cyclic-gmp and Starvation

Generally, the second messenger bis-(3'-5')-cyclic dimeric GMP (c-di-GMP) regulates the switch between motile and sessile lifestyles in bacteria. Here, we show that c-di-GMP is an essential regulator of multicellular development in the social bacterium Myxococcus xanthus. In response to starvation, M. xanthus initiates a developmental program that culminates in formation of spore-filled fruiting bodies. We show that c-di-GMP accumulates at elevated levels during development and that this increase is essential for completion of development whereas excess c-di-GMP does not interfere with development. MXAN3735 (renamed DmxB) is identified as a diguanylate cyclase that only functions during development and is responsible for this increased c-di-GMP accumulation. DmxB synthesis is induced in response to starvation, thereby restricting DmxB activity to development. DmxB is essential for development and functions downstream of the Dif chemosensory system to stimulate exopolysaccharide accumulation by inducing transcription of a subset of the genes encoding proteins involved in exopolysaccharide synthesis. The developmental defects in the dmxB mutant are non-cell autonomous and rescued by co-development with a strain proficient in exopolysaccharide synthesis, suggesting reduced exopolysaccharide accumulation as the causative defect in this mutant. The NtrC-like transcriptional regulator EpsI/Nla24, which is required for exopolysaccharide accumulation, is identified as a c-di-GMP receptor, and thus a putative target for DmxB generated c-di-GMP. Because DmxB can be-at least partially-functionally replaced by a heterologous diguanylate cyclase, these results altogether suggest a model in which a minimum threshold level of c-di-GMP is essential for the successful completion of multicellular development in M. xanthus.

Topics: Bacterial Proteins; Cyclic GMP; Escherichia coli Proteins; Gene Expression Regulation, Bacterial; Myxococcus xanthus; Phosphorus-Oxygen Lyases; Spores, Bacterial; Starvation

All terrestrial animals must find a proper level of moisture to ensure their health and survival. The cellular-molecular basis for sensing humidity is unknown in most animals, however. We used the model nematode Caenorhabditis elegans to uncover a mechanism for sensing humidity. We found that whereas C. elegans showed no obvious preference for humidity levels under standard culture conditions, worms displayed a strong preference after pairing starvation with different humidity levels, orienting to gradients as shallow as 0.03% relative humidity per millimeter. Cell-specific ablation and rescue experiments demonstrate that orientation to humidity in C. elegans requires the obligatory combination of distinct mechanosensitive and thermosensitive pathways. The mechanosensitive pathway requires a conserved DEG/ENaC/ASIC mechanoreceptor complex in the FLP neuron pair. Because humidity levels influence the hydration of the worm's cuticle, our results suggest that FLP may convey humidity information by reporting the degree that subcuticular dendritic sensory branches of FLP neurons are stretched by hydration. The thermosensitive pathway requires cGMP-gated channels in the AFD neuron pair. Because humidity levels affect evaporative cooling, AFD may convey humidity information by reporting thermal flux. Thus, humidity sensation arises as a metamodality in C. elegans that requires the integration of parallel mechanosensory and thermosensory pathways. This hygrosensation strategy, first proposed by Thunberg more than 100 y ago, may be conserved because the underlying pathways have cellular and molecular equivalents across a wide range of species, including insects and humans.

Topics: Acid Sensing Ion Channels; Animals; Behavior, Animal; Caenorhabditis elegans; Caenorhabditis elegans Proteins; Cyclic GMP; Epithelial Sodium Channels; Humans; Humidity; Ion Channel Gating; Ion Channels; Mechanoreceptors; Membrane Proteins; Motor Activity; Multiprotein Complexes; Sensory Receptor Cells; Starvation; Thermosensing

Nonalcoholic fatty liver disease (NAFLD) is the most common chronic liver disorder caused by abnormal lipid metabolisms, such as reduced hepatic fatty acid oxidation (FAO), but intracellular control of FAO under physio- and pathological conditions remains largely undefined. Here, we demonstrate that deprivation of Slc7a3a leads to hepatic steatosis in fasted zebrafish as a result of defects in arginine-dependent nitric oxide (NO) synthesis. Fast-induced hepatic steatosis in slc7a3a-null mutants can be rescued by treatments with NO donor, cyclic guanosine monophosphate analog, adenosine-monophosphate-activated protein kinase (AMPK) activator, or peroxisome proliferator-activated receptor alpha (PPAR-α) agonist. In contrast, inhibitors of NO synthases, AMPK, or soluble guanylate cyclase and liver-specifically expressed dominant negatives of peroxisome proliferator-activated receptor-gamma coactivator 1 alpha and PPAR-α are sufficient to induce hepatic steatosis in fasted wild-type larvae. Moreover, knockdown of Slc7a3 in mice or SLC7A3 in human liver cells impaired AMPK-PPAR-α signaling and resulted in lipid accumulation under fasting or glucose starvation, respectively.. These findings have revealed a NO-AMPK-PPAR-α-signaling pathway that is crucial for the control of hepatic FAO in vertebrates.

Topics: Amino Acid Transport Systems, Basic; AMP-Activated Protein Kinases; Animals; Cell Line; Cyclic GMP; Fasting; Fatty Liver; Humans; Lipid Metabolism; Liver; Mice; Mutation; Nitric Oxide; Phenotype; PPAR alpha; Starvation; Transcription Factors; Zebrafish; Zebrafish Proteins

Food searching strategies of animals are key to their success in heterogeneous environments. The optimal search strategy may include specialized random walks such as Levy walks with heavy power-law tail distributions, or persistent walks with preferred movement in a similar direction. We have investigated the movement of the soil amoebae Dictyostelium searching for food. Dictyostelium cells move by extending pseudopodia, either in the direction of the previous pseudopod (persistent step) or in a different direction (turn). The analysis of approximately 4000 pseudopodia reveals that step and turn pseudopodia are drawn from a probability distribution that is determined by cGMP/PLA2 signaling pathways. Starvation activates these pathways thereby suppressing turns and inducing steps. As a consequence, starved cells make very long nearly straight runs and disperse over approximately 30-fold larger areas, without extending more or larger pseudopodia than vegetative cells. This 'win-stay/lose-shift' strategy for food searching is called Starvation Induced Run-length Extension. The SIRE walk explains very well the observed differences in search behavior between fed and starving organisms such as bumble-bees, flower bug, hoverfly and zooplankton.

Topics: Animals; Cyclic GMP; Dictyostelium; Feeding Behavior; Phospholipases A2; Signal Transduction; Starvation

Fluid transport was measured gravimetrically in vivo in the jejunum, ileum and colon of fed, fasting (four days) and undernourished (50 % of control food intake for 21 days) gerbils (Gerbillus cheesmani). The effects of luminal enterotoxin Escherichia coli STa (50 ng/ml) and luminal 8-bromo-cyclic GMP (cGMP 1 mM) on fluid transport across jejunum, ileum and colon were also assessed. Fasting and undernourishment reversed the normal basal fluid absorption measured in fed ileum and colon into secretion. Neither fasting nor undernourishment had any effect on jejunal basal fluid absorption. In jejuna, ilea and colons of fed animals as well as in jejuna from fasting and undernourished gerbils STa (50 ng) reversed the normal absorptive "tone" to secretion but it had no significant effects on fluid secretion in either the ileum or colon from fasted gerbils. STa increased significantly the fluid secretion in ileum from undernourished gerbils. Luminal cGMP had no effect on basal absorptive tone in the jejunum of fed and fasted gerbils, but reversed absorption into secretion in the jejuna from undernourished gerbils. In the ilea taken from fed animals the small basal absorption was reversed to secretion by luminal cGMP. Although cGMP produced no significant changes in fluid secretion in the ilea taken from fasted gerbils, yet it caused a significant increase in those from undernourished gerbils. In the colon taken from fed animals cGMP decreased the basal fluid absorption significantly, but it had no significant effect on fluid secretion in the colon of fasted or undernourished gerbils. We conclude that fasting and undernourishment have no significant effects on fluid transport across the gerbil jejunum but reversed basal absorption in the fed ileum and colon into secretion. cGMP mimic the effects of STa in the jejunum taken from undernourished gerbils, in the ileum obtained under the three nutritional states and in the colon taken from fasting animals.

Topics: Animals; Bacterial Toxins; Cyclic GMP; Enterotoxins; Escherichia coli Proteins; Fasting; Female; Gerbillinae; Intestinal Mucosa; Intestinal Secretions; Male; Starvation

The effect of luminal enterotoxin Escherichia coliSTa on fluid transport across the jejunum and ileum of fed, starved (72 h) and chronically undernourished (50% control food intake for 21 days) rats was assessed in vivo using a gravimetric technique. Dose-response curves for net fluid secretion (stimulated-basal (30 min)-1) activated by 5,50 and 500 ng ml-1 STa were obtained for jejuna and ilea from fed, starved and chronically undernourished rats. Compared to the fed rats, both dietary deprivations showed enhanced net fluid secretion at 500 ng ml-1, but only the jejunum from chronically undernourished rats showed significantly enhanced secretion at 5 and 50 ng ml-1. Net fluid secretory responses to a standard dose of STa (500 ng ml-1) were monitored in the jejuna and ilea of fed, starved and chronically undernourished rats at 30, 60, 90 and 120 min. The pattern in jejuna and ilea from fed rats was very different; the jejunal secretion over 30 min was transient, but ileal secretion increased continuously to a maximum at 120 min. In both jejunum and ileum from starved rats, the secretory response to STa at 30 min was significantly greater than that in fed rats and subsequently remained near this level. In the jejunum from chronically undernourished rats, the net fluid secreted in response to STa was greatly enhanced at 30 and 60 min, but not at 90 min. The ileal response was significantly greater than that in the fed rats at 30 and 120 min. Luminal procaine (10 mM) selectively increased fluid absorptive tone in the jejunum from fed rats, reduced STa-induced fluid secretion in the ileum from undernourished rats, but had no effect on STa in the jejunum from undernourished rats or in the jejunum or ileum from starved rats. Luminal 8-bromo-cyclic GMP (1 mM) had no effect on basal absorptive tone in the jejunum of fed or chronically undernourished rats, but enhanced the secretory tone of jejunum from starved rats. In the ileum, however, while 8-bromo-cyclic GMP enhanced secretory tone in the fed and starved conditions, it still had no action in the chronically undernourished state. The enhanced fluid secretion observed in the jejunum and ileum of the starved and undernourished rats compared to the fed rats supports previous in vitro findings of an increased electrogenic secretion induced by STa in these dietary deprivations. The fluid hypersecretion could be a cause of the increased severity of diarrhoea often observed in undernourished and starved humans.

Topics: Animals; Bacterial Toxins; Body Fluids; Cyclic GMP; Diet; Enterotoxins; Escherichia coli Proteins; Ileum; Intestinal Mucosa; Jejunum; Male; Nutrition Disorders; Procaine; Rats; Rats, Wistar; Starvation

Membrane potentials of the cellular slime mold Dictyostelium discoideum were monitored after chemotactic stimulation by measuring the distribution of the lipophilic cation tetraphenylphosphonium. Stimulation with the chemoattractant cAMP induces a transient membrane hyperpolarization which reaches its most negative value between 1-3 min after stimulation. This hyperpolarization is consistent with the opening of potassium channels. Measurements in streamer F mutant cells reveal that cGMP likely plays a role in the regulation of the cAMP-induced hyperpolarization.

Topics: Chemotaxis; Cyclic AMP; Cyclic GMP; Dictyostelium; Diethylstilbestrol; Membrane Potentials; Mutation; Potassium; Quinine; Starvation

Cyclic AMP induces postaggregative differentiation in aggregation competent cells of Dictyostelium by interacting with cell surface cAMP receptors. We investigated the transduction pathway of this response and additional requirements for the induction of postaggregative differentiation. Optimal induction of postaggregative gene expression requires that vegetative cells are first exposed to 2-4 hr of nanomolar cAMP pulses, and subsequently for 4-6 hr to steady-state cAMP concentrations in the micromolar range. Cyclic AMP pulses, which are endogenously produced before and during aggregation, induce full responsiveness to cAMP as a morphogen. The transduction pathway from the cell surface cAMP receptor to postaggregative gene expression may involve Ca2+ ions as intracellular messengers. A cAMP-induced increase in intracellular cAMP or cGMP levels is not involved in the transduction pathway.

Topics: Adenylyl Cyclases; Antigens, Fungal; Calcium; Cell Aggregation; Cell Differentiation; Cyclic AMP; Cyclic GMP; Dictyostelium; Enzyme Activation; Gene Expression Regulation; Guanylate Cyclase; Ornithine Decarboxylase; Phosphorylases; Starvation

Mucosa isolated from the proximal third of the small intestine of infant rats had much lower cyclic nucleotide concentrations (expressed both per unit wet weight and per unit DNA content) than those determined in the intestinal wall. The steady-state concentrations of both cyclic AMP and cyclic GMP in jejunum showed dramatic increases during the first 5 d post partum. Another increase in cyclic nucleotide concentrations was observed in the isolated mucosa between d 15 and 21. Starvation for 24 h always resulted in lower intestinal cyclic nucleotide concentrations than those of the fed littermates. This effect was more pronounced in younger animals and more evident for cyclic AMP values. Three-week-old rats fed a high carbohydrate diet for 24-48 h exhibited more pronounced elevations in the concentrations of cyclic nucleotides from the jejunal mucosa than did rats fed a high fat diet.

Topics: Aging; Animals; Animals, Newborn; Cyclic AMP; Cyclic GMP; Dietary Carbohydrates; Dietary Fats; DNA; Intestinal Mucosa; Jejunum; Rats; Rats, Inbred Strains; Starvation

Islets isolated from fed rats released insulin in response to glucose, 8-bromoguanosine 3',5'-cyclic monophosphate (8-Br-cGMP) and 8-bromoadenosine 3',5'-cyclic monophosphate (8-Br-cAMP), but not to 8-bromoinosine 3',5'-cyclic monophosphate. Starving rats for 48 h significantly diminished insulin release from islets in response to these agents, and lowered endogenous levels of cGMP and cAMP. The analogs of cGMP and cAMP potentiated the glucose response in a dose-dependent manner in islets from starved rats, whereas in fed rat islets the cyclic nucleotide analogs did not potentiate glucose-stimulated insulin release. Sodium nitroprusside, which enhances endogenous cGMP levels in islets, also enhanced the glucose response in islets from starved rats. Mannoheptulose inhibited glucose and 8-Br-cGMP-stimulated insulin release, but not 8-Br-cAMP-stimulated release. These results suggest that the impaired glucose response of islets from starved animals is in part due to diminished levels of cyclic nucleotides, and that the role(s) of cGMP in insulin secretion may include enhancement of glucose metabolism.

Topics: Analysis of Variance; Animals; Cyclic AMP; Cyclic GMP; Glucose; In Vitro Techniques; Insulin; Insulin Secretion; Male; Mannoheptulose; Nucleotides, Cyclic; Rats; Rats, Inbred Strains; Starvation

Feeding rats in diet high in glucose has been demonstrated to inhibit the induction of many enzymes, block the action of glucocorticoids, and, in general, appears to result in decreased cyclic AMP activity. We found that glucose feeding depresses both messenger RNA (mRNA) and non-mRNA synthesis. Electron microscopic examination of the nucleus revealed that glucose feeding decreases the granular component of liver cell nucleoli. It only slightly decreases liver cyclic AMP levels, but produces a sixfold elevation in levels of the cyclic AMP antagonist, cyclic GMP. Administration of bromocyclic GMP, like glucose feeding, depresses mRNA synthesis, but does not simulate the effect of the carbohydrate on nuclear morphology. In addition, glucose feeding halves liver inorganic phosphate and triples ATP levels. Phosphorylation of nuclear proteins, however, remains unaltered. Despite the antagonism between glucose feeding and glucocorticoid activity, the former compound did not change the binding of dexamethasone to liver nuclei.

Topics: Adenosine Triphosphate; Animals; Cell Nucleolus; Cell Nucleus; Cyclic AMP; Cyclic GMP; Dexamethasone; Female; Glucose; Liver; Microscopy, Electron; Organophosphorus Compounds; Phosphates; Rats; Receptors, Glucocorticoid; Starvation; Transcription, Genetic

Topics: Amylases; Animals; Bile Ducts; Catheterization; Cholecystokinin; Chromatography, Ion Exchange; Cyclic AMP; Cyclic GMP; Gastrointestinal Hormones; Intestines; Male; Pancreas; Peptides; Pilocarpine; Radioimmunoassay; Radioligand Assay; Rats; Secretin; Starvation; Time Factors

Topics: Animals; Bucladesine; Cyclic GMP; Diazoxide; Drug Interactions; Epinephrine; Inosine Nucleotides; Insulin; Insulin Antagonists; Insulin Secretion; Male; Nucleotides, Cyclic; Pancreas; Rats; Starvation; Stimulation, Chemical; Theophylline; Thymine Nucleotides; Time Factors; Tolbutamide