On the presence and character of the nucleolus in the spermatid and fecundant spermatozoons of heteropachyloidellus robustus roew

Summary The presence of the nucleolus and the nucleolonema in the spermatozoons of a specie of arachnides (Heteropachyloidellus robustus Roew.) is shown in this paper. This fact confirms the thesis supported in a previous work that the nucleolonema is a permanent cell structure.During the maturative process of the spermatid the nucleolus does not disappear, and only its morphology changes. After proving that the intranuclear body found in the spermatozoon is really the nucleolus, and after showing that this body is present in the fecundative spermatozoons removed from the female spermathecae, it is concluded that the nucleolus is carried to the egg together with the chromosomic material.     

On the action of ribonuclease in salivary gland cells of drosophila

Summary It was shown that ribonuclease degrades the nucleolus in actively metabolizing cells. It does this without inhibiting RNA synthesis in the puffs and the nucleolus organizer. DNA synthesis still continues before or after puff formation, while amino acid incorporation is inhibited before the puffs are formed, indicating pre-existence of proteins involved in the process of puff formation.Dedicated to Professor H. Bauer on the occasion of his 60th birthday. — Research sponsored jointly by the U. S. Atomic Energy Commission under contract with the Union Carbide Corporation, the International Laboratory of Genetics and Biophysics, Naples, Italy (Partially supported by Euratom), and the Whitman Laboratory, University of Chicago, Chicago, Illinois.Supported by NTH Postdoctoral Fellowship 2-F 2–6 M-17, 187-02.     

Autoradiographische Untersuchungen zur funktionellen und pathologischen Kernschwellung in der Rattenleber nach Fütterung von Thioacetamid

Summary In the liver parenchyma of rats a markedly increase of the nuclear volume occurs after the application of thioacetamide. According to autoradiographic and morphological criterions this nuclear enlargement can be separated into a functional and a pathological swelling. In case of the functional swelling the rate of incorporation for H³-phenylalanine and H³-cytidine in the karyoplasm — as a measurement for protein and ribonucleic acid metabolism — is proportional to the nuclear volume. But in case of the pathological swelling the incorporation of these precursors is very much lower. In the functional swollen as well as in normal nuclei the volumetrical nucleolus/karyoplasm ratio can serve as an expression of the synthetic activity of both nuclear components. Yet this relation does not exist in pathological swollen nuclei. Therefore the functional swelling of the nucleus in comparison with the normal synthetic activities of the nucleus (karyoplasm + nucleolus) is an increased stage of function with completely preserved nuclear productivity. The pathological swelling however is a decreased state of function with a decompensated nuclear capacity.

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Die autoradiographischen Arbeiten wurden im Institut für Medizinische Isotopenforschung der Universität Köln (Leiter: Prof. Dr. W. Maurer) durchgeführt, für die Synthese der tritiierten Aminosäure danken wir Herrn Dr. Dr. K. Hempel.

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Die Arbeit lag 1963 der Medizinischen Fakultät der Universität Würzburg als Teil einer Habilitationsschrift vor. Sie wurde durch Mittel des Bundesministeriums für Wissenschaftliche Forschung ermöglicht.     

Cytochemical and ultrastructural studies of ribonucleoprotein containing structures in oocytes of Acheta domesticus

Summary Two distinct types of ribonucleoprotein containing structures are found in oocytes of the house cricket, Acheta domesticus, a large secondary or accessory nucleolus and many small primary nucleoli. The secondary nucleolus increases in size during oocyte development and is similar in appearance to the nucleolus of somatic cells. The primary nucleoli are intimately associated with a large, extrachromosomal DNA containing body. The DNA body is no longer visible in nuclei of late diplotene stage cells when the primary nucleoli are dispersed within the nucleoplasm. Both types of nucleoli contain cytochemically detectable RNA and acid protein, little or no DNA and basic protein, and particulate structures similar to but smaller than cytoplasmic ribosomes.The authors acknowledge the technical assistance of Miss Celeste Malinoski and Mrs. Marcia Andrews. This work was supported by a U.S.P.H.S. grant, number GM-16440-01 and grants number L-16 and J-1 from the Health Research Services Foundation.     

Formation,transport, and storage of ribonucleic acid containing structures in oocytes of Acheta domesticus (Orthoptera)

Summary Oocyte development of Acheta domesticus was investigated morphologically and cytochemically. The studies demonstrated a size decrease and final disappearance of a large extrachromosomal DNA body in the nuclei as the cells proceeded through the diplotene stage of meiosis. The body was surrounded by fascicles of RNA containing material. This material remained within the nuclei in individual packets after the DNA body was no longer detectable. An active nucleo-cytoplasmic migration of RNA was seen prior to the disappearance of the DNA body. After the disappearance of the body very little migration was detected. Evidence was presented to demonstrate the ribosomal nature of this migratory RNA. The RNA packets remaining in nuclei of cells arrested in the diplotene stage of oogenesis functioned as storage depots for ribosomal RNA.The authors acknowledge the technical assistance of Miss Agnes Cralley. Dr. D. Ammermann, Zoologisches Institut, Tübingen, W. Germany, provided the animals used in this study. — This work was supported in part by a Health Research Services Foundation Grant No. J-1 and in part by U.S.P.H.S. Grant No. 7-FZCA-23,971-O1A1.     

Analysis of nucleolus organizer regions (NORs) in mitotic and polytene chromosomes ofPhaseolus coccineus by silver staining and Giemsa C-banding

Chromosomes with active nucleolus organizer regions (NORs) were visualized in root tip metaphases ofPhaseolus coccineus using the silver staining technique. A mean number of 5.5 Ag-NORs per cell was observed in 54 cells from eight plants. In the endopolyploid nuclei of the suspensor the silver technique did not demonstrate the reported specificity for nucleolus organizer activity, because there was usually pale staining of nucleoli and preferential staining of heterochromatic regions in the polytene chromosomes including pericentromeric material, telomeres and NORs. The mean number of NORs per nucleolus as detected by this method was 5.8 (28 nucleoli analysed). Using a modified preparation technique, giant chromosomes stained pale, but nucleoli of suspensor cells displayed darkly silver staining internal domains, each of which originating from a nucleolus organizer.—Giemsa C-banding of endopolyploid suspensor nuclei revealed C-positive nucleolus organizers with darkly staining intranucleolar fibrils. The latter were frequently involved in inter-NOR associations. In 34 nucleoli analysed, the mean number of Giemsa C-positive NORs per nucleolus was 6.0.Dedicated to Professor Dr.Lothar Geitler on the occasion of his 80th birthday.     

Der DNS-Gehalt von Kotyledonen und Hypokotyl des Senfkeimlings (Sinapis alba L.) bei der phytochromgesteuerten Photomorphogenese

Zusammenfassung Der DNS-Gehalt und somit auch die Zellzahl von Kotyledonen und Hypokotyl des Senfkeimlings sind zwischen 36 und 60 Std nach Aussaat sowohl im Dunkel als auch im Dunkelrot weitgehend konstant. Die durch Phytochrom bewirkte Zunahme des RNS-und Proteingehaltes (Weidner u. Mitarb., 1965) muß deshalb als eine RNS-bzw. ProteinZunahme pro Zelle aufgefaßt werden. Dieser Befund stützt die Vorstellung einer differentiellen Genaktivierung durch P₇₃₀ (z.B. Mohr, 1966).—Die weitgehend konstante Zellzahl von Hypokotyl und Kotyledonen während des von uns verwendeten Experimentierzeitraumes ist eine wichtige Voraussetzung für die Vergleichbarkeit biochemischer Daten, z.B. bei kinetischen Studien (vgl. Mohr, 1966).

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The DNA contents of cotyledons and hypocotyl of the mustard seedling (Sinapis alba L.) during phytochrome-mediated photomorphogenesis

Summary DNA contents and accordingly cell numbers of cotyledons and hypocotyl of the mustard seedling were virtually constant during the experimental period (between 36–60 hours after sowing) in the dark as well as under the influence of P₇₃₀, the active phytochrome (Table).—Therefore the phytochromemediated increase of the RNA and protein contents (Weidner et al., 1965) must be understood as an increase of RNA and protein per cell. This fact is in agreement with the conception of differential gene activation mediated by P₇₃₀ (Mohr, 1966). The virtually constant DNA contents during the period of time which is regularly used for experimentation on photomorphogenesis in our laboratory (36–60 hours after sowing; Mohr, 1966) is an important prerequisite for comparing biochemical data under the point of view of differential gene activation, e.g. in kinetical studies in the dark and under continuous far-red light.

     

Partial inactivation of wheat nucleolus organisers by the nucleolus organiser chromosomes from Aegilops umbellulata

Two pairs of chromosomes (1U and 5U) in Aegilops umbellulata possess ribosomal RNA genes. This has been proven by studying wheat plants into which 1U and 5U chromosomes have been introduced separately. These plants have more ribosomal RNA genes than the recipient wheat plants and additional clusters of rDNA when examined by in situ hybridisation. The repeating rDNA unit in Aegilops umbellulata is longer than most of the units in the wheat variety Chinese Spring, the additional DNA probably being in the non-transcribed spacer. This was determined from restriction endonuclease maps of rDNA. In Chinese Spring plants possessing 1U or 5U chromosomes, the largest nucleoli are formed on 1U or 5U chromosomes and the wheat nucleolus organisers form micronucleoli. This is not because the nucleolus organisers on chromosomes 1U and 5U have many more rRNA genes than wheat nucleolus organisers. It is suggested that the Aegilops umbellulata nucleolus organisers are dominant over those of wheat because they compete more effectively for some limiting factor. — The partial inactivation of the wheat nucleolus organisers by chromosomes 1U or 5U does not result in a reduced total nucleolus volume in root tip or pollen mother cells, because of the compensation by the nucleolus organisers of chromosomes 1U or 5U. The amount of RNA in seedlings is not markedly affected by the partial inactivation of the wheat nucleolus organisers.     

Three-dimensional architecture of the higher plant nucleolonema disclosed on serial ultrathin sections

Summary— The three-dimensional architecture of the nucleolonema of Vicia faba has been studied by applying a silver impregnation technique to serial ultrathin sections. This technique disclosed lateral and transverse segments of the nucleolonema which were heavily impregnated with silver. The lateral profiles of the nucleolonema segments were classified into three main categories; a segment made up of one to several rod-like filaments (type I); a ladder-like segment consisting of two parallel and of transverse filaments (type II); and a last type constructed from two parallel filaments (type III). Tracing of the lateral segments through serial sections has indicated that type I first appears, then either type II or III and finally type I reappears at the corresponding sites on sections. Types II and III remained constant in width, about 1.0 μm, along their longitudinal axes whereas the width of type I was significantly smaller than that of the two former. The lateral filaments of both types II and III showed heterogeneity in width on account of the presence of knobs intermittently distributed along them. The thickness of these knobs was about 0.35 μm. Combining the observations on serial ultrathin sections and the morphometrical data it is very probable that the elementary structure of the nucleolonema is a 0.35-μm thick filament that tightly coils up into a solenoid structure with a thickness of approximately 1.0 μm. This model can explain the appearance of open- and closed-argyrophilic rings in serial sections since transverse segments of the solenoid are expected to show the argyrophilic rings. The elementary filament of the nucleolonema solenoid was sometimes loosened. Judging from our cytochemical data at the electron microscope level, some argyrophilic proteins appear to reside in the axial space of the solenoid but both DNA and RNA were not detectable in this space.     

Multiplication of nucleolar fibrillar centres and absence of rDNA amplification in mouse ooctye during meiotic prophase I

During meiotic prophase I the nucleolus of the mouse oocyte assumes a reticulate structure of ‘nucleolonema’ type. This change coincides with the appearance of several secondary fibrillar centres. The number of these centres at diplotene (97–113), largely exceeds that of nucleolar organizers (4c DNA = 20 NORs). The quantitatative analysis of autoradiographs after hybridization in situ with -³H-uridine labelled rRNA, enabled us to demonstrate that the multiplication of the fibrillar centres in mouse oocyte nucleolus during meiotic prophase I is not the result of an amplification of the rDNA. The number of silver grains in pachytene and diplotene nuclei was twice that counted for somatic cell and oogonium nuclei (2c DNA).     

Control of stem-gall of coriander by fungicides

Summary The stem-gall caused byProtomyces macrosporus Ung. is a common disease of coriander which is an important spice crop of India and other countries. For the first time, it has been possible to obtain at least a partial field control of this disease, through seed treatment with thiram at the dose of 0.25 kg per 100 kg of seed. There are indications that better control is possible by combining seed and soil treatments with thiram. Seed treatment with TCNA — an experimental fungicide — may prove to be effective at a dose of 0.5 kg per 100 kg of seed.The authors are grateful to Dr.N. K. A. Rao, Dean, College of Agriculture and Dr.R. L. Paliwal, Director of Experiment Station, for their help. The Junior authors thank U. S. Agency for International Development (University of Illinois) for the award of graduate traineeships.     

Protein- und RNS-Gehalt des Hypokotyls beim stationären Wachstum im Dunkeln und unter dem Einfluß von Phytochrom (Keimlinge von Sinapis alba L.)

Zusammenfassung Das Wachstum des Hypokotyls wurde an Restkeimlingen ohne Kotyledonen (Abb. 1) untersucht. Die Wachstumsgeschwindigkeit ist in dem von uns untersuchten Zeitraum sowohl im Dunkeln als auch unter dem Einfluß von P₇₃₀ (Dauer-Dunkelrot) praktisch konstant. Obgleich sich die Wachstumsgeschwindigkeiten im Dunkeln und im Dauer-Dunkelrot um den Faktor 4 unterscheiden, hat das Dunkelrot keinen signifikanten Einfluß auf den Gesamt-Proteingehalt des Hypokotyls (bzw. der durchschnittlichen Hypokotylzelle). Der Proteingehalt nimmt im Dunkeln und im Licht kontinuierlich ab. Auch der Gesamt-RNS-Gehalt zeigt innerhalb des Versuchszeitraums eine Abnahme, die unter dem Einfluß von Dunkelrot früher einsetzt als im Dunkeln. — Man kann aus den Daten der vorliegenden Arbeit schließen, daß nur ein kleiner Teil des Gesamt-Proteins und der Gesamt-RNS einer Zelle mit dem Zellwachstum unmittelbar in Verbindung gebracht werden kann.

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Protein and RNA contents of the hypocotyl during steady state growth lengthening in the dark and under the influence of phytochrome (seedlings of sinapis alba L.)

Summary Inhibition of hypocotyl lengthening by phytochrome can be regarded as a prototype of a negative photoresponse. The hypothesis has been advanced (Schopfer, 1967) that negative photoresponses are the consequence of a differential gene repression which is exerted by P₇₃₀, the active phytochrome. This hypothesis is mainly based on experiments with specific inhibitors of RNA- and protein synthesis. —The present paper is part of an experimental program which has been designed to check this hypothesis.—Continuous irradiation with standard far-red has been used to establish a virtually stationary concentration of P₇₃₀ over the whole period of experimentation (36–60 hours after sowing). To correlate more strictly the growth response of the hypocotyl with molecular changes in this organ the axis system without cotyledons has been used (Fig. 1). Even under these conditions the growth rate of the hypocotyl is nearly constant in light (continuous far-red) and dark during the whole period of experimentation (36–60 hours after sowing) (Fig. 2, 3). It is known from earlier experiments that cell division in the hypocotyl are very rare during this period and that there is virtually no increase in the DNA contents of the organ during the period of our experimentation (Weidner, 1967). Obviously the number of cells per hypocotyl is virtually constant between 36 and 60 hours after sowing. Organ (i.e. hypocotyl) lengthening is nearly exclusively due to cellular lengthening.—If we follow the protein contents of the hypocotyl we find (Fig. 4) that the total protein of the organ decreases steadily in spite of the fact that the organ grows at a constant rate. There is no significant difference in protein contents between dark-grown and far-red grown systems although the growth rates differ by a factor of 4 (Fig. 2, 3).—The situation is some-what different with respect to total RNA (Fig. 5). The RNA contents eventually decrease in far-red as well as in dark-grown systems but the decrease is significantly faster in the far-red treated systems than in the dark controls.—It is concluded that only a very small part of the total RNA and total protein of a cell can be related to the control of cellular growth. Changes in bulk RNA and bulk protein obviously do not necessarily reflect changes in the growth rate or growth capacity of an organ or a cell.

     

Ultrastructural localization of RNA in cryptomonads

Summary In a previous study, DNA was localized in cells of two cryptomonads,Pyrenomonas sp. andCryptomonas ovata, by use of immuno-gold technique. Of particular interest was the ultrastructural localization of DNA in the nucleomorph, supposed to be a vestigial nucleus of a former endosymbiont [Hansmann Pet al. (1986) Eur J Cell Biol 42: 152–160]. In the present paper, distribution of RNA in the same two organisms is reported. RNA was detected by the specific and very sensitive RNase-gold method. RNA could be demonstrated in all of the four plasmatic compartments of cryptomonad cells (cytoplasm, periplastidal compartment, mitochondrion, and plastid), although the amounts differed greatly in the respective compartments. In the nucleus, the condensed chromatin and the nucleolus were preferentially labeled. Intense labeling could also be found over the fibrillogranular region of the nucleomorph. This fact lends strong support to the supposition that the fibrillogranular body represents the structural and functional equivalent of a nucleolus and thus again supports the hypothesis that the nucleomorph represents a vestigial eukaryotic nucleus. InPyrenomonas sp., gold-particle density over the nucleolus and the fibrillogranular body was quantitatively evaluated in order to compare their respective RNA synthesizing activities. Labeling density over the nucleolus was found to be 2.7 times higher and thus, on account of its greater volume, the nucleolus may contain 17 times more RNA than the fibrillogranular body of the nucleomorph.Abbreviations BSA bovine serum albumin - ER endoplasmic reticulum - GA glutaraldehyde - SSC standard saline citrate - SSCB SSC containing BSA     

Histologische,histochemische und experimentelle Studien am Subkommissuralorgan von Anuren (Mit Hinweisen auf den Epiphysenkomplex)

Ohne ZusammenfassungHerrn Professor Dr. Dr. H. Becher, Münster, gewidmet.Mit Unterstützung durch die Deutsche Forschungsgemeinschaft und den Marburger Universitätsbund (1958–1960).Einige elektronenmikroskopische Befunde wurden gemeinsam mit Frl. Dr. M. v. Harnack am Anatomischen Institut der Universität Kiel (Direktor: Prof. Dr. W. Bargmann) gewonnen. — Für technische Assistenz danke ich meiner Frau, Herrn cand. med. F. Wolff und Herrn cand. med. E. Börner.     

Karyometric and cytophotometric study of hepatocyte nuclei of frogs exposed to cold and prolonged starvation

Summary Prolonged starvation of Rana pipiens at room temperature results in a decrease in the volume of the hepatocyte nuclei from that of recently fed individuals. A converse increase in the volume of the liver nuclei over the normal was noted in frogs exposed to low temperature for 10 days. The present work was conducted in an effort to determine which nuclear fraction is responsible for the observed changes in nuclear size.The amount of DNA, RNA and protein-bound sulfhydryl groups per nucleus was measured cytophotometrically and was found to be independent of changes in nuclear size. Total nuclear protein content, on the other hand, was found to vary in proportion to nuclear volume. It was concluded that the variation in nuclear size resulting from starvation and cold-treatment is due to differences in non-chromosomal protein content and cannot be attributed to changes in ploidy.Dedicated to Prof. Berta Scharrer in honor of her 60th Birthday. — Supported by U.S. Public Health Service Grant HD-1499-04 to Dr. Cowden.Work completed while a Postdoctoral Trainee under U.S. Public Health Training Grant to the Department of Pathology, University of Florida, NIH-GM-1142-03.Supported by a Career Development Award from the National Institute of Child Health and Human Development, K 3-HD-6176-04.     

A phasic-tonic proprioceptor in the telson of the crayfish Procambarus clarki (Girard)

Summary A proprioceptor in the telson of the crayfish Procambarus clarki was found, which is made up of about five units. These show range fractionation, i.e. different threshold distributions and their response to bending of the telson has a transient and a stationary component. The peak frequencies reached during the period of the transient component were shown to increase with increasing bending velocity and had — with the exception of one case — the highest values when extension (contrary to flexion) was imposed. The position characteristic discharge frequencies of the stationary component and their variation with different telson positions were considerably smaller. The receptor signals movement and position to the central nervous system and is therefore called a phasictonic one.I am grateful to Prof. Dr. T. H. Bullock for his hospitality and interest in this work and also want to thank Mr. John W. Thorson for many constructive discussions. This work was aided by grants from the U.S. Public Health Service, National Science Foundation and Office of Naval Research to Prof. Dr. T. H. Bullock.     

Pollenanalytische Untersuchungen zur Frage der natürlichen Waldgrenze und des natürlichen Fichtenanteils im Stuhleckgebiet (Fischbacher Alpen)

Ohne ZusammenfassungDie Anregung zur vorliegenden Arbeit verdanke ich Herrn Prof. Dr.Hannes Mayer. Bei der Probengewinnung unterstützte mich Herr Dipl.-Ing. Dr.Kurt Zukrigl, der mich auf die beiden Moore hinwies und mir auch Angaben über die gegenwärtigen Bewaldungsverhältnisse — zum Teil auf Grund seiner noch unveröffentlichten Vegetationsaufnahmen — in dankenswerter Weise zur Verfügung stellte.     

Some aspects of the Feulgen reaction in situ

Summary Cytological observations combined with studies on absorption spectra of Feulgen stained normal and lipid — extractet HeLa and ehrlich-Lettré mouse ascites cells were performed after fixation of the cells as well in neutral formaldehyde as in Serra fixative. The effects of formaldehyde treatment of the stained cells to substitute all the free amino groups of DNA bond pararosaniline molecules, were also studied. The results obtained by using DNA samples containing 2% protein and relatively free from protein, led to the conclusion that after acid hydrolysis for a short period purines in DNA become splitted and these released aldehydes react with one or two amino groups of pararosaniline, a triphenylmethane dye (according to the arrangement of purines and pyrimidines in the helices). Some protein molecules also take part in the reaction and substitute some of the free amino groups of DNA bound pararosaniline. Peulgen stained cells fixed in Serra fixative show an absorption maximum at 546–550 m. Under appropriate conditions, as in cells fixed in formaldehyde, other substances e.g. phospholipids and lipoproteins interfere with the reaction by substituting most of the free amino groups of DNA bound pararosaniline molecules. It has been argued that in histochemical reactions monosubstituted pararosaniline molecules should be coloured and further substitution of free amino groups of pararosaniline, bound in DNA helices, does not change the intensity of the colour, but gives a shift in the wavelength of the absorption spectra.It has been suggested that the differential response of the nucleoli to the Feulgen-reaction, depending on whether the cells were fixed in formaldehyde or in Serra fixative, may be due to the formation of a protecting shield around the finely distributed intranucleolar chromatin strands, when formaldehyde is being used. After this fixation lipoproteins and other lipids, present in a relatively high percentage and closely associated with the intranucleolar chromatin strands, are especially well preserved.Evidences have been put foreward in support of the amino alkylsulfonic acid theory of Rumpf (1935) and Hörmann et al. (1958) whereas the amino sulfinic acid theory to explain the Schiffs reaction (Wieland and Scheuing, 1921) was shown not to be in agreement with our results.On leave from the Department of Botany, Calcutta University, 35, Ballygunge Circular Road, Calcutta-19, India; on a fellowship from the German Academic Exchange Service.