CHROMOSOME MICROMANIPULATION : III. Spindle Fiber Tension and the Reorientation of Mal-Oriented Chromosomes

Kinetochore reorientation is the critical process ensuring normal chromosome distribution. Reorientation has been studied in living grasshopper spermatocytes, in which bivalents with both chromosomes oriented to the same pole (unipolar orientation) occur but are unstable: sooner or later one chromosome reorients, the stable, bipolar orientation results, and normal anaphase segregation to opposite poles follows. One possible source of stability in bipolar orientations is the normal spindle forces toward opposite poles, which slightly stretch the bivalent. This tension is lacking in unipolar orientations because all the chromosomal spindle fibers and spindle forces are directed toward one pole. The possible role of tension has been tested directly by micromanipulation of bivalents in unipolar orientation to artificially create the missing tension. Without exception, such bivalents never reorient before the tension is released; a total time "under tension" of over 5 hr has been accumulated in experiments on eight bivalents in eight cells. In control experiments these same bivalents reoriented from a unipolar orientation within 16 min, on the average, in the absence of tension. Controlled reorientation and chromosome segregation can be explained from the results of these and related experiments.     

Unique Protein Moieties for 30S and 50S Ribosomes of Escherichia coli

Each major component of the proteins of 30S ribosomes from Escherichia coli was compared with the proteins of 50S ribosomes. The comparisons were done by using polyacrylamide gel electrophoresis in urea with differentially labeled proteins. The data show that no major protein is common to both ribosomes.     

Aspects of the life cycle of marine nematodes

Summary 1. Life cycles of approximately one month are recorded for laboratory maintained cultures of chromadorid, monhysterid and oncholaimid nematodes.2. Amphimictic and parthenogenetic reproduction occurs in the species investigated. Reproduction inMonhystera parelegantula is by parthenogenesis.3. The chromadoridChromadorina epidemos and the oncholaimidViscosia macramphida are able to reproduce by either amphimixis or parthenogenesis.4.Acanthonchus cobbi, Chromadora macrolaimoides andEuchromadora gaulica are amphimitically reproducing species.

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Aspekte des Lebenszyklus mariner Nematoden

Kurzfassung Bei einigen marinen Nematoden, die in Biscayne Bay (Florida) gesammelt und im Labor gezüchtet worden sind, vollzieht sich der gesamte Lebenszyklus ungefähr innerhalb eines Monats. BeiAcanthonchus cobbi, Chromadora macrolaimoides undEuchromadora gaulica ist zweigeschlechtliche Fortpflanzung festgestellt worden.Monohystera parelegantula vermehrt sich parthenogenetisch. Ein möglicherweise durch Umwelteinflüsse bedingter Wechsel von bisexueller und parthenogenetischer Fortpflanzung tritt beiChromadora epidemos undViscosia macramphida auf.

     

Studies on marine fungal-nematode associations and plant degradation

Summary 1. Studies of the broad-leafed turtle grass,Thalassia testudinum König, have revealed a diverse range of fungal infestation different in generic composition and dynamics of attack from that found on submerged wood. Certain of the fungi, notably the AscomyceteLindra thalassiae, initiate considerable degradation of leaf tissue and show a developmental cycle in nature related to the physiological state of the host plant.2. Use of fungal-cellulose mats as a trapping substrate has been extremely effective for discernment of ecologically significant shifts in nematode concentrations, especially those of the omnivorous species,Metoncholaimus scissus.3. Patterns of activity ofM. scissus, as well as those of various foliicolous nematodes, suggest that loci of organic material, such as fungal infested leaves and decaying plant tissue, significantly affect biological activity of these animals.4. Laboratory analysis of degraded cotton cellulose filters show a striking incidence of fungal reproduction of the ascomycetous fungusLulworthia, along with development of a considerable associated nematode fauna, especially species ofViscosia (V. macramphida) andLeptolaimus (L. plectoides). Successional patterns in nematode development are noted with continued degradation of the cotton cellulose matrix.

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Studien über marine Pilz-Nematoden-Assoziationen und Pflanzendegradation

Kurzfassung Untersuchungen am SeegrasThalassia testudinum König haben ergeben, daß sich hier Pilzinfektionen hinsichtlich der Komposition der beteiligten Gattungen und der Dynamik des Befalls von den am untergetauchten Holz festgestellten Infektionen unterscheiden. Bestimmte Pilze, insbesondere der AscomycetLindra thalassiae, leiten eine erhebliche Degradation des Blattgewebes ein und zeigen einen Entwicklungszyklus, welcher in Beziehung steht zum physiologischen Zustand der Wirtspflanze. Die Anwendung von Pilz-Zellulose-Matten als Einfangsubstrat war außerordentlich erfolgreich für das Erkennen ökologisch signifikanter Verschiebungen in den Nematodenkonzentrationen, insbesondere bei der omnivoren ArtMetoncholaimus scissus. Die Aktivitätsmuster vonM. scissus — ebenso wie die verschiedener foliicolöser Nematoden — deuten darauf hin, daß pilzinfizierte und zerfallende Pflanzenteile in entscheidendem Maße die biologische Aktivität dieser Tiere beeinflussen. Laboratoriumsanalysen degradierter Wollzellulosefilter lassen eine überraschend starke Vermehrung des AscomycetenLulworthia erkennen und gleichzeitig die Entwicklung einer beachtlichen Fauna assoziierter Nematodenarten, insbesondere vonViscosia macramphida undLeptolaimus plectoides. Im Verlaufe der weiteren Degradation der Wollzellulosematrix kommt es bei der Nematodenfauna zu entsprechenden Sukzessionen.

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Contribution No. 768 from the Institute of Marine Science, University of Miami, Miami, Florida, and from the Canada Department of Agriculture, Ottawa, Canada. This work was supported at the IMS by grant GM 12842 from the National Institutes of Health, USA.     

Growth of epithelium from a preneoplastic mammary outgrowth in response to mammary adipose tissue

Summary We investigated the effects of conditioned media derived from mouse mammary fat pads on the proliferation of CL-S1 cells, an epithelial cell line originally isolated from a preneoplastic mammary outgrowth line. Cell proliferation in vitro in serum-free defined medium was compared to that in this medium conditioned using intact mammary fat pad pieces or isolated fat pad adipocytes. Culture medium was conditioned by incubating the conditioning material in defined culture medium for 24 h at 37°C. Conditioned medium induced CL-S1 proliferation as much as 10- to 20-fold above the minimal levels of growth in control cultures after 13 d of culture. The growth-stimulatory factor(s) had an apparent molecular weight of greater than 10 kDa. This growth-stimulatory activity was both heat and trypsin stable. Because the role of adipose tissue is to store and release lipids, we next tested whether lipids are released during medium conditioning. The lipid composition of the fat pad conditioned medium was characterized using both thin layer and gas liquid chromatography. These lipid analyses indicated that the fat pad pieces released significant amounts of fatty acids and phospholipids into the medium during the conditioning period. The free fatty acid composition included both saturated and unsaturated molecules, and about 80% of the total fatty acids consisted of palmitate, stearate, oleate, and linoleate. These same fatty acids were a structural component of the majority of phospholipid found in the medium. The addition of palmitate or stearate to defined medium had no effect or was inhibitory for CL-S1 proliferation, depending on the concentration used. Defined medium supplemented with oleate, arachidonate, or linoleate induced CL-S1 proliferation, and the inhibitory effects of palmitate and stearate were overcome by addition of oleate and linoleate. These data indicate that both unsaturated and saturated fatty acids are released from intact adipose cells of the mouse mammary fat pad and that fatty acids can influence the growth of prenoplastic mouse mammary epithelium. Thus, unsaturated fatty acids, perhaps in conjunction with other substances released simultaneously, are candidate molecules for the substances that mediate the effect of adipose tissue on growth of epithelium. This work was supported in part by a grant from the American Institute for Cancer Research; grant CA 46885 from the National Institutes of Health, Bethesda, MD; and by State of Washington initiative 171.     

Oxidant inhibition of epithelial active sodium transport

The purpose of this study was to quantify the effects of extracellularly generated partially reduced oxygen species on active sodium (NA⁺) transport across the ventral toad skin, a well-studied epithelium. Sections of skin from decapitated toads were mounted in an Ussing chamber, bathed on both sides with electrolyte solution containing 500 μM xanthine and bubbled continuously with room air. The tissues were short-circuited, and short circuit current (Isc) and tissue resistance (R_t were monitored continuously with an automatic voltage clamp apparatus. Fifteen mU/ml of xanthine oxidase (XO), either purchased from Calbiochem or purified from cream, were instilled in either the apical (mucosal) or basolateral (serosal) baths at t = 0 and T = 10 min. Hydrogen peroxide (H₂O₂) concentrations increased to 200 μM within the first 20 min and then decreased, reaching a value of 40 μM by 60 min. Mean [H₂O₂] was 90 μM. Instillation of XO in the apical bath resulted in a large decrease in Isc and an increase in R_t, their values being 43% and 160% of their corresponding controls 85 min after the first instillation. Addition of superoxide dismutase and catalase completely prevented these changes. Instillation of XO in the basolateral bath had no effect. Similar physiological responses were obtained using the Calbiochem XO or the purified XO, which contained no measurable protease activity. It was concluded that extracellularly generated partially reduced oxygen species may interfere with active Na⁺ transport by possibly damaging apical Na⁺ channel proteins.