Evidence that positional information is used to establish the prestalk-prespore pattern in <i>Dictyostelium discoideum</i>
 aggregates

Krefft, Marianne; Voet, Ludwig; Gregg, John R.; Mairhofer, Helga; Williams, Keith L.

doi:10.1002/j.1460-2075.1984.tb01784.x

Cited by 86 publications

(38 citation statements)

References 23 publications

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“…They are also consistent with the results of Takeuchi and co-workers, who used antibodies against a carbohydrate moiety common to prespore proteins ITakeuchi 1963;Tasaka et al 1983;Takemoto et al 1985;Gomer et al 1986a). The localized developmental pattern of SP60 expression reveals the initial patterning of prespore cells and is consistent with the involvement of spatial determinants in this process and with the results of Krefft et al (1984) for the D19 gene product.…”

Section: Discussionsupporting

confidence: 84%

“…This is also consistent with earlier two-dimensional gel results examining the expression of SP60 in dissected anterior and posterior portions of migrating slugs and mature spores (Devine et al 1983;Morrissey et al 1984). A similar pattern is seen for the prespore gene DI9, which encodes a protein found in the slime sheath surrounding the migrating slug, through the use of monoclonal antibodies and lacZ fusion constructs (Krefft et al 1984;Dingermann et al 1990). Untransformed cells (see…”

Section: Spatial Expression Of Fusion Gene Productssupporting

confidence: 88%

“…Cells starved early in the cell cycle (S and early G2; there is no detectable G~) preferentially differentiate into prestalk cells, whereas those starved in mid and late G2 differentiate into prespore cells (Weijer et al 1984;Gomer and Firtel 1987b). After the pseudoplasmodium has formed, cAMP, adenosine, NH4 +, and the morphogen DIF all play roles in regulating gene expression, patterning, and subsequent differentiation (see Mehdy et al 1983;Chisholm et al 1984;Williams et al 1984;Mehdy and Firtel 1985;Schaap and van Driel 1985;Gomer et al 1986b;Oyama and Blumberg 1986;Schaap and Wang 1986;Wang et al 1988).…”

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confidence: 99%

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A spatial gradient of expression of a cAMP-regulated prespore cell-type-specific gene in Dictyostelium.

Haberstroh¹,

Firtel²

1990

Genes Dev.

125

109

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Previously, we identified a class of genes in Dictyostelium that are prespore cell-type specific in their expression in the multicellular aggregate and are inducible by cAMP acting through cell-surface cAMP receptors. In this paper, we report the cloning and analysis of the regulatory regions controlling the expression of one such gene that encodes a spore coat protein, SP60. By use of a fusion of the firefly luciferase gene and Escherichia coli lacZ [expresses [3-galactosidase (~-gal)], we have identified cis-acting regions required for proper spatial and temporal expression in multicellular aggregates and for cAMP induction in shaking cell culture. Deletion analysis suggests that a CA-rich element (CAE) and surrounding sequences present three times within the 5'-flanking sequence are required for proper regulation. SP60-IacZ fusions that include all three of these regions express lacZ only in the posterior -85% of migrating slugs (prespore zone). Studies show that SP60 is expressed during mid to late aggregation, and SP60-lacZ-positive cells are spatially localized as a doughnut-shaped ring within the forming aggregate. Cells within the skirt that surrounds the aggregate and that are still migrating into the aggregate do not stain. Sequential 5' deletions of CAEs and surrounding regions affect the expression level of SP60-luciferase in response to developmental signals and cAMP, as well as the spatial pattern of SP60-IacZ. Deletion of the first (most 5') of these regions restricts the spatial expression of SP60-IacZ fusions to the anterior of the prespore zone. When both the first and second regions are removed, the expression level drops, and the staining is restricted to the prespore/prestalk boundary. Furthermore, the staining pattern that is seen with these two deletions is present as a gradient from anterior to posterior within the prespore zone. Deletion of all three regions results in a loss of both cAMP and developmentally induced expression. These results suggest the presence of a gradient within the prespore zone that differentially affects the activity of promoters containing different numbers of response elements.

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Section: Discussionsupporting

confidence: 84%

Section: Spatial Expression Of Fusion Gene Productssupporting

confidence: 88%

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confidence: 99%

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A spatial gradient of expression of a cAMP-regulated prespore cell-type-specific gene in Dictyostelium.

Haberstroh¹,

Firtel²

1990

Genes Dev.

125

109

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“…Figure 1A shows the typical pattern of labelling of untreated slug cells with monoclonal antibody MUD1, which specifically labels the surface of prespore cells (9). Note that both FALS and LI90 are appreciable for these unpermeabilised cells.…”

Section: Resultsmentioning

confidence: 99%

Detergent treatment of dictyostelium discoideum cells allows examination of internal cell type‐specific antigens by flow cytometry

Bernstein

Browne

et al. 1988

Cytometry

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Monoclonal antibodies are used extensively in flow cytometry to identify subpopulations of cells differing in surface antigens. Conventional studies on living cells do not allow analysis of internal antigens, because antibody molecules do not pass through an intact plasma membrane. It is important for developmental studies on Dictgostelium discoideum that not only surface but also internal antigens be analysed. Here techniques are reported that make possible such studies by permeabilising cells with mild detergent treatments using digitonin. Over the past ten years there has been a revolution in culture cells that it is possible to permeabilise them so the understanding of the differentiation of lymphocyte that antibodies have access to internal antigens for flow subpopulations. The isolation of monoclonal antibodies cytometric measurements (14). that recognise antigenic determinants exposed at the In this report, studies that optimise techniques for surface of blood cell subpopulations and their applica-analysing internal antigens of D. discoideurn slug cells tion in flow cytometry has hastened this understanding are described. With mild detergent treatments, it is pos-(15). We have developed techniques for disaggregating sible to analyse internal antigens of unfixed cells. The the cells of the cellular slime mold Dictyostelium discoz-techniques outlined here are simple and rapid. We use deum slug, which is one of the simplest multicellular digitonin to permeabilise unfixed cells and to demoneukaryotes, and analysing them by flow cytometry (16). strate the effectiveness of this technique using two preOur studies centre on the differentiation of this organ-spore-specific internal antigens. ism, to produce two predominant cell types, the prespore and prestalk cells (16). In this way, cell surface antigens MATERIALS AND METHODS that identify prespore (9) or prestalk (10) cells have been Materials discovered.Digitonin (80% pure) was from Calbiochem. DeterIn recent work antigens that are cell type-specific but gents Triton X-100 and Nonidet P-40 were from Sigma. not found at the cell surface have been discovered (Alex-Monoclonal antibodies MUD1 (71, MUD3 (171, and ander et al., submitted). While internal antigens can be MUD102 (Smith, unpublished) raised against D. discoistudied after sorting the cells on the basis of cell surface markers, it would be more convenient to be able to detect internal antigens, directly in mixed-cell popula-'This research was supported by a NH & MRC (Australia) p a n t to tions, in the Same way that cell surface antigens are KLW. RLB was on sabbatical leave from San Francisco State studied by flow cytometry. Since antibodies cannot pen-u~~~~~r e p r i n t requests to Keith Williams, School of Biological etrate intact flow cytometry precludes such studies. However, it has been shown in tissue 2109.

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“…Studies from several laboratories have suggested that prestalk and prespore cells arise in a spatially random manner throughout the aggregate and that sorting results from cell-type specific alterations in adhesion (Abe et al, 1994;Nicol et al, 1999). However, it has also been proposed that pattern can be generated by position-dependent differentiation of the different cell types (Krefft et al, 1984;Wolpert, 1981). The positional information model states that the value related to a position in a coordinate system determines cell differentiation (Wolpert, 1989;Wolpert and Szathmary, 2002).…”

Section: Research Articlementioning

confidence: 99%

The cell adhesion molecule DdCAD-1 regulates morphogenesis through differential spatiotemporal expression inDictyostelium discoideum

et al. 2011

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SUMMARYDuring development of Dictyostelium, multiple cell types are formed and undergo a coordinated series of morphogenetic movements guided by their adhesive properties and other cellular factors. DdCAD-1 is a unique homophilic cell adhesion molecule encoded by the cadA gene. It is synthesized in the cytoplasm and transported to the plasma membrane by contractile vacuoles. In chimeras developed on soil plates, DdCAD-1-expressing cells showed greater propensity to develop into spores than did cadA-null cells. When development was performed on non-nutrient agar, wild-type cells sorted from the cadA-null cells and moved to the anterior zone. They differentiated mostly into stalk cells and eventually died, whereas the cadA-null cells survived as spores. To assess the role of DdCAD-1 in this novel behavior of wild-type and mutant cells, cadA-null cells were rescued by the ectopic expression of DdCAD-1-GFP. Morphological studies have revealed major spatiotemporal changes in the subcellular distribution of DdCAD-1 during development. Whereas DdCAD-1 became internalized in most cells in the post-aggregation stages, it was prominent in the contact regions of anterior cells. Cell sorting was also restored in cadA -slugs by exogenous recombinant DdCAD-1. Remarkably, DdCAD-1 remained on the surface of anterior cells, whereas it was internalized in the posterior cells. Additionally, DdCAD-1-expressing cells migrated slower than cadA -cells and sorted to the anterior region of chimeric slugs. These results show that DdCAD-1 influences the sorting behavior of cells in slugs by its differential distribution on the prestalk and prespore cells.

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Evidence that positional information is used to establish the prestalk-prespore pattern in Dictyostelium discoideum aggregates

Cited by 86 publications

References 23 publications

A spatial gradient of expression of a cAMP-regulated prespore cell-type-specific gene in Dictyostelium.

A spatial gradient of expression of a cAMP-regulated prespore cell-type-specific gene in Dictyostelium.

Detergent treatment of dictyostelium discoideum cells allows examination of internal cell type‐specific antigens by flow cytometry

The cell adhesion molecule DdCAD-1 regulates morphogenesis through differential spatiotemporal expression inDictyostelium discoideum

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