Die Regeneration des Plasmalemms vonPhysarum polycephalum

Zusammenfassung Die Regeneration des Plasmalemms vonPhysarum polycephalum erfolgt nach experimenteller Ruptur sowohlbei Protoplasmatropfen als auch bei isoliertem Protoplasma durch Membranvesikulationsprozesse. Verantwortlich für die Neubildung des Plasmalemms sind verschiedene protoplasmatische Vakuolen (Schleimvakuolen, Nahrungsvakuolen). Unmittelbar nach experimenteller Entfernung des Plasmalemms kommt es zu einer Ansammlung von Vakuolen in einer 5–10 tiefen Region an der Oberfläche des nackten Protoplasten. Durch successive Konfluation dieser Vesikel entstehen zunächst einige gro\e, parallel zur Protoplasmaoberfläche abgeflachte Vakuolen, die schlie\lich zu einer einzigen, das gesamte Protoplasma allseitig umschlie\enden Vakuole verschmelzen. Diedistale Vakuolenmembran zerfällt in einzelne, unterschiedlich gro\e Areale, die zusammen mit degenerierenden Bestandteilen des Protoplasmas abgesto\en werden. Dieproximale Vakuolenmembran wird zur neuen Oberfläche des überlebendenPhysarum-Protoplasmas. Der gesamte Vorgang der Membran-Neubildung erfolgt innerhalb von 5–6 sec und scheint unabhängig von der Natur des umgebenden Milieus zu verlaufen. Trotz der Tatsache, da\ innerhalb der ersten Phase des Regenerationsprozesses das Protoplasma kurzfristig nackt war, d.h. ohne membranöse Begrenzung existierte, lä\t sich keine Einwanderung von partikulären Substanzen des Au\enmediums in das überlebende Protoplasma nachweisen, wie durch Versuche mit dem MarkierungsmittelAerosil gezeigt werden konnte.

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The regeneration of the plasmalemma of Physarum polycephalum

Summary After experimental rupture, the plasmalemma of protoplasmic droplets and isolated protoplasm ofPhysarum polycephalum regenerates by means of membrane vesiculation. Various (mucus- and food-) vacuoles are responsible for the new formation of the plasmalemma. Immediately after removal of the old plasma membrane, many vacuoles accumulate below the boundary between the protoplasm and the surrounding medium in a region of 5–10 depth. Later some large vacuoles, which are strongly flattened parallel to the surface of the protoplasm, originate by means of successive confluence of these single vesicles. The large vacuoles finally fuse to a single vacuole which surrounds the whole protoplast. Thedistal membrane of this vacuole disintegrates together with degenerating protoplasmic components. Theproximal membrane of the vacuole represents the new surface of the surviving protoplasm. The process of regeneration of the plasmalemma takes 5–6 sec and seems not to depend on the kind of medium surrounding the protoplasm. Inspite of the fact that during a part of the first phase of regeneration, the cytoplasm is naked (i.e. without a membranous boundary), there is no migration of particulate substances from the outer medium into the surviving protoplasm, as shown by experiments with the marker substanceaerosil.

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Frau B. Koeppen, Fräulein I. Kletschke und Frau M. Sauernheimer danken wir für technische Hilfe.     

Zahl und Verteilung antennaler Sensillen bei der Honigbiene (Apis mellifera L.)

Zusammenfassung An Totalpräparaten der Antennengeißel von Arbeiterin und Drohne vonApis mellifera carnica wurden Zahl und Verteilung aller Sensillen und Setae ermittelt. Dabei ließen sich anhand des cuticularen Baues folgende Sensillentypen unterscheiden: S. placodeum, S. ampullaceum, S. coeloconicum, S. campaniforme und 5 Haarsensillen S. trichodeum A, B1, B2, C, D, sowie 4 Setatypen (A 1–3, B), die wahrscheinlich nicht innerviert sind. Die Benennungen der Sensillen wurde den bisher gebrauchten Bezeichnungen gegenübergestellt. Sensillenzahl und -Verteilung, Sinneszellzahl und Funktion der Sensillen wurden anhand von Literaturangaben zusammengestellt und diskutiert. Bemerkenswert ist der starke Dimorphismus zwischen Arbeiterin und Drohne in der relativen Sensillenzahl für die einzelnen Sensillentypen und in der Gesamtzahl der Sinneszellen. So sind bei der Arbeiterin die wahrscheinlich olfaktorischen S. trichodea A und die mechanorezeptorischen S. trichodea B 1 wesentlich stärker vertreten. Die Drohne hat keine S. basiconica und im übrigen wesentlich mehr S. placodea als die Arbeiterin. Insgesamt hat die Drohne eine ca. 2-fach größere Geißeloberfläche und etwa 5-mal soviele Sinneszellen wie die Arbeiterin. Die Arbeiterinnengeißel hat auf ihrer Rückseite eine porenplattenfreie Zone, die dicht mit nichtinnervierten Setae besetzt ist. Bei der Drohne findet man stattdessen eine porenplattenärmere Zone mit einer geringeren Zahl von Setae. Charakteristische Verteilungsmuster bestehen auch für alle anderen Sensillen und Setae.

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Number and distribution of the sensilla on the antennal flagellum of the honeybee (Apis mellifera L.)

Summary Number and distribution of sensilla and setae on the antennal flagellum of the honeybeeApis mellifera carnica were determined on whole antennal preparations. The following types of sensilla were distinguished according to their cuticular structure: Sensillum placodeum, S. ampullaceum, S. coeloconicum, S. basiconicum, S. campaniforme and 5 hair sensilla S.trichodeum A, B1, B2, C, D, as well as 4 types of probably non-innervated setae (A1–3, B). The names used here for the different types were compared with the previously used terms. Number and distribution of sensilla, the number of sensory cells and the function of the sensilla were discussed with respect to the data available from the literature. There is a notable dimorphism between the worker and drone with respect to the relative number of sensilla of each type and to the total number of sensory cells. The worker has far more of the presumably olfactory S. trichodea A and of the mechanoreceptive S. trichodea B1. The drone lacks the S. basiconica and has far more S. placodea than the worker. The flagellum surfarce of the drone is twice as large as that of the worker and has 5 times as many sensory cells. The worker flagellum has a poreplate-free zone on the side facing the head which is densely packed with non-innervated setae. In the corresponding zone the drone has a lower density of poreplates than elsewhere on its antennal flagellum. All other sensilla and setae are also characteristically distributed.

     

Abolition of Intrinsically Bent DNA Structure Components in AT Clusters by Netropsin Interaction; Titration Viscometric Investigations

Abstract

It is argued that the enhancement of the apparent DNA contour length by the specifically binding non-intercalating drug netropsin (Nt) (Reinert et al., NAR 9,2335, 1981) at very low Nt/DNA-phosphate ratios essentially is the result of an abolition of periodically arranged intrinsic helix bends in A · T rich tracts of base pairs.

In the preceding paper the existence of pronounced DNA tertiary structure components has been postulated for (two species of) natural eukaryotic DNA. The resulting model suggests local apparent solenoid-related DNA tertiary structure components at high sodium ion concentration c_s, partly/totally molten out at 45/60 C. With decreasing c_s the tertiary structure components have been found to be gradually reduced, at least below c_s = 0.010 M, as titration viscometrically revealed by a gradual rise of the apparent DNA contour length (Reinert et al., JBSD 9, 537, 1991).

Hence, we performed titration viscometric analyses about Nt interaction with calf thymus DNA (ctDNA) at c_s = 0.075 M, 0.010 M and 0.004 M Na⁺. The concomitant DNA conformational changes are quantitatively described in terms of the relative changes of both DNA persistence length and hydrodynamically operative apparent DNA contour length for the three first resolved interaction modes below a Nt/DNA-P ratio of 0.03.

These experiments, together with previous respective analyses at c_s = 0.20 M Na⁺ and different temperatures (I.e.), suggest that those DNA sites binding Nt most strongly predominantly are responsible for the formation of solenoid-related DNA tertiary structure components. Most probably these are A tract-containing sequences. As the essential factor for their apparent elongation effect at low Na⁺ concentrations, a gradual alteration of the number of base pairs per helix turn seems to occur below c_s = 0.010 M Na⁺ and, concomitantly, a change in phasing between intrinsic helix bends and helix screw.     

Multimode Interaction of Hoechst 33258 with Eukaryotic DNA; Quantitative Analysis of the DNA Conformational Changes

Abstract

The interaction of the minor groove binding ligand Hoechst 33258 (Hoe) with natural DNA was investigated by high resolution titration rotational viscometry. Analysis of the concomitant DNA conformational changes was performed with two DNA samples of sufficiently different molar mass M, at 4°C, 22°C and 40°C, for Hoe/DNA-P ratios below r = 0.02. In this narrow r range several interaction modes could be resolved. The measured conformational changes were quantified in terms of relative changes of both apparent DNA persistence length, Δa/a, and hydrodynamically operative DNA contour length, ΔL/L. Δa/a(r) primarily is a measure of ligand-induced DNA helix stiffening, but both, Δa/a(r) and ΔL/L(r), generally depend also on ligand binding induced DNA bending or DNA unbending. The essential difference obviously is that Δa/a(r) is influenced by the randomly distributed helix bends and ΔL/L(r) by phased ones. The measurements performed at different temperatures deliver informations about existence and temperature dependent abolition of intrinsic helix curvature.

Both Hoe and netropsin (Nt) prefer binding to AT rich DNA segments, which are candidates for intrinsic DNA helix bends. But our data for Hoe interaction with calf thymus DNA (ctDNA) show characteristic differences to those for Nt-ctDNA interaction. Especially for Hoe, the mode of highest affinity is saturated already at a ligand concentration of roughly 1 nM (r = 0.0015 Hoe/DNA-P). It exhibits an unusually strong temperature dependence of the conformational DNA response. A Hoe-Nt competition experiment shows that Hoe binding to the sites of the very first Hoe mode is almost unaffected by bound Nt. But Hoe binding to the sites of the following Hoe modes does not occur due to the competition with Nt. Thus this mode of strongest Hoe-DNA interaction reflects a unique mechanism, possibly of high relevance for gene regulatory systems.     

The antennal benzoic acid receptor cell of the female silk moth <Emphasis Type="Italic">Bombyx mori</Emphasis> L.: structure–activity relationship studies with halogen substitutes

Studies on structure–activity relationships were carried out to characterize the response specificity of the benzoic acid cell of the female of the moth Bombyx mori by means of single sensillum electrophysiological recordings. We demonstrated that this cell type responds best to a natural key substance (benzoic acid) and has similar response profiles for less effective compounds, including various halogen substitutes of benzoic acid, benzaldehyde and other derivates of the key compound. Using different halogen substitutes (F, Cl, Br, I), we showed that the cellular response decreases with increasing atomic size of the substitute and that halogen substitutes were most effective in the meta-position. Thus, m-fluor benzoic acid was even more effective than benzoic acid. These results indicate that a critical feature of the stimulus molecule is the inductive effect generated by the halogen substitutes. Increasing the atomic size of the halogen substitute impairs the recognition of the molecule by the receptor cell, possibly due to steric effects. Decreasing the electron density in the aromatic ring improves the receptor response. The benzoic acid receptor cell can be considered as specialist despite not being involved in pheromone detection as it responds maximally to a key substance and has similar response profiles for less effective compounds.