Cell differentiation during fruiting body formation in polysphondylium pallidum

1979 ◽  
Vol 35 (1) ◽  
pp. 203-215
Author(s):  
D.H. O'Day
Keyword(s):  
Cell Differentiation ◽  
Fruiting Body ◽  
Neutral Red ◽  
Immunofluorescent Staining ◽  
Stalk Cell ◽  
Fruiting Body Formation ◽  
Body Formation ◽  
Polysphondylium Pallidum ◽  
Slime Moulds ◽  
Cellular Slime

The spatial pattern of cellular differentiation was studied during fruiting body formation in Polysphondylium pallidum using 3 different staining methods: Calcofluor fluorescence (cellulose accumulation), neutral red (prestalk cells) and immunofluorescence (prespore cells). Neutral-red staining revealed the existence of a clear prestalk region which becomes evident during aggregation and continues throughout culmination. Immunofluorescent staining demonstrated that cells in the prestalk region gradually lose their presporeness (fluorescence) as they are transformed into differentiated stalk cells. Calcofluor staining revealed that stalk cell differentiation begins during the mid-aggregation phase and that the mode of formation of the main stalk and the side branches differs slightly in morphology. Calcofluor staining also demonstrated the development, during aggregation, of a thick cellulosic girdle with lateral tubular extensions which surround the aggregation streams. The above results are discussed in terms of our present knowledge about differentiation and morphogenesis in cellular slime moulds.

Cellular differentiation and pattern formation in the absence of morphogenesis in the cellular slime mould Polysphondylium pallidum: evidence for a biochemical tip (organizer) in submerged aggregates

1981 ◽  
Vol 27 (9) ◽  
pp. 924-936 ◽  
Author(s):  
Gary D. Paterno ◽  
Danton H. O'Day
Keyword(s):  
Cell Differentiation ◽  
Cellular Differentiation ◽  
Periodic Acid ◽  
Neutral Red ◽  
Stalk Cell ◽  
Periodic Acid Schiff ◽  
Slime Mould ◽  
Polysphondylium Pallidum ◽  
Cellular Slime ◽  
Roller Tube Culture

When amoebae of Polysphondylium pallidum WS320 are placed in nonnutrient buffer in roller tube culture they form spherical or ellipsoidal aggregates. At first the aggregates demonstrate a "loose" morphology but by 12 h, with the formation of a cellulose-containing, peripheral sheath, they become "tight" aggregates. At this time stalk differentiation begins. Using various methods for the resolution of prespore (ultrastructure, spore antigen immunofluorescence, periodic acid – Schiff staining) and prestalk (ultrastructure, alkaline phosphatase histochemistry, neutral red staining, Calcofluor fluorescence) cell localization, the pattern of cell differentiation in submerged aggregates was shown to be essentially identical to that of normal pseudoplasmodia. Furthermore, using a cAMP bioassay it was revealed that the submerged aggregates, while devoid of a morphological tip, do possess a biochemical tip which is correlated with sites of neutral red staining and stalk cell differentiation. As a result of these studies, an earlier argument that the tip of the pseudoplasmodium is not essential for the establishment of pattern or in the "organization" of cellular differentiation during slime mould development is contradicted.

Patterns of alkaline phosphatase activity during alternative developmental pathways in the cellular slime mold, Polysphondylium pallidum

1973 ◽  
Vol 51 (2) ◽  
pp. 301-310 ◽  
Author(s):  
Danton H. O'Day ◽  
David W. Francis
Keyword(s):  
Alkaline Phosphatase ◽  
Phosphatase Activity ◽  
Alkaline Phosphatase Activity ◽  
Fruiting Body ◽  
Slime Mold ◽  
Cellular Slime Mold ◽  
Fruiting Body Formation ◽  
Body Formation ◽  
Polysphondylium Pallidum ◽  
Cellular Slime

The enzyme alkaline phosphatase (EC 3.1.3.1) was studied during axenic growth, microcyst differentiation and fruiting body formation in the cellular slime mold Polysphondylium pallidum. The enzyme activity decreases during growth and microcyst differentiation but increases during fruiting body formation where it is localized in prestalk cells. Two major isozymes exist for the enzyme and these change qualitatively and quantitatively during multicellular development. Beryllium was found to be a potent inhibitor of the slime mold phosphatase. When beryllium was added to growing cells or cells undergoing fruiting body formation markedly reduced alkaline phosphatase activity was detectable in the cells but growth and development were unaffected. The results are discussed in relation to other work on the cellular slime molds.

In vitro stalk cell differentiation in wild-type and ‘slugger’ mutants of Dictyostelium discoideum

Development ◽  
1993 ◽  
Vol 118 (2) ◽  
pp. 523-526 ◽  
Author(s):  
K. Inouye ◽  
J. Gross
Keyword(s):  
Dictyostelium Discoideum ◽  
Fruiting Body ◽  
Cell Maturation ◽  
Stalk Cell ◽  
Wild Type ◽  
Fruiting Body Formation ◽  
Body Formation ◽  
Close Relationship ◽  
Long Time

In ‘slugger’ mutants of Dictyostelium discoideum, aggregates of cells remain for an abnormally long time in the migratory phase under conditions where wild-type aggregates form fruiting bodies. In the present work, we have examined the relationship between the defect in fruiting body formation in these mutants and their ability to form mature stalk cells. We dissociated anterior cells from slugs of the mutants and their parents and tested their ability to form stalk cells when incubated at low density in the presence of (1) the stalk cell morphogen Differentiation Inducing Factor-1 (DIF-1) together with cyclic AMP, or (2) 8-Br-cAMP, which is believed to penetrate cell membrane and activate cAMP- dependent protein kinase (PKA). Most of the mutants were markedly defective in forming stalk cells in response to DIF-1 plus cAMP, confirming a close relationship between fruiting body formation and stalk cell maturation. On the other hand, many of these same mutants formed stalk cells efficiently in response to 8-Br-cAMP. This supports evidence for an essential role of PKA in stalk cell maturation and fruiting body formation. It also indicates that many of the mutants owe their slugger phenotype to defects in functions required for optimal adenylyl cyclase activity.

Participation of the cell surfaces in determining the developmental courses in the cellular slime mould Dictyostelium purpureum

Development ◽  
1978 ◽  
Vol 48 (1) ◽  
pp. 153-160
Author(s):  
M. Saito ◽  
K. Yanagisawa
Keyword(s):  
Fruiting Body ◽  
Developmental Courses ◽  
Fruiting Bodies ◽  
Cell Surfaces ◽  
Fruiting Body Formation ◽  
Con A ◽  
Body Formation ◽  
Dictyostelium Purpureum ◽  
Cellular Slime ◽  
Type Strains

Dictyostelium purpureum S5 and S6, mating type strains, form fruiting-bodies in a monoclonal culture, but produce macrocysts in a mix culture. The effects of Concanavalin A (Con A) on both fruiting-body formation and macrocyst formation, and changes of Con Amediated cell agglutinability during development were studied. It was found that Con A inhibits macrocyst formation but not fruiting-body formation, and that macrocyst-forming cells are much more susceptible to Con A agglutination than are fruiting-body-forming cells during the aggregation stages. When fruiting-body-forming cells are treated with either trypsin or α-chymotrypsin, their Con A agglutinability is enhanced to the same extent as that of macrocyst-forming cells. It was also found that when S6 cells are treated with proteases they sometimes produce normal macrocysts even in a monoclonal culture. The results obtained in these experiments showed that the surface properties of fruitingbody- forming cells and macrocyst-forming cells are different, and that the cell surface might play an important role in determining the two developmental courses.

Colchicine induces multiple axis formation and stalk cell differentiation in Dictyostelium discoideum

Development ◽  
1978 ◽  
Vol 47 (1) ◽  
pp. 195-206
Author(s):  
Danton H. O'Day ◽  
Antony J. Durston
Keyword(s):  
Cell Differentiation ◽  
Dictyostelium Discoideum ◽  
Immunofluorescent Staining ◽  
Stalk Cell ◽  
Axis Formation ◽  
Cellular Slime Mould ◽  
Slime Mould ◽  
Long Time ◽  
Cellular Slime

Colchicine is shown to have several effects on the development of the pseudoplasmodia of the cellular slime mould Dictyostelium discoideum At concentrations of 0·01 M and above culmination was prevented, while differentiation of cells into stalk cells occurred at the rear of cell masses. Essentially all cells transformed into stalk cells when slugs were left on colchicine agar for a long time. At concentrations of 0·01 M normal slug architecture was maintained while above 0·025 M pseudoplasmodia reorganized into multiple mounds. Each of these mounds developed an apparently normal discrete tip which was devoid of prespore cells as shown by immunofluorescent staining. The same effects were observed in growing cultures and in regulating slugs treated with colchicine. The data are consistent with the ideas that microtubules are involved in the maintenance of slug architecture and in the differentiation of stalk cells. The modes by which these intracellular structures may operate in these functions are discussed.

The presence and location of sporopollenin in fruiting bodies of the cellular slime moulds

1984 ◽  
Vol 66 (1) ◽  
pp. 297-308
Author(s):  
Y. Maeda
Keyword(s):  
Cell Membrane ◽  
Biological Significance ◽  
Spore Wall ◽  
Fruiting Bodies ◽  
Fruiting Body Formation ◽  
Body Formation ◽  
Slime Moulds ◽  
Inner Surface ◽  
Cellular Slime ◽  
Cellular Slime Moulds

The presence of an acetolysis-resistant polymer (sporopollenin) in the cellular slime moulds is demonstrated. This polymer is located on the stalk sheath of fruiting bodies as a bundle of fine fibrils (4-5 nm diameter). The location and structure of sporopollenin in spores are shown to vary considerably, depending upon the species. In Polysphondylium violaceum spores, sporopollenin is composed of fine spicules (4-5 nm in diameter, 25–50 nm long) that cover both the outermost layer of spore wall and the inner surface of the cell membrane. The sporopollenin of Dictyostelium discoideum spores is located preferentially close to the inner surface of the cell membrane, forming a mass of electron-opaque fine granules (4-5 nm in diameter). D. mucoroides spores, however, appear not to possess a tight network of sporopollenin, since they were less resistant to acetolysis than those of the other species. The biological significance of the results is discussed with special reference to fruiting body formation.

scholarly journals A WD40 Repeat Protein Regulates Fungal Cell Differentiation and Can Be Replaced Functionally by the Mammalian Homologue Striatin

2004 ◽  
Vol 3 (1) ◽  
pp. 232-240 ◽  
Author(s):  
Stefanie Pöggeler ◽  
Ulrich Kück
Keyword(s):  
Cell Differentiation ◽  
Fruiting Body ◽  
Differentiation Process ◽  
Wild Type ◽  
Fruiting Body Formation ◽  
Wd40 Repeat ◽  
Body Formation ◽  
Repeat Protein ◽  
Type Protein ◽  
Wild Type Protein

ABSTRACT Fruiting body development in fungi is a complex cellular differentiation process that is controlled by more than 100 developmental genes. Mutants of the filamentous fungus Sordaria macrospora showing defects in fruiting body formation are pertinent sources for the identification of components of this multicellular differentiation process. Here we show that the sterile mutant pro11 carries a defect in the pro11 gene encoding a multimodular WD40 repeat protein. Complementation analysis indicates that the wild-type gene or C-terminally truncated versions of the wild-type protein are able to restore the fertile phenotype in mutant pro11. PRO11 shows significant homology to several vertebrate WD40 proteins, such as striatin and zinedin, which seem to be involved in Ca2+-dependent signaling in cells of the central nervous system and are supposed to function as scaffolding proteins linking signaling and eukaryotic endocytosis. Cloning of a mouse cDNA encoding striatin allowed functional substitution of the wild-type protein with restoration of fertility in mutant pro11. Our data strongly suggest that an evolutionarily conserved cellular process controlling eukaryotic cell differentiation may regulate fruiting body formation.

scholarly journals Loss of the Polyketide Synthase StlB Results in Stalk Cell Overproduction in Polysphondylium violaceum

2020 ◽  
Vol 12 (5) ◽  
pp. 674-683
Author(s):  
Takaaki B Narita ◽  
Yoshinori Kawabe ◽  
Koryu Kin ◽  
Richard A Gibbs ◽  
Adam Kuspa ◽  
...  
Keyword(s):  
Fruiting Body ◽  
Polyketide Synthase ◽  
Stalk Cell ◽  
Marker Genes ◽  
Cell Type ◽  
Fruiting Body Formation ◽  
Knock Out ◽  
Body Formation ◽  
Group 4 ◽  
Basal Disk

Abstract Major phenotypic innovations in social amoeba evolution occurred at the transition between the Polysphondylia and group 4 Dictyostelia, which comprise the model organism Dictyostelium discoideum, such as the formation of a new structure, the basal disk. Basal disk differentiation and robust stalk formation require the morphogen DIF-1, synthesized by the polyketide synthase StlB, the des-methyl-DIF-1 methyltransferase DmtA, and the chlorinase ChlA, which are conserved throughout Dictyostelia. To understand how the basal disk and other innovations evolved in group 4, we sequenced and annotated the Polysphondylium violaceum (Pvio) genome, performed cell type-specific transcriptomics to identify cell-type marker genes, and developed transformation and gene knock-out procedures for Pvio. We used the novel methods to delete the Pvio stlB gene. The Pvio stlB− mutants formed misshapen curly sorogens with thick and irregular stalks. As fruiting body formation continued, the upper stalks became more regular, but structures contained 40% less spores. The stlB− sorogens overexpressed a stalk gene and underexpressed a (pre)spore gene. Normal fruiting body formation and sporulation were restored in Pvio stlB− by including DIF-1 in the supporting agar. These data indicate that, although conserved, stlB and its product(s) acquired both a novel role in the group 4 Dictyostelia and a role opposite to that in its sister group.

Differentiation in roller tube cultures of wild-type and two cyclic AMP mutant strains of the cellular slime mould Polysphondylium pallidum

1982 ◽  
Vol 28 (10) ◽  
pp. 1143-1149
Author(s):  
Gary D. Paterno ◽  
Danton H. O'Day
Keyword(s):  
Cyclic Amp ◽  
Fruiting Body ◽  
Weight Fraction ◽  
Stalk Cell ◽  
Wild Type ◽  
Submerged Cultures ◽  
Slime Mould ◽  
Polysphondylium Pallidum ◽  
Exogenous Agents ◽  
Cellular Slime

Submerged cultures of wild-type Polysphondylium pallidum (WS320) undergo a developmental sequence in which cells agglutinate and form tight aggregates within which extensive stalk and some spore differentiation occurs. Development of submerged cultures of P. pallidum bears many similarities to fruiting body cultures except that differentiation occurs in the absence of morphogenesis. Here we extend the results of an earlier study of submerged cultures of P. pallidum WS320 (Paterno and O'Day. 1981. Can. J. Microbiol. 27: 924–936) by showing that these cultures respond to several exogenous agents (cyclic AMP, lithium chloride, ammonium chloride, colchicine, and concanavalin A) in the same way as slime mould fruiting body cultures. However, two mutants abnormal in cyclic AMP production which complement to form fruiting bodies on agar plates could not form normal submerged culture aggregates when mixed together. Complementation tests with mutant and wild-type cells also failed. The inability of the mutants (PN507 and PN518) to complement in submerged cultures suggests that their fruiting complementarity may be based on a morphogenetic event. A low molecular weight fraction from wild-type cells could enhance development and stalk cell differentiation in WS320 and one mutant, PN507, but not in PN518. Together these data reveal that submerged cultures can be utilized to test the effects of extracellular factors on development and used as a source for the isolation of factors that regulate cellular differentiation.

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