Coordination of fast and slow rhythmic neuronal circuits.
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Interactions among rhythmically active neuronal circuits that oscillate at different frequencies are important for generating complex behaviors, yet little is known about the underlying cellular mechanisms. We addressed this issue in the crab stomatogastric ganglion (STG), which contains two distinct but interacting circuits. These circuits generate the gastric mill rhythm (cycle period, approximately 10 sec) and the pyloric rhythm (cycle period, approximately 1 sec). When the identified modulatory projection neuron named modulatory commissural neuron 1 (MCN1) is activated, the gastric mill motor pattern is generated by interactions among MCN1 and two STG neurons [the lateral gastric (LG) neuron and interneuron 1]. We show that, during MCN1 stimulation, an identified synapse from the pyloric circuit onto the gastric mill circuit is pivotal for determining the gastric mill cycle period and the gastric-pyloric rhythm coordination. To examine the role of this intercircuit synapse, we replaced it with a computational equivalent via the dynamic-clamp technique. This enabled us to manipulate better the timing and strength of this synapse. We found this synapse to be necessary for production of the normal gastric mill cycle period. The synapse acts, during each LG neuron interburst, to boost rhythmically the influence of the modulatory input from MCN1 to LG and thereby to hasten LG neuron burst onset. The two rhythms become coordinated because LG burst onset occurs with a constant latency after the onset of the triggering pyloric input. These results indicate that intercircuit synapses can enable an oscillatory circuit to control the speed of a slower oscillatory circuit, as well as provide a mechanism for intercircuit coordination. (+info)
1.3 kilobases of the lung type I cell T1alpha gene promoter mimics endogenous gene expression patterns during development but lacks sequences to enhance expression in perinatal and adult lung.
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The T1alpha gene is one of few markers for the type I cell phenotype in the adult mammalian lung. Type I cells form a large, thin epithelial layer that facilitates gas exchange and transport of fluids between the air spaces and capillaries. The T1alpha gene has a complex pattern of developmental expression in lung and brain; in vitro studies indicate that expression is regulated in part by thyroid transcription factor 1, forkhead proteins, and Sp1/Sp3 proteins. To explore the mechanisms that confine T1alpha expression in intact adult animals to alveolar type I and choroid plexus epithelial cells, we generated mice bearing a 1.3-kb T1alpha promoter-chloramphenicol acetyltransferase (CAT) gene. In situ hybridization and RNase protection assays show that the 1.3-kb promoter confers a pattern of CAT expression that largely matches the endogenous T1alpha in embryos and mid-term fetuses in lung and central nervous system. However, the 1.3-kb promoter lacks elements important for perinatal up-regulation of T1alpha in the lung and maintenance of that expression in the adult lung and brain. The final adult pattern of T1alpha expression may be directed by elements outside the 1.3-kb fragment, perhaps those 5' to the 1.3-kb fragment as we show herein, or in 3' and intronic regions. Dev Dyn 1999;215:319-331. (+info)
Development of Babesia gibsoni in the midgut of larval tick, Rhipicephalus sanguineus.
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Studies were made on the development of Babesia gibsoni in the midgut of the larval tick, Rhipicephalus sanguineus. Six hr after repletion, merozoites of B. gibsoni, freed from erythrocytes, were observed in the midgut contents of the tick. After that, within 24 hr, those merozoites were transformed into the ring-forms which were relatively large, 2-3 microns in diameter. Later, the ring forms developed into the spherical forms which were subelliptical in shape and 4-6 microns in diameter. Within 2-4 days, the elongated forms, 5-8 microns in length, were found. At this time, some of the binucleated fusion form has assumed a form intermediate between the spherical and elongated-forms. About 5-6 days after repletion, large round or elliptic zygotes, 8-10 microns in diameter, were observed in the tick gut. (+info)
Development of Babesia gibsoni in the midgut of the nymphal stage of the tick, Rhipicephalus sanguineus.
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Studies were made on the development of Babesia gibsoni in the midgut of the nymphal stage of the tick, Rhipicephalus sanguineus. Six hr after repletion, merozoites of B. gibsoni, free of erythrocytes, were observed in the midgut contents of the ticks. After that, within 24 hr, those merozoites were transformed into ring-forms which were relatively large ring 1-2 microns in diameter. Later, the ring forms developed into spherical forms which were somewhat elliptical in shape and 3-4 microns in diameter. Within 2-4 days, bizarre forms (5-6 microns in diameter) developed into elongated forms (5-6 microns in length). About 5-6 days after repletion, large round or elliptic zygotes (7-9 microns in diameter) were observed in the ticks gut. (+info)
Enteral IGF-I enhances fetal growth and gastrointestinal development in oesophageal ligated fetal sheep.
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Infants with upper gut atresia often have impaired intrauterine growth and gut function. IGF-I is important in fetal growth and is contained in amniotic fluid. We therefore wanted to test the hypothesis that IGF-I infused into fetal gut would reverse the effects of an upper gut obstruction on gut structure and growth in fetal sheep. At 90 days gestation fetuses (n=6 per group) underwent oesophageal ligation, followed by continuous infusion of IGF-I (1-8 microgram/day) or saline into the gut beyond the ligation until 137 days. Controls underwent sham ligation only. Oesophageal ligation tended to reduce fetal body and organ weights. IGF-I treatment prevented this reduction and increased body length and spleen weight above those of controls. The decrease in bowel wall thickness induced by oesophageal ligation was also prevented by IGF-I treatment. Amniotic fluid IGF-I concentrations did not change over gestation and were higher in the IGF-I treated group. No change in fetal plasma IGF-I concentrations were detectable. We conclude that enterally administered IGF-I may enhance fetal growth and gut development in utero and that IGF-I in amniotic fluid may play a physiological role in gut development in the fetus. (+info)
Induction of protease activity in Vibrio anguillarum by gastrointestinal mucus.
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The effect of gastrointestinal mucus on protease activity in Vibrio anguillarum was investigated. Protease activity was measured by using an azocasein hydrolysis assay. Cells grown to stationary phase in mucus (200 microg of mucus protein/ml) exhibited ninefold-greater protease activity than cells grown in Luria-Bertani broth plus 2% NaCl (LB20). Protease induction was examined with cells grown in LB20 and resuspended in mucus, LB20, nine-salts solution (NSS [a carbon-, nitrogen-, and phosphorus-free salt solution]), or marine minimal medium (3M) ( approximately 10(9) CFU/ml). Induction of protease activity occurred 60 to 90 min after addition of mucus and was >/=70-fold greater than protease activity measured in cells incubated in either LB20 or 3M. Mucus was fractionated into aqueous and chloroform-methanol-soluble fractions. The aqueous fraction supported growth of V. anguillarum cells, but did not induce protease activity. The chloroform-methanol-soluble fraction did not support growth, nor did it induce protease activity. When the two fractions were mixed, protease activity was induced. The chloroform-methanol-soluble fraction did not induce protease activity in cells growing in LB20. EDTA (50 mM) inhibited the protease induced by mucus. Upon addition of divalent cations, Mg(2+) (100 mM) was more effective than equimolar amounts of either Ca(2+) or Zn(2+) in restoring activity, suggesting that the mucus-inducible protease was a magnesium-dependent metalloprotease. An empA mutant strain of V. anguillarum did not exhibit protease activity after exposure to mucus, but did grow in mucus. Southern analysis and PCR amplification confirmed that V. anguillarum M93 contained empA. These data demonstrate that the empA metalloprotease of V. anguillarum is specifically induced by gastrointestinal mucus. (+info)
Evidence for involvement of gut-associated denitrifying bacteria in emission of nitrous oxide (N(2)O) by earthworms obtained from garden and forest soils.
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Earthworms (Aporrectodea caliginosa, Lumbricus rubellus, and Octolasion lacteum) obtained from nitrous oxide (N(2)O)-emitting garden soils emitted 0.14 to 0.87 nmol of N(2)O h(-1) g (fresh weight)(-1) under in vivo conditions. L. rubellus obtained from N(2)O-emitting forest soil also emitted N(2)O, which confirmed previous observations (G. R. Karsten and H. L. Drake, Appl. Environ. Microbiol. 63:1878-1882, 1997). In contrast, commercially obtained Lumbricus terrestris did not emit N(2)O; however, such worms emitted N(2)O when they were fed (i.e., preincubated in) garden soils. A. caliginosa, L. rubellus, and O. lacteum substantially increased the rates of N(2)O emission of garden soil columns and microcosms. Extrapolation of the data to in situ conditions indicated that N(2)O emission by earthworms accounted for approximately 33% of the N(2)O emitted by garden soils. In vivo emission of N(2)O by earthworms obtained from both garden and forest soils was greatly stimulated when worms were moistened with sterile solutions of nitrate or nitrite; in contrast, ammonium did not stimulate in vivo emission of N(2)O. In the presence of nitrate, acetylene increased the N(2)O emission rates of earthworms; in contrast, in the presence of nitrite, acetylene had little or no effect on emission of N(2)O. In vivo emission of N(2)O decreased by 80% when earthworms were preincubated in soil supplemented with streptomycin and tetracycline. On a fresh weight basis, the rates of N(2)O emission of dissected earthworm gut sections were substantially higher than the rates of N(2)O emission of dissected worms lacking gut sections, indicating that N(2)O production occurred in the gut rather than on the worm surface. In contrast to living earthworms and gut sections that produced N(2)O under oxic conditions (i.e., in the presence of air), fresh casts (feces) from N(2)O-emitting earthworms produced N(2)O only under anoxic conditions. Collectively, these results indicate that gut-associated denitrifying bacteria are responsible for the in vivo emission of N(2)O by earthworms and contribute to the N(2)O that is emitted from certain terrestrial ecosystems. (+info)
BRK/Sik expression in the gastrointestinal tract and in colon tumors.
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Clones encoding the breast tumor kinase BRK were isolated from a normal human small intestinal cDNA library that was screened with the cDNA encoding the mouse epithelial-specific tyrosine kinase Sik. Although BRK and Sik share only 80% amino acid sequence identity, Southern blot hybridizations confirmed that the two proteins are orthologues. Sik was mapped to mouse distal chromosome 2, which shows conservation of synteny with human chromosome 20q13.3, the location of the BRK gene. BRK expression was examined in the normal gastrointestinal tract, colon tumor cell lines, and primary colon tumor samples. Like Sik, BRK is expressed in normal epithelial cells of the gastrointestinal tract that are undergoing terminal differentiation. BRK expression also increased during differentiation of the Caco-2 colon adenocarcinoma cell line. Modest increases in BRK expression were detected in primary colon tumors by RNase protection, in situ hybridization, and immunohistochemical assays. The BRK tyrosine kinase appears to play a role in signal transduction in the normal gastrointestinal tract, and its overexpression may be linked to the development of a variety of epithelial tumors. (+info)