Das Bewegungsverhalten der Coelomzellen vonPsammechinus miliaris bei der Wundheilung (Echinodermata)

Zusammenfassung 1. Das Bewegungsverhalten der Coelomzellen des EchinoidenPsammechinus miliaris Gmel. wird an kleinen von Stacheln, Füßchen und Pedicellarien befreiten Stellen der Skelettoberfläche in Periproctnähe untersucht.2. Aus dem freiliegenden Bälkchenwerk treten Coelomzellen aus, von denen nur die rotbraunen Amoebocyten auf dem hellen Kalkuntergrund im Auflicht (Ultropak;E. Leitz) sichtbar sind.3. Nach einigen Stunden ist die Wundfläche mit einer dicken rötlichen Zellmasse bedeckt, dem primären Wundverschluß. Außer den Coelomzellen enthält der Wundverschluß noch verschieden große Kalkpartikel, die vom Abschleifen der Versuchsstelle herrühren.4. Bei direkter Beobachtung ist weder an den rotbraunen Amoebocyten noch am Wundverschluß die geringste Bewegung zu erkennen.5. Zeittransformation (Zeitraffung [Z.R.] auf 1/240 und 1/480) zeigt die mit erheblicher Ortsverlagerung und Metabolie verbundene Bewegung der allein wahrehmbaren rotbraunen Amoebocyten auf der Wundfläche. Im scheinbar in Ruhe befindlichen Wundverschluß findet eine ständig hin- und herwogende Bewegung der Zell-Kalkamsse statt.6. Bereits nach 6 bis 7 Stunden ist das Operationsfeld völlig geglättet; die Lücken im Kalkskelett sind kaum noch zu erkennen infolge der neu eingebauten Kalkelemente. Die eigentlichen Heilungsvorgänge (Wiederherstellung der Feinstruktur des Kalkskelettes) erfolgen unterhalb des primären Wundverschlusses, sind also nicht der Beobachtung zugängig.7. Wird der primäre Wundverschluß im ganzen vorsichtig abgehoben und zerzupft, so kann das Bewegungsverhalten der entstandenen kleinen und großen Aggregate im Durchlicht unter Z.R. untersucht werden.8. Die im zerriebenen Explantat erhaltenen Coelomzell-Aggregate aller Größen weisen erhebliche Ortsveränderungen auf; oft breiten sie sich langsam aus unter Auswanderung zehlreicher randlich liegender Zellen. An den Außenzonen mittlerer und großer Aggregate werden plasmatische Netze sichtbar, die ständig ihre Gestalt und Maschenweite ändern.9. Diese Plasma-Netze bilden die Grundlage der Aggregate; ihre Kontraktionen und Dilatationen bewirken die Ortsverlagerungen der Aggregate (Netzbildende Coelomzellen;Kuhl 1937).10. Wenigzellige Aggregate vereinigen sich in den allermeisten Fällen, sobald ein gewisser Abstand überbrückt ist. Mittlere und große Aggregate gehen häufig eine Verbindung ein; meist werden vorher lockere Coelomzell-Brücken hergestellt. In manchen Fällen gleiten die Aggregate aneinander vorbei.11. Im polarisierten Licht lassen sich bei gekreuzten Nicols die ersten kleinen neugebildeten Kalkkristalle in den skelettbildenden Coelomzellen (= netzbildenden Zellen) nachweisen.12. Der Verschluß kleiner Kratzwunden im noch dünnen primären Wundverschluß (die Kratzer dringen bis zur abgeschliffenen Skelettoberfläche vor) wird unter Z.R. im Ultropak-Auflicht untersucht. Die Ergebnisse am explantierten Wundverschluß im Durchlicht führen zum Verständnis der Bewegungsvorgänge im ungewohnten Auflicht.13. Im zunächst verwirrenden Bewegungsgeschehen (die auffälligen rotbraunen Amoebocyten haben bei der Wundheilung keine Funktion) fallen die durch die Operationsnadel herausgerissenen kleinen Kalktrümmer auf; sie werden passiv durch die Plasmanetze bewegt, gelangen auch zufällig in die Kratzer und werden an den Rändern durch neugebildetes Kalkmaterial festgelegt oder eingebaut. Aus der Tiefe der Kratzer können lose Kalkpartikel heraufbefördert werden; auch diese werden häufig eingebaut. Die entstehenden Kalkbrücken werden schließlich untereinander verbunden und dadurch die kleine Wunde verschlossen. Das eingebaute Kalkmaterial zeigt auch unter starker Z.R. keine passive Bewegung mehr.14. In seltenen Fällen kann der Vorgang des schubweisen Aufsteigens der skelettbildenden Zellen aus dem Panzer und ihre Zusammenballung im Z.R.-Laufbild beobachtet werden.15. Ob der Einbau von herausgerissenem Kalkmaterial temporär oder dauernd ist, muß noch geprüft werden.

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The locomotory behaviour of coelom cells ofPsammechinus miliaris (Echinodermata) during wound-healing

In the sea urchinP. miliaris application of time lapse photography allows a study of the very slow movements of coelom cells during the healing process of small wounds on the surface of the calcareous skeleton near the periproct. For observation and time lapse photography LEITZ-Ultropak objectives were used (incident light). Ambulacral feet, spines and pedicellaria were removed, and the animal was fixed in three places in a ring of plexiglass by means of three little screws, which touched the equator of skeleton. The rate of time transformation was 1/240 to 1/480. The film reveals the behaviour of coelom cells, which move out the skeleton to the surface of the small experimental region. Within several hours the white polished surface is covered with hundreds of red-brown amoebocytes; only these are visible on the white lime-ground; they have no function in the healing process, which takes place below the surface of the primäre Wundverschluß and therefore cannot be observed. There are three main types of coelom cells: red-brown amoebocytes, körnchenführende Zellen (white amoebocytes) and leucocytes (netzbildende or skelettbildende Zellen); the flagellated cells may be neglected here. In order to be able to study the behavior of the three main types of coelom cells, the primäre Wundverschluß, i. e. the total cell-covering of the wound, is removed and torn into microscopic fragments. These are studied (time lapse) under normal optical conditions (transmitted light). The slides show many aggregates of different sizes, single cells and little calcareous concrements torn off the skeleton. The aggregates, even the big ones, exhibit slow locomotion and change their positions considerably. If the distance of two aggregates becomes small enough, they fuse. In these cases a loose cell bridge between the two aggregates is formed. Sometimes no union occurs, although the distance is very small. Even big aggregates suddenly show considerable contractions if spreading has preceded. All movements and place changing of cell-aggregates are caused by contractions and dilatations of the plasmatic network which forms the cellular basis. Little wounds in the newly built Wundverschluß scratched with a lancet, heal within several hours. Time lapse shows passive movements of small calcareous fragments, which by chance sometimes enter the small wounds, where they help and accelerate the closing of the injury. The fragments are fixed on the edge of the wound by newly produced lime. Skeleton building coelom cells (netzbildende Coelomzellen) come up in batches from the depth of the sea urchin's skeleton; each cell contains lime crystals.

     

Age-related difference in bioenergetics of lung and heart mitochondrial from rats exposed to ozone

Bioenergetics of isolated lung and heart mitochondria from adult and aged rats were examined in the presence of glutamate (NAD-linked substrate) or succinate + rotenone (FAD-linked substrate) following ozone exposure (3.0 ppm, 8 hr). In controls, several differences were observed between adults and aged in both organ preparations. Following exposure, all bioenergetic parameters were decreased significantly in lung preparations from both adult and aged rats. In heart mitochondria, the respiration rates in state 3 and in uncoupled state, and the ADP/O ratio were decreased significantly in both exposed age groups. The respiratory control ratio (RCR) was decreased significantly only in the aged exposed rats. These results suggest that acute exposure to high levels of ozone alters energy production in both lung and heart mitochondria of adult and aged rats.     

Poly- and Monoclonal Antibodies Against Recombinant Rat Brain Calbindin D-28K Were Produced to Map Its Selective Distribution in the Central Nervous System

Many processes in the CNS depend on calcium. The calcium signal is transduced into an intracellular response via Ca2(+)-binding proteins, including calbindin D-28K. In many laboratories, polyclonal antibodies against chicken intestinal calbindin D-28K have been used to study its localization in the brain (normal and degenerated) of various species, including humans, but some of these antisera cross-reacted with other proteins, including calretinin. We purified recombinant rat brain calbindin D-28K to raise antisera in rabbits and purified a recombinant rat-chicken calbindin D-28K hybrid protein to immunize mice for the generation of monoclonal antibodies. These antisera were highly specific for calbindin D-28K, as demonstrated by two-dimensional Western blotting analysis. Immunohistochemical analyses combined with in situ hybridization studies demonstrated that calbindin D-28K in the Purkinje cells of the cerebellum is independent of vitamin D. The antibodies described here will be important tools for studying the regulation of expression of calbindin D-28K and its biological function in the brain and in the PNS.     

Nicotine and cannabinoids as adjuncts to neuroleptics in the treatment of Tourette syndrome and other motor disorders

Animal studies suggest nicotine and cannabinoids may significantly enhance the therapeutic value of neuroleptics in motor disorders. This was recently demonstrated in humans by the finding that chewing nicotine gum produced striking relief from tics and other symptoms of Tourette syndrome not controlled by neuroleptic treatment alone. It appears that the use of nicotine or cannabinoids may greatly improve the clinical response to neuroleptics in motor disorders.     

Psychophysiological responsivity on a laboratory stress task: Methodological implications for a stress-muscle hyperactivity pain model

A stress-muscle hyperactivity-pain (SMP) model has been proposed to explain the etiology of certain musculoskeletal pain disorders. According to this model, subjects should show physiological arousal during periods of stress relative to periods of rest. In a test of this prediction, 31 subjects performed a reaction time task that has been used in previous laboratory studies. Multiple psychophysiological variables were monitored during initial and final 10-minute baselines, during performance on nine 2-minute reaction time tasks, and during 36-second rest intervals following each of the 2-minute tasks. Results showed small but statistically significant differences generally supporting the SMP model when masseter EMG was averaged over time periods of 12 seconds to 2 minutes, but not when masseter EMG was averaged over 10- to 18-minute blocks. These results demonstrated the importance of carefully selecting time intervals for analysis. Additional analyses that compared TMD with symptom-free subjects revealed small differences in EMG that supported the SMP model. Analyses of EMG over shorter time intervals also showed, however, that masseter EMG increased during the 36-second rest interval following performance on a 2-minute stress task; this result suggested that a modification of the SMP model may be necessary.This research was supported in part by Grant 2 S06RR08038-17 funded by the National Institutes of Health.     

Sheep Linkage Mapping: Nineteen Linkage Groups Derived from the Analysis of Paternal Half-Sib Families

Nineteen linkage groups containing a total of 52 markers have been identified in the sheep genome after typing large paternal half-sib families. The linkage groups range in size from 2 markers showing no recombination to a group containing 6 markers covering approximately 30 cM of the sheep genome. Thirteen of the groups have been assigned to a sheep chromosome. Three groups contain markers from bovine syntenic groups U2, U7 and U29, and one other group contains a marker that has been mapped only in humans. The remaining three groups are unassigned. This information will provide a useful foundation for a genetic linkage map of sheep.     

Planktivory in benthic nototheniid fish in McMurdo Sound,Antarctica

Synopsis Four species of nototheniid fish were sampled from below the sea ice near Cape Armitage, McMurdo Sound:Pagothenia borchgrevinki from just below the ice 1.5 km offshore,Trematomus bernacchii, Trematomus hansoni andTrematomus centronotus from off the bottom in about 20 m of water near the shore. Scale worms and isopods were conspicuous non-planktonic prey, and present in the three benthic fish species. The planktonic pteropod molluscLimacina helicina was numerically common in all four species of fish. The planktonic hyperiid amphipodHyperiella dilatata was also found in all fish species. WhereasP. borchgrevinki is planktivorous in accord with its pelagic habit, theTrematomus spp. clearly also feed on plankton from the water column.T. hansoni is particularly planktivorous, taking smaller copepods thanP. borchgrevinki.