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Responses of the ciliates Tetrahymena and Paramecium to external ATP and GTP.

ABSTRACT: The unicellular ciliates Paramecium and Tetrahymena are the simplest eukaryotic cells to show reliable depolarizing responses to micromolar concentrations of external ATP and GTP. Their simplicity allows for combined analysis of swimming behavior, electrophysiology, receptor binding, behavioral mutant and drug screens as well as molecular genetic approaches such as RNAi and gene knockouts experiments. ATP and GTP are depolarizing chemorepellents in both ciliates, producing measurable receptor potentials and Ca(2+)-based action potentials that are correlated with jerking behaviors called avoiding reactions (AR). GTP also causes repetitive continuous ciliary reversals (CCR) and oscillating plateau depolarizations in Paramecium. Both ciliates show high affinity, saturable external binding of (32)P-GTP and (32)P-ATP but GTP does not compete for ATP binding and vice versa. Chemosensory adaptation occurs after continued exposure (15 min) to these ligands, producing a loss of external binding and forward swimming. However, cells adapted to ATP still bind and respond to GTP and GTP-adapted cells still bind and respond to ATP. This, combined with pharmacological analyses, suggests that there are two separate receptor systems: A metabotropic ATP receptor pathway and a different, novel GTP receptor pathway. A Paramecium mutant (ginA) lacks the GTP-induced oscillating depolarizations but does show AR in GTP, unveiling isolated GTP-receptor potentials for study. An ecto-ATPase is also present that may be involved in inactivation of ATP and GTP signals. Gene knockout experiments are currently underway to determine the roles of the ecto-ATPase and a putative 7-transmembrane spanning receptor in these responses.

SUBMITTER: Hennessey TM

PROVIDER: S-EPMC2096533 | biostudies-literature | 2005 Jun

REPOSITORIES: biostudies-literature

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Responses of the ciliates Tetrahymena and Paramecium to external ATP and GTP.

Hennessey Todd M TM

Purinergic signalling 20050317 2

The unicellular ciliates Paramecium and Tetrahymena are the simplest eukaryotic cells to show reliable depolarizing responses to micromolar concentrations of external ATP and GTP. Their simplicity allows for combined analysis of swimming behavior, electrophysiology, receptor binding, behavioral mutant and drug screens as well as molecular genetic approaches such as RNAi and gene knockouts experiments. ATP and GTP are depolarizing chemorepellents in both ciliates, producing measurable receptor po ...[more]

PMID: 18404496

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Project description:BackgroundConsiderable attention has focused on how selection on dispersal and other core life-history strategies (reproductive effort, survival ability, colonization capacity) may lead to so-called dispersal syndromes. Studies on genetic variation in these syndromes within species could importantly increase our understanding of their evolution, by revealing whether traits co-vary across genetic lineages in the manner predicted by theoretical models, and by stimulating further hypotheses for experimental testing. Yet such studies remain scarce. Here we studied the ciliated protist Tetrahymena thermophila, a particularly interesting organism due to cells being able to transform into morphs differing dramatically in swim-speed. We investigated dispersal, morphological responses, reproductive performance, and survival in ten different clonal strains. Then, we examined whether life history traits co-varied in the manner classically predicted for ruderal species, examined the investment of different strains into short- and putative long-distance dispersal, while considering also the likely impact of semi-sociality (cell aggregation, secretion of 'growth factors') on dispersal strategies.ResultsVery significant among-strain differences were found with regard to dispersal rate, morphological commitment and plasticity, and almost all core life-history traits (e.g. survival, growth performance and strategy), with most of these traits being significantly intercorrelated. Some strains showed high short-distance dispersal rates, high colonization capacity, bigger cell size, elevated growth performance, and good survival abilities. These well performing strains, however, produced fewer fast-swimming dispersal morphs when subjected to environmental degradation than did philopatric strains performing poorly under normal conditions.ConclusionStrong evidence was found for a genetic covariation between dispersal strategies and core life history traits in T. thermophila, with a fair fit of observed trait associations with classic colonizer models. However, the well performing strains with high colonization success and short-distance dispersal likely suffered under a long-distance dispersal disadvantage, due to producing fewer fast-swimming dispersal morphs than did philopatric strains. The smaller cell size at carrying capacity of the latter strains and their poor capacity to colonize as individual cells suggest that they may be adapted to greater levels of dependency on clone-mates (stronger sociality). In summary, differential exposure to selection on competitive and cooperative abilities, in conjunction with selective factors targeting specifically dispersal distance, likely contributed importantly to shaping T. thermophila dispersal and life history evolution.

Project description:Understanding how and why individual movement translates into dispersal between populations is a long-term goal in ecology. Movement is broadly defined as 'any change in the spatial location of an individual', whereas dispersal is more narrowly defined as a movement that may lead to gene flow. Because the former may create the condition for the latter, behavioural decisions that lead to dispersal may be detectable in underlying movement behaviour. In addition, dispersing individuals also have specific sets of morphological and behavioural traits that help them coping with the costs of movement and dispersal, and traits that mitigate costs should be under selection and evolve if they have a genetic basis. Here, we experimentally study the relationships between movement behaviour, morphology and dispersal across 44 genotypes of the actively dispersing unicellular, aquatic model organism Tetrahymena thermophila. We used two-patch populations to quantify individual movement trajectories, as well as activity, morphology and dispersal rate. First, we studied variation in movement behaviour among and within genotypes (i.e. between dispersers and residents) and tested whether this variation can be explained by morphology. Then, we addressed how much the dispersal rate is driven by differences in the underlying movement behaviour. Genotypes revealed clear differences in terms of movement speed and linearity. We also detected marked movement differences between resident and dispersing individuals, mediated by the genotype. Movement variation was partly explained by morphological properties such as cell size and shape, with larger cells consistently showing higher movement speed and higher linearity. Genetic differences in activity and movement were positively related to the observed dispersal and jointly explained 47% of the variation in dispersal rate. Our study shows that a detailed understanding of the interplay between morphology, movement and dispersal may have potential to improve dispersal predictions over broader spatio-temporal scales.

Project description:The F-type ATP synthase complex is a rotary nano-motor driven by proton motive force to synthesize ATP. Its F(1) sector catalyzes ATP synthesis, whereas the F(o) sector conducts the protons and provides a stator for the rotary action of the complex. Components of both F(1) and F(o) sectors are highly conserved across prokaryotes and eukaryotes. Therefore, it was a surprise that genes encoding the a and b subunits as well as other components of the F(o) sector were undetectable in the sequenced genomes of a variety of apicomplexan parasites. While the parasitic existence of these organisms could explain the apparent incomplete nature of ATP synthase in Apicomplexa, genes for these essential components were absent even in Tetrahymena thermophila, a free-living ciliate belonging to a sister clade of Apicomplexa, which demonstrates robust oxidative phosphorylation. This observation raises the possibility that the entire clade of Alveolata may have invented novel means to operate ATP synthase complexes. To assess this remarkable possibility, we have carried out an investigation of the ATP synthase from T. thermophila. Blue native polyacrylamide gel electrophoresis (BN-PAGE) revealed the ATP synthase to be present as a large complex. Structural study based on single particle electron microscopy analysis suggested the complex to be a dimer with several unique structures including an unusually large domain on the intermembrane side of the ATP synthase and novel domains flanking the c subunit rings. The two monomers were in a parallel configuration rather than the angled configuration previously observed in other organisms. Proteomic analyses of well-resolved ATP synthase complexes from 2-D BN/BN-PAGE identified orthologs of seven canonical ATP synthase subunits, and at least 13 novel proteins that constitute subunits apparently limited to the ciliate lineage. A mitochondrially encoded protein, Ymf66, with predicted eight transmembrane domains could be a substitute for the subunit a of the F(o) sector. The absence of genes encoding orthologs of the novel subunits even in apicomplexans suggests that the Tetrahymena ATP synthase, despite core similarities, is a unique enzyme exhibiting dramatic differences compared to the conventional complexes found in metazoan, fungal, and plant mitochondria, as well as in prokaryotes. These findings have significant implications for the origins and evolution of a central player in bioenergetics.

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Responses of the ciliates Tetrahymena and Paramecium to external ATP and GTP.

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Responses of the ciliates Tetrahymena and Paramecium to external ATP and GTP.

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