D-Glucose Transport in Cultured Cells of Neural Origin: The Membrane as Possible Control Point of Glucose Utilization

Abstract: The function of plasma membrane as control point of glucose metabolism has been studied in confluent monolayer of C1300 neuroblastoma (N2A) and glioma (C6) cells. In neuroblastoma, steady state intracellular glucose concentration reached the extracellular levels, while intracellular contents in C6 glioma cells remained very low. In C6 glial cells the amount of glycogen as source of energy was much higher than that found in C1300 neuroblastoma cells. Influx rates of D-glucose in C6 glioma cells were only half those found in neuroblastoma cells. During the influx period (0-40 s) the transport of glucose in these cells did not exceed the phosphorylation rate, whereas a steady, time-dependent increase in glucose content was observed in neuroblastoma cells. While glucose uptake in neuroblastoma cells seems to be regulated at the level of phosphorylating enzymes, the control point in C6 glioma is believed to be membrane transport.     

Unusual behaviour of SPO1 DNA with respect to restriction and modification enzymes recognizing the sequence 5''-G-G-C-C

Summary SPO1 DNA contains only 5 cleavage sites for restriction enzymes which recognize and cleave the sequence 5-G-G-C-C (HaeIII or BsuR). Fragments of SPO1 DNA cloned in E. coli to substitute 5-hydroxymethyluracil (HMU) by thymine (T) remain resistant to HaeIII indicating that this unexpectedly small number of cleavages by HaeIII is not correlated with the presence of HMU in the normal phage DNA. It was previously shown that SPO1 is neither subject to B. subtilis R restriction (Trautner et al., 1974) nor modification in vivo (Günthert et al., 1975). We now show that SPO1 DNA can however be restricted and modified in vitro.     

The use of phenylmethylsulfonyl fluoride in the study of catabolite inactivation and repression in intact cells of Saccharomyces cerevisiae

Catabolite inactivation of fructose-1,6-bisphosphatase, isocitrate lyase, phosphoenolpruvate carboxykinase and malate dehydrogenase in intact cells could be prevented by phenylmethylsulfonyl fluoride added 40 min prior to the addition of glucose. Protein synthesis, fermentative and respiratory activity and catabolite repression were not affected. Elimination of catabolite inactivation by the addition of PMSF revealed that catabolite repression started at different times for different enzyme.Abbreviation PMSF phenylmethylsulfonyl fluoride     

Zur Digestion bei Pseudomicrothorax dubius Mermod Nahrungsvakuolen-Vesikulation (Ciliophora) im Anschluß an die Phagocytose

Zusammenfassung Der Schlinger Pseudomicrothorax dubius ingestiert innerhalb von 1–2 min ein großes Volumen fädiger Blaualgen. Die Nahrung ist unmittelbar nach dieser rapiden Phagocytose in einer einzigen, sehr großen Vakuole eingeschlossen, die fast den ganzen Ciliaten ausfüllt. Im Verlaufe der folgenden Stunde vesikuliert diese große Nahrungsvakuole über Zwischenstufen zu einer Vielzahl von Vakuolen mit 1–2 m Durchmesser. Gleichzeitig erfolgt eine Kondensierung des Vakuoleninhaltes. Erst zu diesem Zeitpunkt setzt die Verdauung der Nahrung ein, wie an Hand von zahlreichen Dictyosomen belegt wird, die nun in unmittelbarer Nähe der Nahrungsvakuolen nachzuweisen sind. Durch die Vesikulation der großen Nahrungsvakuole in kleinere Einheiten sowie durch die Kondensierung der Nahrung wird bewirkt, daß die über Lysosomen in die Nahrungsvakuolen abgegebenen Verdauungsenzyme optimal eingesetzt werden. Nach Beendigung der Verdauung liegen viele leere Vakuolen vor, die durch eine stark gefaltete Kontur gekennzeichnet sind. Diese Vakuolen gehen allem Anschein nach wieder in den Membranhaushalt der Zelle ein.

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On the digestion in Pseudomicrothorax dubius Mermod (Ciliophora) vesiculation of the food vacuole following phagocytosis

Summary The gulper Pseudomicrothorax dubius ingests a large volume of filamentous blue-green algae within 1–2 min. Immediately after this rapid phagocytosis, the food is enclosed in a single, extremely large food vacuole, which fills up the ciliate almost entirely. During the following hour this giant food vacuole vesiculates. Finally numerous small vacuoles are present, 1–2 m in diam. Simultaneously the content of the vacuoles is noticeably condensed. At this time the digestion of the food starts as is indicated by numerous dictyosomes, which now surround the periphery of the food vacuoles. Due to both, the prior vesiculation of the food vacuole and the condensation of the food, the digestive enzymes can act very effectively. After 6–8 hours, when the digestion of the food is finished, numerous empty vacuoles are found. Each is characterized by a highly irregular, convoluted outline. Apparently these vacuoles are eventually recycled to the membrane pool of the cell.

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Für umsichtige und sorgfältige technische Assistenz danke ich Frl. A. Rüskens. Die Deutsche Forschungsgemeinschaft unterstützte diese Untersuchung durch die Sachbeihilfe Ha 818/7     

Regeneration of NADH with immobilized systems of alanine dehydrogenase and hydrogen dehydrogenase

Summary An alternative approach to the regeneration of coenzymes using immobilized hydrogen dehydrogenase (hydrogenase) is described. Hydrogenase isolated from Alcaligenes Eutrophus was immobilized to porous glass particles and used in combination with alanine dehydrogenase for formation of alanine, while the NADH consumed was regenerated by molecular hydrogen. Different physical arrangements of the two enzymes were compared. Alanine was conveniently assayed with a specially designed enzyme thermistor method.     

Involvement of smooth and coated vesicles in the function of the contractile vacuole complex of some cryptophycean flagellates

The contractile vacuole complex of cryptophycean flagellates comprises the contractile vacuole, a pore and a vesicular spongiome. A minority of spongiome vesicles bear a 15-nm coat on the cytoplasmic surface of the membrane. The coat superficially resembles a clathrin coat. The majority of vesicles are smooth surfaced. Both types of vesicles are found at the same time. Smooth vesicles can be seen in profile suggesting vesicle-vesicle and vesicle-vacuole fusion. It is suggested that smooth vesicles are involved in the segregation of fluid from the cytoplasm and in filling the vacuole. Coated elements exist only as independent vesicles and as coated pits in the contractile vacuole membrane. There is no evidence of fusion of coated vesicles. It is suggested that coated vesicles function to retrieve specific membrane components from the contractile vacuole.     

Versuche zum angenommenen Kontrastverlust im Gesang der Blaumeise (Parus caeruleus) auf Teneriffa

Zusammenfassung Der Gesang von Blaumeisen-Populationen in Mitteleuropa, Spanien, Marokko und auf Teneriffa wurde miteinander verglichen (Abb. 1). In Süddeutschland und auf Teneriffa wurden Klang-Attrappen-Versuche durchgeführt.1. Der Blaumeisen-Gesang ist in Marokko und auf Teneriffa sehr ähnlich. Er unterscheidet sich vom Blaumeisengesang in Europa (Mitteleuropa und Spanien) in folgenden Eigenschaften (2.–4.):2. Ihm fehlen phrasierte Strophen (Abb. 2 a–c, e, 3, 4 a, b, f, g), also Strophen, in denen dasselbe Element mindestens viermal nacheinander wiederholt wird.3. Er enthält mehr verschiedene Elemente (Abb. 5, 7).4. Die interindividuelle Variation ist größer (Abb. 6, 7).5. Die Unterschiede zwischen dem Blaumeisen-Gesang in Europa und auf Teneriffa lassen sich nicht mit der Kontrast-Betonungs- und der Kontrast-Verlust-Hypothese nachLack &Southern (1949) undMarler (1960) erklären. Die Lernentzugs-Hypothese erklärt sie besser.6. Die Blaumeisen von Teneriffa und Marokko sind sehr nahe miteinander verwandt, aber vermutlich sind sie von den europäischen Blaumeisen artlich nicht getrennt.

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The song of the Blue Tit (Parus caeruleus) on Tenerife: Experiments on a presumed loss of contrast

Summary Songs of Blue Tit populations from Central Europe, Spain, Morocco and Tenerife (fig. 1) were compared. Playback experiments were made in South Germany and on Tenerife.1. Blue Tit songs from Morocco and Tenerife are very similar. They differ from songs of Blue Tits in Europe (Central Europe and Spain) in the following characteristics (2–4):2. Lack of phrased songs (fig. 2 a–c, e, 3, 4 a, b, f, g), i. e. songs with the same notes repeated successively at least four times.3. A greater number of different types of notes (fig. 5, 7).4. Greater interindividual variation (fig. 6, 7).5. Differences between Blue Tit song from Europe and from Tenerife cannot be explained by the hypothesis of contrast reinforcement and loss of contrast, according toLack &Southern (1949) andMarler (1960). The withdrawal of learning hypothesis is a better explanation.6. Blue Tits on Tenerife and Morocco are very closely related to each other but are probably not a different species from the Blue Tits of Europe.

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Für die Übersetzung danken wir FrauR. Jellis und HerrnP. Lundberg.     

Analysis of actin and microfilament-associated proteins in the mitotic spindle and cleavage furrow of PtK2 cells by immunofluorescence microscopy: A critical note

The distribution of actin and the microfilament-associated proteins myosin and tropomyosin was studied in mitotic PtK2 cells. Using fluorescent heavy meromyosin and two different antibodies against actin we have found no evidence for increased accumulations of actin in the mitotic spindle but have found increased levels of actin in the cleavage furrow and the contractile ring. Short, thin microfilament pieces remain detectable in the cytoplasm throughout mitosis. Purified antibodies against myosin and tropomyosin also revealed no increased levels of these proteins in the spindle region, although both proteins were found in the contractile ring and areas of the cytoplasm close to the intercellular bridge. These data are in agreement with functional and ultrastructural studies involving a role for actin and microfilament-related proteins in cytokinesis. They do not support models in which microfilament-related proteins are assumed to be a major constituent of the mitotic spindle.     

Freeze-fracture study of taste bud pores in the foliate papillae of the rabbit

Summary In freeze-fractured specimens of taste buds from the foliate papillae of rabbits, the intercellular spaces are separated from the pore of the taste bud by zonulae occludentes of the tight-type. Below these tight junctions numerous desmosomes are found at irregular intervals. The epithelial cells adjacent to the pore are also joined by single strands of fusion. The microvilli arising from the neck of the type I cells have a high particle density. The microvilli of type II cells and especially the short microvilli of peripherally situated cells have a lower intramembranous particle density. The single microvillus of type III cells has a very large diameter and is longer than the other microvilli. It contains a few larger intramembranous particles and vesicle-like protrusions of the membrane facing the cytoplasm. Transverse fracturing reveals a filamentous fine structure in all microvilli. The physiological implications of these observations are discussed.Supported by grants from the Deutsche Forschungsgemeinschaft (Ja 205/5+6)     

Identification and quantitative determination of the flavin component of soluble hydrogenase from Alcaligeneseutrophus

The flavin component of soluble hydrogenase (hydrogen: NAD⁺ oxidoreductase, EC 1.12.1.2) from $\text{Alcaligenes}\text{eutrophus}$ was identified as FMN by thin layer chromatography in two solvent systems and by binding studies with apoflavodoxin from $\text{Megasphaera}\text{elsdenii}$. The flavin of hydrogenase reacted rapidly with apoflavodoxin with almost complete quenching of the fluorescence at 525 nm. Quantitative determination of FMN was performed by fluorimetric titration with a standardized solution of apoflavodoxin. From the determined FMN content of different enzyme preparations and from the percentage of stimulation of hydrogenase activity by exogenous FMN it is concluded that hydrogenase contains 2 FMN per molecule.