Monoclonal antibodies that depress a specific subset of multiple components of the glutathione-induced response ofHydra

Summary Reduced glutathione evokes a feeding response, the tentacle-ball formation inHydra japonica. This response consists of at least 5 components (R1–R5). We raised 6 monoclonal antibodies (mAbs), each of which depressed a specific subset of these components, and we also examined the immunocytochemical localization of antigens with these mAbs at light microscopic level. The 2 mAbs that depressed R2 and R4 bound to the cnidocils of the desmoneme and the stenotele nematocytes; the 3 mAbs that depressed R5 bound to the apical surface adjacent to the cnidocils of the nematocytes; and the 2 mAbs that depressed R1 and R3 bound to the apical spot structures of unidentified cells in the ectoderm.Together with the specificity of the action of the mAbs on the behavioral response, the correspondence between the effects on the response and the structures visualized with these mAbs suggests that these structures include components of the receptor-effector system relevant to chemoreception.     

Ectonucleotidase Activities Associated with the Olfactory Organ of the Spiny Lobster

The olfactory system of the Florida spiny lobster, Panulirus argus, has olfactory receptors that are excited by the purine nucleotides AMP, ADP, and ATP. These receptors reside on chemosensory neurons that are contained within aesthetasc sensilla on the lateral filaments of the antennules. Also associated with the lobster's olfactory system are ectonucleotidase activities that dephosphorylate excitatory nucleotides, resulting in the production of the nonstimulatory nucleoside adenosine. Our studies of the 5'-ectonucleotidase, ecto-ADPase, and ecto-ATPase activities of this olfactory system showed that each activity was characterized by Michaelis-Menten kinetics; Michaelis constants ranged from 6.9 to 33.5 microM, and maximum velocities ranged from 2.5 to 28.8 fmol/sensillum/s. Evidence that AMP dephosphorylation may serve as an inactivation process was shown by the close correlation between the kinetics of 5'-ectonucleotidase activity and the periodicity of olfactory sampling. Decreased magnesium ion concentration or increased calcium ion concentration resulted in increased ecto-ATPase activity; this activity was insensitive to vanadate ion. Ectonucleotidase activities may have multiple effects on the detection of exogenous nucleotides by a chemosensory system. These effects can be either direct, such as the conversion of an odorant to an inactive compound, or indirect, such as the conversion of an odorant to another compound that can activate or inhibit either receptors or enzymes associated with the system.     

Kinetics of ligand binding to a cluster of membrane-associated receptors

The process of ligand binding to a cluster of membrane-associated receptors is examined theoretically. The theoretical model proposed involves the diffusion of ligands from the solution to the disc-like cluster of receptors on the surface of the spherical cell. When the ligand hits the internal part of the disc-like cluster, it begins to move laterally until it leaves the disc through its outer surface or is bound by one of the receptors inside the disc. If the ligand leaves the cluster, it returns to the solution and hits the disc again after a certain period, etc. According to our model the transition from a diffusion-limited to a reaction-limited process of binding is determined by the dimensionless parameter Dt _c/a ², where D is the lateral diffusion coefficient,t _c is the characteristic time of reaction, anda is the radius of the disc-like cluster. The forward rate constantk _f turns out to be a function of . Comparing the results of our calculations ofk _f with some experimental data we found that agreement is achieved at high , i.e. the process of ligand binding by clustered receptors is predominantly reaction-limited.     

Ca2+ transport and chemoreception in Paramecium

Intracellular Ca²⁺ levels in Paramecium must be tightly controlled, yet little is understood about the mechanisms of control. We describe here indirect evidence that a phosphoenzyme intermediate is the calmodulin-regulated plasma membrane Ca²⁺ pump and that a Ca²⁺-ATPase activity in pellicles (the complex of cell body surface membranes) is the enzyme correlate of the plasma membrane pump protein. A change in Ca²⁺ pump activity has been implicated in the chemoresponse of paramecia to some attractant stimuli. Indirect support for this is demonstrated using mutants with different modifications of calmodulin to correlate defects in chemoresponse with altered Ca²⁺ homeostasis and pump activity.Abbreviations EGTA ethyleneglycol tetra-acetate - ER endoplasmic reticulum - IBMX isobutyl methylxanthine - I _che index of chemokinesis - Mops 3-[N-morpholino] propanesulfonic acid - PEI phosphoenzyme intermediate - STEN sucrose, TRIS, EDTA, sodium chloride - TCA trichloroacetic acid - TRIS tris[hydroxymethyl] aminomethane     

Localization of cholecystokinin-like and calcitonin-like peptides in infant carotid bodies: a light- and electron-microscopic immunohistochemical study

Previous immunohistochemical studies have identified several regulatory peptides in the carotid body chief cells in both humans and animals. These peptides, together with amines, may be important in the modulation of the chemoreflex by the carotid body. We report the localization and distribution of calcitonin and cholecystokinin-like (CCK) immunoreactivity in chief cells of human infant carotid body by light- and electron-microscopic immunohistochemical techniques. Consecutive sections immunostained with calcitonin and/or CCK antibodies revealed positively stained chief cells, both alone and in clusters, scattered throughout the carotid body lobule. Generally more chief cells were positive for calcitonin than for CCK. This was confirmed by quantiative analysis showing that the ratio of calcitonin to CCK immunoreactive cells was consistently >2:1 in all cases studied. There was no apparent correlation between the immunoreactivity for the two peptides and the age, sex, or postmortem interval. Calcitonin-like and CCK-like immunoreactivities were localized electron-microscopically over the dense core granules of the chief cells. Calcitonin and CCK-like peptides in carotid body chief cells may act as neutransmitters or neuromodulators involved in chemoreception.     

Physiological and behavioral effects of chemoreceptors located in different body parts of the swimming crab Callinectes danae

By perfusing their branchial chambers with filtered seawater, we have developed a preparation that allows us to maintain the swimming crab Callinectes danae outside water without any major effects on its cardiac activity. This in turn allowed us to selectively stimulate chemoreceptors located in different body parts, and specifically to discriminate between the receptors located in the branchial chambers and those located in the oral region (mainly in the mouthparts, antennules and antennae). We show that a taurine solution can evoke bradycardia when applied to the oral region or to a combination of the oral region and the branchial chambers. Although the precise localization of the oral region receptors involved remains to be determined, ablation experiments show that the olfactory organs (i.e., the antennules) are not involved. Finally, we show that although stimulating the pereiopods has no effect on the animals' cardiac activity it causes the animals to move, putatively to try to grasp a piece of food, a reaction not evoked by stimulating the gills or the oral regions. Our results lend support to the idea that chemoreceptors located in different parts of the body play different functional roles in decapod crustaceans.     

Studies on the chemoreception of the nemertean, Procephalothrix simulus Iwata, 1952

The feeding responses of Procephalothrix simulus Iwata to the whole-body extracts of different animals (Tubifex sp., Saccocirrus gabrillae) and single compounds (including 17 amino acids, 2 compounds related to amino acids and 2 sugars) were tested, and the response of the nemerteans with and without a frontal organ to stimulus was compared experimentally. Results showed that P. simulus is able to detect food from a distance by chemoreception. It preferred extracts to live prey, and boiled extracts were more attractive than extracts that were not boiled. Of the 21 compounds tested, 6 amino acids (glycine, alanine, isoleucine, phenylalanine, glutamine, asparagine) and betaine induced a feeding response, suggesting that they were possibly the chemical signals in the food location of P. simulus. When the frontal organ was removed, P. simulus apparently lost its selectivity, which indicated that in this nemertean chemoreception for the detection of prey was performed by the frontal organ.     

Ionic mechanisms of depolarizing and hyperpolarizing quinine receptor potentials in Paramecium caudatum

The ionic mechanisms of the depolarizing and the hyperpolarizing quinine receptor potentials in the ciliate Paramecium caudatum were examined by using a behavioral mutant strain. The depolarizing receptor potential was induced by stimulating the anterior end of the specimen, and the hyperpolarizing receptor potential by stimulating the posterior end. The amplitude of both the depolarizing and the hyperpolarizing receptor potentials increased linearly with logarithmic increase in quinine concentration applied. Threshold concentration for inducing the depolarizing receptor potential was lower than that for the hyperpolarizing one. The peak level of the depolarizing receptor potential shifted towards the depolarizing direction with increasing external Ca²⁺ concentration while that of the hyperpolarizing receptor potential shifted in the depolarizing direction with increasing external K⁺ concentration. Under voltage-clamp conditions, the specimen produced an inward current in response to anterior stimulation, and an outward current in response to posterior stimulation. Both the peak inward and the peak outward currents showed a linear relationship with membrane potential. Current-voltage relationships of the receptor currents indicated conductance increase during the application of quinine. The depolarizing quinine receptor potential appears to be produced by an activation of Ca²⁺ channels, and the hyperpolarizing quinine receptor potential by an activation of K⁺ channels. Accepted: 3 October 1997     

Do incubation-induced changes in a lizard's phenotype influence its vulnerability to predators?

Strong evidence affirms that incubation temperatures can influence the phenotype of hatchling reptiles, but few studies have examined the fitness consequences of such modifications. Vulnerability to predation is one plausible way that phenotypic shifts could affect an organism's fitness. We incubated the eggs of three sympatric lizard species at temperatures similar to the thermal extremes of natural nests, and measured several traits that are likely to influence a hatchling's susceptibility to a natural (snake) predator. We also examined the lizards' actual vulnerability to snake predators in direct encounters in the laboratory. Our results show that incubation temperature can affect an individual's date of hatching, morphology, locomotor performance, chemosensory responses to snake scent, and ability to avoid a snake predator during staged laboratory encounters. Incubation temperature did not modify the hatchling's `attractiveness' to snakes (as measured via chemical cues) or its antipredator tactics (propensity to escape predation through fleeing or caudal autotomy). The magnitude and direction of incubation- induced phenotypic shifts varied among the three species (even those with similar life histories, thermoregulatory preferences, and microhabitat requirements), and depended on body temperatures and hatchling age. We conclude that incubation-induced modifications to a lizard's phenotype affect a suite of traits that are likely to influence its vulnerability, and also its actual ability to escape from a predator. This result suggests that incubation regimes can influence organismal fitness via their effects on predator-prey interactions. Received: 21 December 1998 / Accepted: 23 March 1999