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V Nanjundiah

Publications and source records attributed to V Nanjundiah.

14 recordsLinked to original sources

Spatial variation of sequestered calcium in the multicellular stage of Dictyostelium discoideum as assayed by chlortetracycline fluorescence.

We have used chlortetracycline (CTC) as a fluorescent probe to detect the distribution of sequestered calcium in multicellular stages of Dictyostelium discoideum. Tips of late aggregates, slugs and early culminating masses fluoresce very strongly. Most of the fluorescence is intracellular in origin and emanates from a small number of intense punctate sources. The sources correspond in part to autophagic vacuoles vis. neutral-red staining, acidic digestive vesicles, and may also include intracellular organelles; cytoplasmic fluorescence is much weaker in comparison. The level of fluorescence drops in the middle portion of slugs and rises again in the posteriormost region, though not to as high a level as in the tip. This holds good irrespective of whether CTC is applied only in the neighbourhood of the aggregate centre, only in the aggregate periphery, or to the whole aggregate. We infer that there must be a good deal of mixing in the stages leading from aggregation to slug formation; thus the serial order in which cells enter an aggregate does not bear any relation to their ultimate fates. The other implication of our study is that calcium sequestration is much more extensive in prestalk and anterior-like cells than in prespore cells. These findings are discussed with regard to possible implications for pattern formation.

Animals

The evolution of genomic imprinting.

We explore three possible pathways for the evolution of genomic imprinting. (1) Imprinting may be advantageous in itself when imprinted and unimprinted alleles of a locus confer different phenotypes. If a segment of DNA is imprinted in the gametes of one sex but not in those of the other, it might lead to effects correlated with sexual dimorphism. More fundamentally, in certain organisms, sex determination might have evolved because of imprinting. When imprinting leads to chromosome elimination or inactivation and occurs in some embryos but not in others, two classes of embryos, differing in the number of functional gene copies, would result. A model for sex determination based on inequality in the actual or effective copy-number of particular noncoding, regulatory sequences of DNA has been proposed (Chandra, Proc. natn. Acad. Sci. U.S.A. 82. 1165-1169 and 6947-6949, 1985). Maternal control of offspring sex is another possible consequence of imprinting; this would indicate a potential role for imprinting in sex ratio evolution. (2) Genes responsible for imprinting may have pleiotropic effects and they may have been selected for reasons other than their imprinting ability. Lack of evidence precludes further consideration of this possibility. (3) Imprinting could have co-evolved with other traits. For instance, gamete-specific imprinting could lead to a lowered fitness of androgenetic or gynogenetic diploids relative to the fitness of 'normal' diploids. This in turn would reinforce the evolution of anisogamy. The reversibility of imprinting raises the possibility of occasional incomplete or improper erasure.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Computer viruses.

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Software

Periodic stimuli are more successful than randomly spaced ones for inducing development in Dictyostelium discoideum.

Aggregation in the cellular slime mold Dictyostelium discoideum is due to chemotaxis. The chemoattractant, cyclic AMP, is synthesised and released periodically by the cells. Externally applied periodic pulses of cyclic AMP can also induce differentiation in this organism. The present work examines the role of periodicity per se in cyclic AMP-mediated stimulation of cell differentiation. For this purpose we use Agip53, a Dictyostelium mutant which does not develop beyond the vegetative state but can be made to aggregate and differentiate by reiterated applications of cyclic AMP. Importantly, Agip53 cells do not make or release any cyclic AMP themselves even in response to an increase in extracellular cyclic AMP. A comparison of the relative efficiencies of periodic and aperiodic stimulation shows that whereas the two patterns of stimulation are equally effective in inducing the formation of EDTA-stable cell contacts, periodic stimuli are significantly superior for inducing terminal differentiation. This suggests that there must be molecular pathways which can only function when stimulation occurs at regular intervals.

Cyclic AMP

PH oscillations in cell suspensions of Dictyostelium discoideum: their relation to cyclic-amp signals.

Cells of Dictyostelium discoideum known to release cyclic AMP (cAMP) rhythmically in the form of pulses, change with the same period of about 8 min the pH of their medium. The pH is used here as an indicator to investigate the effect of externally added cAMP pulses on the oscillations. Both a temporary increase in amplitude and a permanent phase shift can be induced. The phase-response curve indicates that the period can be increased and decreased by rhythmic stimulation with cAMP pulses.

Cyclic AMP

A theoretical study of the effects of cyclic AMP phosphodiesterases during aggregation in Dictyostelium.

During aggregation the larger Dictyostelium species use cAMP as a chemoattractant and possibly also as a transmitter. In passage from cell to cell, cAMP levels are modulated by diffusion and by enzyme hydrolysis. It appears that the important cAMP-hydrolysing enzyme is a phosphodiesterase bound to the cell membrane, the main roles of which are (1) very fast hydrolysis of cAMP and (2) steepening of spatial cAMP gradients. An extracellular phosphodiesterase has no function, so far as can be conjectured from present data.

3',5'-Cyclic-AMP Phosphodiesterases

A plausible role for a membrane-bound cyclic AMP phosphodiesterase in cellular slime mold chemotaxis.

1. Kinetics of membrane-bound cyclic AMP phosphodiesterase of the cellular slime mold, Dictyostelium discoideum, were studied under two conditions: in the 27 000 times g sediment of cell homogenates (particle-bound phosphodiesterase) and in cell suspensions using external cyclic AMP as a substrate (cell-bound phosphodiesterase). Both methods revealed non-Michaelian kinetics with interaction coefficients less than 1. 2. The membrane-bound phosphodiesterase has a specificity different from that of the cyclic AMP receptor, also present at the cell surface. 3. The membrane-bound enzyme was solubilized by lithium 3, 5-diiodosalicylate and partially purified. In this state the non-linear kinetics were still retained; however, the enzyme was not inhibited by the D. discoideum inhibitor, unlike the cell-bound phosphodiesterase in vivo. This indicates that both enzymes share an inhibitor binding site and that this site is cryptic in the cell-bound state. 4. Production of periodic cyclic AMP pulses by centers, and their relay by other cells, is believed to occur during aggregation. It is suggested that the cell-bound enzyme determines a "time window" significantly smaller than the period of pulsing, and optimizes stimulation of the cyclic AMP receptors in chemotaxis and signal relaying.

Binding Sites

Signal input for a chemotactic response in the cellular slime mold Dictyostelium discoideum.

Drops with different concentrations of 3':5'-cyclic AMP were deposited at various distances from small populations of Dictyostelium discoideum amoebae, and the distances over which 50% of the amoebae drops reacted positively were determined. The linear regression analysis of a bouble logarithmic plot of distance against concentration gives a straight line with a slope of 1/4.25, which value suggests that the amoebae respond to a spatial gradient of cyclic AMP concentration. The threshold value for the signal is 3.6 X 10(-9) M/mm with a sensitivity of measurement of about 1%. These findings are discussed in relation to our present knowledge of cyclic AMP receptors.

Chemotaxis