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J D Gross

Publications and source records attributed to J D Gross.

At least 37 records · Page 2Linked to original sources
PubMed1994

Characterisation of an intracellular Ca2+ pump in Dictyostelium.

Calcium uptake by microsomal membranes from the cellular slime mould Dictyostelium discoideum was measured using Calcium Green-2 as a fluorescent probe of external free Ca2+ concentration. High-affinity Ca2+ uptake was found to be completely inhibited by low concentrations of vanadate, but not by thapsigargin, suggesting that the activity is mediated by a Ca(2+)-ATPase distinct from sarco(endo)plasmic reticulum type of higher animal cells. On sucrose density gradients, Ca2+ uptake distributes with vacuolar proton pump activity and part of the observed Ca2+ uptake is dependent on the pH gradient generated by the vacuolar-type H(+)-ATPase, indicating that the Ca2+ pump is located on both acidic and non-acidic vesicles, possibly derived from the H(+)-ATPase-rich contractile vacuole complex.

Animals↗
PubMed1994

Motion aftereffects with random-dot chequerboard kinematograms: relation between psychophysical and VEP measures.

A random-dot chequerboard kinematogram was used to investigate the effect of motion adaptation both on evoked potentials and on motion aftereffects (MAEs). The experimental paradigm used allowed simultaneous measurement of both variables. Each adaptation period was followed by a series of 5 short test stimuli to which evoked potentials were recorded. Motion aftereffects were observed in the intervals between test stimuli. An inverse relationship between mean N2-P1 amplitude and mean reported MAEs was found as a function of adaptation durations of 1.4, 5.6, and 17.5 s. When the shortest and longest adaptation durations were compared, this relationship held for thirteen of fourteen subjects tested when adaptation-motion and test-motion directions corresponded and for twelve of fourteen subjects when they were opposed. The possibility that the effect of motion adaptation on N2-P1 amplitude was due to local luminance-contrast adaptation is discussed and shown to be unlikely. The suitability of this paradigm for the combined psychophysical and electrophysiological assessment of disturbances in motion perception is discussed.

Adolescent↗
PubMed1994

Developmental decisions in Dictyostelium discoideum.

A few hours after the onset of starvation, amoebae of Dictyostelium discoideum start to form multicellular aggregates by chemotaxis to centers that emit periodic cyclic AMP signals. There are two major developmental decisions: first, the aggregates either construct fruiting bodies directly, in a process known as culmination, or they migrate for a period as "slugs." Second, the amoebae differentiate into either prestalk or prespore cells. These are at first randomly distributed within aggregates and then sort out from each other to form polarized structures with the prestalk cells at the apex, before eventually maturing into the stalk cells and spores of fruiting bodies. Developmental gene expression seems to be driven primarily by cyclic AMP signaling between cells, and this review summarizes what is known of the cyclic AMP-based signaling mechanism and of the signal transduction pathways leading from cell surface cyclic AMP receptors to gene expression. Current understanding of the factors controlling the two major developmental choices is emphasized. The weak base ammonia appears to play a key role in preventing culmination by inhibiting activation of cyclic AMP-dependent protein kinase, whereas the prestalk cell-inducing factor DIF-1 is central to the choice of cell differentiation pathway. The mode of action of DIF-1 and of ammonia in the developmental choices is discussed.

Ammonia↗
PubMed1994

Programmed cell death in Dictyostelium.

Programmed cell death (PCD) of Dictyostelium discoideum cells was triggered precisely and studied quantitatively in an in vitro system involving differentiation without morphogenesis. In temporal succession after the triggering of differentiation, PCD included first an irreversible step leading to the inability to regrow at 8 hours. At 12 hours, massive vacuolisation was best evidenced by confocal microscopy, and prominent cytoplasmic condensation and focal chromatin condensation could be observed by electron microscopy. Membrane permeabilization occurred only very late (at 40-60 hours) as judged by propidium iodide staining. No early DNA fragmentation could be detected by standard or pulsed field gel electrophoresis. These traits exhibit some similarity to those of previously described non-apoptotic and apoptotic PCD, suggesting the hypothesis of a single core molecular mechanism of PCD emerging in evolution before the postulated multiple emergences of multicellularity. A single core mechanism would underly phenotypic variations of PCD resulting in various cells from differences in enzymatic equipment and mechanical constraints. A prediction is that some of the molecules involved in the core PCD mechanism of even phylogenetically very distant organisms, e.g. Dictyostelium and vertebrates, should be related.

Animals↗
PubMed1994

Ammonia hypersensitivity of slugger mutants of D. discoideum.

The weak base ammonia inhibits aggregation and culmination of wild-type amoebae of Dictyostelium discoideum. Here we have examined its effect on a series of 'slugger' mutants previously assigned to 10 complementation groups, and so-called because they remain as slugs for extended periods. We show that the mutants accumulate normal levels of ammonia and hence may be abnormally susceptible to the ammonia they produce. In agreement with this we find that representatives of the slugger complementation groups are hypersensitive to ammonia inhibition at three clearly recognisable morphological stages of development: aggregation, tip formation and culmination. This finding suggests that a common ammonia-sensitive process underlies each of these developmental events.

Amines↗
PubMed1994

Magnetic resonance imaging evidence of a focal pontine ischemia in sudden hearing loss and seventh nerve paralysis.

Sudden sensorineural hearing loss (SSHL) has been associated with many etiologies. However, determining the exact cause of SSHL remains elusive. Recent reports have demonstrated the usefulness of magnetic resonance imaging in identifying the site of lesion in SSHL. This report presents the case of a 68-year-old male who presented with right-sided SSHL associated with right-sided facial paralysis. Magnetic resonance imaging demonstrated a focal area of increased signal on T2-weighted image within the right inferior pons, suggestive of a postischemic edematous focus. Magnetic resonance imaging is becoming important in establishing the etiology of SSHL. Magnetic resonance imaging findings in cases of hearing loss are discussed.

Aged↗
PubMed1993

The target of ammonia action in dictyostelium.

The weak base ammonia is implicated in a number of key processes in Dictyostelium development, notably aggregation and culmination. To determine its intracellular site of action, we compared its biological potency with that of other weak bases. All the bases inhibited these developmental processes effectively, but differed manyfold in potency in accordance with their activity in dissipating pH gradients, as measured by in vivo 31P NMR with pH-sensitive phosphonate probes. These results indicate that weak bases influence development by raising the pH of an intracellular acidic compartment.

Alkalies↗
PubMed1992

ATP-driven Ca2+/H+ antiport in acid vesicles from Dictyostelium.

Amoebae of the cellular slime mold Dictyostelium discoideum possess an extensive and dynamic endomembrane system that includes many types of acidic vacuoles. A light membrane fraction from Dictyostelium, rich in vacuolar-type H(+)-ATPase, has been described [Padh, H., Lavasa, M. & Steck, T.L. (1989) J. Cell Biol. 108, 865-874]. Here, we show that this "acidosomal" fraction also contains a high-affinity vanadate-sensitive Ca2+ uptake activity that is stimulated by the pH gradient formed by the H(+)-ATPase. We attribute this Ca2+ uptake to the presence of a H(+)-countertransporting Ca(2+)-ATPase, pumping Ca2+ into an acidic compartment.

Acid-Base Equilibrium↗
PubMed1991

Potential morphogens involved in pattern formation during Dictyostelium differentiation.

Upon starvation, Dictyostelium amoebae aggregate together and then differentiate into either the stalk or spore cells that, respectively, form the stalk and sorus of the fruiting body. During differentiation, the prestalk and prespore cells become spatially segregated in a clearly defined developmental pattern. Several low molecular weight molecules that influence cell type determination during in vitro differentiation have been identified. The possible role of these molecules as morphogens, responsible for the formation of the developmental pattern, is discussed.

Adenosine↗
PubMed1990

Nested polymerase chain reaction assay for the detection of cytomegalovirus overcomes false positives caused by contamination with fragmented DNA.

The polymerase chain reaction (PCR) technique offers a promising alternative to tissue culture for the rapid and sensitive detection of cytomegalovirus (CMV) infection. However, high levels of background amplification detected in samples containing water but no DNA make interpretation of borderline positive samples extremely difficult and reduce the sensitivity of the assay. The signal from amplification of water or positive samples can be eliminated by DNase treatment, but not by filtration through anisotropic membrane, autoclaving, or ultraviolet irradiation. A lag time of 10 to 12 cycles is observed before the reactions with water will show product formation by liquid hybridization detection. The use of nested PCR eliminates the background and, in serial dilutions of a positive sample, shows a 500- to 1000-fold increase in sensitivity by liquid hybridization detection. We suggest that the background signal is arising from small fragments of DNA, which may be produced by autoclaving viral culture material. Such fragments would escape filtration, and overlapping fragments of DNA can prime one another to form complete mosaic sequences that will then amplify. Nested PCR, appropriately controlled for the number of cycles at each step, should successfully overcome such false positives caused by fragmented DNA, no matter if the contamination occurs at the collection site, in processing, or at the facility performing the test.

Base Sequence↗
PubMed1989

The effect of ammonia on cell-type-specific enzyme accumulation in Dictyostelium discoideum.

We have shown that ammonia inhibits stalk cell formation and promotes spore formation during cyclic AMP induced differentiation of monolayers of sporogenous amoebae (Gross, J.D. et al., Nature (London), 303, 244-245, 1983). Here we show that exposure to ammonia favours the accumulation of prespore- over prestalk-specific enzymes and related products. We conclude that ammonia switches cells from the prestalk to the prespore pathway rather than simply inhibiting stalk cell maturation and hence may act as a morphogen during development.

Acid Phosphatase↗
PubMed1988

Plasma membrane proton pump inhibition and stalk cell differentiation in Dictyostelium discoideum.

The choice of the stalk cell differentiation pathway in Dictyostelium is promoted by an endogenous substance, DIF-1, which is 1-(3,5-dichloro-2,6-dihydroxy-4-methoxyphenyl)-1-hexanone. It is also favoured by weak acids and two inhibitors of the plasma membrane proton pumps of fungi and plants, diethylstilbestrol (DES) and zearalenone, and antagonised by ammonia and other weak bases, which promote spore differentiation. These observations led to the proposal that the choice of differentiation pathway is regulated by intracellular pH. They also prompted the conjecture that DIF-1 itself is a plasma membrane proton pump inhibitor. We report here experiments showing that DIF-1 is not a plasma membrane proton pump inhibitor. We demonstrate that diethylstilbestrol and zearalenone do inhibit the plasma membrane proton pump of Dictyostelium and we show that there is an excellent qualitative and quantitative correlation between the inhibitory activity of these agents, and of a number of other substances, and their ability to divert differentiation from the spore to the stalk pathway. We conclude that inhibition of the plasma membrane proton pump does shift the choice of differentiation pathway in Dictyostelium towards the stalk pathway, but that DIF does not act by this route, and we propose a model for the actions of DIF and plasma membrane proton pump inhibitors in which the differentiation pathway is controlled by the pH of intracellular vesicles rather than by intracellular pH itself. The model invokes a DIF- and proton-activated vesicular chloride channel whose opening permits acidification of the vesicles and lowers cytosolic Ca++ concentration.

Adenosine Triphosphatases↗
PubMed1981

Cell patterning in Dictyostelium.

We summarize studies on stalk and spore cell formation in D. discoideum cell monolayers, aimed at revealing factors involved in controlling the prestalk:prespore pattern in this organism. We propose that there are no cell interactions dependent on cell contact per se. Formation of mature stalk cells from isolated amoebae incubated in a buffered salts medium requires only cyclic AMP and a lipid-like factor (DIF) released by cells developing at high density. In addition, a variety of sporogenous mutants can form spores rapidly and efficiently when incubated at low density in tissue culture dishes containing a similar cyclic AMP and salts medium. In some cases spore formation is improved by the addition of one or other of a variety of protective agents such as bovine serum albumin. Wild-type amoebae at low density form prespore cells under the same conditions. We present some evidence that DIF is the activator of prestalk cell formation in a two-component patterning mechanism of the kind proposed by Wolpert et al. (Symp. Soc. exp. Biol. 25, 391-415 (1971)) and Gierer & Meinhardt (Kybernetik 12, 30-39 (1972)). We also provide data indicating that the role of inhibitor is played by ammonia, an idea first mooted by Sussman & Schindler (Differentiation 10, 1-5 (1978)).

Cell Adhesion↗
PubMed1976

Signal emission and signal propagation during early aggregation in Dictyostelium discoideum.

Waves of chemotactic movement during the early phase of aggregation in Dictyostelium discoideum are of 2 kinds, concentric waves produced by cells that emit cyclic AMP signals spontaneously, and spirals generated by excitations relayed continuously around loops of excitable cells. The period of a spiral wave is the time taken for the excitation to make one complete circuit of the pacemaker loop. We have compared signal emission from the 2 types of source in time-lapse films made at a variety of temperatures. Our results show that spiral waves have a characteristic period length throughout most if not all of the early phase of aggregation, and that the period of concentric waves is generally longer and more variable. Temperature has a pronounced effect on period length and a lesser effect on propagation velocity. We find that each individual wave is propagated at constant velocity over distances of 1-2 cm but that the velocity of successive waves declines. This decline probably reflects some cumulative effect of the chemotactic excitations on the excitable properties of the aggregating cells.

Analysis of Variance↗