuine non-invasive functional indicator for acclimatory changes during the LTR. Here we give supportive data that the Fs/Fm can be also used to monitor the LTR in field experiments in which Arabidopsis plants were grown either under canopies or wavelength-neutral shade. Furthermore our data support the notion that acclimation responses to light quality and light quantity are separate mechanisms. Thus, the long-term response to light quality represents an important and distinct acclimation strategy for improving plant survival under changing light quality conditions.Key words: photosynthetic acclimation, redox control, long-term responses, light quality, Arabidopsis, plant fitness     

Pedigree and marker information requirements to monitor genetic variability

There are several measures available to describe the genetic variability of populations. The average inbreeding coefficient of a population based on pedigree information is a frequently chosen option. Due to the developments in molecular genetics it is also possible to calculate inbreeding coefficients based on genetic marker information. A simulation study was carried out involving ten sires and 50 dams. The animals were mated over a period of 20 discrete generations. The population size was kept constant. Different situations with regard to the level of polymorphism and initial allele frequencies and mating scheme (random mating, avoidance of full sib mating, avoidance of full sib and half sib mating) were considered. Pedigree inbreeding coefficients of the last generation using full pedigree or 10, 5 and 2 generations of the pedigree were calculated. Marker inbreeding coefficients based on different sets of microsatellite loci were also investigated. Under random mating, pedigree-inbreeding coefficients are clearly more closely related to true autozygosity (i.e., the actual proportion of loci with alleles identical by descent) than marker-inbreeding coefficients. If mating is not random, the demands on the quality and quantity of pedigree records increase. Greater attention must be paid to the correct parentage of the animals.     

Changes of the organotypic retinal organization in Borna virus-infected Lewis rats

Retinae of Borna disease virus (BDV)-infected Lewis rats were investigated with emphasis on long-term changes in organotypic tissue organization and glia-neuron relationship. Virus inoculation was attained via intracerebral BDV injection. Following survival times ranging between two and eight months, the retinal thickness was reduced up to one third of that of controls. Photoreceptor segments were completely extinguished and the number of neurons was dramatically reduced. The typical laminar organization of the retina was largely dissolved. Electron microscopy revealed severe spongy degeneration. Large numbers of activated microglia and macrophages were found, both cell types performing very active phagocytosis. The microglial cells expressed an extraordinary phenotype as characterized by large numbers of processes, with some of them penetrating the endfeet of Müller cells and others establishing highly complex interdigitations with vacuolized swellings and endings of neuronal processes. Müller cells were not reduced in number but displayed clear indications of gliosis such as alterations in the immunoreactivity for filament proteins and glutamine synthetase, significantly thickened stem processes, and an altered pattern of K⁺ currents in patch-clamp recordings. These findings demonstrate for the first time long-term neuron-glia interactions in the retina of BDV-infected rats. Moreover, the data contribute to our knowledge on structural and functional alterations accompanying persisting virus infection in the central nervous system.     

De Novo Mutation and Rapid Protein (Co-)evolution during Meiotic Adaptation in Arabidopsis arenosa

A sudden shift in environment or cellular context necessitates rapid adaptation. A dramatic example is genome duplication, which leads to polyploidy. In such situations, the waiting time for new mutations might be prohibitive; theoretical and empirical studies suggest that rapid adaptation will largely rely on standing variation already present in source populations. Here, we investigate the evolution of meiosis proteins in Arabidopsis arenosa, some of which were previously implicated in adaptation to polyploidy, and in a diploid, habitat. A striking and unexplained feature of prior results was the large number of amino acid changes in multiple interacting proteins, especially in the relatively young tetraploid. Here, we investigate whether selection on meiosis genes is found in other lineages, how the polyploid may have accumulated so many differences, and whether derived variants were selected from standing variation. We use a range-wide sample of 145 resequenced genomes of diploid and tetraploid A. arenosa, with new genome assemblies. We confirmed signals of positive selection in the polyploid and diploid lineages they were previously reported in and find additional meiosis genes with evidence of selection. We show that the polyploid lineage stands out both qualitatively and quantitatively. Compared with diploids, meiosis proteins in the polyploid have more amino acid changes and a higher proportion affecting more strongly conserved sites. We find evidence that in tetraploids, positive selection may have commonly acted on de novo mutations. Several tests provide hints that coevolution, and in some cases, multinucleotide mutations, might contribute to rapid accumulation of changes in meiotic proteins.     

Das Orientierungssystem der V?gel IV. Evolution

Zusammenfassung Ein erster Versuch von Bellrose, die Evolution des Orientierungssystems der Vögel zu beschreiben, ging von der Annahme aus, Kompaßorientierung und die Fähigkeit zur Navigation habe sich im Zusammenhang mit dem Vogelzug entwickelt. Kompaßmechanismen sowie die Mosaik- und die Navigationskarte spielen jedoch bereits bei der Orientierung im Heimbereich entscheidende Rollen, müssen sich also dort entwickelt haben unter dem Selektionsdruck, die täglichen Flugwege zu optimieren, vielleicht schon bei den Vorfahren der Vögel.Magnetkompaßorientierung erscheint als der einfachste Orientierungsmechanismus und müßte deshalb an den ältesten Orientierungsstrategien beteiligt gewesen sein. Ein Magnetkompaß ist bei Wirbeltieren weit verbreitet, doch gibt es Hinweise auf unterschiedliche Funktionsprinzipien. Es ist deshalb offen, ob die Vögel ihn von ihren Vorfahren übernommen oder eigenständig entwickelt haben. Das gleiche gilt für den Sonnenkompaß. Die entscheidende Rolle des Magnetkompaß bei der ontogenetischen Entwicklung des Sonnenkompaß läßt eine ähnliche Beziehung bei der phylogenetischen Entwicklung vermuten.Über kurze Entfernungen kann man sich Orientierung durch Wegumkehr allein mit Kompaßmechanismen vorstellen, wobei Umwege integriert werden müssen. Bei dieser Strategie akkumulieren sich jedoch die Fehler; die bei größeren Entfernungen resultierende Ungenauigkeit erzeugte einen Selektionsdruck, der das Benutzen von Ortsinformation begünstigte. Dies führte zur Entstehung der Mosaikkarte, die auf Kompaßorientierung und Landmarken beruht. Sie ist heute als eigenständiger Mechanismus anzusehen, der nach angeborenen Regeln aufgebaut wird. Die Navigationskarte entsteht, indem die gleichen Regeln auf Faktoren mit Gradienten-Charakter angewandt werden; sie hat sich offenbar aus der Mosaikkarte entwickelt. Ob sie eine Sonderentwicklung der Vögel infolge ihrer Flugfähigkeit ist, muß offen bleiben. Da die Vögel die Grundelemente ihres Orientierungssystems wahrscheinlich von ihren Vorfahren übernommen haben, würden wir erwarten, daß diese Mechanismen bei allen Vögel gleich sind bzw. nach den gleichen Regeln erstellt werden.Vorstufen des Vogelzugs waren zunächst ungerichtete Flüge auf der Suche nach günstigeren Bedingungen; in diesem Stadium reichten die vorhandenen Navigationsmechanismen zur Orientierung zwischen den verschiedenen Gebieten aus. Als aus diesen ersten Ortsbewegungen ein regelmäßiger Zug zwischen zwei Regionen wurde, begann sich das Zugprogramm zu entwickeln, wobei sich zunächst eine spontane Richtungstendenz herausbildete. Der Magnetkompaß konnte als erstes Referenzsystem für diese Zugrichtung dienen. Später erhielt die Himmelsrotation ihre entscheidende Bedeutung, wobei die Vögel die Referenzrichtung Süd zunächst aus dem Polarisationsmuster am Tage ableiteten. Im Laufe der Zeit entstanden die differenzierten Zugprogramme mit Richtungsfolgen, steuernden Zeitprogrammen und Triggermechanismen. Die Zugrichtung und Länge der Zugstrecke unterliegen auch weiterhin einer ständigen Selektion, die für optimale Anpassung an die jeweiligen Umweltbedingungen sorgt. Der Übergang vom Tag- zum Nachtzug bereitete keine Probleme, denn die Vögel mußten zunächst keine neuen Orientierungsmechanismen entwickeln, da sich der Magnetkompaß zu jeder Tageszeit einsetzen läßt. Später entstand der Sternkompaß, der in seinen Funktionseigenschaften hervorragend auf die Bedürfnisse von Zugvögeln angepaßt ist und als eigenständige Entwicklung der Nachtzieher angesehen werden muß. Dazu erwarben die Nachtzieher die Fähigkeit, die Information der Himmelsrotation aus der Bewegung der Sterne abzuleiten und direkt auf den Sternkompaß zu übertragen. Da das Zugverhalten bei Vögeln mehrfach unabhängig voneinander entstanden ist, muß man Entsprechendes auch von den Mechanismen der Zugorientierung annehmen. Das bedeutet, daß sich die betreffenden Mechanismen bei den verschiedenen Arten unterschiedlich entwickelt haben könnten, doch ist mit konvergenten Entwicklungen zu rechnen.

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The orientation system of birds — IV. Evolution

Summary In a first attempt to explain the evolution of the avian navigational system, Bellrose suggested that compass mechanisms and the ability for true navigation had developed in connection with migration across increasing distances. Yet birds use compasses, the mosaic and the navigational maps even close to home and for homing. This means that those mechanisms must have developed for orientation within the home range, with the necessity to optimize the everyday flights acting as selective pressure. In view of this, any attempt to reconstruct the evolution of the avian navigational system must start out with the non-flying ancestors of birds.Considering the requirements of orientation by landmarks and by using a compass, compass orientation with the help of the magnetic field appears to be the simplest mechanism; consequently, it must be assumed to belong to the most ancient orientation strategies. The magnetic compass is wide-spread among animals, but it appears to function according to different principles among the various groups of vertebrates so that it is unclear whether birds inherited their magnetic compass from their reptilian ancestors or developed a mechanism of their own. The same is true for the sun compass. The crucial role of the magnetic compass in the ontogenetic development of the sun compass might indicate a similar relationship for the phylogenetic development.Over short distances within the home range, orientation based solely on compass orientation appears possible, using the strategy of route reversal, with non-straight routes being integrated. Since this strategy accumulates errors, it becomes inaccurate over longer distances, thus causing selective pressure to use local site-specific information. This leads to the formation of the mosaic map, a mechanism that includes landmarks as well as compass orientation. Today, the mosaic map of landmarks is a mechanism by itself, established according to innate learning principles that associate information on path integration with site-specific information, thus forming a directionally oriented mental representation of the distribution of landmarks. The navigational map is formed by applying the same principles to factors of the nature of gradients; it thus appears to have developed from the mosaic map. Whether or not it is a special development of birds associated with their flying ability is unclear. Because the birds probably inherited the basic mechanisms of orientation from their ancestors, one would expect these mechanisms to be similar in all birds. For the mechanisms involving learned components, this means that they are established following common rules. Birds improved those mechanisms and adapted them to their specific needs.Migration is assumed to have begun with non-directed search movements for regions offering better conditions. At this stage, the already existing mechanisms of homing were sufficient for navigation between the various areas. When these first movements turned into regular migration between two regions, the migratory program began to evolve, starting out with spontaneous tendencies in a preferred direction. The magnetic compass may have served as first reference system for the migratory direction; later, celestial rotation, indicated by the changing pattern of polarized light during the day, obtained its important role in indicating the reference direction geographic South. In the course of time, sophisticated migration programs with changes in direction, controlling time programs, responses to trigger mechanisms etc. developed. The migratory direction and distance, i.e. the amount of migratory activity, continue to be subject to selective pressure so that birds can respond to the environmental conditions in an optimal way. The transition from daytime migration to night migration did not require new mechanisms, as the magnetic compass can be used at any time of the day. Later, however, the star compass evolved, which is to be considered a special development of night-migrating birds, with its way of functioning well adapted to the specific needs of migrants. Birds also developed the ability to derive information on celestial rotation from the rotating stars at night and to transfer this information directly to the star compass. Since migratory habits evolved many times independently among birds, the same has to be assumed for the specific mechanisms of migratory orientation. This means that they need not necessarily be identical in all bird migrants. We are to expect convergent developments, however, leading to mechanisms of the most suitable type.

     

Avian magnetic compass can be tuned to anomalously low magnetic intensities

The avian magnetic compass works in a fairly narrow functional window around the intensity of the local geomagnetic field, but adjusts to intensities outside this range when birds experience these new intensities for a certain time. In the past, the geomagnetic field has often been much weaker than at present. To find out whether birds can obtain directional information from a weak magnetic field, we studied spontaneous orientation preferences of migratory robins in a 4 µT field (i.e. a field of less than 10 per cent of the local intensity of 47 µT). Birds can adjust to this low intensity: they turned out to be disoriented under 4 µT after a pre-exposure time of 8 h to 4 µT, but were able to orient in this field after a total exposure time of 17 h. This demonstrates a considerable plasticity of the avian magnetic compass. Orientation in the 4 µT field was not affected by local anaesthesia of the upper beak, but was disrupted by a radiofrequency magnetic field of 1.315 MHz, 480 nT, suggesting that a radical-pair mechanism still provides the directional information in the low magnetic field. This is in agreement with the idea that the avian magnetic compass may have developed already in the Mesozoic in the common ancestor of modern birds.     

Effects of upstream deletions on light- and oxygen-regulated bacterio-opsin gene expression in Halobacterium halobium

The bacterio-opsin gene (bop) of Halobacterium halobium is located within a cluster with three other genes. Growth conditions of high light intensity and low oxygen tension induce bop gene cluster expression. To identify putative regulatory factor binding sites upstream of the bop gene, we have compared sequences upstream of the bop gene with the corresponding sequences from two other genes in the bop gene cluster. Conserved sequence motifs were observed which may mediate the effect of high light intensity and/or low oxygen tension on bop gene expression. Based on these motifs, a set of mutants was constructed which contained deletions upstream of the bop gene. These constructs were tested in a host strain where bop gene expression is independent of oxygen regulation and in another strain where it is regulated by oxygen and light. The minimal upstream sequence required for both light- and oxygen-regulated bop gene expression was determined to be 54 bp.     

Aus der Geschichte der Orientierungsforschung

Zusammenfassung Die Arbeit schildert die Entwicklungen auf dem Gebiet der Orientierungsforschung und den Wandel der Ansichten im Laufe der Zeit. Am Anfang stand Mitte des letztenJh. v. Middendorffs Vermutung, Zugvögel würden sich nach einem Magnetkompaß orientieren.Viguier diskutierte die Möglichkeit, verfrachtete Vögel würden anhand des Magnetfelds heimfinden, währendExner undReynaud kinästhetische Orientierungsmechanismen vermuteten. Erste Versuche, diese Hypothesen experimnentell zu bestätigen, blieben erfolglos.Anfang dieses Jh. versuchte man durch Verfrachtungsversuche festzustellen, unter welchen Umständen und über welche Entfernungen sich Vögel orientieren können. Seevögel zeigten sehr gute Heimkehrleistungen auch aus unbekanntem Gelände; die Versuche mit Brieftauben führten dagegen zu dem Schluß, Vögel könnten sich nur anhand bekannter Landmarken orientieren. In den 30er Jahren machten die systematischen Verfrachtungsversuche vonRüppell undWojtusiak jedoch deutlich, daß auch Stare und Schwalben von unbekannten Orten über große Entfernungen mit beachtlicher Geschwindigkeit heimkehren konnten. Die gleichzeitig begonnenen Versuche der Vogelwarte Rossitten und Helgoland zur Analyse des Vogelzugs ergaben, daß das Zugverhalten von den Bedingungen, unter denen die Vögel lebten, und bei sozialen Arten auch von Artgenossen mitbestimmt wird. Wildvögel zeigten bemerkenswerte Orientierungsleistungen, doch deren Grundlage blieb weiterhin offen.In den 40er Jahren führten die Untersuchungen vonHeinroth &Heinroth an Brieftauben abermals zu der Vermutung, die Heimkehr nach Verfrachtung beruhe auf mehr oder weniger systematischem Suchen, bis die Vögel auf bekanntes Gelände treffen; dann würden sie sich nach vertrauten Landmarken orientieren. Diese Ansicht fand weite Verbreitung;Yeagleys Hypothese einer Orientierung nach Magnetfeldparametern und der Coriolis-Kraft konnte sich nicht durchsetzen.Methodische Neuerungen leiteten Anfang der 50er Jahre die systematische experimentelle Analyse der Vögel ein.Kramer beschrieb den Sonnenkompaß und stelle mit dem Karte-Kompaß-Prinzip eine erste umfassende Theorie der Orientierung vor, die davon ausgeht, daß bei jedem Orientierungsvorgang ein externes Referenzsystem — ein Kompaß — beteiligt ist. Die Untersuchungen der Folgezeit führten zu einem recht guten Verständnis der Zugorientierung. Das Heimfinden nach Verfrachtung ist dagegen immer noch ein offenes Problem; keine der derzeit diskutierten Hypothesen kann alle bekannten Phänomene erklären. Es mehren sich jedoch die Hinweise, daß das Navigationssystem sehr variabel und in hohem Maße redundant ist, so daß mit einer einfachen Antwort gar nicht zu rechnen ist.

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On the history of orientation research

Summary This paper describes the developments and changes of ideas in the field of bird orientation research, which began in the middle of last century when v.Middendorff first suggested magnetic compass orientation in migrating birds. Still in the 19th century, two different hypotheses were proposed to explain homing.Viguier suggested that the birds made use of the spatial distribution of magnetic parameters, whereasExner andReynaud assumed that the birds were able to trace the route of the outward journey kinaesthetically. Attempts to obtain experimental support for either theory were unsuccessful.The first three decades of this century were characterized by displacement experiments designed to learn under what conditions and over what distances birds were able to return. The results obtained with seabirds indicated much more sophisticated navigational abilities than the experiments with homing pigeons, which led authors to believe that birds oriented by following familiar landmarks and could only return from unknown sites if they by chance reached a familiar region. This view became widely accepted when it was presented in the review articles ofWarner andStresemann.In the 1930s, however, systematic displacement experiments with wild birds changed the general views on orientation completely.Rüppell andWojtusiak showed that Starlings and Swallows could return from unknown sites over large distances with considerable speed. At the same time, the Vogelwarten (bird banding stations) Rossitten and Helgoland initiated programs to obtain more information on the nature of migratory orientation. The results indicated that migratory behavior can be modified by the conditions under which the birds live and, in social species, by conspecifics.Drost's experiment with Sparrowhawks gave the first indications that young birds and adult birds use different orientation mechanisms. Remarkable navigational abilities were demonstrated in wild birds, yet the mechanisms they used, were still unknown.The ideas on orientation changed again in the 1940s, whenHeinroth &Heinroth reported the resuls of their study on the orientation of homing pigeons. They suggested that pigeons, when released in an unfamiliar area, searched until they meet familiar terrain and then oriented by landmarks known from previous flights.Grifffin, whose attempts to interfer with homing by depriving displaced birds of magnetic and kinaesthetic information had failed, held similar views. A new global hypothesis byYeagley, suggesting that homing pigeons made use of a grid map formed by isolines of Coriolis force and vertical intensity of the magnetic field, was not accepted.In 1950, methodological innovations byKramer andMatthews brought new impetus to orientation research: It became possible to measure the directional tendencies of caged migrants, which allowed a systematic analysis of migratory orientation in the laboratory. At the same time, the vanishing bearings of displaced birds were found to be closely related to the home direction, which opened up new possibilities in pigeon homing.Kramer, who had described the sun compass as the first known orientation mechanism in birds, also contributed substantially to the theoretical framework of orientation by his map and compass-concept which states that an external reference system — a compass — is always involved in determining the home direction.The following period of experimental analysis began with a controversis on the role of the sun. Experimental evidence did not support sun navigation, but rather indicated that the sun was used as a compass. The analysis of the birds' orientation system during the last 30 years led to a fairly good understanding of migratory orientation. The processes of homing, however, are still not well understood. Many of the old concepts and new ones like olfactory orientation have been discussed, without any one hypothesis being able to explain all known phenomena. Recent findings suggest that there may not be one simple answer to the question of how birds navigate, as the birds' navigational system turns out to be redundant and highly variable.