Hydra's feeding response: Effect of GABAB ligands on GSH-induced electrical activity in the hypostome of H. vulgaris.

The feeding response in the Cnidarian, hydra, consists of mouth opening, tentacle writhing, and the cessation of pacemaker-controlled tentacle and body contractions. The behavior can be induced by reduced glutathione (GSH), contained in body fluids that leak from prey impaled by hydra's cnidocysts. Mouth, tentacle, and body-contraction behavior is carried out by hydra's ectodermal and endodermal epitheliomuscular cells. Here, we present the first evidence of GSH-induced electrical activity in the hypostome and its modification by GABA and GABAB ligands. The 'heads' of hydra were ablated and the tentacles removed. Suction electrodes, positioned on the mouth, recorded electrical activity produced by GSH, contained either within the electrode, or in the surrounding bath, the mouth being shielded. Recorded impulses were characterized, according to size and temporal pattern, as small, medium and (large) pacemaker impulses. GSH applied in the bath caused a frequency increase of small and medium impulses and a decrease in pacemaker bursts. The changes in frequencies of medium and pacemaker bursts, though not obviously affected by GABA, were counteracted by blocking GABAB inhibition with phaclofen. Only the highest concentration of GSH applied at the mouth potentially decreased pacemaker frequency and potentially increased medium impulses, without affecting small impulses. GABA caused a significant increase in small and medium impulses relative to GSH which was counteracted by baclofen and/or baclofen plus phaclofen. The results indicate that considerable GSH-receptor circuitry is located in hypostomal tissue proximal to hydra's mouth, and substantiate GABA and GABAB inhibition within the neuroeffector network of the feeding response.

The two nerve rings of the hypostomal nervous system of Hydra vulgaris-an immunohistochemical analysis.

In Hydra vulgaris, physiological and pharmacological evidence exists for a hypostomal circumferential neuro-effector pathway that initiates ectodermal pacemaker activity at tentacular-hypostomal loci coordinating body and tentacle contractions. Here, we describe an ectodermal nerve ring that runs below and between the tentacles, and an anti-GABAB receptor antibody-labeled ring coincident with it. The location of this ring is consistent with the physiology of the hypostomal pacemaker systems of hydra. We also describe a distally located, ectodermal ring of nerve fibers that is not associated with anti-GABAB receptor antibody labeling. The neurites and cell bodies of sensory cells contribute to both rings. The location of the distal ring and its sensory cell neurites suggests an involvement in the behavior of the mouth. Between the two rings is a network of anastomosing sensory and ganglion cell bodies and their neurites. Phase contrast, darkfield, and antibody-labeled images reveal that the mouth of hydra comprises five or six epithelial folds whose endoderm extensively labels with anti-GABAB receptor antibody, suggesting that endodermal metabotrobic GABA receptors are also involved in regulating mouth behavior.

Extraocular spectral photosensitivity in the tentacles of Hydra vulgaris.

Previous electrophysiological studies on the cnidarian Hydra vulgaris have shown that hydra have a highly developed and specific photoresponse despite their lack of any structure recognizable as a traditional photoreceptor. In an effort to identify the site of hydra's photoreceptors, we recorded extracellularly from single excised tentacles and from ablated hypostomes lacking tentacles in absolute darkness and during exposure to light of various wavelengths. During recording, after an initial period of absolute darkness, tentacles or hypostomes were exposed to light from 450nm to 600nm, red, and white light. Exposure to light caused a change in the pattern and frequency of impulses in the tentacles that varied with color. The number of large tentacle pulses (TPs) increased at 550 and 600nm relative to darkness, whereas the number of small tentacle pulses (STPs) tended to decrease in 500nm light. Impulse frequency was significantly different among the different wavelengths. In addition to bursts of tentacle contraction pulses, long trains of pulses were observed. A change in lighting caused a switch from bursting to trains or vice versa. In contrast to excised tentacles, no change in electrical activity was seen in ablated hypostomes at any of the wavelengths relative to each other or relative to darkness. These results indicate that isolated tentacles can distinguish among and respond to various colors across the visible spectrum and suggest that electromagnetic information is transmitted from the tentacles to the hypostome where it may be integrated by the hypostomal nervous system, ultimately contributing to hydra's photoreceptive behavior.

Exposure to the steroid hormone 20-hydroxyecdysone modulates agonistic interactions in male Homarus americanus.

In this study we present evidence that 20-hydroxyecdysone (20E) affects agonistic behavior in male American lobsters and that male and female animals differ in their response to the hormone. Thirty-minute staged fights were conducted between large males exposed either to artificial seawater (ASW) or 20E and small, anosmic opponents. The nephropores of both combatants were blocked. Fights were videotaped and quantitatively analyzed for aggressive, defensive and avoidance behaviors using an ethogram in which behaviors are ranked according to aggressiveness. Unlike female lobsters, exposing male lobsters to 20E did not increase their aggressive behavior; however, there was a marginally significant trend toward an increase in defensive behaviors with a lower aggressive content than in their ASW-exposed counterparts. The opponents of 20E-exposed animals performed significantly more aggressive behaviors than their counterparts. In fights with 20E-exposed animals, the overall aggressive intensity of the fight was increased and the animals performed a greater number of avoidance behaviors. Unlike the effects of 20E on females, where exposure to 20E caused an increase in overall agonistic arousal, males only exhibited a change in frequency of their behaviors. These findings suggest that while 20E affects both males and females in agonistic encounters, the nature of the effect is different for the two sexes.

Nematocyst discharge in Hydra vulgaris: Differential responses of desmonemes and stenoteles to mechanical and chemical stimulation.

The effects of mechanical and chemical stimulation on desmoneme and stenotele discharge in Hydra vulgaris were quantified, in situ, in isolated tentacles and single tentacles with attached hypostomes. Cnidocils of desmonemes and stenoteles were stimulated 24h after tentacle ablation. Single impacts with maximum calculated forces of 3.7x10(-3) to 3.8x10(-2) Newtons were delivered by piezoelectrically-driven glass capillary probes. Video analysis revealed that desmonemes discharged at forces of 3.7x10(-3)N; stenoteles required forces of 1.9x10(-2)N. Desmonemes not directly stimulated discharged if another desmoneme was adequately stimulated; the effect was carried through to at least two adjacent battery cells. Tentacles responded to desmoneme stimulation, by bending at the stimulation site. These findings imply afferent excitatory pathways between nematocytes onto other nematocytes and myonemes. Locomotory nematocysts (atrichous isorhizas) discharged only in hypostome-attached tentacles contacting a substrate; desmonemes and stenoteles did not discharge during substrate attachment, implying differential neuronal inhibition. At low concentrations, bath-applied mucin, a prey-associated glycosylated protein, lowered desmoneme and stenotele firing thresholds, abolishing the force dose-dependency in stenoteles, and allowing them to discharge at previously below threshold forces. At higher concentrations, mucin inhibited discharge, suggesting an involvement in prey-induced feeding inhibition.

Glutamatergic transmission in hydra: NMDA/D-serine affects the electrical activity of the body and tentacles of Hydra vulgaris (Cnidaria, Hydrozoa).

Previous electrophysiological studies on the early-evolved metazoan Hydra vulgaris provided evidence that glutamate, acting through alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) and kainate receptors, affects hydra's pacemaker systems; immunocytochemical studies showed that N-methyl-d-aspartate (NMDA) receptors were present in hydra tentacles; behavioral studies demonstrated that NMDA/d-serine affected mouth opening induced by reduced glutathione, and with AMPA/kainate, discharge of nematocysts. In this study, extracellular recordings were made from the tentacle and peduncle of hydra during bath application of NMDA and d-serine (both at 1 x 10(-5) mol l(-1) to 1 x 10(-9) mol l(-1)) in the presence of 1 x 10(-7) mol l(-1) AMPA or kainate. NMDA/d-serine produced a significant increase in tentacle activity, increasing the rate of tentacle pacemaker pulses (TPs) at 1 x 10(-7) mol l(-1), and small, behaviorally uncorrelated tentacle pulses (SUTPs) at 1 x 10(-5) mol l(-1). The NMDA antagonist, d-2-amino-5-phosphonopentanoic acid (D-AP5), counteracted the effects. NMDA/d-serine (1 x 10(-7) mol l(-1)) also caused a potentially significant (trend) decrease in the rate of small, behaviorally uncorrelated electrical body pulses (SUBPs) and rhythmic potentials (RPs). The effect was counteracted by D-AP5. The ectodermal contraction burst (CB) pacemaker system was unaffected by NMDA/d-serine. Our results indicate that glutamate, acting on NMDA/AMPA-kainate receptors, may cause opposing effects on the coordinating systems of tentacle and body-exciting the tentacle effectors and potentially causing an inhibition in the body column.

NMDA and GABA B receptors are involved in controlling nematocyst discharge in hydra.

The role of chemical neurotransmission in nematocyst discharge was investigated by stimulating the cnidocils of nematocysts in ablated tentacles of Hydra vulgaris with a piezoelectrically-driven glass probe, in the presence of selected neurotransmitters. Acetylcholine, dopamine, epinephrine, glycine, and serotonin (10(-4), 10(-6), 10(-8) M) per se, did not alter stenotele and desmoneme discharge. gamma-Amino-butyric acid (GABA) significantly increased desmoneme discharge when the cnidocil of another desmoneme in the same or adjacent battery cell complex was stimulated without affecting the discharge rates of the directly stimulated desmonemes or stenoteles. Baclofen (GABA(B) agonist) mimicked the increase; its antagonist, phaclofen, counteracted it. GABA(A) agonists and antagonists did not alter discharge rates. Glutamate caused a dose-dependent increase in the discharge rate of directly stimulated stenoteles; distant stenotele and desmoneme discharge rates were unaffected. Kainate, AMPA, and NMDA, per se, did not alter discharge rates. Co-administration of NMDA and kainate mimicked glutamate's effects. AMPA plus NMDA increased discharge rates. DAP-5 (NMDA antagonist) and CNQX, (kainate/AMPA antagonist) counteracted the increase. The findings suggest that metabotropic GABA is involved in recruiting desmonemes by disinhibiting those previously inhibited, and that the NMDA/kainate-AMPA mechanism regulating Ca(++) entry in higher neuroeffector systems is an early-evolved process, which, in hydra, modulates nematocyst discharge.

Cnidarian chemical neurotransmission, an updated overview.

The ultrastructural, histochemical, immunocytochemical, biochemical, molecular, behavioral and physiological evidence for non-peptidergic and peptidergic chemical neurotransmission in the Anthozoa, Hydrozoa, Scyphozoa and Cubozoa is surveyed. With the possible exception of data for the catecholamines and peptides in some animals, the set of cumulative data - the evidence from all methodologies - is incomplete. Taken together, the evidence from all experimental approaches suggests that both classical fast (acetylcholine, glutamate, GABA, glycine) and slow (catecholamines and serotonin) transmitters, as well as neuropeptides, are involved in cnidarian neurotransmission. Ultrastructural evidence for peptidergic, serotonergic, and catecholaminergic synaptic localization is available, but the presence of clear and dense-cored synaptic vesicles also suggests both fast and slow classical transmission. Immunocytochemical studies, in general, reveal a continuous, non-localized distribution of neuropeptides, suggesting a neuromodulatory role for them. Immunocytochemical and biochemical studies indicate the presence of glutamate, GABA, serotonin, catecholamines (and/or their receptors), RFamides, nitric oxide and eicosanoids in cnidarian neurons and tissues. Gene sequences for peptidergic preprohormones have been reported; putative gene homologies to receptor proteins for vertebrate transmitters have been found in Hydra. Behavioral and physiological studies implicate classical transmitters, neuropeptides, eicosanoids and nitric oxide in the coordination of the neuroeffector systems.

Pacemaker activity in hydra is modulated by glycine receptor ligands.

In the mammalian central nervous system, the neurotransmitter, glycine, acts both on an inhibitory, strychnine-sensitive receptor (GlyR) and an excitatory, strychnine-insensitive site at the NMDA receptor. Here we present electrophysiological evidence that the strychnine-sensitive glycine agonists, glycine and taurine, and the antagonist, strychnine, affect the endodermal rhythmic potential (RP) system and that the ectodermal contraction burst (CB) pacemaker system is modulated by glycine and strychnine in hydra. The RP and CB pacemaker systems are responsible for the respective elongation and contraction of hydra's body column. Activity of the CB system, quantified by the rate of contraction bursts (CBs), the number of pulses per contraction burst (P/CB), and the duration of bursts, was decreased by glycine. Glycine, coadministered with the strychnine-insensitive glycine site blocker, indole-2-carboxylic acid (I2CA), decreased RPs but not CBs or P/CB. The effect was mimicked by taurine. Strychnine increased the duration of RP production, and decreased CB duration. The effect of glycine with I2CA was counteracted by strychnine. The results support the idea that a vertebrate-like GlyR may be involved in modulating activity of the endodermal RP system and suggest that a glycine site on an NMDA receptor may be involved in the CB system.

Immunocytochemical evidence for an NMDA1 receptor subunit in dissociated cells of Hydra vulgaris.

We have previously reported immunocytochemical, biochemical, behavioral, and electrophysiological evidence for glutamatergic transmission through (+/-)-alpha-amino-3-hydroxy-5-methyl-isoxazole-4-propionic acid (AMPA)/kainate receptors in hydra. We now report specific localization of the N-Methyl-D-aspartic acid receptor subunit 1 (NMDAR1) in epithelial, nerve, nematocytes, and interstitial cells of hydra. Macerates of tentacle/hypostome pieces of Hydra vulgaris were prepared on agar-coated slides, fixed with buffered formaldehyde/glutaraldehyde, and fluorescently labeled with monoclonal antibodies against mammalian NMDAR1. Negative controls omitted primary antibody. Digital images were recorded and analyzed. Specific localized and intense labeling was found in ectodermal battery cells, other epithelial cells, nematocytes, interstitial cells, and sensory and ganglionic nerve cells, and in battery cells was associated with enclosed nematocytes and neurons. The labeling of myonemes was more diffuse and less intense. In nerve and sensory cells, punctate labeling was prominent on cell bodies. These results are consistent with our earlier evidence for glutamatergic neurotransmission and kainate/NMDA regulation of stenotele discharge. They support other behavioral and biochemical evidence for a D-serine-sensitive, strychnine-insensitive, glycine receptor in hydra and suggest that the glutamatergic AMPA/kainate-NMDA system is an early evolved, phylogenetically old, behavioral control mechanism.

GABA and glutamate receptors are involved in modulating pacemaker activity in hydra.

The effects of gamma-amino butyric acid (GABA) and glutamate, their ionotropic agonists and antagonists on hydra's ectodermal and endodermal pacemaker systems were studied. GABA decreased ectodermal body contraction bursts (CBs) and the number of pulses in a burst (P/CB) and endodermal rhythmic potentials (RPs); tentacle pulses (TPs) were not affected. The GABA(A) agonist, muscimol, and the benzodiazepine receptor agonist, diazepam, mimicked the effects of GABA on the endodermal system. The GABA(A) antagonist bicuculline counteracted GABA's effects. Low concentrations of glutamate increased CBs and RPs. Higher concentrations required concanavalin A (Con A) to produce the same effect on CBs and P/CB. TPs were increased by high concentrations of glutamate and kainate. The ionotropic glutamate agonist, alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) also required Con A to increase CBs and RPs. The effects of AMPA were antagonized by 6-nitro-7-sulfamoylbenzo[f]quinoxaline-2,3-dione (NBQX), which, per se, decreased CBs. The results indicate that GABA and glutamate, acting on their ionotropic receptors, modify the impulses of hydra's pacemaker systems. On the whole GABA decreased the outputs of both ectodermal and endodermal impulse generating systems, while glutamate increased them.

20-hydroxyecdysone causes increased aggressiveness in female American lobsters, Homarus americanus.

Lobsters become transiently more aggressive before ecdysis. This aggressiveness accompanies an increase in hemolymph titers of 20-hydroxyecdysone (20-HE). Combats between intermolt female lobsters, injected with premolt levels of 20-HE, and larger, saline-injected opponents were videotaped. Aggressive, defensive, and avoidance behaviors were ranked according to aggressiveness in a Rank of Aggression hierarchy, which included opponent-directed and (nonopponent) redirected behaviors. Treated animals performed more and more highly aggressive behaviors than saline-injected controls. Opponents of treated animals performed fewer aggressive behaviors than saline-injected control opponents. Controls performed more defensive behaviors than treated animals, when redirected behaviors were considered. Differences in avoidance behaviors among the four types of combatants were not significant. The total aggressive content was the same in treated and control fights, but the interactions between combatants in the two fights were significantly different. Treated animals were equally as aggressive and defensive as their opponents; controls were relatively less aggressive and more defensive than their opponents. These results correlate with molt-cycle variations in behavior, 20-HE titers, and the effects of 20-HE and molt-differentiated hemolymph on the claw opener muscle. They suggest that 20-HE orchestrates intrinsic, cellular, and nuclear events that produce the molt-cycle transformations in agonistic behavior and aggressive state of lobsters.

Molt-related and size-dependent differences in the escape response and post-threat behavior of the American lobster, Homarus americanus.

Videotaped recordings of adult lobsters of different molt stages were analyzed. The escape response of adults was compared with that of juveniles recorded in an earlier study. Juvenile lobsters always respond to a threat with escape behavior irrespective of their molt stage, but in adults the probability of eliciting a response was a function of molt stage: more hard-shelled (intermolt stage C) and (premolt stage D) animals tailflipped than did soft-shelled (postmolt stages A and B) animals. The number, frequency, and duration of tailflips, and the average distance swum by animals in each molt stage were measured for the entire escape response, for the initial power swim, and for the subsequent swims. These measurements were used to compute several parameters: velocity, acceleration, force, and work; average distance traveled in a tailflip for each kilogram of body weight (distance/kg/tailflip); and average distance traveled for each bodylength (distance/bodylength). Among adults, intermolt (stage C) lobsters traveled significantly farther and faster than postmolt animals (stages A and B). Among juveniles, late postmolt (stage B) animals traveled farther. Among adults, although the total number of tailflips and the duration of the response were not significantly different among molt stages, the number of tailflips/second (frequency) and distance traveled/kg/tailflip were greater for intermolt animals. In juvenile intermolts, however, frequency and distance/kg/tailflip were markedly lower than in the premolt stages. Although values were lower than intermolts and premolts, postmolt adults sustained their swimming frequency, distance/kg/tailflip, and distance/bodylength for the entire escape distance (as did postmolt juveniles). These parameters then dropped off sharply for both adult and juvenile intermolt and premolt animals in the second half of the escape distance. Post-threat behaviors reveal that stage D animals have the highest aggression index and often attack the presented stimulus, whereas stage A animals are the least likely to approach the stimulus and typically back away in a non-aggressive posture. Thus, although effects of the molt cycle on adult and juvenile escape behavior are similar in some ways, other physical characteristics of adults, such as weight, allometry, and physiology, seem to become important in determining the likelihood of escape behavior and the characteristics of the escape swim in each molt stage.

Time in residence affects escape and agonistic behavior in adult male american lobsters.

Acquisition and retention of a shelter by a lobster are two of the variables that play a role in lobster agonistic interactions. Since shelter procurement and retention are important for lobster survival, behaviors related to this activity frequently outrank other daily behaviors (e.g., searching for food). Here, we examine the effects of time in residence on the parameters of the escape response of the American lobster, Homarus americanus. Adult male intermolt lobsters (Stage C4) were placed in an experimental tank for three different time periods (one hour, 24 hours, and 48 hours). The probability of eliciting an escape response was inversely related to the time spent in the tank. Eighty percent of the animals in residence for 1 h tailflipped in response to a threat, whereas only 14% of the animals in residence for 48 h tailflipped. There were also significant changes in some of the parameters of the escape response among animals in residence for 24 h compared to those in residence for 1 h. The number of tailflips and the distance traveled were reduced, although frequency, velocity, acceleration, force, and work factors were not significantly different. Furthermore, with increased time in residence, lobsters switched from an avoidance or escape-prone behavior to an aggressive-prone behavior. Most of the animals in residence for 48 h approached and attacked a threat-stimulus rather than fleeing from it. On an empirically defined "index of aggressiveness," in which various behaviors were numerically ranked from least aggressive (0) to most aggressive (6), animals residing in the tank for 1 h had an average index value of 0.1 compared to a value of 5.0 for animals in residence for 48 h. These findings are consonant with the suggestion that lobsters that have occupied a given space for an extended period of time take possession of the site and defend it instead of fleeing when threatened with a threat-inducing stimulus; it supports the idea that shelter retention increases aggressiveness and diminishes avoidance behaviors.

Comparative Analysis of Escape Behavior in Male, and Gravid and Non-Gravid, Female Lobsters.

Few studies exist in which the parameters of a single behavior have been quantitatively compared for male and female lobsters. Here, we have examined the effects of sex and gravidity on the parameters of the escape behavior of the American lobster, Homarus americanus, elicited by a visual threat. Both non-gravid females and male lobsters readily tailflipped in response to the stimulus, but gravid females failed, with one exception, to initiate a swim, even when stimulus strength was increased. Although the total distance swum by males and non-gravid females was not statistically different, males covered more ground in the initial power swim and during the subsequent swims than did non-gravid females. Males swam for a longer time, performing more tailflips, than females. Relative to their length and weight, males swam a greater distance at each stroke during the initial power swim and the subsequent swims, although, females might have compensated by swimming at a higher frequency. There were no significant differences in swimming velocity or acceleration, nor in the calculated force or work performed by the two sex classes (male and non-gravid females). Therefore, apart from egg-bearing, which severely inhibits the escape response, it remains to be seen whether the subtle physiological and anatomical sexual dimorphism that produces longer and more swim strokes in males but higher frequency tailflips in females results in the same chances of survival for the sexes.

Chemoreceptive control of feeding processes in hydra.

Cnidarians are the simplest metazoans to exhibit satiety after feeding. When hydra are fed to repletion, they close their mouths and cease to capture prey. As feeding stops, contractions of the tentacles and body column increase. Our earlier experiments showed that a gel chromatographic fraction of prey substances inhibits prey capture. We now present evidence that the same fraction reduces the duration of mouth opening induced by reduced glutathione (GSH) and inhibits the binding of GSH to its putative receptor. The fraction also induces column contractions which are similar to those normally seen in sated animals. Prey substances, of unfractionated homogenate, also induce post-feeding tentacle contractions similar to those seen in sated animals. Gut distention does not appear to induce behavior associated with satiety. Therefore, these experiments suggest that chemoreception of prey substances induce satiety in hydra.

Partial characterization and detergent solubilization of the putative glutathione chemoreceptor from hydra.

Feeding behavior in hydra is initiated by the association of glutathione (GSH) with a putative external chemoreceptor. In the present study, the binding of [35S]GSH to hydra membranes has been characterized. Nondisplaceable [35S]GSH binding which compromised previous analyses [Grosvenor, W., Bellis, S., Kass-Simon, G., & Rhoads, D. (1992) Biochim. Biophys. Acta (in press)] was eliminated by treating membranes with an inhibitor of GSH metabolism, borate in combination with L-serine. The specific binding which was not inhibited by borate/serine demonstrated many of the characteristics expected of a ligand/receptor interaction. The binding was rapid, reversible, and saturable. A Scatchard analysis of saturation isotherms indicated a dissociation constant (KD) of 3.4 microM, a value which is in good agreement with concentrations of glutathione which are known to induce feeding behavior. Hydra membranes were detergent-solubilized with 10 mM 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS), 100 mM KCl, and 10% glycerol. The soluble fraction contained 40% of the original saturable, reversible GSH binding activity. The KD for GSH binding to the solubilized preparation was estimated as 2.7 microM, a valuable which is not appreciably different from the KD for binding to intact membranes. The fidelity of GSH binding in the solubilized preparation suggests that this preparation will be useful in further characterization of the putative glutathione chemoreceptor.

Chemoreception in hydra: specific binding of glutathione to a membrane fraction.

Specific binding of reduced [35S]glutathione (GSH) was measured using a crude membrane fraction of Hydra vulgaris (attenuata). The specific binding shows both rapid displaceable and nondisplaceable components. Rapid displaceable binding accounted for 20% of the total specific binding. Data from saturation binding experiments indicates half maximal total specific binding occurs at 2 microM GSH which is similar to reported EC50 values from behavioral experiments. Calcium is required for displaceable binding of GSH to the putative receptor. Oxidized glutathione (GSSG), an antagonist of the GSH-activated feeding response, and S-methylglutathione (GSM), an agonist of the feeding response, inhibit the binding of radiolabeled GSH to the putative receptor. Glutamate, a putative competitive antagonist of the GSH-activated feeding response in hydra, does not inhibit the specific binding of radiolabeled GSH to the receptor and must therefore block the feeding response by a mechanism other than competitive inhibition.

Suspected chemoreceptors in coelenterates and ctenophores.

Chemoreceptors in coelenterates and ctenophores have not been identified with certainty. Among prospective chemoreceptive cells are the sensory nerve cells, the cnidocyst-bearing cnidocytes, and the epitheliomuscular cells that are likely to be involved in feeding or aggression. Both behaviors are mediated by coordinated chemical and mechanical reception. This is reflected in the close apposition of putative chemo- and mechanoreceptors. Among the structures that have been designated as likely chemo- and/or mechanoreceptors are stereocilia, kinocilia, and/or microvilli which are universally present on all the putative chemoreceptor complexes, while gland cells and mucous secretions are prevalent. Evidence that the actin-containing stereocilia are chemically modulated mechanoreceptors is presented for several forms.

Chemoreception in Hydra vulgaris (attenuata): initial characterization of two distinct binding sites for L-glutamic acid.

To elucidate the relationship between L-glutamic acid and the putative chemoreceptor for glutathione, binding of L-[3H]glutamate to a crude membrane fraction from Hydra vulgaris (attenuata) has been characterized. The binding of L-[3H]glutamate was rapid, reversible and saturable. A Scatchard analysis of the specific binding revealed values of 10 microM for the dissociation constant (Kd) and 170 pmol/mg for the maximal capacity of binding sites (Bmax). A maximum of 65% of the specific L-[3H]glutamate binding was inhibited by the chemostimulatory peptide, glutathione. This glutathione-sensitive glutamate binding presumably represents the association of glutamate with a putative chemoreceptor which modulates feeding behavior in hydra. The remaining 35% of the specific L-[3H]glutamate binding may be due to a second class of glutamate binding sites which is insensitive to glutathione. The identification of glutathione-insensitive glutamate binding is the first indication of a putative glutamate receptor, which may mediate an action independent of the glutathione-induced feeding response. The glutathione-insensitive and glutathione-sensitive sites must have similar affinities for glutamate since these sites were indistinguishable by Scatchard analysis. A preliminary characterization of the glutathione-insensitive site, performed in the presence of saturating levels of glutathione, revealed inhibition of glutathione-insensitive glutamate binding by kainate and quisqualate, but not by N-methyl-D-aspartate. A glutathione-insensitive L-[3H]glutamate binding suggests that kainate and alpha-aminoadipate may be selective ligands for the glutathione-insensitive and glutathione-sensitive glutamate binding sites, respectively.