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Impedance signature of pharyngeal gaseous reflux.

BACKGROUND: Pharyngeal impedance changes induced by various pharyngeal reflux events have not been characterized. OBJECTIVES: To characterize pharyngeal impedance changes induced by participant-perceived belching events. METHODS: We systematically evaluated pharyngeal impedance and pH changes related to 453 belch events in 11 gastroesophageal reflux disease, 10 reflux attributed-laryngitis patients and 16 controls. RESULTS: Of 453 belch events, 362 were analyzable. Of these, 72% occurred within 10 s, 93% within 20 s, 99% within 30 s and 100% within 40 s of the time that participants marked a belch event. In 15% impedance changes in the pharynx preceded, in 12% they were simultaneous and in 73% they occurred after the start of the impedance change in the proximal esophagus. Time interval between the two events ranged between 0.4+/-0.03 and 0.7+/-0.1 s. In all, there were three types of belch-induced impedance changes: (a) impedance increase, (b) impedance decrease and (c) multiphasic. Twenty percent of impedance events associated with belching had less than 50% change from baseline, whereas in 51% changes exceeded or were equal to 50%. Among events with a drop in pharyngeal impedance, only two satisfied the criteria for the liquid reflux event. CONCLUSIONS: Pharyngeal ventilation of gastric gaseous content seems to have a unique impedance signature. During pharyngeal gas reflux events, impedance changes may start before or after proximal esophageal changes. Belching may induce negative pharyngeal changes that do not meet the criteria for liquid reflux. These findings need to be taken into consideration in the analysis of pharyngeal reflux events.

Adult↗

Frequency-dependent taste-rejection by avian predation may select for defence chemical polymorphisms in aposematic prey.

Chemically defended insects advertise their unpalatability to avian predators using conspicuous aposematic coloration that predators learn to avoid. Insects utilize a wide variety of different compounds in their defences, and intraspecific variation in defence chemistry is common. We propose that polymorphisms in insect defence chemicals may be beneficial to insects by increasing survival from avian predators. Birds learn to avoid a colour signal faster when individual prey possesses one of two unpalatable chemicals rather than all prey having the same defence chemical. However, for chemical polymorphisms to evolve within a species, there must be benefits that allow rare chemical morphs to increase in frequency. Using domestic chicks as predators and coloured crumbs for prey, we provide evidence that birds taste and reject proportionally more of the individuals with rare defence chemicals than those with common defence chemicals. This indicates that the way in which birds attack and reject prey could enhance the survival of rare chemical morphs and select for chemical polymorphism in aposematic species. This is the first experiment to demonstrate that predators can directly influence the form taken by prey's chemical defences.

Animals↗

Avian predators taste-reject aposematic prey on the basis of their chemical defence.

Avian predators learn to avoid defended insects on the basis of their conspicuous warning coloration. In many aposematic species, the level of chemical defence varies, with some individuals being more defended than others. Sequestration and production of defence chemicals is often costly and therefore less defended individuals enjoy the benefits of the warning signal without paying the full costs of chemical production. This is a fundamental theoretical problem for the evolutionary stability of aposematism, since less defended individuals appear to be at a selective advantage. However, if predators sample aposematic prey and selectively reject individuals on the basis of their chemical investment, aposematism could become evolutionarily stable. Previous research aimed at testing whether birds can use taste to discriminate between palatable and unpalatable prey has been confounded by other experimental factors. Here, we show that birds can taste and reject prey entirely on the basis of an individual's level of chemical defence and more importantly, they can make decisions on whether or not to consume a defended individual based upon their level of chemical investment. We discuss these results in relation to the evolution of aposematism, mimicry and defence chemistry.

Adaptation, Physiological↗

Visualization of satellite cells in living muscle fibres of the frog.

Satellite cells were visualized in living muscle fibres of the frog. Single fibres or bundles consisting of a few fibres were isolated after treatment with collagenase, and viewed under the light microscope. Subsequent electron microscopy of identified cells confirmed that they were satellite muscle cells. Under the light microscope, satellite cells appear as fusiform cells, tapering into long fine processes usually orientated parallel to the muscle fibre axis. Horseradish peroxidase injected into the muscle fibre was not transferred to the satellite cells.

Animals↗

Junctional and extrajunctional membrane channels activated by GABA in locust muscle fibres.

Iontophoretic application of GABA to voltage-clamped locust muscle fibres has demonstrated the presence of both extrajunctional and junctional GABA receptors. Extrajunctional GABA receptors are distinct from extrajunctional glutamate receptors which also occur in these muscle fibres. Inward GABA currents are nonlinearly dependent on membrane potential. Analysis of membrane current noise produced by iontophoretic GABA application shows that for junctional and extrajunctional GABA receptors the mean channel lifetime is 3-4 ms and the single-channel conductance is approximately 22 pS at - 80 mV (T = 21 degrees C). The mean lifetime as previously demonstrated for glutamate-sensitive excitatory channels in locust muscle fibres.

Animals↗

Block of glutamate-activated synaptic channels by curare and gallamine.

Excitatory junctional currents (e.j.cs) and glutamate-activated currents have been examined in voltage-clamped locust muscle fibres exposed to curare or gallamine. Although these drugs have little action on channel kinetics at the resting potential, there is an increasingly pronounced effect at hyperpolarized levels. In the presence of curare (5-100 microM), fibres held at hyperpolarized potentials showed e.j.cs with an initial 'fast component' followed by a 'slow tail'. In many fibres, hyperpolarization beyond -50 mV decreased the amplitude of the peak synaptic current; the decay time constant of the fast component was decreased by hyperpolarization while the time constant of the slow component was increased. Iontophoretic application of brief pulses of glutamate also produced two-component glutamate currents in these conditions. Gallamine was considerably more effective than curare, markedly altering the decay time and amplitude of the e.j.c. and the glutamate current at 1-5 microM. Its effects appeared qualitatively similar to those of curare. The observations are consistent with the idea that curare and gallamine produce a transient block of glutamate-activated synaptic channels.

Animals↗

Inhibitory synaptic currents in voltage-clamped locust muscle fibres desensitized to their excitatory transmitter.

Inhibitory junctional currents (i.j.c.s) have been examined in locust muscle fibres to give properties of GABA-channels activated by the neurally released transmitter. A nerve-muscle preparation is described which has proved suitable for voltage-clamp analysis of inhibitory transmission. I.j.c.s were recorded from fibres in which excitatory synapses had been desensitized with glutamate, to abolish excitatory junctional currents. This procedure had no apparent effect on inhibitory channel properties. The time constant of decay of the i.j.c. was 7.7 +/- 0.3 ms, slightly exceeding the time constant of the membrane noise induced by externally applied GABA. Peak i.j.c. conductance decreased with hyperpolarization. I.j.c.s showed measurable fluctuations permitting an estimate of the mean size of the quantal events composing the i.j.c. Their mean size coincided with the spontaneously occurring miniature inhibitory junctional currents that could be directly recorded in some fibres. The inhibitory nerve-impulse released an average of 35 transmitter packets at sites distributed along the muscle fibre length. Since each m.i.j.c. produced a current of about 0.6 nA (at Vm = -80 mV, ECl = -40 mV) the single quantum of inhibitory transmitter opens 600-1000 postsynaptic chloride channels. This is roughly three to four times the number of channels opened by the excitatory transmitter packet at glutamate synapses in the same fibres.

Animals↗

Ion channels activated by L-glutamate and GABA in cultured cerebellar neurons of the rat.

Glutamate and GABA-receptor channels were investigated in explants of rat cerebellum grown in cell culture. The patch-clamp technique was used to examine neurons under whole cell clamp and the properties of channels were derived by analysis of glutamate and GABA-evoked current noise. In addition, single channel currents activated by glutamate were recorded from isolated outside-out patches of membrane. We found evidence for at least two types of glutamate receptor-channels in cerebellar cells. Some neurons exhibited a channel of 50 pS conductance with a Lorentzian noise spectrum of 5.9 ms time constant. Single channels were readily resolved both in whole cell clamp and excised patches. Other neurons possessed low conductance channels which produced two component spectra. Estimates of the single channel conductance gave a value of about 140 fS. GABA channel noise obtained from these cells was also fitted by two component spectra which gave single channel conductance of 16 pS.

Animals↗

Single-channel and whole-cell currents evoked by acetylcholine in dissociated sympathetic neurons of the rat.

Single acetylcholine-activated channels have been recorded from neurons dissociated from the sympathetic chain of 17-21 day old rats. The mean single channel conductance is 35 pS in normal medium containing 1 mM calcium, and 51 pS in the absence of calcium. The measured current amplitudes are about five times more variable than at the frog endplate, at least in part because the current, while the channel is open, is much noisier than when it is shut. Single activations of the receptor by acetylcholine (ACh) produce a burst of openings; the distribution of the burst length has two components, the longer of which is of primary importance in synaptic transmission. Whole-cell currents, in response to ACh (up to 30 microM), show strong inward rectification with no outward current being detectable. This phenomenon is similar whether the intracellular ion is sodium or cesium, whether or not divalent cations are present, and whether or not atropine is present. Nevertheless, outward single-channel currents (of normal conductance) are detectable in isolated outside-out patches.

Acetylcholine↗

On the kinetics of large-conductance glutamate-receptor ion channels in rat cerebellar granule neurons.

Ion channels activated by glutamate, aspartate, and N-methyl-D-aspartate (NMDA) have been investigated in outside-out patches from cultured cerebellar granule neurons of the rat. Openings of these channels occur in bursts, within which the individual openings are separated by brief shuttings or gaps. The shut-time distributions obtained with each agonist were fitted with four exponential components. The briefest two components were considered as 'gaps within bursts'. Their mean time-constants were: glutamate, 58.0 microseconds and 592 microseconds; aspartate, 31.3 microseconds and 644 microseconds; NMDA, 40.5 microseconds and 903 microseconds. Distributions of burst durations were fitted with three exponential components. The mean time-constants obtained for the longest two components were: glutamate, 1.33 ms and 10.5 ms; aspartate, 2.15 ms and 10.3 ms; NMDA, 2.42 ms and 10.5 ms. Evidence is given that these two components of burst duration reflect the gating kinetics of 50 pS openings and not the fact that each agonist produces openings to more than one conductance level. Not only do openings occur in bursts, but these bursts were observed to occur in clusters, which can be hundreds of milliseconds long. We discuss the relation between the kinetics of single-channel openings observed in patches and the spectral components detected in whole-cell current noise.

Animals↗

Excitatory amino acid receptor-channels in Purkinje cells in thin cerebellar slices.

Glutamate receptors of the N-methyl-D-aspartate (NMDA) and non-NMDA type serve different functions during excitatory synaptic transmission. Although many central neurons bear both types of receptor, the evidence concerning the sensitivity of cerebellar Purkinje cells to NMDA is contradictory. To investigate the receptor types present in Purkinje cells, we have used whole-cell and outside-out patch-clamp methods to record from cells in thin cerebellar slices from young rats. At a holding potential of -70 mV (in nominally Mg(2+)-free medium, with added glycine) NMDA caused a whole-cell current response which consisted of a dramatic increase in the frequency of synaptic currents. In the presence of tetrodotoxin (TTX) and the gamma-aminobutyric acidA (GABAA) receptor antagonist bicuculline, spontaneous synaptic currents and responses to NMDA were inhibited. In a proportion of cells a small polysynaptic response to NMDA persisted, which was further reduced by the non-NMDA receptor antagonist 6-cyano-2,3-dihydro-7-nitroquinoxalinedione (CNQX). The non-NMDA glutamate receptor agonists kainate (KA), quisqualate (QA) and s-alpha-amino-3-hydroxy-5-methyl-4-isoazolepropionic acid (s-AMPA), evoked large inward currents due to the direct activation of receptors in Purkinje cells. NMDA applied to excised membrane patches failed to evoke any single-channel currents, whereas s-AMPA and QA caused small inward currents accompanied by marked increases in current noise. Spectral analysis of the s-AMPA noise in patches gave an estimated mean channel conductance of approximately 4 pS.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Sound improves visual discrimination learning in avian predators.

Aposematic insects use warning colours to deter predators, but many also produce odours or sounds when attacked by a predator. One possible role for these additional components is that they promote the association between the warning colour and the non-profitability it signals, thus reducing the chance of future attacks from visually hunting predators. This experiment explicitly tests this idea by looking at the effects of sound on a visual discrimination task. Young domestic chicks were trained to look for food rewards under coloured paper cones scattered in an experimental arena. In a subsequent visual discrimination task, they learned to discriminate between rewarded and non-rewarded hats on the basis of colour. Half the chicks performed this task in silence, whilst the other half had a tone played when they attacked non-rewarded hats. The presence of the tone improved the speed of colour discrimination learning. This demonstrates that there could be a selective advantage for aposematic coloured insects to emit sounds when attacked, since avian predators will learn to avoid their coloration more quickly. The role of psychological interactions between signal components in receivers is discussed in relation to the evolution of multimodal displays.

Acoustic Stimulation↗

Competition for male reproductive investment elevates testosterone levels in female dunnocks, Prunella modularis.

In many songbirds, females occasionally sing in contexts of high female-female competition. Testosterone may be involved in the activation of song, because testosterone implants elicit female song in many species with rare female song. A possible mechanism for the hormonal control of female song is provided by the challenge hypothesis, which predicts a rise in testosterone in response to aggressive interactions during socially unstable situations. We tested this by comparing faecal testosterone levels in polygynandrous and monogamous female dunnocks. In groups with two to three females (polygynandry and polygyny) males provide less help at each nest than in groups with a single female (monogamy and polyandry). Polygynandrous and polygynous females are aggressive towards one another and attempt to expel rivals. Polygynandrous females had significantly higher testosterone levels than monogamous females. Competition between females that was induced by removal of males caused testosterone levels to rise. Further, female testosterone levels were correlated with the rate of 'tseep' calls, which are produced during aggressive encounters between females. Finally, polygynandrous and polygynous females sang significantly more than monogamous females. To the best of our knowledge, these results provide the first experimental support for the challenge hypothesis in female birds, and suggest that testosterone can regulate facultative female song in songbirds.

Animals↗

The importance of pattern similarity between Müllerian mimics in predator avoidance learning.

Müllerian mimicry, where unpalatable prey share common warning patterns, has long fascinated evolutionary biologists. It is commonly assumed that Müllerian mimics benefit by sharing the costs of predator education, thus reducing per capita mortality, although there has been no direct test of this assumption. Here, we specifically measure the selection pressure exerted by avian predators on unpalatable prey with different degrees of visual similarity in their warning patterns. Using wild-caught birds foraging on novel patterned prey in the laboratory, we unexpectedly found that pattern similarity did not increase the speed of avoidance learning, and even dissimilar mimics shared the education of naive predators. This was a consistent finding across two different densities of unpalatable prey, although mortalities were lower at the higher density as expected. Interestingly, the mortalities of Müllerian mimics were affected by pattern similarity in the predicted way by the end of our experiment, although the result was not quite significant. This suggests that the benefits to Müllerian mimics may emerge only later in the learning process, and that predator experience of the patterns may affect the degree to which pattern similarity is important. This highlights the need to measure the behaviour of real predators if we are to understand fully the evolution of mimicry systems.

Adaptation, Physiological↗

Avian psychology and communication.

The evolution of animal communication is a complex issue and one that attracts much research and debate. 'Receiver psychology' has been highlighted as a potential selective force, and we review how avian psychological processes and biases can influence the evolution and design of signals as well as the progress that has been made in testing these ideas in behavioural studies. Interestingly, although birds are a focal group for experimental psychologists and behavioural ecologists alike, the integration of theoretical ideas from psychology into studies of communication has been relatively slow. However, recent operant experiments are starting to address how birds perceive and respond to complex natural signals in an attempt to answer evolutionary problems in communication. This review outlines how a psychological approach to understanding communication is useful, and we hope that it stimulates further research addressing the role of psychological mechanisms in signal evolution.

Adaptation, Psychological↗

Tasting the difference: do multiple defence chemicals interact in Müllerian mimicry?

Müllerian mimicry, where two unpalatable species share a warning pattern, is classically believed to be a form of mutualism, where the species involved share the cost of predator education. The evolutionary dynamics of Müllerian mimicry have recently become a controversial subject, after mathematical models have shown that if minor alterations are made to assumptions about the way in which predators learn and forget about unpalatable prey, this textbook case of mutualism may not be mutualistic at all. An underlying assumption of these models is that Müllerian mimics possess the same defence chemical. However, some Müllerian mimics are known to possess different defence chemicals. Using domestic chicks as predators and coloured crumbs flavoured with either the same or different unpalatable chemicals as prey, we provide evidence that two defence chemicals can interact to enhance predator learning and memory. This indicates that Müllerian mimics that possess different defence chemicals are better protected than those that share a single defence chemical. These data provide insight into how multiple defence chemicals are perceived by birds,and how they influence the way birds learn and remember warningly coloured prey. They highlight the importance of considering how different toxins in mimicry rings can interact in the evolution and maintenance of Müllerian mimicry and could help to explain the remarkable variation in chemical defences found within and between species.

Adaptation, Physiological↗

Kinetics, tissue specificity and pathological changes in murine rotavirus infection of mice.

Mice that did not contain antibodies to rotavirus were orally infected with murine rotavirus (EDIM strain) and observed over 7 days. As judged by ELISA, only the small intestine was infected, not the colon. The infection was biphasic, viral antigen peaks being observed at 48 h and approximately 120 h post-infection. Clinically evident diarrhoea was maximal at 72 h. Virus in the upper, middle and lower regions of the small intestine was mainly tissue-associated; most virus was found in the middle small intestine. Two peaks (48 h and 120 h post-infection) of virus antigen were observed in the colon, but these corresponded to luminal, not tissue-associated viral antigen. Only enterocytes in the upper two-thirds of villus epithelia were infected as judged by fluorescent-antibody analysis and transmission electron microscopy. Scanning electron microscopy revealed morphological appearances not hitherto correlated with the progress of the infection: villus tips were convoluted, corresponding to the shedding of virus-infected cells but the lower regions of infected villi were shrunken and considerably narrowed compared to tips.

Animals↗

Expression of active yeast pyruvate decarboxylase in Escherichia coli.

We have shown by appropriate modification of the translational signals and using the strong T7 RNA polymerase promoter phi 10, that a cloned Saccharomyces cerevisiae pyruvate decarboxylase gene (pdc1) can be expressed in Escherichia coli. This protein, which migrated as a single band on SDS-polyacrylamide gels, was found to have a subunit molecular mass of approximately 62 kDa, similar to that of the enzyme produced by yeast. Polyclonal antibodies raised against purified yeast pyruvate decarboxylase recognized this bacterially produced protein. We found that this recombinant enzyme is active, indicating that the homotetramer encoded by the pdc1 gene is functional.

Base Sequence↗