The precursor of the F1β subunit of the ATP synthase is covalently modified upon binding to plant mitochondria

We present evidence for a unique covalent modification of a nuclear-encoded precursor protein targeted to plant mitochondria. We investigated the early events of in vitro import for the mitochondrial precursor of the ATP synthase F₁ subunit from Nicotiana plumbaginifolia (pF₁) into plant mitochondria. When pF₁ of 59 kDa was incubated with mitochondria isolated from different higher-plant species, a band of 61 kDa was generated. The 61 kDa protein was a covalently modified form of the 59 kDa pF₁. The modification was dependent on the 25 amino acid long N-terminal region of the presequence of pF₁. The modification was catalysed by an enzyme located in the outer mitochondrial membrane which was specific for higher plants and could not be washed off from the membrane by urea, KCl or EDTA. The modification was ATP- and Ca²⁺-dependent, but it was not affected by inhibitors of protein kinases. No inhibition of the modification was observed with phosphatase, methylation or acylation inhibitors. The modification occurs prior to translocation through the mitochondrial outer membrane. Inhibition of the modification process does not affect the import of the precursor protein, hence precursor modification was not a prerequisite for import. Both the modified and the unmodified pF₁ proteins were strongly associated with the mitochondrial outer membrane.     

Isolation and characterization of β-glucosidases from Aspergillus nidulans mutant USDB 1183

Two extracellular -glucosidases (cellobiase, EC 3.2.1.21), I and II, from Aspergillus nidulans USDB 1183 were purified to homogeneity with molecular weights of 240,000 and 78,000, respectively. Both hydrolysed laminaribiose, -gentiobiose, cellobiose, p-nitrophenyl--L-glucoside, phenyl--L-glucoside, o-nitrophenyl--L-glucoside, salicin and methyl--L-glucoside but not -linked disaccharides. Both were competitively inhibited by glucose and non-competitively (mixed) inhibited by glucono-1,5-lactone. -Glucosidase I was more susceptible to inhibition by Ag⁺ and less inhibited by Fe²⁺ and Fe³⁺ than -glucosidase II.     

Bewegungsstudien an Anatinen

Zusammenfassung Vorliegende Arbeit ist eine Weiterführung derLorenz'schen Bewegungsstudien an Anatinen aus dem Jahre 1941, fortgesetzt an Mischlingen zwischen den dort beschriebenen Arten. Die sich dabei ergebenden Befunde machten eine erneute Untersuchung der Elternarten notwendig. Außerdem wurden einige Arten beobachtet, deren Verhalten noch nicht untersucht worden war. Fragestellung und Begründung werden in der Einleitung gegeben.Im zweiten Abschnitt werden einige der vonLorenz gemachten Beobachtungen berichtigt. So zeigten einige der Kreuzungen mitbahamensis, daß die vonLorenz bei eben dieser Art als Kurzhoch-werden bezeichnete Bewegungsweise dem Ab-auf anderer Schwimmenten homolog ist. Ebenso ist die eine der beiden vonLorenz als Kurzhoch-werden bezeichneten Verhaltensweisen des Krickerpels als Ab-auf zu deuten. Der Gruß desflavirostre-Erpels wurde auch bei weiblichen Tieren gesehen. BeiAnas acuta wurde ein Kinnheben festgestellt, das sich in der Form stark vom Kinnheben beiplatyrhynchos unterscheidet. Als neue Verhaltensweisen wurden u. a. das Haltungannehmen und das Tendieren beimflavirostre-Erpel beschrieben.Im dritten Abschnitt werden einige Verhaltensweisen und ihre Funktion diskutiert und der Versuch gemacht, eine Motivationsanalyse zu geben.(Zeichnungen vonHermann Kacher)     

Purification and characterization of flavone-specific β-glucuronidase from callus cultures of Scutellaria baicalensis Georgi

-Glucuronidase from callus cultures of Scutellaria baicalensis Georgi was purified to apparent homogeneity by fractionated ammonium-sulfate precipitation and chromatography on diethylaminoethyl-cellulose, hydroxylapatite and baicalin-conjugated Sepharose 6B. A 650-fold purification was obtained by this purification system. When subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis the purified protein migrated as a single band with a molecular mass of 55 kDa. We determined that the native enzyme has a molecular mass of 230 kDa using gel-filtration chromatography. These results suggested that the enzyme exists as a homotetramer composed of four identical 55-kDa subunits. The enzyme showed a broad pH optimum between 7.0 and 8.0. The K _m values were 9 M, 10 M, 30 M and 40 M for luteolin 3 -O--d-glucuronide, baicalin, wogonin 7-O--d-glucoronide and oroxlin 7-O--d-glucuronide, respectively. The enzyme was most active with flavone 7-O--d-glucuronides.Abbreviations BA N⁶-benzyladenine - 2,4-D 2,4-dichlorophenoxyacetic acid - pI isoelectric point - R _t retention time     

Flufenamic acid as an inducer of mitochondrial permeability transition

To assess the mechanism by which mitochondrial permeability transition (MPT) is induced by the nonpolar carboxylic acids, we investigated the effects of flufenamic acid (3-trifluoromethyl diphenylamine-2-carboxylic acid, FA) on mitochondrial respiration, electrical transmembrane potential difference (), osmotic swelling, Ca²⁺ efflux, NAD(P)H oxidation and reactive oxygen species (ROS) generation. Succinate-energized isolated rat liver mitochondria incubated in the absence or presence of 10 M Ca²⁺, 5 M ruthenium red (RR) or 1 M cyclosporin A (CsA) were used. The dose response-curves for both respiration release and dissipation were nearly linear, presenting an IC₅₀ of approximately 10 M and reaching saturation within 25-50 M, indicating that FA causes mitochondrial uncoupling by a protonophoric mechanism. Within this same concentration range FA showed the ability to induce MPT in energized mitochondria incubated with 10 M Ca²⁺, followed by dissipation and Ca²⁺ efflux, and even in deenergized mitochondria incubated with 0.5 mM Ca²⁺. ADP, Mg²⁺, trifluoperazine (TFP) and N-ethylmaleimide (NEM) reduced the extent of FA-promoted swelling in energized mitochondria by approximately one half, whereas dithiothreitol (DTT) slightly enhanced it. NAD(P)H oxidation and ROS generation (H₂O₂ production) by mitochondria were markedly stimulated by FA; these responses were partly prevented by CsA, suggesting that they may be implicated as both a cause and effect of FA-induced MPT. FA incubated with mitochondria under swelling assay conditions caused a decrease of approximately 40% in the content of protein thiol groups reacting with 5,5-dithiobis(2-nitrobenzoic acid) (DTNB). The present results are consistent with a ROS-intermediated sensitization of MPT by a direct or indirect FA interaction with inner mitochondrial membrane at a site which is in equilibrium with the NAD(P)H pool, namely thiol groups of integral membrane proteins.     

Direct somatic embryogenesis through thin cell layer culture in Panax ginseng

Direct somatic embryos were differentiated on cotyledon transverse Thin Cell Layers (tTCLs) of Panax ginseng after 9 weeks in the Murashige and Skoog basal (MS) medium containing 2,4-d (5M). When MS medium containing 2,4-d (5M) was used for seedling pretreatment and for tTCLs culture, somatic embryos were observed 2 weeks earlier, i.e. after 7 weeks of culture. On the tTCLs from seedlings pretreated with 2,4-d (5M) combined with benzyladenine and zeatin at 0.1 M (BZ), somatic embryos were observed after 6 weeks of culture and the percentage of embryogenesis was higher (62%) than when 2,4-d was used alone for pretreatment (40%). Similar results were also obtained from pretreatment with combinations of 2,4-d (5M) and thidiazuron (TDZ) (0.01, 0.1M). When a combination of 2,4-d (5M) and BZ (0.1M) was used both for seedling pretreatment and for tTCLs culture, both somatic embryos and shoots were observed after only 3 weeks. As the concentration of BZ increased, the percentage of somatic embryogenesis decreased but the percentage of organogenesis increased. Similar responses were obtained with a combination of 2,4-d (5M) and TDZ (0.01M). On the medium containing both NAA (0.3M) and BZ (1M), globular- and heart- stage embryos developed after 4 weeks of culture into cotyledonary-staged embryos which remained dormant after a short elongation of the embryo axis. The importance of seedling pretreatment by growth substances in enhancing somatic embryogenesis is reported.Abbreviations BA 6-benzyladenine - BZ combination of BA and zeatin - 2,4-d 2,4-dichlorophenoxyacetic acid - MS medium Murashige and Skoog basal medium - NAA a-naphthaleneacetic acid - TDZ thidiazuron - tTCLs transverse thin cell layers - TCL longitudinal thin cell layer     

Modulation of the serotonin-activated K+ channel by G protein subunits and nucleotides in rat hippocampal neurons

In hippocampal neurons, 5-hydroxytryptamine (5-HT) activates an inwardly rectifying K⁺ current via G protein. We identified the K⁺ channel activated by 5-HT (K_5-HT channel) and studied the effects of G protein subunits and nucleotides on the K⁺ channel kinetics in adult rat hippocampal neurons. In inside-out patches with 10 m 5-HT in the pipette, application of GTP (100 m) to the cytoplasmic side of the membrane activated an inwardly rectifying K⁺ channel with a slope conductance of 36±1 pS (symmetrical 140 mm K⁺) at –60 mV and a mean open time of 1.1±0.1 msec (n=5). Transducin activated the (K_5-HT) channels and this was reversed by -GDP. Whether the K_5-HT channel was activated endogenously (GTP, GTPS) or exogenously (), the presence of 1 mm ATP resulted in a 4-fold increase in channel activity due in large part to the prolongation of the open time duration. These effects of ATP were irreversible and not mimicked by AMPPMP, suggesting that phosphorylation might be involved. However, inhibitors of protein kinases A and C (H-7, staurosporine) and tyrosine kinase (tyrphostin 25) failed to block the effect of ATP. These results show that G activates the G protein-gated K⁺ channel in hippocampal neurons, and that ATP modifies the gating kinetics of the channel, resulting in increased open probability via as yet unknown pathways.     

Techniques for the quantitative study of mutation in plant viruses

Summary Hardly any other virus is chemically and ultramicroscopically as well known as TMV. It is not possible to perform genetic recombinations with this object. The phenomenon of mutation is, however, known and an analysis of the dosis-effect relationship was possible by using the characters chlorotic versus necrotic primary symptoms. Taking into account the phenomenon of interference (mutual exclusion), i.e., comparing the induced mutation frequency with that of a control virus sample diluted to the same level of infectivity, on can perform quantitative analyses. In this way the first chemical mutagensis in the test tube was demonstrated 10 years ago with nitrous acid as mutagenic agent. The criticism raised byBawden to the first publication ofMundry andGierer was already inappropriate at that time. In the meantime it has been demonstrated byWittmann-Liebold andWittmann through analysis of amino acid exchanges in spontaneous mutants and in those isolated after incubation with HNO₂ that the difference between spontaneous and induced mutants demanded byBawden, which cannot be postulated for symptoms in plants, lies, as expected, in amino acid exchanges of the protein coat.

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Zusammenfassung Kaum ein anderes Virus ist chemisch und ultramikroskopisch so gut bekannt wie das TMV. Rekombinations-Genetik ist nicht möglich. Das Phänomen der Mutation ist aber bekannt, und eine Analyse der Dosis-Effekt-Beziehung wurde möglich durch Benutzung der Symptomcharaktere chlorotische versus nekrotische Primärsymptome. Bei Berücksichtigung des Phänomens der Interferenz (mutual exclusion), d. h. wenn man die induzierte Mutationsrate mit der auf gleiche Infektiosität durch Verdünnen der Viruslösung gebrachten als Kontrolle vergleicht, kann eine quantitative Analyse durchgeführt werden. So wurde vor 10 Jahren die erste Chemomutagenese im Reagenzglas mit salpetriger Säure als mutagenes Agens nachgewiesen. Die an der ersten Veröffentlichung vonMundry undGierer vonBawden geäußerte Kritik war schon damals unzutreffend. Inzwischen ist durch die Analyse der Aminosäureaustausche von spontanen und nach Inkubation mit HNO₂ isolierten Mutanten vonWittmann-Liebold undWittmann gezeigt worden, daß die vonBawden geforderte Verschiedenheit spontaner und induzierter Mutanten, die für Symptome an den Pflanzen nicht postuliert werden kann, in den Aminosäureaustauschen des Hüllproteins wie zu erwarten vorhanden ist.

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This paper was a first written for Methods in Virology, Academic Press. The editors and the author did not come to an agreement in the question of citation ofBawden's criticism to the work ofMundry andGierer 1958. It is published here on the occasion of the 10th anniversary of the first chemomutagenesis in the test tube.     

Tetramethyl Rhodamine Methyl Ester (TMRM) is Suitable for Cytofluorometric Measurements of Mitochondrial Membrane Potential in Cells Treated with Digitonin

A new method for cytofluorometric analysis of mitochondrial membrane potential has been developed by using TMRM as a cationic, mitochondrial selective probe. The method is based on limited treatment of cultured cells with digitonin which permeabilises the plasma membrane and leaves mitochondria intact. The resulting signal of TMRM-stained cells thus represents only the probe accumulated in mitochondria. Fibroblasts and cybrids were used as a model cell systems and optimal conditions for digitonin treatment and staining by TMRM were described. The TMRM signal collapsed by valinomycin, KCN and antimycin A and FCCP titration was used to gradually lower and characterise the stability of . The method is suitable for sensitive measurement of in different types of cultured cells.     

Cellular localization of glycoside hydrolases inBacteroides fragilis

The localizations of six glycosidases produced byBacteroides fragilis—-glucosidase, -glucosidase, -galactosidase, -galactosidase, -N-acetylglucosaminidase, and -l-fucosidase—were studied. Cell fractions and cell extracts were obtained by Triton X-100 release, by disruption by freeze-pressing and sonication, and by osmotic release. Isoelectric focusing of a cytoplasmic and of a Triton X-100 extract of the cell wall fraction was performed and revealed differences in the relative distribution of differently charged forms of -N-acetylglucosaminidase. -Galactosidase and alkaline phosphatase were used as cytoplasmic and periplasmic markers, respectively. It is concluded that inB. fragilis -glucosidase is periplasmic, -l-fucosidase and -galactosidase are cytoplasmic, and -n-acetylglucosaminidase is cell associated and bound to the cell envelope by hydrophobic interactions. -Glucosidase and -galactosidase are localized cytoplasmically and/or located in the cell envelope.     

“Balbiani's body” in the Oocyte of the golden hamster

Summary At a particular stage in the development of the ovarian oocyte in the golden hamster, massive juxtanuclear accumulations of mitochondria grouped in a specific pattern have been observed. This phenomenon is described and its relationship to the Balbiani's body or yolk nucleus of the light microscopists is discussed.Supported by U.S.P.H.S. Post-doctoral Fellowship 5 F 2 HD-25, 190-02.I wish to thank Professor R. E. Coupland for his continued interest in this work and for his helpful comments.     

High frequency somatic embryogenesis in Coffea canephora

Leaf explants of Coffea canephora (P. ex Fr.) produced a friable yellow callus when they were cultured on a conditioning basal medium with 2.2 M 2,4-D, 2.4 M IBA and 9.8 M 2iP for 4 weeks then on an induction basal medium with 4.4 M 2,4-D and 17.8 M BA for 10 weeks. This calus could be maintained by means of regular subcultures or it could give rise to somatic embryos depending on the culture medium. Cytological studies documented somatic embryogenesis and embryo development.Abbreviations BA 6-benzyladenine - 2,4-D 2,4-dichlorophenoxyacetic acid - IBA indole-3-butyric acid - 2iP 2-isopentenyladenine - MS Murashige & Skoog medium - NAA -naphthaleneacetic acid - NPR nucleoplasmic ratio - PGR plant growth regulator     

Delineation of sodium-stimulated amino acid transport pathways in rabbit kidney brush border vesicles

Summary We have confirmed previous demonstrations of sodium gradient-stimulated transport ofl-alanine, phenylalanine, proline, and -alanine, and in addition demonstrated transport of N-methylamino-isobutyric acid (MeAIB) and lysine in isolated rabbit kidney brush border vesicles. In order to probe the multiplicity of transport pathways available to each of these¹⁴C-amino acids, we measured the ability of test amino acids to inhibit tracer uptake. To obtain a rough estimate of nonspecific effects, e.g., dissipation of the transmembrane sodium electrochemical potential gradient, we measured the ability ofd-glucose to inhibit tracer uptake.l-alanine and phenylalanine were completely mutually inhibitory. Roughly 75% of the¹⁴C-l-alanine uptake could be inhibited by proline and -alanine, while lysine and MeAIB were no more effective thand-glucose. Roughly 50% of the¹⁴C-phenylalanine uptake could be inhibited by proline and -alanine; lysine was as effective as proline and -alanine, and the effects of pairs of these amino acids at 50mm each were not cumulative. MeAIB was no more effective thand-glucose. We conclude that three pathways mediate the uptake of neutral,l, -amino acids. One system is inaccessible to lysine, proline, and -alanine. The second system carries a major fraction of thel-alanine flux; it is sensitive to proline and -alanine, but not to lysine. The third system carries half the¹⁴C-phenylalanine flux, and it is sensitive to proline, lysine, and -alanine. Since the neutral,l, -amino acid fluxes are insensitive to MeAIB, we conclude that they are not mediated by the classicalA system, and since all of thel-alanine flux is inhibited by phenylalanine, we conclude that it is not mediated by the classicalASC system.l-alanine and phenylalanine completely inhibit uptake of lysine. MeAIB is no more effective thand-glucose in inhibiting lysine uptake, while proline and -alanine appear to inhibit a component of the lysine flux. We conclude that the¹⁴C-lysine fluxes are mediated by two systems, one, shared with phenylalanine, which is inhibited by proline, -alanine, andl-alanine, and one which is inhibited byl-alanine and phenylalanine but inaccessible to proline, -alanine, and MeAIB. Fluxes of¹⁴C-proline and¹⁴C-MeAIB are completely inhibited byl-alanine, phenylalanine, proline, and MeAIB, but they are insensitive to lysine. Proline and MeAIB, as well as alanine and phenylalanine, but not lysine, inhibit¹⁴C--alanine uptake. However, -alanine inhibits only 38% of the¹⁴C-proline uptake and 57% of the MeAIB uptake. We conclude that two systems mediate uptake of proline and MeAIB, and that one of these systems also transports -alanine.     

Production of β-fructofuranosidase byAspergillus japonicus

A newly isolated strain, MU-2, which produces very high -fructofuranosidase activity, was identified asAspergillus japonicus. For enzyme production by the strain, sucrose at 20% (w/v) was the best carbon source and yeast extract at 1.5 to 3% (w/v) the best nitrogen source. Total enzymatic activity and cell growth were at maximum after 48 h, at 1.57×10⁴ U/flask and 0.81 g dry cells/flask, respectively. The optimum pH value of the enzymatic reaction was between 5.0 and 5.5 and the optimum temperature 60 to 65°C. The enzyme produced 1-kestose (O--d-fructofuranosyl-(21)--d-fructofuranosyl -d-glucopyranoside) and nystose (O--d-fructofuranosyl-(21)--d-fructofuranosyl-(21)--d-fructofuranosyl -d-glucopyranoside) from sucrose by fructosyl-transferring activity. The strain was found to be very useful for industrial production of -fructofuranosidase.     

Fragmenta mycologica

Summary Castellani's water culture method for microscopic fungi was re-examined and confirmed in its every detail. It is an ideal method for, at least, the smaller culture collection, in order to avoid continuous, short term subculturing. Benedek's suggestion for establishment of a mycotheca preserved in chlorallactophenol on the analogy of the herbarium of phanerogamists, may fill a long felt gap in mycological laboratories.

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Zusammenfassung Castellani's Wasserkultur-methode für mikroskopische Pilze wurde einer Kontrolluntersuchung unterzogen. Sie konnte in all ihren Einzelheiten bestätigt werden. Sie ist eine ideale Methode, wenigstens für kleine Kultursammlungen, um eine fortgesetzte, kurz-fristige Überimpfung zu vermeiden. Benedek's Vorschlag für die Einrichtung einer Mykotheka in Chlorallactophenol in Analogie mit dem Herbarium für Blütenpflanzen, mag eine lang gefühlte Lücke im mykologischen Laboratorium beseitigen.

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Résumé La méthode deCastellani, la culture en d'eau des champignons microscopiques, a été ré-examinée. Elle pouvait être confirmée en tous les détails. C'est une méthode idéale, au moins, pour la collection de cultures de petite dimension, pour éviter les transferts continus et d'intervalle courte.La suggestion deBenedek pour établir une mycothèque en chlorallactophénol, par analogie avec l' herbier pour les phanérogames peut remplir une lacune, existante pour long temps, aux laboratoires mycologiques.

     

The in vitro production of an anthocyanin from callus cultures of Oxalis linearis

Callus growth and the production of anthocyanins were sustained on the salts and vitamins of Murashige and Skoog. Callus growth was stimulated at a concentration of 8–32 M -naphthaleneacetic acid (NAA) and 2,4-dichlorophenoxyacetic acid (2,4-d). Benzyladenine (BA) and zeatin at 8 M inhibited callus growth whereas isopentenyladenine (iP) stimulated callus growth. NAA repressed anthocyanin production with an increase in NAA from 8–32 M. Anthocyanin synthesis was promoted by an increase in 2,4-d from 0.5 to 2 M and decreased thereafter up to a concentration 32 M 2,4-d. A concentration of 8 M BA, thidiazuron and zeatin, respectively stimulated pigment production. Sucrose stimulated callus growth at 60 mM and pigment production at 120–360 mM.Abbreviations BA 6-benzyladenine - 2,4-d 2,4-dichlorophenoxyacetic acid - NAA -naphthaleneacetic acid - iP isopentenyladenine - TZ thidiazuron-N-phenyl-N-1,2,3-thiadiazol-5-yl-urea - Bu-HCl Butanol-2N HCl - BAW Butanol-acetic acid-water     

The neurosecretory system of the adult Calliphora erythrocephala

Summary In continuation of previous light microscopical investigations using darkfield microscopy of the living cells and sections stained with paraldehyde-fuchsin, an electron microscopical study of the medial neurosecretory cells (m.n.c.) of virgin females of Calliphora has been performed. The neurosecretory material consists of elementary granules corresponding in quantity to the amount of secretory material found by the two other methods in flies of the same age and kept on the same diet. The majority of the cells (m.n.c. I) contain granules measuring c. 2000–3000 Å, while fewer cells (m.n.c. II) show a smaller granular diameter (c. 1000–1500 Å). Due to the Tyndall effect the elementary granules are visible when using darkfield microscopy.The granules were seen to be pinched off from the Golgi complexes. These are numerous and well-developed, except in the less active m.n.c. I of the six days old sugar-flies. The reticulum and mitochondria are described. Axoplasmic channels were observed in the m.n.c. I, probably corresponding to structures found by Wigglesworth (1959 and 1960) in other insect neurons with another technique.The fine structure of the giant neurons and the vacuolated cells has been studied, the observations supporting the conclusions of M. Thomsen in a light microscopical study (1965). Lacunae in the ramifying glia are interpreted as belonging to the glial lacunar system described by Wigglesworth (1960).Dedicated to the memory of Ernst Scharrer (1905–1965), pioneer in the study of neurosecretion.We are grateful to the Carlsberg Foundation and the State General Scientific Foundation for financial support.     

Pleiotropic transport mutants of Escherichia coli lack porin, a major outer membrane protein.

Summary Four pleiotropic transport mutants of Escherichia coli B/r with decreased affinity for the uptake of most nutrients were found to lack a major outer membrane protein of 36,500 daltons (porin) previously shown to produce transmembrane diffusion channels in in vitro reconstitution experiments. Consequent decrease in outer membrane permeability was confirmed by measuring the transmembrane diffusion rate of 6-aminopenicillanic acid. Quantitative considerations on the porin-dependent permeability of the outer membrane show that (a) there may be very large differences in the actual rates of penetration, even among the permeable substances and (b) the numbers of porin molecules present in wild type cells is several orders of magnitude higher than that necessary for the uptake of rapidly diffusing substrates such as glocose from ordinary culture media. The absence of porin and the pleiotropic transport defect were always contransduced, and the mutation was mapped at 73.7 min between aroB and malT by P1 transduction. When revertants able to grow on low concentrations of lactose were selected, in addition to true revertants suppressor strains with increased amounts of non-porin membrane proteins were isolated.This paper corresponds to paper XVI of the series dealing with the bacterial outer membrane from the laboratory of H.N. The preceding paper in the series is Nikaido, Bavoil, and Hirota, J. Bacteriol., in press     

Zur Entstehung des Golgi-Apparates. Elektronenmikroskopische Untersuchungen an Spermatiden von Eisenia foetida (Annelidae)

Zusammenfassung An Spermatiden von Eisenia foetida wurde die Zelldifferenzierung zu Beginn der Spermiohistogenese untersucht. Die mit der zentralen, kernlosen Nährmasse, dem Cytophor, ein Syncytium bildenden Spermatiden besitzen zu Anfang der Zelldifferenzierung Vesikel, die im peripheren Kernbereich und im Lumen der perinucleären Zisterne liegen. Die Zellmembran am Verbindungsstück zum Cytophor trägt einen fuzzy coat, die Spermatidenmembran weist deutliche coated vesicles auf. Der Golgi-Apparat entsteht am prospektiven Schwanzende durch Auswachsen der ersten vesikelbildenden Lamelle aus der Kernmembran, die zuvor typische morphologische Veränderungen durchmacht. Die weiteren Lamellen eines Dictyosoms bilden sich durch Umfaltung der zuvor entstandenen. Anschließend werden an der Kernmembran die Mitochondrien differenziert. Die Genese der Cristae und der äußeren Mitochondrienmembran bleibt problematisch. Im Cytophor findet man annulierte Lamellen, degenerierte Golgi-Apparate und vesikelhaltige, abgeschnürte Zisternen des ER. Diese Formationen werden im Zusammenhang mit einer Reduktion des Cytoplasmas von den Spermatiden an das Cytophor abgegeben.

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On the origin of the golgi-apparatus. electron microscopical observations on spermatids of Eisenia foetida (Annelidae)

Summary The early cytodifferentiation is studied in developing spermatids of Eisenia foetida. The spermatids are attached to a central, anucleate mass of cytoplasm, the cytophore. They contain vesicles in the periphery of their nuclei and the perinuclear space. The cell membrane of the connecting bridge to the cytophore is covered with a fuzzy coat. The spermatid membrane is provided with coated vesicles. The Golgi apparatus develops at the prospective tailside by outgrowth of a vesicle producing lamella from the nuclear membrane, which before has undergone typical structural changes. Further dyctiosomal lamellae are formed by folding of already existing lamellae. Finally the mitochondria adhering to the nuclear membrane differentiate. The genesis of the cristae and the outer mitochondrial membrane remains problematical. In the cytophore there are annulate lamellae, degenerated dictyosomes and vesicle-containing cisternae of the endoplasmic reticulum. These formations are transferred from the spermatids to the cytophore, thus reducing the volume of their cytoplasm.

Herrn Prof. Dr. med. J. Staubesand danke ich für wertvolle Hinweise und Kritik.     

The role of β-alanine in the biosynthesis of nitrate byAspergillus flavus

d-Serine (0.05m) inhibited nitrification byAspergillus flavus in media containing either peptone, aspartate,a-alanine or -alanine as the sole nitrogen source. A similar inhibition was observed in an aspartate + peptone medium, but nitrate was formed in a -alanine + peptone medium in the presence of the inhibitor. Exceptionally high yields of nitrate were obtained in the -alanine + peptone medium. In replacement cultures,d-serine inhibited nitrification of aspartate but not of -alanine. Manometric studies indicated that aspartate was decarboxylated byA. flavus and that the reaction was inhibited byd-serine. In view of these results, it is suggested that aspartate is a precursor and -alanine is an intermediate in nitrification by this fungus.