Induction by acid load of the maturation of prestalk cells in Dictyostelium discoideum

Development ◽  
1988 ◽  
Vol 104 (4) ◽  
pp. 669-681
Author(s):  
K. Inouye
Keyword(s):  
Cell Differentiation ◽  
Dictyostelium Discoideum ◽  
Cell Maturation ◽  
Stalk Cell ◽  
Body Construction ◽  
Acid Load ◽  
Cellular Slime ◽  
Plasma Membrane Proton Pump

During the process of fruiting body construction in the cellular slime mould Dictyostelium discoideum, prestalk cells become mature stalk cells in a well-controlled manner. To identify the natural inducer of stalk cell maturation, substances known to induce stalk cell differentiation under in vitro conditions, and some other related compounds, were examined for their effects in vivo on migrating slugs, the precursor structures of the fruiting bodies. Among these substances, addition of weak acids such as CO2, and addition followed by removal of weak bases such as NH3, strikingly induced the maturation of prestalk cells in situ in slugs. On the other hand, inhibitors of the plasma membrane proton pump did not efficiently induce the maturation of prestalk cells in intact slugs. Differentiation inducing factor (DIF), an endogenous inducer of prestalk differentiation, seemed to be an even poorer inducer of stalk cell maturation when applied to intact slugs. The activities of these substances in inducing stalk cell maturation showed a good correlation with their effects on the cytoplasmic pH (pHi) of prestalk cells; the larger the pHi drop, the stronger the induction of stalk cell maturation, suggesting a requirement for a pHi decrease for the maturation of prestalk cells. Based on these results, it was proposed that stalk cell differentiation, which is induced by DIF, is blocked halfway during normal development by (an) agent(s) that prevent(s) the decrease in pHi.

scholarly journals Secreted Cyclic Di-GMP Induces Stalk Cell Differentiation in the Eukaryote Dictyostelium discoideum

2015 ◽  
Vol 198 (1) ◽  
pp. 27-31 ◽  
Author(s):  
Zhi-hui Chen ◽  
Pauline Schaap
Keyword(s):  
Cell Death ◽  
Cell Differentiation ◽  
Dictyostelium Discoideum ◽  
Autophagic Cell Death ◽  
Stalk Cell ◽  
Signal Molecule ◽  
Major Life ◽  
Content Type

Cyclic di-GMP (c-di-GMP) is currently recognized as the most widely used intracellular signal molecule in prokaryotes, but roles in eukaryotes were only recently discovered. In the social amoebaDictyostelium discoideum, c-di-GMP, produced by a prokaryote-type diguanylate cyclase, induces the differentiation of stalk cells, thereby enabling the formation of spore-bearing fruiting bodies. In this review, we summarize the currently known mechanisms that control the major life cycle transitions ofDictyosteliumand focus particularly on the role of c-di-GMP in stalk formation. Stalk cell differentiation has characteristics of autophagic cell death, a process that also occurs in higher eukaryotes. We discuss the respective roles of c-di-GMP and of another signal molecule, differentiation-inducing factor 1, in autophagic cell deathin vitroand in stalk formationin vivo.

Ammonia depletion and DIF trigger stalk cell intact Dictyostelium discoideum slugs

Development ◽  
1989 ◽  
Vol 105 (3) ◽  
pp. 569-574 ◽  
Author(s):  
M. Wang ◽  
P. Schaap
Keyword(s):  
Cell Differentiation ◽  
Cyclic Amp ◽  
Dictyostelium Discoideum ◽  
Stalk Cell

The differentiation-inducing factor, DIF, was induce stalk cell differentiation in Dictyostelium incubated as submerged monolayers. We investigated the regulates the differentiation of stalk cells in the was found that in migrating or submerged slugs DIF cell differentiation, which is most likely due to the antagonist. Cyclic AMP and ammonia were earlier antagonists in vitro. We show here that ammonia, but an antagonist for DIF-induced stalk cell can induce stalk cell differentiation when ammonia are enzymically depleted. However, depletion of cAMP increase the efficacy of DIF. We propose that the cell differentiation during early culmination may be drop in ammonia levels inside the organism.

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.

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.

Synthesis of a ribosome-bound translatable poly(A)− mRNA during spore germination in Dictyostelium discoideum

1989 ◽  
Vol 35 (5) ◽  
pp. 573-577
Author(s):  
S. Ramagopal
Keyword(s):  
Dictyostelium Discoideum ◽  
Spore Germination ◽  
Gel Electrophoresis ◽  
Rna Synthesis ◽  
In Vitro Translation ◽  
Cellular Slime Mold ◽  
Free System ◽  
Cellular Slime

A distinct poly(A)− RNA sedimenting around 10–12S was identified during spore germination in Dictyostelium discoideum. Activated spores were labeled with [3H]uracil and the poly(A)− RNA was purified from ribosomal particles for analysis. In the spore swelling stage, 40 to 50% of the newly synthesized poly(A)− RNA was 10–12S RNA. This fraction diminished to one-half or one-fourth depending on the labeling period at the stage of amoeba emergence. The 10–12S RNA was associated with both monosomes and polysomes in vivo. Translation in a wheat germ cell-free system and gel electrophoresis demonstrated that the 10–12S RNA coded for a number of polypeptides, some of which were also represented among the in vitro products of poly(A)+ RNA. However, there were seven unique polypeptides (37.5, 28.2, 27.5, 23, 17.7, 17, and 14.2 kilodaltons) encoded exclusively by 10–12S RNA.Key words: cellular slime mold, RNA synthesis, development, poly(A)− mRNA, in vitro translation.

scholarly journals The proximal pathway of metabolism of the chlorinated signal molecule differentiation-inducing factor-1 (DIF-1) in the cellular slime mould Dictyostelium

1995 ◽  
Vol 306 (3) ◽  
pp. 735-743 ◽  
Author(s):  
P Morandini ◽  
J Offer ◽  
D Traynor ◽  
O Nayler ◽  
D Neuhaus ◽  
...  
Keyword(s):  
Side Chain ◽  
Stalk Cell ◽  
Reaction Products ◽  
Signal Molecule ◽  
Slime Mould ◽  
Phenolic Ring ◽  
Cellular Slime ◽  
Cytochrome P 450

Stalk cell differentiation during development of the slime mould Dictyostelium is induced by a chlorinated alkyl phenone called differentiation-inducing factor-1 (DIF-1). Inactivation of DIF-1 is likely to be a key element in the DIF-1 signalling system, and we have shown previously that this is accomplished by a dedicated metabolic pathway involving up to 12 unidentified metabolites. We report here the structure of the first four metabolites produced from DIF-1, as deduced by m.s., n.m.r. and chemical synthesis. The structures of these compounds show that the first step in metabolism is a dechlorination of the phenolic ring, producing DIF metabolite 1 (DM1). DM1 is identical with the previously known minor DIF activity, DIF-3. DIF-3 is then metabolized by three successive oxidations of its aliphatic side chain: a hydroxylation at omega-2 to produce DM2, oxidation of the hydroxy group to a ketone group to produce DM3 and a further hydroxylation at omega-1 to produce DM4, a hydroxyketone of DIF-3. We have investigated the enzymology of DIF-1 metabolism. It is already known that the first step, to produce DIF-3, is catalysed by a novel dechlorinase. The enzyme activity responsible for the first side-chain oxidation (DIF-3 hydroxylase) was detected by incubating [3H]DIF-3 with cell-free extracts and resolving the reaction products by t.l.c. DIF-3 hydroxylase has many of the properties of a cytochrome P-450. It is membrane-bound and uses NADPH as co-substrate. It is also inhibited by CO, the classic cytochrome P-450 inhibitor, and by several other cytochrome P-450 inhibitors, as well as by diphenyliodonium chloride, an inhibitor of cytochrome P-450 reductase. DIF-3 hydroxylase is highly specific for DIF-3: other closely related compounds do not compete for the activity at 100-fold molar excess, with the exception of the DIF-3 analogue lacking the chlorine atom. The Km for DIF-3 of 47 nM is consistent with this enzyme being responsible for DIF-3 metabolism in vivo. The two further oxidations necessary to produce DM4 are also performed in vitro by similar enzyme activities. One of the inhibitors of DIF-3 hydroxylase, ancymidol (IC50 67 nM) is likely to be particularly suitable for probing the function of DIF metabolism during development.

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