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.

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.

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.

Spots and stripes: the patterning spectrum in the cellular slime mould Polysphondylium pallidum

Development ◽  
1989 ◽  
Vol 105 (2) ◽  
pp. 323-333
Author(s):  
J.G. McNally ◽  
E.C. Cox
Keyword(s):  
Pattern Formation ◽  
Specific Antigen ◽  
Spherical Cell ◽  
The Novel ◽  
Cellular Slime Mould ◽  
Slime Mould ◽  
Size And Shape ◽  
Polysphondylium Pallidum ◽  
Cellular Slime ◽  
Pattern Determination

Whorls of the cellular slime mould Polysphondylium pallidum originate as spherical cell masses that during normal morphogenesis produce tips only at equidistant positions around their equator. We have observed a series of new patterns in whorls that differ from normal whorls only in that they are larger or more elongated. Among the novel patterns found were arrays of tips distributed fairly regularly over the whole whorl surface, as well as striped patterns detected at earlier stages with a tip-specific antigen. These altered patterns demonstrate that a whorl's size and shape are by themselves important factors in pattern determination. We have compared the range of observed patterns to those predicted by a variety of different theories. We find that while no one theory can account in detail for all of our observations, predictions based on Turing's scheme of pattern formation come the closest.

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.

Evidence that elevated intracellular cyclic AMP maturation in Dictyostelium

Development ◽  
1989 ◽  
Vol 105 (4) ◽  
pp. 753-759 ◽  
Author(s):  
R.R. Kay
Keyword(s):  
Cyclic Amp ◽  
Fruiting Body ◽  
Normal Development ◽  
Environmental Influences ◽  
Intracellular Camp ◽  
Spore Coat ◽  
Transduction Pathway ◽  
Wild Type ◽  
Type Strains ◽  
Spore Maturation

Spore maturation occurs during normal development in when environmental influences induce a migrating slug a fruiting body. As the amoeboid prespore cells turn spores there is a burst of enzyme accumulation, epimerase, and at a later stage the exocytosis of of the spore coat. Evidence is presented here that triggered by an elevated intracellular cAMP number of rapidly developing (rde) mutants, whose cAMP been investigated previously, are shown to be able to submerged monolayers, whereas wild-type strains are of these mutants are best explained by a derepression transduction pathway utilizing intracellular cAMP. direct, it is shown that the permeant cAMP analogues

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.

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