Studies on the fine structure of puffs in Sciara coprophila

Fine structure of RNA and DNA puffs of Sciara coprophila was studied during late developmental stages of the fourth larval instar. In RNA puffs the predominant structure seen seems to be a diffuse, lampbrush-like thread or threads sectioned in a variety of planes. The thread is composed of filamentous and granular material. Three types of RNA puffs, each with a slightly different morphology, are found. In their development DNA puffs pass through a precise sequence of stages, each with its distinct morphologic and metabolic characteristics. At the initial and final stages, when much of the puff chromatin is in the compacted state, DNA puffs resemble condensed chromosomal bands. In contrast, at stages when most chromatin is diffuse, DNA puffs share many structural characteristics of RNA puffs. Most of the expanded puff area is permeated by lampbrush-like threads composed of fibrils and granules. RNA and DNA puffs were compared with respect to granule size and distribution by means of electron micrographs of known magnification. The results of the statistical analysis show that: 1) The coefficient of variation (C.V.) of the method of measurement falls between 5 and 7%. 2) There is a fluctuation in granule sizes within each puff with a C.V. of 24–26%. 3) The average granule diameter is 238 Å for DNA puffs and 310 Å for RNA puffs; the difference is statistically significant. 4) The variation in mean granule size in a sample of DNA puffs is rather small (C.V. 12%), while the variation in granule size between different RNA puffs is somewhat larger (C.V. 20%). 5) The relative spread of granule sizes in DNA puffs is more restricted than that in RNA puffs. It is evident then that, on the average, DNA puff granules are smaller and more uniform than granules found in RNA puffs.     

Ultrastructural localization of cell wall teichoic acids in Streptococcus faecalis by means of concanavalin A

Some teichoic acids are known to be partially substituted by α-D-glucopyranosyl residues such as the teichoic acids of Streptococcus faecalis NCIB 8191. They will, therefore, bind specifically the phytohemagglutinin concanavalin A. Concanavalin A labelled with mercury or colloidal gold coated with concanavalin A has been used to mark isolated cell walls in order to localize the teichoic acids at the ultrastructural level. Besides these two direct marking techniques, the indirect concanavalin A-peroxidase technique (localization of peroxidase by the diaminobenzidine method followed by postosmication) has been applied to thin sections of premarked cells. All three methods gave almost identical results, namely, a dense and homogeneous distribution of the cell wall teichoic acids. In control experiments total inhibition was achieved in the presence of methyl-α-D-mannopyranoside. After trichloroacetic acid or alkali extraction of the teichoic acids from isolated walls no marking could be detected.     

Aspekte der Siebröhren-Differenzierung bei Monocotylen

Zusammenfassung Ausgehend vom nachmeristematischen Zustand einer prospektiven Siebröhre machen Kern, Tonoplast, ER, Dictyosomen und Ribosomen wÄhrend der Zelldifferenzierung folgende Umwandlungen durch:DerKern erfÄhrt in seiner Matrix eine zunehmende Substanzverarmung, an deren Ende seine vollstÄndige Auflösung steht. In der Kernmembran sind lokale Erweiterungen des intrazisternalen Raumes charakteristisch für den Beginn der degenerativen VerÄnderungen. Stets bleibt die Kernhülle jedoch in lockerem Kontakt mit dem ER; vermutlich geht sie zum Abschlu\ der Kernauflösung in das Membransystem des ER ein.In AbhÄngigkeit von der Rückbildung des Zellsaftraums zerfÄllt derTonoplast zunÄchst in zisternenartige Abschnitte, um spÄter in der ausdifferenzierten Siebröhre ganz zu verschwinden. Wie an verschiedenen Beispielen abzulesen ist, werden diese VerÄnderungen offensichtlich auf dem Höhepunkt der protoplasmatischen Differenzierung eingeleitet.Das gleichmÄ\ig granulÄr-zisternaleEndomembransystem junger Siebelemente — hÄufig charakterisiert durch spiralig aufgereihte Ribosomen — wandelt sich mit zunehmender Hydratisierung des Protoplasten in tubulÄre bis vesikulÄre Elemente um und geht zum anderen über verschiedene Zwischenstufen in gitterartige Membranstrukturen ein.Die in der jungen Siebröhre zahlreichen, polar gebautenDictyosomen tragen mit der Produktion von Golgi-Vesikeln zum Wandaufbau bei. Im Laufe der Zelldifferenzierung werden in steigendem Ma\e auch coated vesicles abgegeben, deren Bedeutung noch unklar ist. In der ausdifferenzierten Siebröhre sind weder Dictyosomen noch die von ihnen abgegebenen Vesikel nachzuweisen. Dieses Schicksal teilen sie mit denRibosomen, die gleichfalls nur in jungen Siebelementen anzutreffen sind und dort zu Polysomen vereint, hÄufig in spiraliger oder helixförmiger Anordnung vorkommen.

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Aspects of sieve-tube differentiation in monocotyledons

Summary Subsequent to the postmeristematic state of a future sieve-tube its nucleus, tonoplast, endoplasmic reticulum, dictyosomes, and ribosomes undergo the following transformations:Thenucleus shows a gradually declining ground substance until at last it entirely disintegrates. Local enlargements of the intracisternal space of the nuclear membrane indicate the beginning of the degenerative phase. During all these alterations as well as in the meristematic state the nuclear envelope is in loose contact with the endoplasmic reticulum. We suppose that it finally becomes part of a complex ER system. Subject to the disorganization of the central cell-sap room thetonoplast initially removing from the parietal protoplast later on completely disappears. According to our results these transformations are obviously introduced at the climax of protoplasmatic differentiations.The uniform rough surfaced cisternal ER of young sieve elements—often characterized by spirally arranged ribosomes—changes during sieve-tube ontogeny to tubular or vesicular elements and partly fuses to lattice-like membrane structures passing several other stages of membrane condensation.Polardictyosomes that are numerous in young sieve-tubes produce coated vesicles besides smooth golgi-vesicles.Ribosomes are abundant in young elements, too, they are often found as polysomes of helical or spiral arrangement. Neither dictyosomes nor ribosomes have been seen in differentiated sieve-tubes.

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Teil einer Habilitationsschrift der Math.-Naturw. FakultÄt Bonn.     

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.     

Cytological and autoradiographic studies in Sciara coprophila salivary gland chromosomes

Summary Morphological and metabolic changes on the salivary chromosomes of Sciara coprophila were followed during the later half of the fourth larval instar.Cytological maps were prepared for five successive stages from mid-fourth instar to the prepupal stage. These maps, which constitute a revision of those published earlier by Crouse, summarized our cytological findings and were the basis for studies on DNA replication of these chromosomes.Similar to earlier studies in Chironomidae, differences in the puffing pattern were noted between the anterior and the posterior portions of the salivary gland. The most striking difference was noted in region 2B on chromosome III which produces a large puff only in nuclei from the anterior part of the gland. Other autosomal puffs, although present in both parts of the gland, showed constant differences in size.An increase in the number of bands from mid-fourth to late fourth instar was observed. The new bands are all of the light-staining kind.In Sciara the puffed area may include a large number of bands in addition to the bands which originated the puff. The maximal extent of puffs was determined in terms of chromosomal map regions and the number of bands subject to obliteration.In the autoradiographic experiments use was made of H³-thymidine as DNA precursor. The aim of these studies was to detect any asynchronies in the replication time of bands. In fact, marked differences in the relative rates of uptake of H³-thymidine of a number of bands in a certain proportion of chromosomes have been observed, while others showed uniform incorporation. Since these latter were found with higher frequency the period of uniform labeling must comprise a larger part of the replication cycle then the periods of localized labeling. To assess the validity and constancy of the observed patterns of unequal incorporation, a semiquantitative analysis was carried out. It showed that the bands showing localized uptake may be separated into two broad groups. In one of these groups are the centromere regions and certain chromosomal ends, which are presumably heterochromatic. The other group comprises most of the puff sites and bulbs. Since late replication is characteristic of heterochromatin, we assumed that bands of the former group (C) replicate late in the cycle, while puffs and bulbs start replication early, and the period of equal labeling is intermediate. Other intermediate labeling patterns were observed and are described.It is known that in the fourth instar from two to three DNA replications occur in the salivary gland nuclei, the last of which coincides with puffing. Several stages may be distinguished in the puffing process based on morphology and rates of isotope uptake of the puffs. The first sign of puffing is a very high rate of incorporation at puffs. It is maintained throughout this last DNA synthesis period and only declines when all other chromosomal regions have ceased to replicate. A pattern of high and exclusive uptake at the heterochromatic sites (pattern C) was never observed in this replication; instead puffs are the last regions to terminate DNA synthesis.These results are discussed in relation to several current problems, such as, asynchronous DNA replication, the problem of metabolic DNA, and the concept of the heterochromatic state.Submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy, in the Faculty of Pure Science, Department of Zoology, Columbia University, New York. This work has been supported by U.S. Public Health Training Grant No. 2Tl-GM-216-05; partial support has been received also from Grants GB 42 and G-14043 from the National Science Foundation to Dr. H. V. Crouse.