Ingestion of sugar diets correlates with spike activity in labellar chemosensilla of the flesh-fly, Neobellieria (= Sarcophaga ) bullata

Abstract .Female 2-day-old Neobellieria (= Sarcophaga ) bullata (Parker) (Diptera: Sarcophagidae) were exposed to different concentrations of sucrose, glucose and fructose in a single-choice potometer, and the volume ingested in the first hour was measured. Nerve spike activity in response to the same sugars was recorded from medium labellar taste hairs of similar flies by tip-recording. Two classes of chemosensory cells responded to sucrose, glucose and fructose. Cell 1 showed an increasing spike activity with sugar concentration, whereas cell 2 did not; cell 1 was identified as the 'sugar cell'.
For both spike activity in cell 1 and feeding, sucrose was the most stimulatory sugar. The dose–response curves for glucose and fructose crossed over at about 200 m m . At higher concentrations, glucose was more stimulatory for both cell 1 and for feeding, and at lower concentrations, fructose. The pattern of spike activity supports a separate location on the sensory cells of receptors for pyranose and fructose forms of sugar. The strong correlation between volume ingested and spike activity indicates that sugar feeding is controlled by sensory input from the 'sugar' cells of labellar chemosensilla. Moreover, the results suggest that the flies do not distinguish between these sugars except by apparent 'sweetness'.     

A direct comparison of electrophysiological and behavioural taste responses in the blowfly

Behavioural and electrophysiological taste responses in theblowfly (Calliphora vicina) are measured in successive experiments,using the same flies twice. Inter-individual variations in behaviourare at least partly due to differences in the functioning ofthe tarsal taste hairs; flies with low behavioural thresholdvalues have taste hairs firing with higher rates. The percentageof taste hairs responding with spike trains is not important.Behavioural responses are predominantly influenced by the ‘best’firing hair. The most effective of the parameters describingspike trains seems to bef_t, the firing rate after an ‘infinite’length of stimulation time.     

Response characteristics of rat taste cells to potassium benzoate

The rat taste cells responded to K-benzoate solutions higher than the threshold concentrations (0.03-0.3 M) with a depolarizing receptor potential, but they responded to K-benzoate lower than the thresholds with a hyperpolarizing receptor potential. In either depolarizing or hyperpolarizing receptor potentials the rise time decreased with increasing amplitude, but the fall time increased with increasing amplitude. During generation of either depolarizing or hyperpolarizing receptor potentials the input resistance of taste cells decreased with increasing amplitude. Application of the mixtures of various concentrations of NaCl and 0.05 M K-benzoate resulted in a reduction of receptor potential amplitude, as compared with that evoked by application of NaCl alone. It is concluded that a depression of gustatory neural impulse frequency by low concentrations of K-benzoate is mainly due to the hyperpolarizing receptor potential of taste cells elicited by the K-benzoate solutions.     

Electrophysiological responses of galeal contact chemoreceptors of Apis mellifera to selected sugars and electrolytes

Using conventional electrophysiological methods, the galeal sensilla chaetica of the honey bee, Apis mellifera, responded linearly to the log of solute concentrations of sucrose, glucose, fructose, NaCl, KCl, and LiCl but not to CaCl₂ or MgCl₂, which failed to give consistent responses. These sensillae had much higher firing rates for sugar than salt solutions; their relative responses to lower concentrations being NaCl < KCl < LiCl ? fructose < glucose ? sucrose. At higher concentrations NaCl < LiCl < KCl ? glucose < fructose ? sucrose. Four different spike types were seen. The first type had the highest amplitude and resulted from sugar stimulation. The second type had a lower height and occurred in the first 30 sec of salt stimulation. A third type with the lowest height appeared with those of the second type after prolonged stimulation with KCl. A fourth type with a high amplitude resulted from mechanical stimulation. The sensilla adapted to sugar solutions linearly to the logarithm of time and non-linearly to the log of salt concentrations. Glucose-fructose mixed-sugar solution effected synergism of response while sucrose solutions caused inhibition when mixed with glucose and/or fructose. Responses of the sensilla to mechanical stimulation showed phasic-tonic characteristics. None of the sensilla tested responded to water.     

Simultaneous chemical and electrical stimulation of labellar taste hairs of the blowfly Calliphora vicina

When recording from the tip of insect taste hairs, responses to chemical stimulation may be influenced by electrical currents, such as the preamplifier's input bias current. The effect of electrical currents on firing frequency of the salt receptor cell to KCl and NaCl stimulation was determined in labellar ‘aboral’ and ‘adoral’ taste hairs of the blowfly Calliphora vicina. Negative currents always decreased spike frequency, whereas positive currents either increased it, or did not change it significantly. Spike frequency changed less than 1% per 5 × 10^?11 A.A consistent picture of the electrophysiology of blowfly taste hairs is given. It includes a distal pore, present in the dendrite-free lumen of the hair. It abandons the concept of a generator current that transmits excitation from the distal, chemoreceptive part of the taste cell dendrite to the action potential generator in or near the taste cell body. The experimental results are interpreted on the basis of this picture. It is concluded that the ‘electrophoretic effect’ of the electrical current is very small. Thus, the measured effect should mainly be due to a ‘direct effect’ of electrical current on electrically excitable structures in the salt receptor cell, particularly in its dendrite.     

Multiple sensitivity of single taste cells of the frog tongue to four basic taste stimuli

The electrical response of the taste cells of the frog fungiform papillae to four fundamental taste solutions (NaCl, acetic acid, quinine-HCl and sucrose) was studied by using the intracellular recording technique. The average value of resting membrane potential was 22.5 mV, inside negative. Each of the four taste solutions applied to the tongue produced a slow depolarizing potential, the receptor potential, on which no spike potential was superimposed. The amplitude of the receptor potentials increased linearly as a function of the logarithm of the concentration of the stimulus. Amplitudes of depolarizations to a given taste stimulation varied from one cell to another even within a single taste bud. Most of the cells responded to more than two of the four basic taste solutions. Sensitivity patterns in terms of the number of effective solutions and the relative effectiveness of different kinds of solutions were variable among cells. Statistical analysis suggests that at the receptor membranes of the taste cells, the sensitivities for the four basic stimuli are independent and random.     

Transduction ion channels directly gated by sugars on the insect taste cell

Insects detect sugars and amino acids by a specialized taste cell, the sugar receptor cell, in the taste hairs located on their labela and tarsi. We patch-clamped sensory processes of taste cells regenerated from the cut end of the taste hairs on the labelum of the flashfly isolated from the pupa approximately 20 h before emergence. We recorded both single channel and ensemble currents of novel ion channels located on the distal membrane of the sensory process of the sugar receptor cell. In the stable outside-out patch membrane excised from the sensory processes, we could repeatedly record sucrose-induced currents for tens of minutes without appreciable decrease. An inhibitor of G-protein activation, GDP-beta-S, did not significantly decrease the sucrose response. These results strongly suggested that the channel is an ionotropic receptor (a receptor/channel complex), activated directly by sucrose without mediation by second messengers or G protein. The channel was shown to be a nonselective cation channel. Analyses of single channel currents showed that the sucrose-gated channel has a single channel conductance of approximately 30 pS and has a very short mean open time of approximately 0.23 ms. It is inhibited by external Ca(2+) and the dose-current amplitude relation could be described by a Michaelis-Menten curve with an apparent dissociation constant of approximately 270 mM. We also report transduction ion channels of the receptor/channel complex type directly gated by fructose and those gated by L-valine located on the sensory process.     

Depression of gustatory receptor potential in frog taste cell by parasympathetic nerve-induced slow hyperpolarizing potential

Parasympathetic nerve (PSN) innervates taste cells of the frog taste disk, and electrical stimulation of PSN elicited a slow hyperpolarizing potential (HP) in taste cells. Here we report that gustatory receptor potentials in frog taste cells are depressed by PSN-induced slow HPs. When PSN was stimulated at 30 Hz during generation of taste cell responses, the large amplitude of depolarizing receptor potential for 1 M NaCl and 1 mM acetic acid was depressed by approximately 40% by slow HPs, but the small amplitude of the depolarizing receptor potential for 10 mM quinine-HCl (Q-HCl) and 1 M sucrose was completely depressed by slow HPs and furthermore changed to the hyperpolarizing direction. The duration of the depolarizing receptor potentials depressed by slow HPs prolonged with increasing period of PSN stimulation. As tastant-induced depolarizing receptor potentials were increased, the amplitude of PSN-induced slow HPs inhibiting the receptor potentials gradually decreased. The mean reversal potentials of the slow HPs were approximately -1 mV under NaCl and acetic acid stimulations, but approximately -14 mV under Q-HCl and sucrose stimulations. This implies that when a slow HP was evoked on the same amplitude of depolarizing receptor potentials, the depression of the NaCl and acetic acid responses in taste cells was larger than that of Q-HCl and sucrose responses. It is concluded that slow HP-induced depression of gustatory depolarizing receptor potentials derives from the interaction between gustatory receptor current and slow hyperpolarizing current in frog taste cells and that the interaction is stronger for NaCl and acetic acid stimulations than for Q-HCl and sucrose stimulations.     

Impulse frequency and action potential amplitude in labellar chemosensory neurones of Drosophila melanogaster

The electrophysiological responses of water, salt and sugar receptors in the labellar chemosenory hair of Drosophila were investigated. In contrast to the responses of large flies such as blowfly and fleshfly, spike height changed in parallel with the spike frequencies in all the three kind of receptors, and at the same time, the spike also changed in shape: when the receptor potential was small, the spike was small and biphasic, but when the receptor potential was large, the spike was large and monophasic. These phenomena are consistently explained by assuming that antidromic conduction of spikes in the distal process of the receptor cells is blocked due to inactivation of Na channels by the depolarizing receptor potential.     

Detection of alkaloids and carbohydrates by taste receptor cells of the galea of gypsy moth larvae, Lymantria dispar (L.)

Gypsy moth larvae are polyphagous feeders. The electrophysiological responses of the medial and lateral styloconic sensilla to four secondary compounds (e.g., alkaloids), two carbohydrates, and one inorganic salt were examined using an extracellular tip-recording method. In the medial sensillum, one taste receptor cell responded to the alkaloids, strychnine, caffeine, nicotine, and aristolochic acid (i.e., deterrent-sensitive cell), while another, responded to the sugar alcohol and inositol (inositol-sensitive cell). In both medial and lateral sensilla, two taste receptor cells in each sensillum responded minimally and sporadically to 30?mM potassium chloride (KCl) (i.e., KCl-sensitive cells); one cell produced much larger amplitude action potentials than the other. In the medial sensillum, only the large-amplitude KCl-sensitive cell exhibited an increased firing rate with increasing salt concentration. When binary mixture experiments were conducted, it was confirmed that the large-amplitude KCl-sensitive cell and the deterrent-sensitive cell in the medial sensillum were one in the same cell. Only a single cell in the lateral sensillum responded to the sugar, sucrose (sucrose-sensitive cell). The temporal dynamics of responses of the deterrent-sensitive, sucrose-sensitive, and inositol-sensitive cells were compared. Concentration?Cresponse data were obtained for the deterrent-sensitive cell to various alkaloids, as well as to KCl.     

The effects of amiloride on the labellar taste receptor cells of the fleshfly Boettcherisca peregrina

Amiloride is known to inhibit the taste response of vertebrates to salt by blocking the amiloride-sensitive sodium channel. In this study, we investigated electrophysiologically the effect of amiloride on the taste response of the fleshfly Boettcherisca peregrina. When 0.5 mM amiloride was included in taste solutions, the response of the salt receptor cell (salt response) to sodium chloride (NaCl) was not depressed but those of the sugar receptor cell (sugar responses) to sucrose, glucose, fructose, l-valine (l-Val) and l-phenylalanine (l-Phe) were strongly depressed. An inhibitory effect of amiloride on the concentration-response relationship for both sucrose and l-Phe was clearly revealed, but not at high concentrations of sucrose. After pretreatment of a chemosensory seta with 0.15 mM amiloride for 10 min, the salt response to NaCl was not affected. On the other hand, the sugar responses to sucrose, fructose, l-Val and l-Phe were depressed just after amiloride pretreatment. The sugar response to adenosine 5’-diphosphate (ADP) mixed with 0.5 mM amiloride was not depressed, but the response to ADP alone was depressed after amiloride pretreatment. It was therefore observed that amiloride depressed the responses to all stimulants that react with each of the receptor sites of the sugar receptor cell.     

Protein-sensitive elements of the labellar sensillas of Musca domestica flies

Using electrophysiological technique of registration of impulse activity in chemoreceptive cells of the labellar sensillae of the housefly, it has been demonstrated that taste hairs are not uniform in their properties. They differ from each other by the set of receptive elements which exhibit different sensitivity, range of selectivity and pattern of impulse activity. It was shown that albumen solution (10(-5) M) evokes the activity in 1--2 cells of a sensilla which are classified as water and sugar receptors. Among these receptors, protein-sensitive and protein-insensitive cells may be distinguished. Considering the inhomogeneity of sugar receptor sites, it was suggested that chemo receptive membranes in most sensitive to protein cells contain more numerous fructose receptive sites, that glucose ones.     

Intracellular responses of antennal chordotonal sensilla of the American cockroach

The responses of mechanoreceptor neurons in the antennal chordotonal organ have been examined in cockroaches by intracellular recording methods. The chordotonal organ was mechanically stimulated by sinusoidal movement of the flagellum. Stimulus frequencies were varied between 0.5 and 150 Hz. Receptor neurons responded with spike discharges to mechanical stimulation, and were classed into two groups from plots of their average spike frequencies against stimulus frequency. Neurons in one group responded to stimulation over a wide frequency range (from 0.5 to 150 Hz), whereas those in a second group were tuned to higher frequency stimuli. The peak stimulus frequency at which receptor neurons showed maximum responses differed from cell to cell. Some had a peak response at a stimulus frequency given in the present study (from 0.5 to 150 Hz), whereas others were assumed to have peak responses beyond the highest stimulus frequency examined. The timing for the initiation of spikes or of a burst of spikes plotted against each stimulus cycle revealed that spike generation was phase-locked in most cells. Some cells showed phase-independent discharges to stimulation at lower frequency, but increasing stimulus frequencies spike initiation began to assemble at a given phase of the stimulus cycle. The response patterns observed are discussed in relation to the primary process of mechanoreception of the chordotonal organ.     

Taste cell responses in the polyphagous arctiid, Grammia geneura: towards a general pattern for caterpillars

The caterpillars of Grammia geneura are polyphagous as individuals. Electrophysiological responses of its medial and lateral galeal styloconic sensilla to 21 amino acids, 6 carbohydrates, 10 chemically diverse plant secondary compounds and two inorganic salts were examined. In the medial sensillum, a single cell responded to 8 amino acids, 3 carbohydrates, and the iridoid, catalpol, which is present in a favored hostplant. In the lateral sensillum, one cell responded to amino acids and another to fructose. Two cells in each sensillum responded to secondary compounds and it is suggested that the same cells are stimulated by inorganic salts. There was no evidence of a separate salt-sensitive cell. Phenylalanine stimulated a deterrent cell in the medial sensillum and was behaviorally deterrent. Some essential amino acids did not stimulate any cells and it is suggested that a small number of amino acids (sometimes non-essential) may serve as indicators of nutrient quality. Sugars probably serve as the primary phagostimulants because they are in relatively high concentrations in plants. It is proposed that taste receptor cells should be categorized primarily by their behavioral effects as phagostimulatory or deterrent, rather than their specific ranges of responsiveness. This would emphasize the basic similarities across taxa.     

Variability in blowfly taste responses

Summary An attempt was made to quantify the various sources of variability in the responses of taste cells on the leg of the blowflyCalliphora, and to discover which processes can be held responsible for these sources of variability. Variability increases the proportion of misclassification of response, and therefore seriously hinders attempts to unravel neural coding. Recommendations are formulated for a better experimental procedure.Ten flies were used, 10 hairs on each left frontal leg were stimulated 16 times, resulting in 1600 spike trains. The sources of variability investigated are: (1) differences between flies (effect of individual fly); (2) differences between taste hairs of the same type (effect of hair topology); (3) the moment of stimulation after amputation of the leg (amputation effect).Additionally, the unexplained residual variability is quantified.A 3-way analysis of variance was used. The results provide a strong argument to carry out experiments with one and the same fly: interindividual variability constitutes almost 50% of the total variation (Tables 1a, 2a, b).It might be advantageous to restrict ourselves to cell responses from a single tarsomere: hair topology constitutes about 6% of the total variation (Tables 1a, 2a, b).Recordings should be made during a short period — 10 to 30 min — after amputation. Presumably this period can be enhanced by preventing the evaporation of water from the open wound. The reason is that amputated legs show a decrease in response values with time. The contribution to the total variation is more prominent in B-hairs, but remains below 5% (Tables 1a, 2a, b).Residual variability amounts to about 40% of the total variability ¯f (Tables 1a, 2a, b). Fluctuations take place faster than can be detected with intervals of 10 min. Phenomena such as residual variability, non-responsiveness, delayed responses and irregular responses probably have a common causal factor; changes in geometry of hair tip and lumen are a possible cause of these phenomena.     

The sensitivity of the labellar sugar receptors of Phormia regina in relation to feeding

The hypothesis that behavioural feeding threshold of P. regina modulates and is modulated by the sensitivity of the labellar contact chemoreceptors is not supported by the examination of individual flies.Sensitivity (number of impulses fired in the first second of stimulation) of the sugar receptors in selected labellar hairs of flies of known age was recorded periodically on a fixed schedule for up to 72 hr. Experimental flies were hand fed 2.0 M fructose once every 24 hr. No correlations between feeding or age and receptor sensitivity were found. Statistically significant changes in firing frequency did occur but were unrelated to feeding and age.The proboscis extension response, on which behavioural threshold measures are based, is known to be triggered by the first few sugar receptor spikes of sufficient frequency. Neither age nor feeding significantly affected the number or frequency of impulses during the first 50 msec of stimulation.     

Diverse roles for the Drosophila fructose sensor Gr43a

The detection of nutrients, both in food and within the body, is crucial for the regulation of feeding behavior, growth, and metabolism. While the molecular basis for sensing food chemicals by the taste system has been firmly linked to specific taste receptors, relatively little is known about the molecular nature of the sensors that monitor nutrients internally. Recent reports of taste receptors expressed in other organ systems, foremost in the gastrointestinal tract of mammals and insects, has led to the proposition that some taste receptors may also be used as sensors of internal nutrients. Indeed, we provided direct evidence that the Drosophila gustatory receptor 43a (Gr43a) plays a critical role in sensing internal fructose levels in the fly brain. In addition to the brain and the taste system, Gr43a is also expressed in neurons of the proventricular ganglion and the uterus. Here, we discuss the multiple potential roles of Gr43a in the fly. We also provide evidence that its activation in the brain is likely mediated by the neuropeptide Corazonin. Finally, we posit that Gr43a may represent only a precedent for other taste receptors that sense internal nutrients, not only in flies but, quite possibly, in other animals, including mammals.     

The Effects of Amino Acids on the Labellar Hair Chemosensory Cells of the Fly

The effects of amino acids on the labellar hair chemosensory cells were examined with two kinds of flies (the fleshfly, Boettcherisca peregrina, and the blowfly, Phormia regina). As a result of this examination, the effects of amino acids were divided into four main classes. Amino acids in class 1 did not stimulate any chemoreceptor cell. Amino acids in class 2 inhibited nonspecifically the discharges from three kinds of chemosensory cells. Amino acids in class 3 stimulated the salt receptor cell. Amino acids in class 4 stimulated the sugar receptor cell. A possibility that a fourth neuron in the labellar hair chemosensory cell might be a protein or an amino acid receptor cell was eliminated.     

Diverse roles for the Drosophila fructose sensor Gr43a

The detection of nutrients, both in food and within the body, is crucial for the regulation of feeding behavior, growth, and metabolism. While the molecular basis for sensing food chemicals by the taste system has been firmly linked to specific taste receptors, relatively little is known about the molecular nature of the sensors that monitor nutrients internally. Recent reports of taste receptors expressed in other organ systems, foremost in the gastrointestinal tract of mammals and insects, has led to the proposition that some taste receptors may also be used as sensors of internal nutrients. Indeed, we provided direct evidence that the Drosophila gustatory receptor 43a (Gr43a) plays a critical role in sensing internal fructose levels in the fly brain. In addition to the brain and the taste system, Gr43a is also expressed in neurons of the proventricular ganglion and the uterus. Here, we discuss the multiple potential roles of Gr43a in the fly. We also provide evidence that its activation in the brain is likely mediated by the neuropeptide Corazonin. Finally, we posit that Gr43a may represent only a precedent for other taste receptors that sense internal nutrients, not only in flies but, quite possibly, in other animals, including mammals.