Isolation and visualisation of alkali stable protein/DNA complexes

Distinct polypeptides, 54,000–68,000 daltons in size, are alkali-stably bound to eukaryotic DNA. DNA fragments several hundred base pairs in length associated with these polypeptides are preferentially retained on glass fibre filters from solutions containing 1 M sodium chloride. About 50 percent of the protein/DNA complexes present in total DNA are retained on filters together with about 2 percent of the DNA. This preferential binding is demonstrated (a) by the ratio of ³H and ³⁵S radioactivity retained on filters after filtration of DNA from [³H]thymidine and L-[³⁵S]methionine labelled cells, (b) radioiodination of the material retained on filters and passing filters respectively and (c) by electron microscopical visualisation of the polypeptide component in the complexes after chemical modification with dinitrofluorobenzene (DNFB) followed by incubation with dinitrophenyl (DNP) specific antibodies.     

Activation of human T lymphocytes by 12-O-tetradecanoylphorbol-13-acetate: role of accessory cells and interaction with lectins and allogeneic cells

Stimulatory effects of the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) on human T lymphocytes have been investigated. TPA was found to stimulate highly purified T cells (obtained by a three-step isolation procedure involving plastic adherence, nylon wool passage and Ig-anti-Ig column passage) in the absence of accessory cells (stimulation index of 5 to 10), whereas phytohemaglutinin (PHA) and concanavalin A (Con A) did not. This response was, however, increased by the addition of autologous adherent cells. Addition of TPA, but not adherent cells, induced T-cell proliferation in response to the nonmitogenic lectin, wheat germ agglutinin (WGA), while both adherent cells and TPA restored T-cell proliferation to mitogenic lectins such as PHA and Con A. Furthermore, TPA greatly increased the mixed-lymphocyte response of purified T cells to otherwise nonstimulating allogeneic cells such as T lymphocytes or tumor cells from some patients with chronic lymphocytic leukemia. These results suggest that TPA can directly act on human T cells to render them reactive to a variety of stimuli.     

New species of Dicliptera (Acanthaceae) from the Guianas

Two new species,Dicliptera knappii from Guyana andD. granvillei from French Guiana, are described, illustrated, and compared with their closest relatives.     

Paarungssystem und Fortpflanzungsstrategie der Beutelmeise (Remiz p. pendulinus)

Zusammenfassung Paarungssystem und Fortpflanzungsstrategie der Beutelmeise wurde in 10 Jahren in Süddeutschland und Österreich untersucht. Der Saisonablauf für und wird in einem Schema zusammengefaßt. Die Brutpflege übernimmt i. allg. nur ein Altvogel (bei ca. 60 % aller Brutnester , bei 10 % ). Unklar ist, wie die Abstimmung erfolgt, welcher Altvogel das Gelege betreut. Offenbar herrscht eine Konfliktsituation zwischen den Geschlechtern, die sich in zahlreichen Gelegeaufgaben niederschlägt (ca. 30 % aller Brutnester). Die Verhaltensweisen betreuen bzw. verlassen sind für und nicht gleichmäßig über die Brutsaison verteilt. Die Mediane für die Betreuung durch das und die Aufgabe der Nester liegen dicht beieinander, betreuen die Nester erst wesentlich später. In Illmitz/Österreich nahm die Gelegegröße mit fortschreitender Jahreszeit ab. Die Gelegegröße der aufgegebenen Gelege war kleiner als die zeitgleich zu erwartende Gelegegröße der betreuten Gelege. Wenn das abwandert, komplettiert es das Gelege offensichtlich nicht; damit wird für das zurückbleibende erfolgreiche Brutpflege möglich. Beide Geschlechter verfolgen unterschiedliche Strategien, um einen möglichst guten Fortpflanzungserfolg zu erreichen. Beim besteht die ganze Brutsaison hindurch Bereitschaft, mehrere Nester zu bauen und möglichst viele zu haben. Gegen Ende der Brutsaison, wenn nur noch wenige brutbereite verfügbar sind, übernehmen die u. U. selbst die Brutbetreuung. betreuen zu Beginn der Brutsaison komplette Gelege selbst. Bei rund 30 % der ist aber schon zu Beginn der Brutsaison die Bereitschaft vorhanden, das Gelege mit unvollständiger Gelegegröße nicht selbst zu betreuen und die Verantwortung hierfür dem zu überlassen. Zum Geschlechterkonflikt kommt es, wenn beide Strategien hinsichtlich Abwandern kollidieren; er läßt sich mit Hilfe der Spieltheorie erklären. Wegen der großen Wanderungen während der Brutperiode haben die Angaben zum Verpaarungsstatus der einzelnen Vögel Minimalcharakter. Mindestens 25 % aller und 12 % aller waren polygam. Daß während der ganzen Brutsaison keine Partner haben, ist nachgewiesen. Der Bruterfolg polygamer war signifikant größer als der von mit nur einem Brutnest. Polygame hatten größere Flügel als die übrigen; auch deutet sich an, daß alte erfolgreicher sind als junge. Eine Polygynierate von bis zu 60 % dürfte realistisch sein; die Polyandrierate dürfte bei ca. 30 % liegen. Polyandrisches Verhalten ist aber nicht immer mit einer Steigerung der Reproduktionsrate verbunden. Besonders zu Beginn der Brutsaison bedeutet die Abwanderung des mit hoher Wahrscheinlichkeit die Aufgabe des Nestes, da die in dieser Phase die Brutbetreuung in der Regel nicht übernehmen. Einzelne waren in der Lage, nacheinander zwei Bruten alleine großzuziehen.

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Mating system and strategy of reproduction in Penduline TitRemiz p. pendulinus

Summary From 1978 to 1987, 477 nests of the Penduline Tit were checked in Bavaria and Austria. Adult birds were marked individually with colour rings.In northern Bavaria, the major part of Penduline Tits started building nests in April and May (median date: May 10). As early as end of May a large number was leaving the breeding grounds. The population in the riverine forests of the river March/Austria basically followed the same cycle. On the other hand, birds breeding at Lake Neusiedl/Austria arrived as late as the beginning of June (median date for start of nestbuilding: June 23). A generally valid seasonal pattern for both sexes see fig. 1, 2. Penduline Tits exhibit uniparental care. Most of the nests (about 60 %) were attended by females, about 10 % by male and about 30 % were deserted, because neither the female nor the male decided to incubate (tab. 1).Early in the year, clutches were reared by the female or abandoned; preferably at the end of the breeding season males incubated. A significant relation between clutch size and season could be found at Illmitz/Austria. If average clutch size for female, male or no care are compared with the average clutch size of the respective five-day periods in which the medians of the three categories were found, a significantly smaller clutch size was apparent in abandoned nests. Due to extensive migratory behaviour during the breeding season, a calender effect of clutch size was not calculated for the Upper Main/Bavaria. However, clutch size in nests incubated by females exceeded the size of the abandoned ones. The incubation of smaller clutches by males in the same area is hard to explain. Insufficient data and/or the lack of potential mates as a consequence of early migration may account for it. If a female decides to abandon a nest she evidently leave the clutch incomplete, as if to give the male a commission to care for the young. Thereby, females economize on their reserves in order to be able to produce additional clutches with other males.To achieve optimal reproductive success both sexes follow different strategies. Males: During the whole breeding season males are able to construct a large number of nests to attract as many females as possible. At the end of the breeding season, when only few females are available, some of the males care for their broods themselves. Females: At the beginning of the breeding season, most females are ready to incubate complete clutches. But already at this time, approximately 30 % of the females desert incomplete clutches so as trying to give commission for brood care to the male.The battle of the sexes arises, if both sexes abandon the clutch. This conflict may probably be explained by the game theory. Due to the migrations during breeding season, an exact documentation of the mating status of individual birds was limited. Polygamy occured at least in 25 % of the males and 12 % of the females (tab. 4). A number of unpaired males was also evident during the whole breeding season. Polygynous males reached higher breeding success (tab. 8), and were larger (wing-length) as compared to the other males. Older males may be more successful than younger. According to the number of males leaving nests, the rate of polygyny might be up to 60 % and, due to the number of clutches abandoned by females, approx. 30 % of them are polyandrous. However, polyandrous behaviour was not linked to an increased reproduction rate. Especially at the beginning of the breeding season, most of the nests deserted by females were completely abandoned. Some females were able to raise two broods in one season.

     

Enhanced Rate of Expression and Biosynthesis of Neuropeptide Y After Kainic Acid-Induced Seizures

Recent studies have shown marked increases in brain content of neuropeptide Y (NPY) after seizures induced by intraperitoneal injection of kainic acid and after pentylenetetrazole kindling in the rat. We have now investigated possible changes in the rate of biosynthesis of NPY after kainic acid treatment, by using pulse-labeling of the peptide and by determining prepro-NPY mRNA concentrations. For pulse labeling experiments, [3H]tyrosine was injected into the frontal cortex, and the incorporation of the amino acid into NPY was determined after purifying the peptide by gel filtration chromatography, antibody affinity chromatography, and reversed-phase HPLC. At 2 and 30 days after kainic acid treatment, the rate of tyrosine incorporation was enhanced by approximately 380% in the cortex. In addition, concentrations of pre-pro-NPY mRNA were determined in four different brain areas by hybridization of Northern blots with a complementary 32P-labeled RNA probe 2, 10, 30, and 60 days after kainic acid treatment. Marked increases were observed in the frontal cortex (by up to 350% of controls), in the dorsal hippocampus (by 750%), and in the amygdala/pyriform cortex (by 280%) at all intervals investigated. In the striatum only a small, transient increase was observed. The data demonstrate increased expression of prepro-NPY mRNA and an enhanced rate of in vivo synthesis of NPY as a result of seizures induced by the neurotoxin kainic acid.     

Regulation and Glucocorticoid-Independent Induction of Lipocortin I in Cultured Astrocytes

Stimulation of prostaglandin (PG) release in rat astroglial cultures by various substances, including phorbol esters, melittin, or extracellular ATP, has been reported recently. It is shown here that glucocorticoids (GCs) reduced both basal and stimulated PGD2 release. Hydrocortisone, however, did not inhibit ATP-, calcium ionophore A23187-, or tetradecanoyl phorbol acetate (TPA)-stimulated arachidonic acid release, and only TPA stimulations were affected by dexamethasone. GC-mediated inhibition of PGD2 release thus appeared to exclude regulation at the phospholipase A2 (PLA2) level. Therefore, the effects of GCs on the synthesis of lipocortin I (LC I), a potent, physiological inhibitor of PLA2, were studied in more detail. Dexamethasone was not able to enhance de novo synthesis of LC I in freshly seeded cultures and failed to increase LC I synthesis in 2-3-week-old cultures. It is surprising that LC I was the major LC synthesized in those cultures, and marked amounts accumulated with culture time, reaching plateau levels at approximately day 10. In contrast, LC I was barely detectable in vivo. This tonic inhibition of PLA2 is the most likely explanation for unsuccessful attempts to evoke PG release in astrocyte cultures by various physiological stimuli. GC receptor antagonists (progesterone and RU 38486) given throughout culture time reduced LC I accumulation and simultaneously increased PGD2 release. Nonetheless, a substantial production of LC I persisted in the presence of antagonists. Therefore, LC I induction did not seem to involve GC receptor activation. This was confirmed in serum- and GC-free brain cell aggregate cultures. Here also a marked accumulation of LC I was observed.(ABSTRACT TRUNCATED AT 250 WORDS)     

Perception of the auxin signal at the plasma membrane of tobacco mesophyll protoplasts

Auxin-induced variations of transmembrane potential difference have been shown to be a useful tool for analyzing hormone sensitivity in tobacco protoplasts. Using this technique, we demonstrated that protoplasts derived from wild-type, an auxin-resistant mutant and Agrobacterium-rhizogenes transformed plants differed widely in the sensitivity of their electrical response to naphthalene acetic acid. We have used different antibodies, raised to auxin binding proteins (ABP) from maize coleoptiles, or to the axr1 gene product (ABP1), to test whether changes in auxin sensitivity can be correlated with the presence of tobacco proteins immunologically related to this ABP. Titrations indicated that 0.4 nM anti-ABP IgG inhibited 50% of the auxin-specific response of wild-type protoplasts, whereas 0.04 nM or 4 nM anti-ABP IgG were necessary to inhibit the response of mutant and transformed protoplasts, respectively, to the same extent. On wild-type protoplasts, blocking part of the immunoreactive sites with anti-ABP antibodies resulted in a decrease in auxin sensitivity of the electrical response (0.4 nM anti-ABP IgG inducing a 10–fold decrease), whereas addition of maize ABP increased this auxin sensitivity (1 pM ABP1 raised the sensitivity more than 1000–fold). The results obtained suggest that the auxin sensitivity detected by our assay system correlates with the amount of tobacco proteins immunologically related to the axr¹ gene product from maize. A hypothesis accounting for the presence of these proteins at the external surface of tobacco protoplasts and for the effects of hetero-logous maize ABP on auxin sensitivity is proposed.