Die Beziehung zwischen Mitochondrienmorphologie und Aktivitätsdauer verschiedener Flugmuskelfasern von Locusta migratoria L.

Zusammenfassung Die indirekten und direkten Flugmuskelfasern der Wanderheuschrecke enthalten parallel und voneinander getrennt verlaufende Myofibrillen. In den interfibrillären Räumen liegen zahlreiche, säulenförmig aneinandergereihte Mitochondrion, ein dichtes endoplasmatisches Retikulum und Tracheolen. In Höhe der A-I-Band-Grenzen befindet sich ein charakteristisches retikuläres System, dessen Queranordnung als Diade zu bezeichnen ist. Die chromatinreichen Kerne liegen peripher dicht unter dem Sarkolemm.Sowohl bei den indirekten als auch bei den direkten Flugmuskelgruppen kommt es im Verlaufe des Dauerfluges zu Transformationen der Mitochondrien, die sich in einer erheblichen Schwellung und in einer Umwandlung der Cristae zu Tubuli äußern. Diese Umwandlung wird als intramitochondriale Kompensation aufgefaßt, um das Verhältnis von Oberfläche der inneren membranösen Phase zum Volumen der inneren wäßrigen Phase — d. h. die spezifische Oberfläche — möglichst konstant zu halten. Eine Vesikulation der Tubuli könnte der morphologische Ausdruck für eine Dekompensation sein, da dann der Zusammenhang der äußeren wäßrigen Phase verlorengeht.Die Mitochondrien verschiedener Muskeln reagieren nicht synchron auf die Dauerbelastung, sondern ausgesprochen belastungs- und funktionsspezifisch: Zunächst transformieren sich die Mitochondrien der indirekten, dann erst die der direkten Flugmuskelgruppen, und der anfängliche Schwirrflug geht während fortschreitenden Dauerfluges in einen Flatterflug über. Die unterschiedlichen Erschöpfungszeitpunkte der indirekten Flugmuskeln werden durch die jeweils spezifische Funktion und die ontogenetische Entwicklung erklärt.     

Harnsäure als zusätzliche stickstoffquelle Für larven von Lasioderma Serricorne F.

Zusammenfassung Larven von Lasioderma serricorne F. sind in der Lage, bei Unterdosierung des Caseins in synthetischen Diäten Guanin, Xanthin, Harnsäure und Allantoin als zusätzliche N- Quellen zu verwerten. Die Bedeutung für den Proteinhaushalt der Larven Bowie die Beteiligung der Symbionten werden diskutiert.     

Ein Beitrag zum histochemischen Verhalten der Kern-DNS in Interphase und Mitose,dargestellt durch kombinierte Säurehydrolyse und Akridinorangefluorochromierung

Zusammenfassung Aufbauend auf vorangegangenen Arbeiten über die unterschiedliche Darstellung von DNS durch Kombination einer Säurehydrolyse mit einer Akridinorange-fluorochromierung, sollte untersucht werden, ob sich, damit unter standardisierten Bedingungen Unterschiede der DNS an alkoholfixierten Kernen bei Tumoren, im Laufe der Mitose sowie bei Kernen nekrotischer Zellen darstellen lassen.Ein rascherer Fluoreszenzumschlag zum Langwelligen bereits nach kurzer Hydrolyse (1 min, n/HCl, 37° oder 60°) war zu beobachten: bei allen nekrotischen Zellkernen, bei den Kernen eines Teiles der untersuchten Tumoren (in 45 von 85 Fällen), innerhalb eines Interphasekernes im Heterochromatin und in allen Fällen an den Mitosechromosomen. Unter Berücksichtigung unserer heutigen Vorstellungen über den Bindungsmechanismus des Akridinorange an die Nukleinsäuren des Zellkernes und über die Veränderungen der DNS-Molekülstruktur nach Säurehydrolyse kommen als morphologisches Substrat für den Fluoreszenzumschlag in Betracht: eine geringere sekundärstrukturelle Festigkeit des DNS-Moleküls, ein geringerer Polymerisationsgrad und ein erhöhter Gehalt an ribonukleaseresistentem DNS-RNS-Komplex.Unabhängig von der noch problematischen Deutung der Befunde ist die angewandte Methode in der Lage, Unterschiede in der Struktur der Nukleinsäuren dieser Zellkerne zur Darstellung zu bringen, die sich bisher einem Nachweis entzogen haben.

---

Summary Based on previous papers on the variable demonstration of DNA by means of a combination of acid hydrolysis and conjugation with acridine orange, a study was conducted to investigate if under standardized conditions alcohol fixed nuclei of tumours show differences in their DNA during mitosis and necrosis. A rather sudden change of the fluorescence to the long wave region after short hydrolysis (1 min, n/HCl, 37° or 60°) was observed: in all necrotic cell nuclei; in the nuclei of some of the tumours (45 out of 85); in the heterochromatine of an interphase nucleus, and in all cases of chromosomes in mitosis. Taking into consideration the current knowledge on the linking mechanism of acridine orange to nucleic acids of the cell nucleus, and on changes of the molecular structure of DNA following acid hydrolysis the morphologic substrate responsible for the change of the fluorescence could be: a lesser secondary structural stability of the DNA molecule; a lower degree of polymerisation and a higher level of a ribonuclease resistant DNA-RNA complex.Although it is rather difficult to interprete the findings, it can be said that the method described reveals differences in the structure of certain cell nuclei, which so far have not been demonstrable.

---

---

Einige Ergebnisse dieser Arbeit wurden auf der 4. Arbeitstagung der Arbeitsgemeinschaft Morphologie in der DDR am 2./3. Oktober 1964 in Leipzig vorgetragen und sind Teil einer Habilitationsschrift, die dem Senat der Medizinischen Akademie Carl Gustav Carus, Dresden, vorgelegen hat.     

A comparison of sperm morphology and silver nitrate staining characteristics in the domestic ferret and the black-footed ferret

Ejaculated sperm from the domestic ferret (Mustela putorius furo) and the black-footed ferret (Mustela nigripes) were compared for differences in morphological abnormalities and argentophilic protein distribution. Thawed domestic ferret sperm was also compared to fresh sperm to determine whether there were any effects on cell morphology due to cryopreservation. There were statistically significant differences between the two species of ferret in two of the categories scored. The domestic ferret had a higher frequency of cells that were bent in the midpiece and in the principal piece, and a higher frequency of headless and tailless cells when compared to the black-footed ferret. There were no statistically significant differences in cell morphology between the fresh and cryopreserved ejaculates of the domestic ferret employing a standard egg yolk cryoextender. Silver nitrate staining distribution was different between the two species in both the head and tail region.     

Explanation and prediction in vegetation science

A definition of vegetation science is given, spanning 6 levels of integration and stressing the interrelations among them. The problems of realism are discussed. The selection of levels is related to the adequate correspondence between conceptualization and research aims. Pattern and process are introduced as the central concepts of vegetation science. The perception of reality is dependent on the spatial and temporal scale chosen. The concept of noise is discussed in relation to stochasticity and randomness of events. Traces of essentialism are found both in classification of communities and habitat ecology. Classification is important, particularly the coexistence of alternative classification approaches. Organicism as a basis of vegetation research is rejected because the organismic view is inadequate on higher integration levels. The concept of function is redefined in a non-teleologic way.Present vegetation ecological research is inductivistic. One possible alternative to inductivism is falsificationism. The major domain of this approach is hypothesis testing, which will become more important. Progress can only be reached by a maximum degree of communication among scientific individuals.Predictive ecology is partly based on historic explanation, partly on complementary approaches. Characters of vegetation worthwhile to be predicted are listed and the necessary requirements for vegetation science to become predictive are discussed. A major requirement is the development of succession and life-history theory. A further elaboration of the individualistic concept will be a main task of vegetation science in the near future.     

Single chloride channels in endosomal vesicle preparations from rat kidney cortex

Summary Endocytotic vesicles from rat kidney cortex, isolated by differential centrifugation and enriched on a Percoll gradient, contain both an electrogenic H⁺ translocation system and a conductive chloride pathway. Using the dehydration/rehydration method, we fused vesicles of enriched endosomal vesicle preparations and thereby made them accessible to the patch-clamp technique. In the fused vesicles, we observed Cl^– channels with a single-channel conductance of 73±2 pS in symmetrical 140mm KCl solution (n=25). The current-voltage relationship was linear in the range of –60 to +80 mV, but channel kinetic properties dependended on the clamp potential. At positive potentials, two sublevels of conductance were discernible and the mean open time of the channel was 10–15 msec. At negative voltages, only one substate could be resolved and the mean open time decreased to 2–6 msec. Clamp voltages more negative than –50 mV caused reversible channel inactivation. The channel was selective for anions over cations. Ion substitution experiments revealed an anion permeability sequence of Cl^–=Br^–=I^–>SO ₄ ^2– F^–. Gluconate, methanesulfonate and cyclamate were impermeable. The anion channel blockers 4,4-diisothiocyanatostilbene-2,2-disulfonic acid (DIDS, 1.0mm) and 5-nitro-2-(3-phenylpropylamino)-benzoic acid (NPPB, 0.1mm) totally inhibited channel activity. Comparisons with data obtained from radiolabeled Cl^–-flux measurements and studies on the H⁺ pump activity in endocytotic vesicle suspensions suggest that the channel described here is involved in maintenance of electroneutrality during ATP-driven H⁺ uptake into the endosomes.     

Fusion of cultured dog kidney (MDCK) cells: I. Technique,fate of plasma membranes and of cell nuclei

Summary The evaluation of the intracellular signal train and its regulatory function in controlling transepithelial transport with electrophysiological methods often requires intracellular measurements with microelectrodes. However, multiple impalements in epithelial cells are hampered by the small size of the cells. In an attempt to avoid these problems we fused cells of an established cell line, Madin Darby canine kidney cells, originally derived from dog kidney, to giant cells by applying a modified polyethylene glycol method. During trypsin-induced detachment from the ground of the petri dish, individual cells grown in a monolayer incorporate volume and mainly lose basolateral plasma membrane by extrusion. By isovolumetric cell-to-cell fusion, spherical giant cells are formed within 2 hr. During this process a major part of the individual cell plasma membranes is internalized. Over three weeks following cell plasma membrane fusion degradation of single cell nuclei and cell nuclear fusion occurs. We conclude that this experimental approach opens the possibility to investigate ion transport of epithelia in culture by somatic cell genetic techniques.     

Localization of the photoreceptor gene ROM1 to human chromosome 11 and mouse chromosome 19: sublocalization to human 11q13 between PGA and PYGM.

Rom-1 is a retinal integral membrane protein that, together with the product of the human retinal degeneration slow gene (RDS), defines a photoreceptor-specific protein family. The gene for rom-1 (HGM symbol: ROM1) has been assigned to human chromosome 11 and mouse chromosome 19 by Southern blot analysis of somatic cell hybrid DNAs. ROM1 was regionally sublocalized to human 11p13-11q13 by using three mouse-human somatic cell hybrids; in situ hybridization refined the sublocalization to human 11q13. Analysis of somatic cell hybrids suggested that the most likely localization of ROM1 is in the approximately 2-cM interval between human PGA (human pepsinogen A) and PYGM (muscle glycogen phosphorylase). ROM1 appears to be a new member of a conserved syntenic group whose members include such genes as CD5, CD20, and OSBP (oxysterol-binding protein), on human chromosome 11 and mouse chromosome 19. Localization of the ROM1 gene will permit the examination of its linkage to hereditary retinopathies in man and mouse.     

Physiological Events in Clostridium acetobutylicum during the Shift from Acidogenesis to Solventogenesis in Continuous Culture and Presentation of a Model for Shift Induction

The pH of continuous cultures of Clostridium acetobutylicum growing at pH 5.6 was allowed to decrease to 4.3 after acid production and thereby to shift the cultures from acetate and butyrate to acetone and butanol formation. Several parameters were determined during the shift. An increase in the intracellular acid concentration to 440 mM was recorded. An excess of undissociated butyric acid but not of acetic acid just before the shift to solventogenesis was followed by a decline in acid production and subsequently by the uptake of acids. The intracellular ATP concentration reached a minimum before the onset of solventogenesis; this presumably reflects the ATP-consuming proton extrusion connected with the increase in the ΔpH from 0.7 to 1.4 units. The pool of NADH plus NADPH exhibited a drastic increase until solventogenesis was induced. The changes in the ATP and ADP and NADH plus NADPH pools during these pH shift experiments were the beginning of a stable metabolic oscillation which could also be recorded as an oscillation of the culture redox potential under steady-state solventogenic conditions. Similar changes were observed when the shift was induced by the addition of butyrate and acetate (50 mM each) to the continuous culture. However, when methyl viologen was added, important differences were found: ATP levels did not reach a minimum, acetoacetate decarboxylase activity could not be measured, and butanol but not acetone was produced. A model for the shift is proposed; it assumes the generation of two signals, one by the changed ATP and ADP levels and the other by the increased NAD(P)H level.