Regulation of stalk and spore antigen expression in monolayer cultures of Dictyostelium discoideum by pH

Development ◽  
1986 ◽  
Vol 96 (1) ◽  
pp. 131-150
Author(s):  
Janice A. Dominov ◽  
Christopher D. Town

The terminal differentiation of Dictyostelium discoideum cells plated as monolayers with cyclic AMP is dramatically affected by developmental buffer conditions. High pH and addition of weak bases induces spore differentiation while low pH and weak acids favour stalk cell formation. In order to analyse the timing and nature of this regulation we have raised and characterized an anti-stalk serum which we have used together with an anti-spore serum to monitor developmental progression in the monolayer system and to detect the phenotypic effects of pH at earlier stages of development. The stalk serum detects both polysaccharide and protein antigens expressed during the terminal stages of normal development. In monolayer culture, the stalk-specific protein antigen appears precociously, while the timing of prespore vacuole appearance is unaffected. Expression of stalk polysaccharide antigens in monolayer cultures occurs as early as 12 h and is localized in a single subset of cells or region of extracellular space within the small cell clumps that form. The effects of pH (and acid/base) on these phenotype-specific antigens can be detected early in development, shortly after their first appearance. In monolayers of wild-type V12 M2 cells, the low pH regimes appear to act more by suppressing the spore than enhancing the stalk pathway, while the high pH regimes both suppress stalk and enhance spore antigen expression. In monolayers of the sporogenous mutant HM29, low pH regimes both enhance stalk antigen and suppress spore antigen expression. These results show that extracellular pH regulates phenotypic expression during a large part of the differentiation process and is not simply restricted to terminal cytodifferentiation.

1969 ◽  
Vol 53 (4) ◽  
pp. 450-470 ◽  
Author(s):  
Akio Shiraishi ◽  
Hiromichi Morita

Reproducible results describing the effects of pH on the response of the labellar sugar receptor of the fleshfly, Boettcherisca peregrina, were obtained. The response to sucrose was independent over a wide range of pH (3.0 to 10.0 for sucrose stimulation), but was inhibited fairly sharply on both sides of this range. Similar results were obtained for monosaccharide stimulation. The receptor was excited on stimulation by water above pH 12.0. The effects of high pH, both inhibitory and excitatory, were affected by the presence of salts. In the presence of 0.5 molar NaCl, for example, the pH-inhibition curve was shifted toward lower pH's by about one pH unit. The effects of low pH, on the other hand, were not affected by salts. Following Dixon's theory, it was concluded that at least five ionizable groups (loosing positive charges above pH 10.5) were located at the receptor site.


1983 ◽  
Vol 13 (4) ◽  
pp. 301-307 ◽  
Author(s):  
Nancy Neave ◽  
André Sobolewski ◽  
Gerald Weeks

2014 ◽  
Vol 960-961 ◽  
pp. 462-468 ◽  
Author(s):  
Yong Qiu ◽  
Hui Wu ◽  
Guang Qian Luo ◽  
Hong Yao

Gaseous oxidized mercury (Hg2+) in the flue gas is soluble in water and can be captured effectively by Wet flue gas desulfurization (WFGD) system. But in some extent Hg0re-emission happens due to the reduction of absorbed Hg2+, and the pH of slurry is an important factor affecting Hg0re-emission. In this study, the theoretical formulas of slurry pH were derived through the conventional solution theory and then were used to evaluate the factors determining the pH of slurry. A series of laboratory experiments were carried out under N2,CO2and O2/N2atmosphere to measure the Hg0re-emission tendency at different pH values. The results show that the higher the pH, the less Hg2+reduced by S(IV), resulting in the decrease of Hg0re-emission. Under N2atmosphere, the Hg0re-emission was mild at pH>4 while it was dramatic at pH<4. Under O2/N2atmosphere, the addition of O2extended the time span of Hg0re-emission at low pH and increased Hg0re-emission unexpectedly in the latter part of the experiments at high pH. CO2atmosphere almost did not affect Hg0re-emission because of its little effect on the slurry pH.


Development ◽  
1988 ◽  
Vol 104 (1) ◽  
pp. 121-127 ◽  
Author(s):  
L. Kwong ◽  
A. Sobolewski ◽  
L. Atkinson ◽  
G. Weeks

Cells from the pseudoplasmodial stage of Dictyostelium discoideum differentiation were dispersed and separated on Percoll gradients into prestalk and prespore cells. The requirements for stalk cell formation in low-density monolayers from the two cell types were determined. The isolated prespore cells required both the Differentiation Inducing Factor (DIF) and cyclic AMP for stalk cell formation. In contrast, only part of the isolated prestalk cell population required both cyclic AMP and DIF, the remainder requiring DIF alone, suggesting the possibility that there were two populations of prestalk cells, one independent of cyclic AMP and one dependent on cyclic AMP for stalk cell formation. The finding that part of the prestalk cell population required only a brief incubation in the presence of DIF to induce stalk cell formation, whilst the remainder required a considerably longer incubation in the presence of both DIF and cyclic AMP was consistent with this idea. In addition, stalk cell formation from cyclic-AMP-dependent prestalk cells was relatively more sensitive to caffeine inhibition than stalk cell formation from cyclic-AMP-independent prestalk cells. The latter cells were enriched in the most anterior portion of the migrating pseudoplasmodium, indicating that there is spatial segregation of the two prestalk cell populations. The conversion of prespore cells to stalk cells took longer and was more sensitive to caffeine when compared to stalk cell formation from cyclic-AMP-dependent prestalk cells.


2010 ◽  
Vol 9 (5) ◽  
pp. 751-761 ◽  
Author(s):  
Wouter N. van Egmond ◽  
Peter J. M. van Haastert

ABSTRACT The Roco family consists of multidomain Ras-GTPases that include LRRK2, a protein mutated in familial Parkinson's disease. The genome of the cellular slime mold Dictyostelium discoideum encodes 11 Roco proteins. To study the functions of these proteins, we systematically knocked out the roco genes. Previously described functions for GbpC, Pats1, and QkgA (Roco1 to Roco3) were confirmed, while novel developmental defects were identified in roco4- and roco11-null cells. Cells lacking Roco11 form larger fruiting bodies than wild-type cells, while roco4-null cells show strong developmental defects during the transition from mound to fruiting body; prestalk cells produce reduced levels of cellulose, leading to unstable stalks that are unable to properly lift the spore head. Detailed phylogenetic analysis of four slime mold species reveals that QkgA and Roco11 evolved relatively late by duplication of an ancestor roco4 gene (later than ∼300 million years ago), contrary to the situation with other roco genes, which were already present before the split of the common ancestor of D. discoideum and Polysphondylium pallidum (before ∼600 million years ago). Together, our data show that the Dictyostelium Roco proteins serve a surprisingly diverse set of functions and highlight Roco4 as a key protein for proper stalk cell formation.


Development ◽  
1993 ◽  
Vol 119 (3) ◽  
pp. 703-710
Author(s):  
L. Blanton R

Stalk formation in Dictyostelium discoideum involves the synthesis of a stalk tube by the prestalk cell population and stalk cell walls by the individual prestalk cells. Cellulose is a major structural component of the stalk tube and stalk cell walls. The DIF-deficient strain HM44 was used to study the events of stalk formation in monolayer cultures. The induction of cellulose synthase activity was shown to require both DIF and cAMP. Microscopical observations of monolayer cultures using the cellulose-indicating fluorochrome Tinopal LPW demonstrated the presence in these cultures of two cellulose-containing materials: the stalk cell walls and an intercellular material found between cells and around cell clumps. The synthesis of intercellular material precedes that of stalk cell walls in induced cultures. Cells committed to stalk cell formation were delayed in doing so if they were switched to medium containing cAMP but no DIF. During this delay the cells synthesized large quantities of the intercellular material. The intercellular material was shown to be microfibrillar, was sensitive to cellulase, and labelled with a colloidal gold-conjugated cellulase. The intercellular material may have the same mode of cellulose synthesis as that involved in stalk tube formation. If so, that mode would be favored by DIF and cAMP in combination, whereas the cellulose synthesis involved in stalk cell wall formation would be DIF-dependent but delayed or repressed by cAMP.


2017 ◽  
Vol 65 (1) ◽  
pp. 50 ◽  
Author(s):  
Muhammad Yousuf Ali ◽  
Ana Pavasovic ◽  
Peter B. Mather ◽  
Peter J. Prentis

Carbonic anhydrase (CA), Na+/K+-ATPase (NKA) and Vacuolar-type H+-ATPase (HAT) play vital roles in osmoregulation and pH balance in decapod crustaceans. As variable pH levels have a significant impact on the physiology of crustaceans, it is crucial to understand the mechanisms by which an animal maintains its internal pH. We examined expression patterns of cytoplasmic (CAc) and membrane-associated form (CAg) of CA, NKA α subunit and HAT subunit a in gills of freshwater crayfish, Cherax quadricarinatus, at three pH levels – 6.2, 7.2 (control) and 8.2 – over 24 h. Expression levels of CAc were significantly increased at low pH and decreased at high pH conditions 24 h after transfer. Expression increased at low pH after 12 h, and reached its maximum level by 24 h. CAg showed a significant increase in expression at 6 h after transfer at low pH. Expression of NKA significantly increased at 6 h after transfer to pH 6.2 and remained elevated for up to 24 h. Expression for HAT and NKA showed similar patterns, where expression significantly increased 6 h after transfer to low pH and remained significantly elevated throughout the experiment. Overall, CAc, CAg, NKA and HAT gene expression is induced at low pH conditions in freshwater crayfish.


1987 ◽  
Vol 65 (8) ◽  
pp. 1798-1801 ◽  
Author(s):  
J. M. Renaud ◽  
R. B. Stein ◽  
T. Gordon

Changes in force and stiffness during contractions of mouse extensor digitorum longus and soleus muscles were measured over a range of extracellular pH from 6.4 to 7.4. Muscle stiffness was measured using small amplitude (<0.1% of muscle length), high frequency (1.5 kHz) oscillations in length. Twitch force was not significantly affected by changes in pH, but the peak force during repetitive stimulation (2, 3, and 20 pulses) was decreased significantly as the pH was reduced. Changes in muscle stiffness with pH were in the same direction, but smaller in extent. If the number of attached cross-bridges in the muscle can be determined from the measurement of small amplitude, high frequency muscle stiffness, then these findings suggest that (a) the number of cross-bridges between thick and thin filaments declines in low pH and (b) the average force per cross-bridge also declines in low pH. The decline in force per cross-bridge could arise from a reduction in the ability of cross-bridges to generate force during their state of active force production and (or) in an increased percentage of bonds in a low force, "rigor" state.


1981 ◽  
Vol 193 (3) ◽  
pp. 1013-1015 ◽  
Author(s):  
G D Jones ◽  
M T Wilson ◽  
V M Darley-Usmar
Keyword(s):  
Low Ph ◽  
High Ph ◽  

1. A low-pH lithium dodecyl sulphate/polyacrylamide-gradient slab-gel system, suitable for electrophoresis, is described, and the migration properties of standard proteins are compared on this and conventional high-pH gels. 2. Cytochrome oxidase may be partially resolved into its component polypeptides. The order of migration of these is, however, dependent on the pH of the gel system.


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