Neurobiology and Behavior

Neurobiology and Behavior, Part 1

The Mollusca

A.O. DENNIS WILLOWS

Department of Zoology and Friday Harbor Laboratories, University of Washington, Friday Harbor, Washington

Table of Contents

Cover image

Title page

The Mollusca

Copyright

Contributors

General Preface

Preface

Contents of Other Volumes

Chapter 1: Behavior of Gastropod Molluscs

Publisher Summary

I Introduction

II The Senses: Chemoreception and Vision

III Feeding

IV Locomotion

V Defensive Behavior

VI Reproductive Behavior

VII Behavioral Hierarchies

VIII Perspective

Chapter 2: Learning in Gastropod Molluscs

Publisher Summary

I Introduction

II Habituation, Dishabituation, and Sensitization in Aplysia californica

III Associative Learning in Aplysia

IV Associative Learning in Hermissenda crassicornis

V Associative Learning in the Land Slug Limax maximus

VI Associative Learning in Pleurobranchaea californica

VII Conclusions and Outlook

Acknowledgments

Chapter 3: Neural Control of Behavior in Gastropods

Publisher Summary

I Introduction

II Simple Behaviors

III Neural Generation of Locomotor Patterns

IV Neural Generation of Feeding Patterns

V Neuronal Control of Autonomic Function

VI Interaction and Modulation of Behavior

VII Summary and Perspectives

Chapter 4: Development and Plasticity of the Molluscan Nervous System

Publisher Summary

I Introduction

II Development

III Plasticity

IV Conclusions and Perspectives

Acknowledgments

Index

The Mollusca

Editor-in-Chief

KARL M. WILBUR

Department of Zoology

Duke University

Durham, North Carolina

Copyright

Copyright © 1985 by Academic Press, Inc.

all rights reserved.

no part of this publication may be reproduced or transmitted in any form or by any means, electronic or mechanical, including photocopy, recording, or any information storage and retrieval system, without permission in writing from the publisher.

ACADEMIC PRESS, INC.

Orlando, Florida 32887

United Kingdom Edition published by

ACADEMIC PRESS INC. (LONDON) LTD.

24–28 Oval Road, London NW1 7DX

Library of Congress Cataloging in Publication Data

(Revised for vol. 8)

Main entry under title:

The Mollusca.

Vol. has imprint: Orlando: Academic Press.

Includes bibliographies and indexes.

Contents: – v. 2. Environmental biochemistry and physiology / edited by Peter W. Hochachka – v. 8.

Neurobiology and behavior, Part 1 / edited by A. O. Dennis Willows.

1. Mollusks–Collected works. I. Wilbur, Karl M.

QL402.M57 1983 594 82-24442

ISBN 0-12-751408-2 (v. 8)

ISBN 0-12-728701-9 (paperback)

printed in the united states of america

85 86 87 88 9 8 7 6 5 4 3 2 1

Contributors

Numbers in parentheses indicate the pages on which the authors’ contributions begin.

Gerald Audesirk(1),     Biology Department, University of Colorado at Denver, Denver, Colorado 80202

Teresa Audesirk(1),     Biology Department, University of Colorado at Denver, Denver, Colorado 80202

Andrew G.M. Bulloch(335),     Department of Medical Physiology, Faculty of Medicine, University of Calgary, Calgary, Alberta, Canada T2N 4N1

Peter A. Getting(269),     Department of Physiology and Biophysics, University of Iowa, Iowa City, Iowa 52240

Ken Lukowiak(95),     Department of Medical Physiology, Faculty of Medicine, University of Calgary, Calgary, Alberta, Canada T2N 4N1

George J. Mpitsos(95)

Mark O. Hatfield Marine Science Center, College of Pharmacy, Oregon State University, Newport, Oregon 97365

Department of Medical Psychology, Oregon Health Sciences University, Portland, Oregon 97201

General Preface

This multivolume work, The Mollusca, had its origins in the mid-1960s with the publication of two volumes entitled Physiology of Mollusca and edited by Wilbur and Yonge. In those volumes, 27 authors collaborated to summarize the status of the conventional topics of physiology as well as biochemistry, reproduction and development, and ecology. Within the past two decades, there has been a remarkable burgeoning of molluscan research generally and with it the development of new fields of investigation. During the same period, several excellent books on molluscs have appeared. However, they do not provide adequate information on the many recent advances or give the breadth of perspective of current knowledge of the phylum. Clearly, there was need for a larger work with a comprehensive treatment of major areas of molluscan research. The Mollusca, as a series of 12 volumes, attempts to fulfill this objective. Even here, practical considerations have meant that certain aspects of molluscan research have not been included.

Each major area is treated by several authors, each reviewing his or her special field. The areas are structure and function, metabolic biochemistry, molecular biomechanics, environmental biochemistry, physiology, ecology, reproduction and development, neurobiology and behavior, and evolution. Throughout, the authors have given emphasis to recent advances and present status of molluscan biology. In so doing, directions of future reseach have become evident.

The Mollusca is intended to serve several disciplines—zoology, biochemistry, physiology, and paleontology. It will prove useful to researchers and to all others with interests in molluscs.

Karl M. Wilbur

Preface

It is doubtful that any animal group has provided more of the critical experimental material needed to get to the fundamental roots of neurophysiology and the neural correlates of behavior than have the molluscs. One can buttress this statement with references to the literature of neuroscience from 1930 to the present, which run the gamut from studies of behavioral-level phenomena all the way down to their molecular underpinnings. It is this dramatic growth of work and understanding that motivated the synthesis of these two volumes (Parts 1 and 2).

At the outset, the extraordinary utility of cephalopod giant nerve fibers that emerged from John Young’s neuroanatomical work of the 1930s proved to be a sine qua non for the experiments and the thinking that led to our present understanding of the roles of sodium and potassium channels as the crucial ones for nerve and muscle electrical activity.

This classical and extraordinarily useful work was critically dependent upon at least three factors: (1) the brilliance of the experiments and their interpretation, (2) the existence of robust cephalopod axons with the dimensions of a pencil lead, and (3) a stroke of great good fortune. The analysis of Hodgkin and Huxley showed that the action potential in squid nerve is driven by a (what now seems relatively simple) duet of sodium and potassium currents. Work in the 1950s and 1970s has since shown that had virtually any other nerve or muscle cell been selected to serve as a model system for these studies, the analysis might have foundered or at least been extensively delayed. Other nerves studied more recently have proven to have multiple (sometimes five or more) interactive ionic channel types that control spiking, repetitive firing, the generation of impulse bursts, and the entry of ionic calcium (as a second messenger) into cells. Once again, the molluscs, especially the gastropods, were critically important. Were it not for the unusually large and robust nerve cell bodies of the gastropods, it is arguable that the diverse roles of these several membrane channel types would not yet be fully understood or appreciated.

The list of mollusc models useful in studies of neurobiology and behavior is quite long. Understanding of the physiology of synaptic transmission would have been set back seriously were it not for the giant synapse in the squid stellate ganglion. At another level, analysis of the neuronal interactions that underlie control of fixed action patterns of behavior is heavily dependent upon the access provided to reidentifiable neurons in the ganglia of opisthobranch molluscs under circumstances that permit simultaneous monitoring and manipulation of behavioral activity. Finally, the aggressive analysis of the cellular and molecular-level mechanisms of both rudimentary and more complex forms of learning is clearly directly contingent upon the ideal experimental features and interesting behavioral capabilities of the opisthobranch Aplysia.

In this volume, I have selected topics that span in a representative way the principal elements of progress in molluscan neurobiology from a much larger list of possibilities. It is painfully obvious that much has been left out. It is especially embarrassing to have ignored the bivalves and to have restricted coverage of cephalopods to a single chapter. Much more could have been reported in both these classes. Neither the scaphopods nor the amphineurans are mentioned—there is little to report yet. This volume, then, is primarily about gastropods.

Recent progress in neurobiology has been dependent in an important way upon the intrinsic interest of the nervous systems and the diverse behavior of molluscs. The work itself has resulted from the efforts of a number of people who acquired their interests in molluscs early in their careers. In that regard, I hope these volumes will be effective in seducing others, particularly graduate students. Finally, I would dedicate these volumes to the memory of Graham Hoyle, who despite a delightful arthropod bias, first encouraged my interest in the neurobiology and behavior of molluscs.

A.O. Dennis Willows

Contents of Other Volumes

Volume 1: Metabolic Biochemistry and Molecular Biomechanics

EDITED BY PETER W. HOCHACHKA

1. Structural Organization, Adaptive Radiation, and Classification of Molluscs

R. Seed

2. Metabolic and Enzyme Regulation during Rest-to-Work Transition: A Mammal versus Mollusc Comparison

P. W. Hochachka, J. H. A. Fields, and T. P. Mommsen

3. Carbohydrate Metabolism in Cephalopod Molluscs

Kenneth B. Storey and Janet M. Storey

4. Carbohydrate Catabolism in Bivalves

Albertus de Zwaan

5. Carbohydrate Metabolism of Gastropods

David R. Livingstone and Albertus de Zwaan

6. Amino Acid Metabolism in Molluscs

Stephen H. Bishop, Lehman L. Ellis, and James M. Burcham

7. Lipids: Their Distribution and Metabolism

Peter A. Voogt

8. Molluscan Collagen and Its Mechanical Organization in Squid Mantle

John M. Gosline and Robert E. Shadwick

9. Molecular Biomechanics of Protein Rubbers in Molluscs

Robert E. Shadwick and John M. Gosline

10. Molecular Biomechanics of Molluscan Mucous Secretions

Mark Denny

11. Quinone-Tanned Scleroproteins

J. H. Waite

Index

Volume 2: Environmental Biochemistry and Physiology

EDITED BY PETER W. HOCHACHKA

1. Repiratory Pigments: Structure and Function

Celia Bonaventura and Joseph Bonaventura

2. Blood Repiratory Properties in Marine Gastropods

Ole Brix

3. Osmolytes and Metabolic End Products of Molluscs: The Design of Compatible Solute Systems

George N. Somero and R. David Bowlus

4. Metal Ions: Metabolic and Toxic Effects

K. Simkiss and A. Z. Mason

5. Developmental and Seasonal Metabolic Activities in Marine Molluscs

Peter A. Gabbott

6. Metabolic and Enzyme Adaptation to Temperature and Pressure

K. H. Hoffmann

7. Molluscan Bioluminescence

Frederick I. Tsuji

8. Biochromy of the Mollusca

Denis L. Fox

9. Biochemical Genetics and Adaptation in Molluscs

Richard K. Koehn

10. Biochemical Interactions between Molluscs and Their Algal and Bacterial Symbionts

Horst Felbeck, James J. Childress, and George N. Somero

Index

Volume 3: Development

EDITED BY N. H. VERDONK, J. A. M. van den BIGGELAAR, AND A. S. TOMPA

1. Gametogenesis

M. R. Dohmen

2. Meiotic Maturation and Fertilization

Frank J. Longo

3. Early Development and the Formation of the Germ Layers

N. H. Verdonk and J. A. M. van den Biggelaar

4. Organogenesis

Beatrice Moor

5. Origin of Spatial Organization

J. A. M. van den Biggelaar and P. Guerrier

6. Morphogenetic Determination and Differentiation

N. H. Verdonk and J. N. Cather

7. The Biochemistry of Molluscan Development

J. R. Collier

8. Physiological Ecology of Marine Molluscan Larvae

B. L. Bayne

Index

Volume 4: Physiology, Part 1

EDITED BY A. S. M. SALEUDDIN AND KARL M. WILBUR

1. Cytology of Muscle and Neuromuscular Junction

Ghislain Nicaise and Jacqueline Amsellem

2. Neuromuscular Transmission and Excitation-Contraction Coupling in Molluscan Muscle

Yojiro Muneoka and Betty M. Twarog

3. Biochemical and Structural Aspects of Molluscan Muscle

P. D. Chantier

4. Locomotion in Molluscs

E. R. Trueman

5. The Mode of Formation and the Structure of the Periostracum

A. S. M. Saleuddin and Henri P. Petit

6. Shell Formation

Karl M. Wilbur and A. S. M. Saleuddin

7. Shell Repair

Norimitsu Watabe

8. Endocrinology

J. Joosse and W. P. M. Geraerts

9. Physiology Energetics of Marine Molluscs

B. L. Bayne and R. C. Newell

Index

Volume 5: Physiology, Part 2

EDITED BY A. S. M. SALEUDDIN AND KARL M. WILBUR

1. Feeding Biology of Gastropods

Alan J. Kohn

2. Feeding and Digestion in Bivalvia

Brian Morton

3. Feeding and Digestion in Cephalopods

Eve Boucaud-Camou and Renata Boucher-Rodoni

4. Circulatory Systems of Gastropods and Bivalves

H. D. Jones

5. Circulation in Cephalopods

Martin J. Wells

6. Ionic Regulation and Water Balance

R. F. Burton

7. Excretion

Arthur W. Martin

8. Molluscan Immunobiology

Christopher J. Bayne

Index

Volume 6: Ecology

EDITED BY W. D. RUSSELL-HUNTER

1. Overview: Planetary Distribution of and Ecological Constraints upon the Mollusca

W. D. Russell-Hunter

2. The Ecology of Deep-Sea Molluscs

J. A. Allen

3. Mangrove Bivalves

Brian Morton

4. Coral-Associated Bivalves of the Indo-Pacific

Brian Morton

5. Reproductive and Trophic Ecology of Nudibranch Molluscs

Christopher D. Todd

6. Physiological and Trophic Ecology of Cephalopods

William C. Summers

7. Physiological Ecology of Freshwater Bivalves

Albert J. Burky

8. Physiological Ecology of Freshwater Prosobranchs

D. W. Aldridge

9. Physiological Ecology of Freshwater Pulmonates

Robert F. McMahon

10. Physiological Ecology of Land Snails and Slugs

Wayne A. Riddle

11. Actuarial Bioenergetics of Nonmarine Molluscan Productivity

W. D. Russell-Hunter and Daniel E. Buckley

12. Ecology of an Invasive Pest Bivalve, Corbicula

Robert F. McMahon

13. Population Genetics of Marine Gastropods and Bivalves

Edward M. Berger

14. Ecology and Ecogenetics of Terrestrial Molluscan Populations

A. J. Cain

15. Life-Cycle Patterns and Evolution

Peter Calow

Index

Volume 7: Reproduction

EDITED BY A. S. TOMPA, N. H. VERDONK, AND J. A. M. van den BIGGELAAR

1. Prosobranchs

Vera Fretter

2. Land Snails (Stylommatophora)

A. S. Tompa

3. Freshwater Snails (Basommatophora)

W. P. M. Geraerts and J. loose

4. Opistobranchs

Michael G. Hadfield and Marilyn Switzer-Dunlap

5. Bivalves

G. L. Mackie

6. Cephalopods

John M. Arnold

Index to Scientific Names

Subject Index

1

Behavior of Gastropod Molluscs

TERESA AUDESIRK and GERALD AUDESIRK,     Biology Department, University of Colorado at Denver, Denver, Colorado

Publisher Summary

Molluscan behavior is extremely diverse, encompassing the relatively limited behavioral repertoire of clams and limpets and the intelligent, highly flexible behaviors of the cephalopods. Gastropods and cephalopods dominate as subjects for molluscan neurobiology for contrasting reasons: (1) cephalopods for the mammal-like intricacy of their brains and behavior and (2) gastropods for their relative simplicity and their large and individually identifiable neurons. This chapter discusses gastropod behaviors, especially those of species commonly used in neurobiology. Light plays an important role in the life of molluscs. For terrestrial forms, bright light can signal heat and dryness, while shade may represent desirable damp shelter. Feeding is stimulated by the detection of food stimuli by the rhinophores, oral veil, anterior foot, or mouth area. Gastropod feeding, like that of more complex organisms, is influenced by the interaction of several variables, including hunger, satiation, quality of food, intensity of the feeding chemostimulus, sensory adaptation, and habituation.

I Introduction

A A Neuroethological Perspective on Behavior

II The Senses: Chemoreception and Vision

A Gastropod Chemoreception

B Visually Mediated Behavior

III Feeding

A The Raspers

B The Browsers

C The Hunters

D Modulation of Feeding

IV Locomotion

A Ciliary Locomotion

B Locomotion by Pedal Waves

C Swimming

D Burrowing

V Defensive Behavior

A Withdrawal

B Defensive Secretions

C Escape Locomotion

VI Reproductive Behavior

A Reproduction in Pulmonates

B Opisthobranchs

VII Behavioral Hierarchies

A Opisthobranchs

B Pulmonates

VIII Perspective

References

I Introduction

Molluscan behavior is extremely diverse, encompassing the relatively limited behavioral repertoire of clams and limpets, as well as the intelligent, highly flexible behaviors of the cephalopods. Needless to say, a single chapter, or indeed an entire volume, cannot begin to do justice to the variety of behaviors displayed by members of the phylum Mollusca. Since this volume is devoted primarily to neurobiology, we have chosen to focus our review on those behaviors which have been, or promise to be, suitable for study at the neural level. The study of ethology, or behavior as performed under natural conditions, has much to offer the field of neuroethology. The neurobiological perspective is that behavior is an emergent property of the intricate interconnections of neurons acting under the influences of hormones and neuromodulators. However, from a broader evolutionary perspective, selective pressure to produce particular adaptive behaviors is the force which has molded the nervous system (Pinsker, 1980).

Gastropods and cephalopods dominate as subjects for molluscan neurobiology for contrasting reasons: cephalopods for the mammal-like intricacy of their brains and behavior, and gastropods for their relative simplicity and their large and individually identifiable neurons. Since the cephalopods are covered in a separate chapter (see Boyle, Chapter 1, Volume 9), we will confine our discussion to gastropod behaviors, especially those of species commonly used in neurobiology. Among the gastropods, members of two subclasses, Pulmonata and Opisthobranchia, predominate as subjects for neuroethological studies. Within these, a few genera stand out: the pulmonates Helix, Helisoma, Limax, and Lymnaea and the opisthobranchs Aplysia, Hermissenda, Navanax, Pleurobranchaea, and Tritonia.

Further, another chapter is devoted specifically to plastic behaviors, especially learning (Chapter 2), and thus, little mention will be made here about changes in behavior brought about by experience. Finally, we will not discuss homeostatic internal behaviors, such as control of respiration, circulation, and digestion, which are covered in other volumes of this series.

A A Neuroethological Perspective on Behavior

Not all behaviors are equally suited for neuroethological analysis. For example, highly flexible behaviors elicited and modified by subtle and often variable external stimuli and internal motivational states are very difficult to analyze at the neural level. The complexity of the behavior would be expected to be reflected in, and indeed to be caused by, highly variable electrophysiological states in the neurons controlling the behavior, making a definitive analysis of which neurons contribute to the behavior, and their mechanisms of action, very difficult. In fact, fluid, optional behaviors, such as mating (which is obligatory over an animal’s life cycle but optional at any given time) tend to be difficult to elicit repeatably in the laboratory, particularly under the conditions required for neuronal recording.

Consequently, most neuroethological studies have concerned behaviors which conform to three criteria: reliability, stereotypy, and robustness.

1. Reliability: The first criterion which a behavior must meet is that it can be reliably evoked in all, or nearly all, animals by the application of well-defined stimuli. As an example, many gastropods, such as the nudibranch Tritonia diomedea (Fig. 25), show escape behavior elicited by contact with specific stimuli, including contact with predators or certain noxious substances (Willows et al., 1973). A T. diomedea will perform its escape swim every time it is touched with the tube feet of the predaceous sea star Pycnopodia, with little habituation during repeated presentations (Abraham and Willows, 1971). If desired, the escape-eliciting stimulus can be quantified in terms of the number of tube feet and duration of contact or of the volume and concentration of noxious chemicals.

Fig. 25 Swimming in Tritonia diomedea is initiated by a vigorous ventral flexion (A) followed by a dorsal flexion (B). [Photos by the authors.]

2. Stereotypy: The simplest behavior to study on the neuronal level is one which is stereotyped, that is, performed in almost exactly the same way every time. Presumably, the neurons controlling such a behavior are all active each time the behavior is performed, and their activity is nearly identical on each repetition. This greatly facilitates identification of participating neurons and their modes of interaction with one another in producing the behavior. The escape swim of T. diomedea again fulfills this criterion very well, with little variability except in number of swim cycles (Abraham and Willows, 1971). The forms of the swim cycles are extremely regular.

3. Robustness: Finally, recording from identifiable neurons of the central nervous system usually requires that the animal be at least partially dissected (but see Pinsker and Eberly, 1982; Parsons et al., 1983). Therefore, to be amenable to neural analysis a behavior must be robust, that is, the animal (or its appropriate parts) must still perform the behavior after dissection to expose the relevant parts of the nervous system. Escape behaviors are particularly stable in this regard. A massively dissected T. diomedea will still swim upon contact with sea star tube feet, salt crystals, or detergents (Audesirk, 1978b; Willows et al., 1973). Indeed, even the isolated nervous system can be stimulated to generate the underlying motor program for swimming (Dorsett et al., 1969).

As the neural analysis of stereotyped behaviors has progressed, an increasing number of investigators have turned to behaviors which ordinarily fulfill these three criteria but which can be modified under appropriate, carefully controlled conditions. In this regard, feeding, locomotion, and withdrawal have thus far proved to be the behaviors of choice. Each of these behaviors, in certain gastropods, has been found to display various degrees of plasticity, including competition with other behaviors, habituation, and both aversive and nonaversive classical conditioning. Behavioral plasticity and the neural mechanisms where known will be discussed by Mpitsos and Lukowiak in Chapter 2.

In this chapter, we will focus on selected gastropod behaviors, particularly as they have been, or might be, used in neuroethological studies. Where possible, we will point out aspects of these behaviors about which our knowledge is incomplete and attempt to identify experiments which would help illuminate them. Since numerous reviews of various aspects of gastropod behavior have appeared previously (referenced throughout), we will emphasize the progress made during what we might call the age of gastropod neuroethology, beginning about 1965.

II The Senses: Chemoreception and Vision

A Gastropod Chemoreception

Although evidence is accruing that a few gastropods rely heavily on vision for locomotory cues (Hamilton, 1977; Hamilton and Winter, 1982), the central role of chemoreception in the gastropod perception of the world is undisputed. Chemical stimuli are used to identify potential food sources, initiate feeding, detect predators and noxious stimuli, identify and locate conspecifics for purposes of aggregation or mating, and synchronize spawning.

Neuroethologists have a particular interest in the investigation of gastropod chemoreception because it represents the initiating stimulus for a variety of rhythmic, stereotyped behaviors such as feeding and locomotion which have proved amenable to analysis at the cellular level. The determination of the complete circuitry underlying such behaviors has been hampered, however, by the location of the primary chemosensory neurons. In all gastropods investigated to date, to our knowledge no primary chemoreceptors with centrally located somata have been identified. In contrast to gastropod mechanoreceptors, which frequently have centrally located cell bodies (Audesirk and Audesirk, 1980a; Byrne et al., 1974; Cobbs and Pinsker, 1978), gastropod chemoreceptors generally, and perhaps exclusively, have peripheral somata, located either within or directly beneath the sensory epithelium or in peripheral ganglia, which innervate sensory structures. The muscularity of the body wall and the small size of these receptors have hampered intracellular recordings which would provide information as to, for example, their specificity of response. Extracellular recordings from the peripherally located tentacle ganglion of the giant African pulmonate Achatina fulica (Chase, 1981), which may contain some primary sensory cell bodies (Chase and Kamil, 1983a), and from nerves distal to the rhinophore and tentacle ganglia of the marine opisthobranch Pleurobranchaea californica, which contain axons of intraepithelial receptors (Bicker et al., 1982b), show the most promise for direct electrophysiological investigation.

Since the mid-1960s considerable progress has been made toward understanding the role of diverse gastropod chemoreceptive organs. In this review, only recent literature will be covered. Readers interested in tracing earlier studies of gastropod chemoreception are directed to reviews by Kohn (1961) and Charles (1966). To help prevent duplication of coverage, the role of chemoreception in nonfeeding behaviors such as reproduction and burrowing will be covered along with those topics later in this chapter. Here we will emphasize the identification of chemoreceptive structures and their role in mediating responses to food stimuli. For more comprehensive coverage, see the review by Croll (1983). Chemoreceptor structure is covered in a review by Dorsett (Chapter 2, Volume 9).

1 Chemoreceptors and Feeding Stimuli

Bovbjerg (1965, 1968), on the basis of work with several widely distributed and relatively omnivorous lymnaeid snails, proposed that there would be no selective pressure on snails favoring the detection of plant food from a distance since they are continuously surrounded by it. They might be expected to evolve the capacity to detect distant chemical stimuli from animal material, however, since it is both relatively rare and of higher nutrient quality. His lymnaeid data have not been challenged; however, investigations using a variety of more selective herbivores (see, e.g., Audesirk, 1975) have disproved this as a general hypothesis, although it may apply to omnivorous species.

a Opisthobranchs.

The herbivorous marine opisthobranch Aplysia californica has been the subject of several investigations because of its utility as a neuroethological system. Preston and Lee (1973), after precisely directing seaweed extract toward various parts of the body, concluded that there were localized areas of chemosensitivity at the tips of the rhinophores and tentacles, and in the area around the mouth (Fig. 1). Their behavioral assay was head movement toward the stimulus, or transition to a food-aroused state, also described by Kupfermann (1974; see Fig. 10). The sensitivity of the rhinophores, tentacles, and lip area to a variety of chemical stimuli has been confirmed by intracellular recordings of central neurons receiving input through nerves innervating these structures (Jahan-Parwar, 1972, 1975; Jahan-Parwar and Fredman, 1976; Fredman and Jahan-Parwar, 1980a).

Fig. 1 The opisthobranch gastropod Aplysia californica. [From Cellular Basis of Behavior by E. R. Kandel. W. H. Freeman and Company. Copyright 1976.]

Fig. 10 Aplysia californica showing appetitive behavior for feeding. [Redrawn from Kupfermann (1974), copyright Academic Press, Inc., cited in "Behavioral Biology of Aplysia" by E. R. Kandel. W. H. Freeman and Company (1979).]

To assess the relative contribution of rhinophores and tentacles to distance food finding, Audesirk (1975) ablated the organs and observed Y-maze performance in A. californica missing rhinophores, tentacles, or both structures. Loss of the rhinophores did not affect performance in the maze as long as the tentacles were intact, while loss of tentacles resulted in a slight decrease in accuracy with rhinophores intact. Loss of both organs significantly reduced ability to make correct responses. Frings and Frings (1965), using an assay similar to that of Preston and Lee, concluded that the rhinophores of A. juliana were not chemosensitive, a puzzling finding in light of the close relationship between the two species, the similar structure of the rhinophores, and the similarity of their food habits. Preston and Lee (1973), Kupfermann (1974), and Audesirk (1975) all reported considerable head waving on the part of animals locating food. The large size of A. californica (up to 3000 g) and the wide spacing of the tentacle tips make them ideal organs for the simultaneous bilateral comparison of stimulus intensity (tropotaxis), a facility which would be enhanced by the use of klinotaxis, which involves lateral movements for the purpose of comparing odors at successive symmetrically placed points in space. Audesirk (1975) observed a reduction in this head waving after tentacle ablation, although the experiment to confirm tropotaxis, unilateral tentacle ablation, has not been reported.

Neither Frings and Frings (1965) nor Preston and Lee (1973) found behavioral responses to the perfusion of food over the osphradia of A. juliana or A. californica, respectively. Jahan-Parwar et al. (1969), using A. californica, recorded from identified neurons in the abdominal ganglion (which innervates the osphradium) in response to a variety of stimuli perfused over the osphradium. They reported excitation of several abdominal ganglion neurosecretory cells by food extracts, including seaweed, shrimp, and oyster. The failure of A. californica to respond behaviorally to perfusion of this organ with food extract implies that these cells are probably not part of the feeding circuitry.

The chemoreceptive organs of the carnivorous marine opisthobranch P. californica have been identified by using both behavioral and electrophysiological assays (Fig. 2). Lee et al. (1974) perfused squid extract over localized areas of the body and observed feeding responses most reliably in response to stimulation of the edge of the oral veil, tentacles, rhinophores, and the proboscis (which is extended during feeding). Extracellular recordings revealed activity in nerves innervating the rhinophores, tentacles, oral veil edge, and anterior foot in response to squid extract perfused over these sites (Lee and Liegeois, 1974). Intracellular recordings from interneurons in the small ganglia which innervate the rhinophores and tentacles were obtained by Bicker et al. (1982b). Many of these neurons showed excitatory responses to food stimuli perfused over the rhinophore or tentacle.

Fig. 2 Chemosensitivity in Pleurobranchaea californica, based on morphological and behavioral data. Upper numbers represent the number of putative chemosensory cells per square millimeter. Lower ratios (from Davis and Mpitsos, 1971) are the proportion of animals (12 animals, one trial per animal) showing consummatory feeding behavior when food stimuli were perfused over the area. Ta, tail; B, back; ME, mantle edge; R, rhinophore; OV, oral veil; M, mouth; L, lips; Tn, tentacle; G, gill; F, foot. [From Davis and Matera (1982), copyright John Wiley and Sons, Inc.]

In the carnivorous marine cephalaspidean opisthobranch Navanax inermis, rhinophores are absent, but the anterior animal is well endowed with chemoreceptive organs (Fig. 3A). On the basis of localized application of prey mucus and careful behavioral observations, Murray and Lewis (1974) concluded that three anterior structures, the anterior lateral folds (ALFs) of the head shield, the labial palps (also called oral tentacles), and the pharyngeal lips, were contact chemoreceptive organs. Trail following (Fig. 3B) was elicited by contact of the ALFs with opisthobranch mucus (the normal prey of Navanax), while prosobranch mucus was ineffective (Paine, 1963). Murray and Lewis (1974) found that prey contact with the labial palps elicits extension of the pharynx, exposing the pharyngeal lips. Contact of these chemoreceptive structures with the prey then elicits rapid expansion of the pharynx, sucking in the prey. The precise function of the paired Hancock’s organs located on the anterior sides of the body is unclear, but they are believed to have both chemosensory and tactile functions (Bullock and Horridge, 1965).

Fig. 3 (A) The opisthobranch Navanax inermis. (B) Navanax (dashed line) follows a trail laid by Bulla (solid line). The reliance of Navanax on chemoreception rather than vision is illustrated by the fact that, at point A, the prey was ignored in favor of following the trail even though the Bulla was only a few centimeters away. [A, From Murray (1971). B, From Paine, (1963).]

The nudibranch oral veil and tentacles are generally agreed to be chemoreceptive (Kohn, 1961), but the role of the rhinophores is less clear and may vary among genera. Agersborg (1922, cited in Kohn, 1961) concluded that rhinophores play no role in distance chemoreception in the nudibranch Hermissenda crassicornis, and his findings have not been challenged in spite of widespread evidence for a chemoreceptive function for rhinophores of other opisthobranchs. The tentacles of H. crassicornis have been implicated in chemoreception through both histological and electrophysiological techniques (Alkon et al., 1978; Alkon, 1980).

The rhinophores of the nudibranch T. diomedea (Fig. 4) have been implicated in positive rheotaxis (Field and Macmillan, 1973), but these results were not supported by Willows (1978), who found that T. diomedea still orient to current in a Y-maze after rhinophore ablation, although they lose their ability to detect food in the maze when rhinophores are removed. The oral veil tips and region surrounding the mouth of T. diomedea have been behaviorally implicated in contact chemoreception of food (Audesirk, 1979b). Excitatory responses from identified central neurons in response to food extract perfused over these areas, as well as the anterior foot, support this finding (Audesirk and Audesirk, 1980a).

Fig. 4 Ventral view of the head of the nudibranch Tritonia diomedea, consuming a sea whip. r, rhinophore; ov, oral veil; l, lips; f, foot. [From Audesirk (1979b).]

b Pulmonates.

The widely distributed freshwater pulmonate Lymnaea stagnalis has been the subject of extensive behavioral and neuroethological investigation. Bovbjerg (1968) found that contact chemoreception in L. stagnalis is mediated by the posterior tentacles, the mouth, and much of the foot as well. The response to crayfish meat was considerably more consistent than that to algae for this omnivorous species in all areas except the mouth, which always responded to both. Distance chemoreception as assayed by using a Y-maze was highly accurate for crayfish meat but nonexistent for food algae, perhaps reflecting the relative availability and desirability of the two types of food in their natural pond habitat. The role of tentacles in feeding was further investigated by Goldschmeding and Jager (1973), who removed the tentacles or severed nerves innervating the lips and tentacles and observed feeding responses to sucrose in the recovered animals. Neither removing both tentacles nor cutting the tentacle nerves caused an apparent reduction in feeding in response to sucrose, while cutting the major nerves to the lips had a clear negative effect.

Electrophysiological evidence for the presence of chemoreceptors within the buccal cavity of the related pulmonate Helisoma trivolvis has been provided by Horowitz and Senseman (1981), who recorded both excitatory synaptic activity on buccal motor neurons in response to internal perfusion of the buccal mass with a variety of phagostimulant solutions, and inhibitory input in response to a behaviorally aversive stimulus (HCL).

In the terrestrial pulmonate slug Limax maximus, chemoreception is used to locate food and assess its palatability (Kittel, 1956, cited in Gelperin, 1983). In electrophysiological investigations, chemical stimuli delivered to the lips have been shown to activate and modulate the feeding motor program (Gelperin et al., 1978; Reingold and Gelperin, 1980), while chemostimuli delivered to the olfactory epithelia of the posterior (superior) tentacles elicit responses in the central cerebral giant neurons which modulate feeding in many gastropods (Egan and Gelperin, 1981). Senseman (1978) demonstrated that the consumption of artificial food (agar plus liquid food) by Ariolimax californicus was in direct proportion to the concentration of the food chemostimuli.

In an earlier study of the Japanese land pulmonate Ezohelix flexibilis, Suzuki (1967) investigated the role of the anterior and posterior tentacles by amputation and observation of subsequent food-finding ability. He concluded that only the posterior tentacles were involved in distance chemoreception. A similar role for the posterior tentacles of the giant African pulmonate A. fulica (Fig. 5) was reported by Chase and Croll (1981), who found that the snails in a T-maze were able to locate food accurately with only their posterior tentacles intact, while removal abolished this ability. Unilateral removal of a posterior tentacle tended to cause circling movements when snails were placed in an open arena with a food source, implicating the use of the tentacles in tropotaxis (simultaneous bilateral comparison of stimulus intensity). This hypothesis was tested and supported (Chase, 1982) by using a tentacular olfactometer (Fig. 6) which allowed each tentacle to experience a different intensity of the same chemostimulus. Snails turned toward the side experiencing the higher intensity. Intracellular recordings from neurons of the small ganglion which innervates each posterior tentacle revealed the presence of cells responsive to specific food odors (Chase, 1981). The anterior tentacles of A. fulica were found to be important for following the trails of conspecifics, a behavior which facilitates aggregation (Chase and Croll, 1981). Both tropotaxis and klinotaxis by head-waving movements have been postulated in the orientation of the pulmonate Biomphalaria glabrata (Townsend, 1973). Head waving is unaffected by posterior tentacle amputation, although food finding is impaired. On the basis of responses to localized application of lettuce extract, which were delayed for 2–4 sec after application to the tentacle tip but were instantaneous when extract was applied at the base, Townsend (1974) hypothesized that the posterior tentacles collect the water at two points and rapidly conduct it down to their bases by using ciliary currents. Such currents can be clearly observed by using a dye suspension.

Fig. 5 The giant African pulmonate Achatina fulica. [From Chase and Croll (1981).]

Fig. 6 Achatina fulica in an olfactometer which is used to present different stimuli to each tentacle and measure the degree of turning in response. [From Chase (1982).]

In pulmonates, the chemosensory role of the osphradium remains in doubt. No evidence for a role in chemoreception was found by Bailey and Benjamin (1968) through intracellular recordings from the osphradial ganglion of Planorbarius corneus, although the size and complexity of the ganglion (Benjamin, 1971) imply an important function for the osphradium. Although Michelson (1960, cited in Townsend, 1973) reported that B. glabrata with cauterized osphradia were unable to locate food in a Y-maze, Townsend (1973) found food localization unaffected by osphradial cautery in this species, leaving the function of the pulmonate osphradium in doubt. There is no convincing evidence that what is called the osphradium in pulmonates is actually homologous to that of prosobranchs (Benjamin, 1971), whose chemosensory function is well established (see the following section).

c Prosobranchs.

The osphradium is found in all prosobranchs, and its chemoreceptive function in this group has been repeatedly demonstrated (Kohn, 1961). Using the carnivorous whelk Buccinum undatum, in which the osphradium is particularly well developed, Bailey and Laverack (1966) recorded from central neurons of the supraintestinal ganglion which innervates it. They recorded the most pronounced activity when the osphradium was perfused with extract of the mussel Mytilus, a prey of B. undatum, and with certain amino acids (see Section II,A,2,a).

A behavioral study of the prosobranch Nassarius reticulatus revealed the presence of contact chemoreceptors on both anterior and posterior tentacles, the anterior border of the foot, and the tip of the siphon (Crisp, 1971). The osphradium was not tested in this investigation.

The boring predatory prosobranch Urosalpinx cinerea preys on a wide variety of intertidal species, including mussels, oysters, and barnacles. In recent studies (Rittschof et al., 1983; Williams et al., 1983) newly hatched snails were exposed to a variety of potential prey odors and to various combinations thereof. Snails innately show different responses to the different odors, and the presence of one odor can modify the snails’ response to a second, admixed odor. Selective Chemotaxis and the integration of various chemical stimuli to produce a behavioral response may help explain the distribution of U. cinerea within the intertidal zone.

2 Responses to Chemically Defined Stimuli

In a number of investigations, well-defined chemical stimuli have been perfused over chemoreceptive organs and their response specificity assayed either by behavioral or electrophysiological means.

a Amino Acids.

Small molecules, and particularly amino acids, are of primary importance in aquatic chemoreception (Bardach, 1975). It appears that among gastropods there is little uniformity of response to amino acids, perhaps not surprisingly, considering the diversity of this class. Aspartic acid appears the most universally attractive, having been reported as stimulatory to herbivores such as A. californica on the basis of both electrophysiological recordings from central neurons and behavioral feeding responses (Jahan-Parwar, 1972, 1975; Jahan-Parwar and Fredman, 1976); Aplysia kurodai and A. dactylomela, on the basis of behavioral responses (Carefoot, 1982); adult (but not juvenile) B. glabrata, on the basis of taxis in an olfactometer (Thomas and Assefa, 1979; Thomas et al., 1980); and to carnivorous species, including B. undatum, on the basis of central recordings in response to osphradial stimulation (Bailey and Laverack, 1963, 1966) and P. californica, on the basis of both feeding behavior and extracellular recordings of the rhinophore nerve (Bicker et al., 1982a). The same species, with the exception of P. californica, were attracted by glutamic acid. Helisoma trivolvis also is attracted to, and will feed on, 100 mM glutamate (Horowitz and Senseman, 1981). These amino acids are released by both plants and animals and are among those found most prevalently in surface water (Bardach, 1975). Carefoot (1982) reports that l-glutamic and l-aspartic acid are among the most abundant free-state amino acids in several common orders of seaweed. MacInnis et al. (1974) showed that both of these were released into the water by adult B. glabrata, and Uhazy et al. (1978) demonstrated their release into the water by lettuce, a preferred food of this snail in the laboratory. The concentrations eliciting a response ranged from a low of 10−8 M for A. californica (Jahan-Parwar, 1975) to 10−1 M for P. californica (Bicker et al., 1982a), where a threshold was not obtained.

Responses to other amino acids are diverse. While A. californica withdrew from presentations of alanine (Jahan-Parwar, 1975), this amino acid was among the most stimulatory to P. californica (Bicker et al., 1982a). Of the 20 amino acids which Carr (1967) found to be present in shrimp, a food of the mud snail Nassarius obsoletus, only glycine elicited food-seeking behavior. This amino acid also stimulates feeding in P. californica (Bicker et al., 1982a). Proline both attracts and elicits feeding in Helisoma at concentrations of 100 mM (Horowitz and Senseman, 1981). Juvenile B. glabrata were attracted to both a wider range and different amino acids than adults of this species (Thomas and Assefa, 1979; Thomas et al., 1980), possibly as a result of their more varied diet. Neither juveniles nor adults discriminated between the d- and l-isomers, although their own body proteins utilize only the l-form. It would be interesting if this lack of discrimination were general in snails, but other investigators have limited their presentations to the l-isomer, for which vertebrates show a distinct preference (Hara, 1976, cited in Thomas et al., 1980).

b Other Organic Compounds.

Carr (1967) analyzed the components of extract of shrimp, a food of the mud snail N. obsoletus, for their ability to elicit food-seeking behavior and found that (in addition to glycine) only lactate showed a significant effect. Shrimp extract itself was more stimulatory than any of its isolated components or any combination of these tested, suggesting that other, as yet unidentified, substances or synergistic combinations are attractive to this species. Chase (1982) found that A. fulica turns toward odors reported to attract insects (butyric acid, hexanoic acid, and 2-octanol) and amyl acetate, a volatile ester in fruits. Aldehydes, which are common volatiles from plants, did not attract this herbivorous species. Both juvenile and adult B. glabrata are more strongly attracted to propionic acid than to any amino acid (Thomas et al., 1980). This substance is present in mucus deposited by the foot gland and may serve as a pheromone rather than a food stimulus.

Carbohydrates are potent phagostimuli for certain herbivorous and omnivorous species. In Lymnaea stagnalis the disaccharides sucrose, maltose, and lactose were highly stimulatory (in that order of effectiveness) and more stimulatory than their constituent monosaccharides (d-fructose, d-glucose, and d-galactose) (Jager, 1971). Senseman (1977), using the slug Ariolimax californicus, and Carefoot (1982), using Aplysia kurodai and Aplysia dactylomela, both reported starch to be an effective phagostimulant.

c Responses to Salt Solutions and Changes in pH and Osmolality.

The role of the osphradium as an osmoreceptor in Aplysia californica, A. fasciata, and A. depilans was investigated by Stinnakre