Struktur und quantitatives Verhalten des perimitochondrialen granulären endoplasmatischen Retikulums der Mäuseleber

Zusammenfassung In normalen Leberzellen der Maus wurde das quantitative Verhalten des perimitochondrialen granulären endoplasmatischen Retikulums untersucht. 74% der Mitochondrien zeigen Beziehungen zum granulären endoplasmatischen Retikulum. Die einzelnen Mitochondrien werden zu 52%19,5 von den Membranen des granulären endoplasmatischen Retikulums bedeckt. Je Mikrometer Membranstrecke sind auf der mitochondriennahen Seite des granulären endoplasmatischen Retikulums 21 und auf der mitochondrienfernen Seite 20 Ribosomen zu finden, was der Zahl im übrigen granulären endoplasmatischen Retikulum entspricht. Die Befunde stellen die Grundlage für Untersuchungen des perimitochondrialen granulären endoplasmatischen Retikulums unter pathologischen Bedingungen dar.

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Structure and quantitative behaviour of the perimitochondrial granular endoplasmic reticulum in the liver cells of the mouse

Summary The perimitochondrial granular endoplasmic reticulum in normal mouse liver cells has been investigated quantitatively. 74% of the mitochondria are in association with the granular endoplasmic reticulum. The individual mitochondrion is covered by the membranes of the granulated E. R. in 52%19.5. The outer surface of the endoplasmic membrane, facing the mitochondrion, is occupied by 21 ribosomes per m; the corresponding surface of the membrane facing the free cytoplasm is occupied by 20 ribosomes per m. These data are in agreement with those of that fraction of the E. R., which is not in association with mitochondria. These findings represent a basis for investigations of the perimitochondrial endoplasmic reticulum under pathological conditions.

     

Geschlechtliche organisation,fortpflanzungsverhalten und ursachen der sexuellen vermehrung von Stenostomum sthenum nov. spec. (Turbellaria,Catenulida)

Certain temperatures and H-Ion concentrations are necessary for the development of male and female reproductive organs. The differentiation of the reproductive system from undifferentiated cells conforms precisely with data on other species of Stenostomum.

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Abkürzungen in den Abbildungen b Bindegewebszelle - c Cilien - cy Cytoplasma - d Darm - de Ductus ejaculatorius - dl Darmlumen - do Dotter - dr Drüsenzellen - lr Lÿckenraum - m Mundöffnung - n Nucleolus - np Nephroporus - o Ovar - p männlicher Genitalporus - pa parenchymatischer Raum - pf periembryonale Flüssigkeit - dse dorsale Epidermis - e dorsolaterale Epidermis - ed extraembryonaler Dotter - ee Epidermiseinstülpung - eh Epidermis +Hautmuskelschlauch - em Embryo - ex Exkretvakuole - f Freßzelle - g Gehirn - ga Gehirnanlagen - gr Granula - h Hautmuskelschlauch - hz Hüzllzelle - kl Versehlußkegel/Klebkegel - in indifferente Zelle - k Kern - kdr Klebdrüsenzelle(n) - kk kollabierter Kanal - kö Körnerkolbenzelle - kp Kopulationsorgan - ku Kufen - kw Körperwand - l Lichtsinnesorgan - li Linsenkörper - lk lichtbrechende Konkremente im Darmgewebe - ph Pharynx - phr Pharynxregion - pr Zentralkanal des Protonephridialsystems - ps Präspermatide - pu Punktfeld - q Zellquartett - r Riechgruben - rh Rhombuszellenband - rhb Rhabditen - rm Radiärmuskelzelle - rhs Rhabditenschleim - rs resorbierende Darmzelle(n) - s Schale - sc Spermatocyten - se Sekretgang - t Tastborste - ta Auflösungsprodukte des Hodens - te Hoden - to Terminalorgane(e) - tz Teilungszone - v Vakuole - w vermutliches Rudiment des weiblichen Genitalporus     

Feinstrukturen der Mikrokörper (Microbodies) des proximalen Nierentubulus

Zusammenfassung Die Feinstrukturen der Mikrokörper des Nierenepithels werden beschrieben und mit denjenigen der Leber-Mikrokörper verglichen. Als besondere Charakteristika der Nieren-Mikrokörper sind eine (nicht kristalline) nucleoide Verdichtung und eigentümliche stabförmige Ausstülpungen (Stäbe) anzusehen. Die Stäbe stellen unterschiedlich lange Zylinder mit einem Durchmesser von 100 nm dar. Im Inneren findet sich eine unmittelbar der Membran anliegende, ring- oder spiralförmig angeordnete, granuläre Struktur (Granula-Durchmesser 50 Å), die in Stablängsschnitten eine Querstreifung vortäuscht. Es wird eine in Phasen ablaufende Bildung der Mikrokörper-Stäbe angenommen: ein in der Matrix entstandener Granula-Zylinder hebt sich aus dem Mikrokörper heraus, wobei die Mikrokörper-Membran entsprechend vorgebuchtet wird, und wächst schließlich zu einem eigenständigen, allseits membranumzogenen Stab aus. Die Möglichkeit, daß die Stäbe von Mikrokörpern abgestoßen werden, wird diskutiert. — Die Segregation von Mikrokörpern in Vakuolen wird nicht als aktive Beteiligung an lytischen Prozessen, sondern als autophagischer Vorgang gedeutet.

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Fine structure of microbodies in proximal tubular epithelium of the kidney

Summary Ultrastructural observations on microbodies in normal proximal tubule cells of the rat kidney are described and compared with microbodies of hepatic parenchymal cells. After fixation in osmium tetroxide with phosphate buffer the special features of the renal microbodies are the non-crystalline nucleoid and protrusions (rods) extending from the main body. These rods are cylindrical in shape having a diameter about 100 nm and are of varying lengths. Inside the limiting membrane are ring- or spiral-like ordered profiles consisting of granules (about 50 Å in diameter) which often appeared as a row of parallel linear densities arranged at approximately right angles to the long axis of the rod. It can be demonstrated that the parallel linear pattern depends on the projection of the granules in the photographic plane. — The findings suggest that the cylindrical structures of granules are formed in the peripheral matrix of microbodies; in a second phase they are lifted outside, in part enveloped with the membrane of the microbody; in this situation, the protrusions are formed. This form of creation would explain the characteristic excentrical (tangential) relation between protrusions and the main body. The observation that rods are often seen apparently isolated in the cytoplasm without visible connection with a microbody is only discussed hypothetically, because of the plane of sectioning. — Microbodies and rods can be identified in cytosegresomes. These investigations were interpreted as an autophagic degradation and not as an active role of the enzymes of microbodies in digestive mechanisms.

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Herrn Prof. Dr. Helmut Ruska zum 60. Geburtstag gewidmet.

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Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.

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Herrn Doz. Dr. W. Thoenes danke ich für wertvolle Hinweise und für die kritische Durchsicht des Manuskriptes.     

Die Bedeutung des Nervensystems für die Regeneration bei den Insekten

Ohne ZusammenfassungHerrn Prof. Dr. O.Pflugfelder zu seinem 60. Geburtstag.     

BIOSYNTHESIS OF COLLAGEN AND NON-COLLAGEN PROTEIN DURING DEVELOPMENT OF THE CHICK CORNEA

The relationship between the rates of increase of corneal protein fractions and incorporation of labeled precursors has been examined during embryonic and early posthatching development of the chick corneal stroma. Non-collagen protein increased gradually from 9 through 20 days of incubation. Collagen accumulated approximately logarithmically through the 19th day, the most rapid rate occurring between 13 and 20 days of incubation. The rates at which labeled amino acids are incorporated into collagen in vivo and in vitro undergo marked changes during the last week of embryonic development, corresponding closely to the rate of collagen accumulation in vivo; whereas incorporation into non-collagen protein changes much less markedly. Changes in the rate of incorporation of precursors into collagen are not due to changes in the rate of conversion of collagen from the soluble to insoluble form, or to changes in the endogenous amino acid pool size. Chick embryo corneal stroma collagen turns over very slowly, if at all. Non-collagen protein turns over more rapidly. An increase in cell number, as indicated by DNA content, does not account for the increased rate of collagen synthesis between the 9th and 16th day of incubation. It is concluded that the observed changes in collagen synthesis reflect changing activities in the individual cornea fibroblasts. These activities are comparable in the intact tissue in vivo and in isolated corneas in vitro.     

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.     

Epidemiological analysis of acute respiratory diseases (ARD) of viral etiology in the Czech Socialist Republic (CSR] and German Democratic Republic (GDR) over the period 1979 to 1984

An analysis is made of the ARD reported in CSR and the GDR over the period July 1st, 1979 to June 30th, 1984. During that time, there were 27,810,000 cases reported in CSR in the framework of ARD epidemiological surveillance, representing 2.67 cases per one inhabitant, whereas in the GDR, the total number of reported ARD was 28,900,000 yielding 1.73 cases per person. However, the GDR reported higher morbidity per one child of preschool age. The authors believe that the differences in the reported incidence of ARD between the two countries are due to differences in the reporting systems and medical officers' activity during an epidemic and in the interim period. Approximately one third of ARD reported annually in the two countries falls to the period of influenza epidemics. The authors also analyze the etiology of the influenza epidemics which affected the two countries in 1980, 1981, 1982, 1983 and 1984. In most seasons, the causative agents and morbidity excesses were different in the two countries. The drift variant B/USSR/100/83, which caused a major epidemy in CSR in 1984, has not to date been implicated in the DGR in the etiology of ARD. The cyclic epidemic due to Mycoplasma pneumoniae occurred in the GDR already in 1979-80, while CSR experienced it a year later. There was a temporal and territorial correlation between the course of A(H1N1) influenza epidemic in the two countries in 1984.     

Scavenging of Hydrogen Peroxide in the Endosperm of Ricinus communis by Ascorbate Peroxidase

The fat-storing endosperm of Ricinus communis L. was found tocontain an ascorbate peroxidase (EC 1.11.1.11 [EC] ), which is nearlyas active as catalase (EC 1.11.1.6 [EC] ) in degradation of hydrogenperoxide (H₂O₂) at its physiological concentrations. This ascorbateperoxidase probably functions together with monodehydroascorbatereductase (EC 1.6.5.4 [EC] ) or dehydroascorbate reductase (EC 1.8.5.1 [EC] )and glutathione reductase (EC 1.6.4.2 [EC] ) to remove the H₂O₂ producedduring the transformation of fat to carbohydrate in the glyoxysomes.The activities of these enzymes as well as the content of ascorbateand glutathione increase parallel to the activities of glyoxysomalmarker enzymes during the course of germination. Inhibitionof catalase by aminotriazole results in increases of the ascorbateperoxidase activity and of the glutathione content. All fourenzymes are predominantly localized in the cytosol of the Ricinusendosperm with low activities found in the plastids and themitochondria. The results suggest, that the ascorbate-dependentH₂O₂ scavenging pathway, which has been shown to be responsiblefor the reduction of photosynthetically derived H₂O₂ in thechloroplasts, operates also in the Ricinus endosperm. (Received June 5, 1990; Accepted July 31, 1990)     

Biosynthesis of digalactosyldiacylglycerol in plastids from 16:3 and 18:3 plants

Intact chloroplasts isolated from leaves of eight species of 16:3 and 18:3 plants and chromoplasts isolated from Narcissus pseudonarcissus L. flowers synthesize galactose-labeled mono-, di-, and trigalactosyldiacylglycerol (MGDG, DGDG, and TGDG) when incubated with UDP-[6-³H]galactose. In all plastids, galactolipid synthesis, and especially synthesis of DGDG and TGDG, is reduced by treatment of the organelles with the nonpenetrating protease thermolysin. Envelope membranes isolated from thermolysin-treated chloroplasts of Spinacia oleracea L. (16:3 plant) and Pisum sativum L. (18:3 plant) or membranes isolated from thermolysin-treated chromoplasts are strongly reduced in galactolipid:galactolipid galactosyltransferase activity, but not with regard to UDP-Gal:diacylglycerol galactosyltransferase. For the intact plastids, this indicates that thermolysin treatment specifically blocks DGDG (and TGDG) synthesis, whereas MGDG synthesis is not affected. Neither in chloroplast nor in chromoplast membranes is DGDG synthesis stimulated by UDP-Gal. DGDG synthesis in S. oleracea chloroplasts is not stimulated by nucleoside 5′-diphospho digalactosides. Therefore, galactolipid:galactolipid galactosyltransferase is so far the only detectable enzyme synthesizing DGDG. These results conclusively suggest that the latter enzyme is located in the outer envelope membrane of different types of plastids and has a general function in DGDG synthesis, both in 16:3 and 18:3 plants.     

Involvement of ferredoxin in desaturation of lipid-bound oleate in chloroplasts

Intact spinach (Spinacia oleracea) chloroplasts, pulse-labeled with [¹⁴C]acetate, desaturate newly formed fatty acids as ester groups of monogalactosyl diacylglycerol in a subsequent chase in the dark. Rupture of pulse-labeled chloroplasts by addition of a detergent solution 3-([3-cholamidopropyl]dimethylammonio)-1-propane sulfonate preserves part of this desaturation activity. Direct addition of different free fatty acids together with appropriate cofactors to detergent-ruptured chloroplasts results in fatty acid labeling of monogalactosyl diacylglycerol. During subsequent incubation these lipid-linked fatty acids are desaturated, i.e. 18:1 to 18:2 and 18:3 and to a small extent also 16:0 to 16:3. The formation of 18:2 was also observed after incorporation of 18:1 into sulfolipid and phosphatidyl choline. Density gradient centrifugation separated a membrane fraction from detergent-ruptured chloroplasts which in the presence of appropriate cofactors incorporated 18:1 and 18:2 into the above-mentioned lipids. In the light, desaturation was dependent on added ferredoxin, whereas in the dark, in addition to ferredoxin NAD(P)H was also required. Preliminary evidence for the involvement of membrane-bound ferredoxin:NADP oxidoreductase (FNR) as a third component of desaturation in the dark was obtained by inhibitor studies including antibodies against FNR. Desaturation of lipid-bound 18:1 and 18:2 resembles stearoyl-ACP desaturation with respect to its requirement of reduced ferredoxin and oxygen.