Occurrence of wall-less cells during synchronous gametogenesis in Chlamydomonas reinhardtii

Synchronous gametogenesis in Chlamydomonas reinhardtii is accompaniedby a round of cell division. Some of the unmated gametes becomenaked at daughter cell liberation. A sporangial wall-lytic enzyme,which is excreted into the medium at zoospore liberation, actson the wall of the gametic cell. (Received August 28, 1980; )     

Studies on the vegetative life cycle of Chlamydomonas reinhardi Dangeard in synchronous culture III. Some notes on the process of zoospore liberation

When Chlamydomonas reinhardi cells liberate zoospores, theyexcrete into the medium a factor(s) which induces zoospore liberationof other cells that are not yet ready to liberate zoosporesby themselves. The "factor" is contained within cells at laterstages of the cell cycle, but its action is suppressed untilthe regular time of zoospore liberation in the cell cycle. (Received November 18, 1974; )     

Spore Discharge in Basidiomycetes: III. Spore Liberation from Narrow Hymenial Tubes

The hymenial tubes of Polyporus betulinus were shown to be approximatelycylindrical,and to have a sporulating hymenium over the greater part oftheir surface. Sporophores were tilted through measured anglesfrom the vertical in the field and in the laboratory, and theeffect on spore liberation was measured by determining the numberof spores liberated from particular tubes in standard time.All tilting caused a reduction in spore liberation. The amountof this reduction was found to compare closely with that predictedfrom calculation of the amount of hymenium lying directly abovethe orifice at various angles of tilt. It is therefore concludedthat liberation from the tube can be accounted for by the initialviolent discharge of the spore and gravitational attractionalone. An addendum corrects an earlier paper (1959) in thisseries. It shows that the variation in density of spore depositscollected from Trametes gibbosa was due to air currents in theapparatus, not to variation in sporulation rate.     

A Light Microscope Study of Carposporophyte Development in Gracilaria verrucosa (Hudson) Papenfuss

Gracilaria verrucosa is a very common marine red alga of Greekcoasts. The diploid carposporophyte which develops attachedto the female gametophyte of Gracilaria is described. The immaturecystocarps are very small while the mature ones are globose,ostiolate and are borne profusely all over the surface of thethallus. The earliest observed fusion cell is small and fusesprogressively with adjacent vegetative cells to form a largemultinucleate cell. From this fusion cell gonimoblast initialsoriginate, dividing further and giving rise to a large numberof gonimoblast cells. The resultant carposporophyte consistsof a basal-central, multinucleate cell surrounded by a conicalor hemispherical mass of gonimoblast cells. Chains or clustersof successively maturing carpospores are borne from the terminalgonimoblast cells. The liberation of mature carpospores takesplace through the ostiole of the cystocarp. The liberated carposporeslack cell walls and are naked in a mucilage mass. Gracilaria verrucosa (Hudson) Papenfuss, Gigartinales, Gracilariaceae, Rhodophyta, carposporophyte, development     

Studies on the vegetative life cycle of Chlamydomonas reinhardi Dangeard in synchronous culture I. Some characteristics of the cell cycle

Cells of Chlamydomonas reinhardi Dangeard were grown synchronouslyunder a 12 hr light-12 hr dark regime. Time courses of nucleardivision, chloroplast division, "apparent cytokinesis" and zoosporeliberation were followed during the vegetative cell cycle inthe synchronous culture. Liberation of zoospores occurred atabout 23–24 hr after the beginning of the light periodat 25°C. Four zoospores were produced per mother cell underthe conditions used. At lower temperatures, the process of zoosporeliberation as well as length of the cell cycle was markedlyprolonged, but the number of zoospores produced per mother cellwas approximately the same. At different light intensities,lengths of the cell cycle were virtually the same, while thenumber of zoospores liberated was larger at higher rather thanat lower light intensities. During the dark period, nuclear division, chloroplast divisionand apparent cytokinesis took place, in diis order, and proceededless synchronously than did the process of zoospore liberation.When the 12 hr dark period was replaced with a 12 hr light periodduring one cycle, the time of initiation as well as the durationof zoospore liberation was litde affected in most cases, whereasnuclear division, chloroplast division and apparent cytokinesiswere considerably accelerated by extended illumination. Whenalgal cells which had been exposed to light for 24 hr were furtherincubated in the light, zoospore liberation started much earlierand proceeded far less synchronously, compared with that under12 hr light-12 hr dark alternation. (Received October 12, 1970; )     

Serial Development of Foliar Gemmae in Tortula (Pottiales, Musci), an Ultrastructural Study

The development and liberation mechanism of foliar gemmae havebeen studied by electron microscopy in two mosses, Tortula latifoliaBruch and Tortula papillosa Wils. The gemmae develop on theadaxial surface of mature leaves from single initial cells onboth the lamina and costa in T. latifolia but only on the costain T. papillosa . Elongation of the initial cell is associatedwith the deposition of a highly extensible new wall whilst theold wall and cuticle in the apical dome rupture. The first divisionis transverse and separates a short basal cell embedded in thefoliar tissue and a distal cell, or gemma primordium, protrudingfrom the leaf surface. Subsequent divisions of the gemma primordiumgive rise to a six-to-eight-celled globose gemma with mucilaginousouter walls. During gemma development the basal cell producesa new wall and elongates again whilst the common wall with thegemma splits apart centripetally along the boundary betweenthe old and new wall in the basal cell; plasmodesmal connectionsare gradually severed and eventually the young gemma remainsconnected to the basal cell only by mucilage. After separationof the first-formed gemma, the basal cell may expand and producea second gemma by the same mechanism. The whole process maybe repeated several times resulting in the formation of a chainof gemmae stuck together by mucilage and which are liberatedonly when the leaves are fully hydrated. Accumulation of abundantlipid deposits in the gemmae after symplasmic isolation reflectsconsiderable photosynthetic autonomy. Abscission; bryophytes; cell wall formation; plasmodesmata; vegetative reproduction     

The Distribution of Daughter Colonies and Cell Numbers in a Natural Population of Volvox aureus Ehrenb

The distribution of cell numbers and daughter colonies was determinedin a population of Volvox aureus from Malham Tarn, North Yorkshireduring summer and autumn. The following ranges were recorded:cell number, 272–2340; colony diameter, 174–520µm; daughter colony number, 2–10 (mode 6). Culturedmaterial gave significantly higher values than these. A strongpositive correlation was found between cell number per colonyand the number of asexual daughters but no correlation was foundbetween the number of daughters and mother colony diameter. The results are discussed with reference to the theoreticalpacking of spheres, which the daughters approximate. The numberof daughters and their size upon liberation was also consideredin relation to grazing pressures and the relative loss of cellscaused by the disintegration of the mother colony. The mostsignificant factor was considered to be the size of the mothercolony which is probably controlled by nutrient supply and temperature. Volvox cell number, daughter colonies     

Sexual process in eight-spored ascus-forming yeast, Schizosaccharomyces japonicus I. Culture medium for the synchronous sexual process

The sexual process of Schizosaccharomyces japonicus consistsof sexual flocculation, zygote formation, eight-spored ascusformation and liberation of spores from the ascus. The culturemedium in which this sexual process took place synchronouslywas prepared. For the completion of the sexual process, glucosewas essential and inorganic salts and vitamins were also required.Elimination of the nitrogen source stimulated the rate of sporeformation. The temporal relationship among the sexual eventswas also elucidated: sexual flocculation, zygote formation,ascus formation and spore liberation occurred at 4, 7, 13 and20 hr, respectively, after transfer to the medium for the synchronoussexual process. (Received May 11, 1978; )     

Studies on the vegetative life cycle of Chlamydomonas reinhardi Dangeard in synchronous culture IV. Effects of 6-methyl purine on the revolution of the cell cycle

Cells of Chlamydomonas reinhardi Dangeard were synchronouslygrown under a 12 hr light-12 hr dark regime. The algal cellcycle under these conditions starts with a light-induced reaction(s)at the beginning of the light period and ends, after a definiteperiod of time (23–24 hr at 25°C), in zoospore liberation.When cells were exposed to 6-methyl purine for short periods(0.5–2.5 hr) at different times during the early and intermediatephases of the cell cycle, it exerted, as an analogue of adenine,two different effects on the revolution of the cell cycle: onea "lengthening" effect seen at its low concentrations in whichthe length of the cell cycle was somewhat prolonged, the othera "return to start" effect at higher concentrations. In thelatter a short exposure of cells to 6-methyl purine broughtthem to the starting point of the cell cycle concurrent withthe abortion of the cycle in process. When 6-methyl purine wasapplied during the later phase of about 1/4 the length of thecell cycle, it casued no effect. Control of the revolution ofthe algal cell cycle by an "adenine-involving reaction(s)" disturbedby this adenine analogue is discussed. (Received September 1, 1975; )     

Development and Liberation of Cauline Gemmae in the Moss Aulacomnium androgynum (Hedw.) Schwaegr. (Bryales): An Ultrastructural Study

The leafy shoots of the mossAulacomnium androgynum form clustersof gemmae borne terminally on long pseudopodial axes. The gemmaearise from single initial cells produced by the activity ofa superficial meristem. Mature gemmae comprise an apical anda basal cell with four to seven cells forming two, sometimesthree, tiers in between. The basal cell is connected to thetip of the pseudopodium by a uniseriate filament consistingof an abscission (tmema) cell and a stalk cell. The first divisionof the initial cell produces a proximal cell and a distal cell.The proximal cell elongates without further division formingthe stalk of the gemma; the distal cell gives rise to a lowerand an upper cell by transverse division. The upper cell dividesrepeatedly by oblique septa forming the apical and middle cellsof the gemma; the basal cell and tmema cell arise from a transversedivision of the lower cell. The first two divisions in gemmadevelopment are highly asymmetrical and exogenous, i.e. preceededby cell expansion. A broad interphase cortical band of microtubulesis associated with intercalary cellular growth during this stage.Subsequent gemma development follows an endogenous pattern withcellular expansion following the completion of proliferativedivisions and involving a conventional system of cortical microtubules.While elongating to about four times its original length withoutdeposition of a distinct new wall the tmema cell undergoes cytoplasmicdegeneration and eventually breaks, causing gemma liberation.The stalk cell elongates about eight-fold and its contents alsodegenerate after gemma liberation. Plasmodesmata in the basaland stalk cells are obliterated by the deposition of additionalwall materials. The highly electron-opaque outer walls of themature gemmae and tips of the stalk cells are water-repellant.The gemmae are dispersed either in water films or by air currents. Abscission; asexual reproduction; bryophytes; morphogenesis; microtubules; ultrastructure     

Antithrombin reduces leukocyte adhesion during chronic endotoxemia by modulation of the cyclooxygenase pathway

Antithrombin (AT) isknown as the most important natural inhibitor of thrombin activity andhas been shown to improve distinct clinical parameters during thecourse of septic (endotoxin)-induced multiple organ dysfunction. Wehypothesized that AT acts by inhibiting leukocyte activation andmicrovascular injury via the promotion of endothelial release ofPGI₂, and therefore, we studied the effects of AT onleukocyte/endothelial cell interaction and microvascular perfusionduring endotoxemia. In a skinfold preparation of Syrian hamsters,severe endotoxemia was induced by repeated administration of endotoxinintravenously [lipopolysaccharide (LPS), Escherichia coli,2 mg/kg] at 0 and 48 h. AT (250 IU/kg) was administered intravenously at 0, 24, and 48 h (n = 6, ATgroup). In control animals (n = 5, control), LPS wasgiven without AT supplementation. By intravital fluorescencemicroscopy, leukocyte-endothelial cell interaction and functionalcapillary density (FCD; measure of capillary perfusion) were analyzedduring a 72-h period after the first LPS injection. AT significantlyattenuated LPS-induced arteriolar and venular leukocyte adherence afterboth the first and the second LPS injection [P < 0.01, measures analysis of variance (MANOVA)]. In parallel, AT waseffective in preventing LPS-induced depression of FCD after the firstand the second LPS administration (P < 0.05, MANOVA).By pretreatment with the cyclooxygenase inhibitor indomethacin(n = 6), effects of AT on leukocyte adherence and FCDwere found completely abolished. Thus our study indicates that ATexerts its beneficial effects in endotoxemia by reducing leukocyte-endothelial cell interaction and microvascular perfusion failure probably via liberation of prostacyclin from endothelial cells.

     

Alteration of the Cell Surface during the Vegetative Cell Cycle of the Unicellular Green Alga Chlamydomonas reinhardtii

Alterations of the cell surface during the vegetative cell cycleof the unicellular green alga Chlamydomonas reinhardtii wereinvestigated using polyclonal antibodies against the purifiedand subsequently deglycosylated insoluble cell wall componentand against a 100 kDa polypeptide of the deglycosylated, chaotrope-solublewall fraction, respectively. Both antibodies recognized epitopeswithin the non-glycosylated domains of a ‘150 kDa’chaotrope-soluble glycoprotein (=GP3B) localized in the outerlayers of the C. reinhardtii cell wall. Immunofluorescence studiesindicated that both antibodies reacted with the surface of ‘late’sporangia (harvested 1 h before liberation of the zoospores),but not with the cell surfaces of released zoospores, growingcells and young sporangia, respectively. After pretreatmentwith aqueous LiCl, however, the cell surfaces of zoospores,growing cells and young sporangia became accessible to theseparticular antibodies. Highly purified preparations of the insolublewall fraction revealed strong immunofluorescence with both antibodiesbut not with the corresponding preimmune sera. Based on thesedata, we concluded that the antigenic sites of the insolubleglycoprotein framework of the C. reinhardtii wall are maskedby LiCl-soluble glycoproteins in single cell stages and youngsporangia, but not or to a lesser extent in the case of themother walls of ‘late’ sporangia. The conclusionwas supported by findings that (I) the multilayered structureof the mother-cell wall was disturbed in ‘late’,but not in young sporangia and that (II) the amounts of chaotropesolublecell wall glycoproteins present in the LiCl-extracts from intactsporangia decreased during ripening of the sporangia. (Received January 10, 1996; Accepted May 27, 1996)     

Ultrastructure of the Sporangiospore of Kickxella alabastrina (Mucorales)

The structure of the sporangiospore of Kickxella alabastrinawas examined by means of carbon replicas, ultrathin sectionsand chemical and physical disintegration. The wall consistsof an outer ornamented, largely amorphous complex and an inner,largely fibrillar complex. The outer complex is disrupted bytreatment in hot dilute alkali whereas the inner complex islargely unaffected. An annulus, similar to that described forCoemansia aciculifera, forms part of the central region of theinner complex and apparently develops prior to liberation ofthe spore from the pseudophialide. A labyrinthiform organelle,attached to the base of the septum which separates the sporefrom the pseudophialide, is described.     

Ca(2+)-dependent activation of Cl(-) currents in Xenopus oocytes is modulated by voltage

Ca²⁺-activatedCl currents (I_Cl,Ca) wereexamined using fluorescence confocal microscopy to monitorintracellular Ca²⁺ liberation evoked by flash photolysis ofcaged inositol 1,4,5-trisphosphate (InsP₃) involtage-clamped Xenopus oocytes. Currents at +40 mV exhibited asteep dependence on InsP₃ concentration([InsP₃]), whereas currents at140 mV exhibited a higher threshold and more graded relationshipwith [InsP₃]. Ca²⁺ levelsrequired to half-maximally activate I_Cl,Ca wereabout 50% larger at 140 mV than at +40 mV, and currents evokedby small Ca²⁺ elevations were reduced >25-fold. Thehalf-decay time of Ca²⁺ signals shortened at increasinglypositive potentials, whereas the decay of I_Cl,Calengthened. The steady-state current-voltage (I-V) relationshipfor I_Cl,Ca exhibited outward rectification withweak photolysis flashes but became more linear with stronger stimuli.Instantaneous I-V relationships were linear with both strongand weak stimuli. Current relaxations following voltage steps duringactivation of I_Cl,Ca decayed with half-times that shortened from about 100 ms at +10 mV to 20 ms at 160 mV. We conclude that InsP₃-mediated Ca²⁺liberation activates a single population of Clchannels, which exhibit voltage-dependent Ca²⁺ activationand voltage-independent instantaneous conductance.

     

Cytological Investigations on Porphyra umbilicalis(L.) Kutz. var. laciniata(Lightf.) J.Ag.

1. Vegetative mitosis of Porphyra umbilicalis var. laciniatais normal, the chromosome number as seen in late prophase beingfive. Special features of this mitosis are the appearance ofseveral stained chromatin segments in early pro-phase and theformation of a crescent-shaped group of chromosomes during metaphaseand anaphase. 2. Genuine monospores have not been seen in the material investigated. 3. Both spores and spermatia arise by repeated division of amother-cell and in identical ways. Evidence is presented toshow that the first division in the mother-cell forming sporesis longitudinal. There is no evidence of reduction divisionin this division, and the chromosome number in the mother-cellis five. 4. The first division in the germinating spore is mitotic andshows five chromosomes. 5. The Conchocelis-phase is haploid throughout and produces‘fertile cell rows’ and ‘plantlets’,but spore liberation has not been observed. It is suggestedthat the ‘plantlets’ may grow out directly intothe leafy phase or may give rise to spores according to prevailingenvironmental conditions. 6. No evidence of sexual reproduction has been obtained in thepresent work and the role of the spermatium in the life-cycleis not clear. It is suggested that further work may profitablybe concerned with investigating the nature of the spermatium.     

The Effect of Various Nitrogen Sources upon the Production of Extracellular Polysaccharide by the Blue-Green Alga Anabaena flos-aquae A-37

Anabaena flos-aquae A-37 was grown in bacteria-free cultureon a variety of nitrogen sources. Extracellular polysaccharideproduction was found to be a part of the normal metabolic processescausing the liberation of excess carbohydrates into the surroundingmedium rather than the result of cellular lysis. Non-adaptedcells supplied with NH₄Cl produced abundant polysaccharide.However, the population did not increase. Cells adapted to NH₄Clproduced smaller quantities of polysaccharide, but the populationdid increase. Polysaccharide production per cell was similaron Mg(NO₃)₂, KNO₃, NaN0₃, NH₄NO₃, and NH₄Cl. It was concludedthat an adequately utilized nitrogen source does not affectpolysaccharide production in A. flos-aquae A-37.     

Alterations of Free Amide and Amino Acid Contents During the Development of Maize Plants, Zea mays L.

Concentrations of free amino acids and amides were measuredin organs of maize plants, Zea mays L. in the period from 14d before pollen liberation until complete seed maturation. Inthis time anthesis took place and only the ovaries of ear 9(numbered from below) developed into seeds. In mature leaf bladesNH₄ ion assimilation had ceased and asparagine and glutaminewere not found there. N redistribution induced the occurrenceof large amounts of aspartate, glutamate and alanine. The amountsof amides in leaf sheaths and stem parts depended on the neighbourhoodof generative parts. The generative plant parts can be distinguishedfrom adjacent vegetative plant parts in concentrations of freeproline or asparagine. Proline occurred in pollen but not inthe empty anthers. Ears had a small, early peak amount of prolinemostly before pollination. Only the ninth ear had a first maximumproline amount after the fifth day of pollen liberation. A secondproline peak in the ears coincided with the period of maximumincrease in d. wt. The occurrence of proline in the generativeorgans relative to metabolic processes inducing fertility orseed maturation is discussed. Zea mays L., amides, amino acids, amino-transferring components, asparagine, glutamine, proline     

Studies on the vegetative life cycle of Chlamydomonas reinhardi Dangeard in synchronous culture V. The transition point for the effect of 6-methyl purine in the cell cycle

By following the nuclear division, chloroplast division, cytokinesisand zoospore liberation of Chlamydomonas reinhardi cells exposedto a high concentration of 6-methyl purine (6-MP), corroborativeevidence was obtained for our previous conclusion that 6-MP-exposedcells are brought to the starting point of a new round of thecell cycle with abortion of the cycle in process ("return-to-start"effect). This effect did not occur after cells had passed acritical stage (transition point) which seemed to be situatedshortly prior to the onset of nuclear division under the conditionsused. When 6-MP was applied to cells after the transition point,it caused an advancing effect on their zoospore liberation.A cycloheximide (CHI)-inhibition step existed shortly alterthe transition point for 6-MP. A model was proposed for theeffects of 6-MP and CHI. (Received August 8, 1977; )     

Sexual process in eight-spored ascus-forming yeast, Schizosaccharomyces japonicus II. Dependency of the sexual process on the carbon source

The carbon-source dependency of the sexual process in Schizosaccharomycesjaponicus was studied. Schiz. japonicus grew well in vegetativemedia containing glucose, sucrose, fructose or raffinose, anddid poorly in one containing mannose. On the other hand, itssexual process proceeded well in sporulation media containingglucose, sucrose or mannose, and was markedly delayed in thosecontaining fructose or raffinose. Neither vegetative growthnor sexual process occurred when non-fermentable carbon sources,such as glycerol, were used. The amount of glucose in the sporulationmedium sufficient for completion of the sexual process varieddepending on the cell-population density. Glucose was requiredfor both zygote and ascus formation but not for spore liberation.Cells were committed to sporulation shortly after the stageof zygote formation. (Received August 3, 1978; )     

SYNCHRONIZATION OF BACTERIAL CULTURE BY PREINCUBATION OF CELLS AT HIGH CELL DENSITY

The growth of Escherichia coli strain B in a liquid medium wasfound to cease at a cell density of 5x10⁹ cells per ml. (Thiscritical concentration is designated as the maximum or M-concentration.)Even cells harvested from the logarithmic growth phase couldnot divide at this or higher cell densities. Investigationson the metabolic activities of such cultures, however, showedthat the synthesis of cellular protein and nucleic acid wastaking place under such circumstances, showing that only someprocess (or processes) particularly related to cell divisionwas suppressed at the critical cell concentration in question. This finding led us to devise a new method of synchronizationof E. coli: cells harvested from a logarithmic phase were preincubatedat the critical concentration of 5x10⁹ cells per ml for 45 minutes,and then diluted 100 times with fresh medium. This led to synchronizationof cell division, as shown by a stepwise multiplication in cellnumber. (Received June 20, 1961; )