Neurobiology and Behavior

Neurobiology and Behavior, Part 2

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

General Preface

Preface

Chapter 1: Neural Control of Cephalopod Behavior

Publisher Summary

I Introduction

II Neuromuscular Organization

III Intrinsic Movements

IV Local Organization of Movement, Lower and Intermediate Motor Centers

V Coordinated Movement, Higher Motor Centers

VI Sensory Systems and Capabilities

VII Complex Behavior

VIII Commentary

Acknowledgment

Chapter 2: Brains to Cells: The Neuroanatomy of Selected Gastropod Species

Publisher Summary

I Introduction

II Topography of the Ganglia

III Neuron Morphology

IV Ganglion Structure

V Neurosecretory Cells

VI Epithelial Receptors

VII Central Sensory Neurons

VII The Gastropod Eye

IX Statocyst

X The Osphradium

XI Perspectives

Chapter 3: Membranes, Gates, and Channels

Publisher Summary

I Introduction

II Selectivity

III Surface Charges

IV Channel Kinetics

V Gating

VI Discussion

Acknowledgment

Chapter 4: Transmitters and Modulators

Publisher Summary

I Introduction

II Serotonin/5-Hydroxytryptamine/Enteramine [3-(2-Aminoethyl)-1H-indol-5-ol]

III Dopamine [β-(3,4-Dihydroxyphenyl)ethylamine; 3-Hydroxytyramine; 5-(2-Aminoethyl)-1,2-benzenediol]

IV Acetylcholine

V Amino Acids

VI Histamine [2-(5-lmidazolyl)ethylamine]

VII Neuropeptides

VIII Summary

Index

The Mollusca

Editor-in-Chief

KARL M. WILBUR

Department of Zoology

Duke University

Durham, North Carolina

Copyright

Copyright © 1986 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 volume 9)

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. 3. Development / edited by N. H. Verdonk, J. A. M. van den Biggelaar, A. S. Tompa – – v. 9, pt. 2. Neurobiology and behavior / edited by A. O. Dennis Willows.

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

QL402.M57 1983 594 82-24442

ISBN 0-12-751409-0 (v. 9)

printed in the united states of america

86 87 88 89 9 8 7 6 5 4 3 2 1

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 what now seems a 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 (sometime 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 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 the chapters of these two volumes (Volumes 8 and 9), I have selected topics that span in a representative way the principal elements of progress in molluscan neurobiology from the 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 focused 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

1

Neural Control of Cephalopod Behavior

PETER R. BOYLE,     Department of Zoology, University of Aberdeen, Aberdeen AB9 2TN, Scotland

Publisher Summary

This chapter discusses the neural control of cephalopod behavior. Cephalopods are among the most intrinsically fascinating invertebrate animals. Generally of large adult size, they are highly mobile and actively predatory on a wide variety of other organisms. The arrangement of the central nervous system is similar in all cephalopods. It is a compact mass surrounding the esophagus and often enclosed in a tough cartilaginous cranium. Many pharmacologically active chemicals, and those with known synaptic potential, have been identified in cephalopod tissues. Separated from the main sources of their innervation, some muscle systems exhibit considerable activity in vitro. This level of intrinsic activity complicates the interpretation of their control by the nervous system. The main circulatory pump in cephalopods—the systemic heart—consists of a single, median ventricle supplied with blood by a pair of lateral auricles. Output from the ventricle is distributed through an anteriorly running dorsal aorta, abdominal and gonadial arteries, respectively. Isolated from the body, the ventricle continues to beat rhythmically and steadily as long as muscle tonus is maintained.

I Introduction

Cephalopods are among the most intrinsically fascinating of invertebrate animals. Generally of large adult size, they are highly mobile and actively predatory on a wide variety of other organisms. With the exception of Nautilus, all of the modern cephalopods belong to the single subclass Coleoidea, which contains approximately 650 species. The basic body form and neurobiological organization is relatively homogeneous throughout the group, which has, nonetheless, proved successful in a wide range of marine habitats.

The familiar cephalopods belong to the orders Sepioidea (cuttlefish and sepiolids), Teuthoidea (squids) and Octopoda (octopuses). Usage of the common terminology is not completely uniform. In North America, members of the family Sepiolidae are frequently referred to as squid, perhaps because the more distinct members of the order Sepioidea, the cuttlefish (family Sepiidae), do not occur there. Of the early cephalopods, which had coiled, external shells, the only survivors are the few species belonging to the genus Nautilus. So little information is available on living nautiluses that, unless it is specifically noted, the generalizations in this chapter apply to the coleoids only. A classification of the genera of cephalopods is to be found in Voss (1977).

Most of our knowledge of the behavior and physiology of cephalopods is derived from relatively few examples. Benthic coastal octopuses, mainly Octopus, and the common European cuttlefish Sepia have both proved adaptable to life in the aquarium and consequently have been the subjects of a great deal of investigation. There is less information about squids because, on the whole, they are pelagic shoaling animals which do not survive well in captivity. Those genera on which major commercial fisheries are based, particularly Loligo, Todarodes and Illex, are the main subjects. Relatives of the inshore and neritic cephalopods, as well as many entire families, live in oceanic water of considerable depth (down to 3000 m). Very little is known about the biology of such animals, which are rarely seen alive and then only on research ships and for short periods of time. Neurobiological and behavioral research, which is essentially based on the living animals, has been heavily biased toward the few species which are both available and amenable to life in captivity. The same constraints have applied also to many other aspects of cephalopod biology (Boyle, 1983b).

Due to their complex behavior patterns, in some respects comparable to those of lower vertebrates, and their highly developed and concentrated nervous system, the cephalopods have been the subject of a great deal of neurobiological work. Much of it has related to various special attributes of the nervous system and may not be directly relevant to the neural control of behavior. The cytology and physiology of the giant fiber system of the squids, for example, have proved to be among the seminal research fields for advances in understanding the basic mechanisms of nervous transmission (for review, see Tasaki, 1982) but will be only briefly mentioned here insofar as they illustrate the mechanism for the squid escape reflex behavior.

Similar, in a way, is the large body of work on the learning capabilities of Octopus. Intended mainly as an approach to understanding the mechanisms of memory, the work has been based on discrimination learning and surgical interference with the brain. The results will be assessed elsewhere in these volumes and have already been the subject of major reviews (Young, 1961a; Wells, 1962, 1966b, 1978; Sanders, 1975). For the cephalopods as a group, this work has resulted in a most detailed knowledge of the anatomy of the nervous system (Young, 1971).

At the level of the neural control of behavior Octopus vulgaris is outstandingly amenable for behavioral work, due to its survival of major brain surgery (Young, 1971, p.641), but the cephalopod nervous system as a whole has proved a difficult physiological preparation. Apart from the giant fiber system, nerve cells are small relative to gastropod brain cells. The central and peripheral ganglia are often firmly sheathed with connective tissue which also permeates the cell masses themselves. The extensive vascularization of these large blocks of tissue is sensitive to damage. The blood vessels are reported to constrict at many points, creating difficulties for artificial perfusion of the preparation, and it is only recently that sustained penetrations of brain cells have been made (Laverack, 1980).

As large, soft-bodied animals, cephalopods present practical difficulties for physiologists in making large preparations. A number of anesthetic agents are effective (Andrews and Tansey, 1981a), but no satisfactory system has yet been developed for chronic maintenance and control of the level of anesthesia. The clear description of movement is also another practical problem. The muscle systems are generally very complex; they interact with a universal skeleton of blood and muscle to permit an unsurpassed range of movement in three dimensions. The musculature acts as the effector of movement as well as providing most of the skeletal support. Correspondingly, it is rarely possible to describe the results of effector action precisely. These several practical problems coupled with the accessibility of relatively few cephalopod types mean that there is rather less physiological literature than might be expected.

The literature on the behavior and biology of cephalopods dates back at least to Aristotle (ca. 300 B.C.). Since 1960 many major reviews in English have appeared (Table I) giving full access to the earlier material (before 1960), which is not consequently included here. Some of these reviews, particularly those by J. Z. Young and M. J. Wells, provide detailed accounts of anatomy and function within the CNS of Octopus and also include many original observations. In an effort to complement the cephalopod literature, rather than duplicate too much of it, a deliberate slant has been imposed in this review toward the peripheral organization of behavior and the lower motor centers. As far as possible, only those reports which bear upon the subject of neural control of behavior will be dealt with here, and only scant mention will be given to other neurobiological topics.

TABLE I

A List of Some Major Reviews in English; Since 1960, of Aspects of Cephalopod Neurobiologya

aSome indication is given of the type of review, whether it has appeared as a book or within a book, and of the major emphasis of its content.

bThis volume contains reviews on a number of relevant topics including contributions from Packard and Hochberg, Gilpin - Brown, Baker, Martin, Muntz, Mauro, Budelmann, Barlow, Messenger, and Young.

II Neuromuscular Organization

A General Organization

The arrangement of the central nervous system is essentially similar in all cephalopods. It is a compact mass surrounding the esophagus and often enclosed in a tough cartilaginous cranium. It is believed to have been formed by a coalescence of cords rather than an accumulation of ganglia (Young, 1971). Peripheral ganglionic masses are located in particular organ systems and serve lower motor and reflex functions. In addition, there are a remarkable number of ancillary tissues (paraneural) associated with the nervous system and having various functions. A list of the principal components of the nervous system is given in Table III, p. 32.

TABLE III

Summary List of the Main Functional Subdivisions of the Cephalopod Nervous System, Based on Octopusa,b

aSource: Young (1971).

bUnpaired lobes indicated (s); older terminology in brackets.

It has been pointed out by Young (1967a) that one feature of outstanding comparative interest is that the cephalopod nervous system is a multichannel network like that of the mammals. This is a design principle in which generally the items of information to be recognized are each enclosed and transmitted in separate channels. The brain of Octopus contains an estimated 1–2 ×1 10⁸ nerve cells. There may be 8 × 10⁶ peripheral receptors (excluding the eyes), the information from which is condensed into 0.5 × 10⁶ channels reaching the brain (compared with about 3 × 10⁶ afferent fibers to the CNS in mammals). There are also very large concentrations of nerve cells peripherally with an estimated 3 × 10⁸ in the nerve cords of the arms alone.

B Central Cell Masses

For Octopus, the last three decades have seen the appearance of an enormous body of histological work, backed by experimental behavior studies, describing the brain and its functional divisions. Young (1971) provides a comprehensive compilation of the neuroanatomy. Comparable anatomical detail is now becoming available for Loligo (Young, 1974, 1976a, 1977a, 1979; Messenger, 1979b), although much of the functional interpretation of the anatomy dates back to the pioneering studies of Boycott (1961) in electrical stimulation of regions of the brain in Sepia.

As might be expected, the brain of Nautilus is not so compact in its organization (Young, 1965a). The supraesophageal part does not show the complex differentiation into lobes characteristic of coleoids, and neither does it have the enormous numbers of small cells characteristic of certain parts of their brains. In these senses it can be considered as anatomically more primitive than the modern cephalopods. There is little comparable detail for cephalopods in general other than those already mentioned by genus, but a review of the available information in the context of evolution is given by Young (1977b).

Quantitative studies on the relative dimensions of the various lobes of the cephalopod brain have been carried out by several authors. Wirz (1959) compared the relative volumes of individual lobes of the brain for 11 species of octopods and 23 species of decapods. This approach was extended by Froesch (1971) to include the newly hatched stages of 10 species. The number and sizes of the nerve cells in all of the main constituent parts of the Octopus nervous system have been estimated by Young (1963a). The relative growth of major sectors of the brain in terms of cell numbers and nucleic acid content has been reported on by Packard and Albergoni (1970).

The functional interpretation of the central cell masses will be dealt with in their respective sections (Table III).

C Peripheral Cell Masses

Early claims that there is a nerve net (Mikhailoff, 1921), a general system of connecting peripheral nerve cells, have been set aside (Alexandrowicz, 1960a; Wilson, 1960). There are, however, dispersed nerve cells at least in some parts of the mantle (Alexandrowicz, 1960a; Gray, 1960; Boyle, 1976), arms (Graziadei, 1964, 1965b), gut (Alexandrowicz, 1928), and cardiac organs (Smith, 1979). Some are certainly sensory but in the gut, for example, a substantial plexus is present which is implicated in motor control.

Very large numbers of nerve cells are grouped into peripheral masses. These ganglia are clearly associated with the motor control of movement in a particular region of the body and are organized basically on the common molluscan pattern of a peripheral rind of cell bodies surrounding a core of neuropile. Nerve cells may also be dispersed along nerves. Along the arms and beneath the attachment of the gills the numbers of nerve cells are such as to justify the term ganglionic chain for these nerve cords. The detailed anatomy, and to some extent the terminology, differs among cephalopods, but the principal groups of peripheral ganglionic masses are listed in the following discussion (based mainly on Octopus).

Brachial ganglionic chain (Figs. 1 and 2). The brachial nerves, the largest and most complex nerves in the body (Young, 1965b, p. 64), arise from the brachial lobes of the brain and travel centrally within each arm. A complete ring of nervous tissue, the interbrachial commissure, links the eight cords close to their origin. It is estimated that these cords constitute by far the greater part of the nervous system of an octopus (Young, 1963 a, p. 249). Opposite the base of each sucker, nerve cells are accumulated, and the swelling so formed is called the sucker ganglion. The cord may be thus demarcated into ganglionic and interganglionic regions, depending on the sucker arrangement. In addition, four thin lateral nerve cords run peripherally in the arm muscle, and there is a small, separate subacetabular ganglion beneath each sucker.

Fig. 1 Sketch plan of an octopus (based on Eledone cirrhosa) to show the approximate layout of the principal ganglionic masses; see text, Section II,C.

Fig. 2 Diagrammatic cross section through the arm of Octopus vulgaris and one of the suckers. Complex arrangements of muscle bundles surround a central canal in which runs the axial nerve cord. Swellings opposite each sucker are known as brachial ganglia. Four intramuscular nerve cords run peripherally; they are connected to each other by an anastomosis of nerve fibers and to the axial nerve cord by defined nerve bundles. Nerves run directly to the sucker from the brachial ganglion and also through the sucker ganglion. [Modified from Graziadei (1965a).]

Stellate ganglion (Fig. 1). The main nerve to the musculature of the mantle, the palliai nerve, arises on each side from the pallio visceral lobe of the brain. It runs into a large stellate ganglion lying on the inner surface of the mantle (Young, 1972). The stellar nerves which arise from the ganglion are distributed throughout the mantle. In the squids this ganglion is the site of the synapse between second- and third-order giant fibers (Young, 1973a).

Inferior buccal ganglion (Figs. 1 and 3). Lying just beneath the esophagus, behind the buccal mass, this bilobed ganglion receives innervation from the superior buccal lobe and is directly responsible for the control of buccal movements (Young, 1965c; Boyle et al., 1979a,b).

Fig. 3 Diagrammatic sagittal section of the buccal mass of Octopus vulgaris showing the main features and the innervation of the muscles. The labial and the salivary-subradular nerves are omitted. [From Boyle et al. (1979b), modeled on Young (1965c).]

Subradular ganglion (Figs. 1 and 3). Embedded within the musculature of the salivary papilla, the paired subradular ganglia are innervated also from the superior buccal lobe (Young, 1965c; Boyle et al., 1979a,b).

Gastric ganglion (Fig. 1). This large, unpaired ganglion lies on the intestine at the point at which it leaves the spiral caecum. It is supplied by the visceral nerve of the brain, and it gives rise to nerves to many parts of the intestine (Young, 1967b).

Fusiform ganglion (Fig. 1). Located symmetrically on the right and left visceral nerves, the fusiform ganglia lie on the ventral surface of the renal sacs. The auricle, lateral vena cavae, and ventricle receive some innervation from nerve branches emerging at this level (Smith and Boyle, 1983).

Cardiac ganglion (Fig. 1). These remarkable ganglia of the octopods lie on the branchial heart of each side (Smith and Boyle, 1983). As well as the normal ganglionic structure, they have an internal pulsatile sac with its own innervation and blood supply. Muscle fibers present in the matrix of the sac cause it to contract rhythmically with the same frequency as the branchial heart (Alexandrawicz, 1963).

Auricular ganglion (Fig. 1). The auricular ganglion is a small ganglionic structure of intermittent occurrence on the auricle of Eledone (Smith and Boyle, 1983) and perhaps in other cephalopods (Alexandrowicz, 1960b; Carlson, 1905).

Branchial ganglionic chain (Fig. 1). Strung out along the base of each gill is a linked chain of ganglionic swellings which are formed along the distal termination of the visceral nerve. The cells, present along the length of the whole trunk, are grouped to a greater extent beneath the pairs of gill lamellae (Young, 1967b; Alexandrowicz, 1960b).

D Nerve Cells

The histology and topography of the nerve cells of Octopus are better known than those of any other animal of comparable nervous complexity. From the viewpoint of experimental neurobiology several features of the cells and their organization combine to pose practical difficulties for physiological work.

1. There are very large numbers of cells, an estimated 5 × 10⁸ in total, of which 3 × 10⁸ are in the brachial ganglia (Young, 1963a). Individual cell somata are not particularly large. The vast majority have nuclei of less than 25 μm in diameter. The relatively small number of large cells (50–60 μm nuclear diameter) cannot be visually located nor their exact positions predicted.

2. As well as a tough sheath of connective tissue, the main ganglionic masses are permeated by blood vessels surrounded by a system of extracellular spaces containing collagen, the gliovascular system (Stephens and Young, 1969). At least two main types of glial cells are present, fibrous and protoplasmic glia. Large nerve cells in the brain may have complex glial sheaths which, interspersed with extracellular channels, invade the nerve cell body to form a trophospongium (Gray, 1969).

3. The largest nerve fibers are about 30 μm in diameter, and the bulk of them in most nerves are less than 10 μm in diameter (Young, 1965b). Categories of nerve fibers of different function are recognized, e.g., somatic sensory fibers, but these cannot always be distinguished histologically; most nerves carry mixed fiber types, and many of the more interesting nerves are short (Young, 1965b). There is no morphological or functional equivalent in octopuses of the giant fiber system of the decapod cephalopods (Martin, 1965, 1969, 1977; Hulet et al., 1980) to mediate a rapid escape response. Synapses all appear to be axoaxonic and not axosomatic.

Clearly, these features of the nervous system militate against the success of the sort of microelectrode work which has proved so fruitful in gastropods.

E Muscles

A few major muscle blocks such as the head retractor and funnel retractor muscles have a strongly organized fiber direction and consequently a single plane of contraction, but these are of relatively little behavioral interest. In the mantle, muscle fibers are arranged in discrete layers in longitudinal, circular, and radial planes (Ward and Wainwright, 1972; Boyle, 1976). The musculature of the buccal mass, particularly that to the beaks, is also formed into muscles of relatively consistent fiber orientation (Boyle et al., 1979a,b). Elsewhere, most of the muscle masses have a more complex organization characteristic of soft-bodied animals (Trueman, 1975), which is described in the older anatomical works.

Very little can be said about muscle fiber types in cephalopods. When individual fibers have been examined ultrastructurally, they always appear to be obliquely striated (Cloney and Rorey, 1968; Florey and Kriebel, 1969;Smith, 1979; Dubas, 1982). Functionally different fiber types are distinguished by the extent of vascularization and mitochondrial density. Within the circular muscle of the mantle of Loligo, Alloteuthis, and Sepia, there is a dichotomy of fiber types organized into inner and outer zones, both of well-vascularized, mitochondria-rich fibers, and a central zone of mitochondria-poor fibers with relatively sparse vascular supply (Bone et al., 1981). The implication of these findings is that it is the inner and outer zones which function continually to provide the rhythmic ventilatory contractions, while the central zone is used only for escape jetting. In the shelled Nautilus, the retraction of head and body into the shell by large head and shell retractor muscles also serves the dual functions of respiration and locomotion (Packard et al., 1980), and it is significant that two fiber types, differing in their mitochondrial densities, are found in these muscles (Hochachka et al., 1978). Striking differences in the disposition of mitochondria relative to the contractile filaments in radial muscles of chromatophores are noted by Dubas (1982). In octopods the filaments occupy the core of the muscle fiber, but in squids they are restricted to the periphery and surround a core of mitochondria.

F Neurotransmitters

Many pharmacologically active chemicals and those with known synaptic potential have been identified in cephalopod tissues. The following list details some of the sites at which various substances are probably present and limited data on their action. It is important to note that the methods of identification, mostly histochemical, often do not separate individual chemicals within a category, e.g., catecholamines.

Dopamine and noradrenaline are present in the brains of Eledone, Octopus, and Sepia (Juorio, 1971, 1972); the posterior salivary glands; visceral tissues (Juorio and Killick, 1973); and blood vessels (Andrews and Tansey, 1983a). Octopamine is also normally present (Juorio and Molinoff, 1971, 1974), and 5-hydroxytryptamine (5-HT, serotonin) is widely distributed in the brain (Juorio, 1972;Tansey, 1979). The presence of cholinesterases and, by implication, cholinergic neurons is demonstrated in the brain of several cephalopods (Loe and Florey, 1966; Chichery and Chichery, 1974). Monoamine fluorescence (dopamine, noradrenaline, octopamine, p-tyramine, and 5-HT) is also demonstrated in the neuropile of many of the peripheral ganglia of octopods (Ducros and Arluison, 1977) and the statocyst (Budelmann and Bonn, 1982a,b). The detailed distribution of aminergic fluorescence in the brain of Octopus has been described by Tansey (1980). There are remarkably high quantities of these biogenic amines localized in the posterior salivary glands and the buccal lobe region of the brain which innervates them (Arluison and Ducros, 1976; Martin and Barlow, 1972;Matus, 1971).

The effects of these and other drugs have been tested on elements of the circulatory system (Johansen and Huston, 1962), chromatophore muscles (Florey and Kriebel, 1969), and preparations of the gut (Andrews and Tansey, 1982). The results are described in Section III. Preparations of muscle from the mantle and fins of Loligo, Alloteuthis, and Sepia respond to l-glutamate by twitching (Bone and Howarth, 1980). Injected via the cephalic aorta into the brain of Octopus (Andrews and Tansey, 1981b), dopamine, noradrenaline, octopamine, γ-aminobutyric acid (GABA), acetylcholine, nicotine, and 5-HT all produced changes in the color or the texture of the animal (Andrews et al., 1981). Of those transmitters known to be present in the brain, dopamine, noradrenaline, and octopamine caused a darkening (chromatophore expansion); acetylcholine caused a paling (chromatophore retraction, due to chromatophore muscle relaxation); and 5-HT provoked a mottled patterning. It is suggested that the experimental effects obtained were due to the central action of the drug on neurons in the brain.

No single neurotransmitter has been specifically associated with an identified synaptic contact or neuromuscular junction.

G Paraneural and Neurosecretory Tissues

Listed in Table III, p. 32, are a number of tissues which are closely related to the nervous system, some of which have known or suspected neurosecretory functions. They are mentioned here because some behavioral control is implicated.

A number of these tissues are associated with the orbit (Fig. 4). Notable is the optic gland, a spherical mass of cells located on the optic stalk of each side and innervated from the subpedunculate lobe at the posterior of the supraesophageal brain mass. A large body of experimental evidence has shown that the optic gland is the source of a gonadotropic hormone in Octopus (Wells and Wells, 1959, 1972a,b, 1975; O’Dor and Wells, 1973, 1975). Sexual maturation is accompanied by profound physiological and behavioral changes. The female stops feeding and seeks a site for egg laying, a process which may be reversed by surgical removal of the optic glands (Wells et al., 1975; Wodinsky, 1977), but no measurable effects on mating behavior have been experimentally demonstrated (Wells and Wells, 1972a). Experimentally the glands may be activated by denervation, and it is generally thought that the subpedunculate lobe normally exerts inhibitory control. The pathways by which the glands are caused to enlarge and secrete in nature are not clear for Octopus. In Sepia it seems that decreasing photoperiod is a primary influence in triggering sexual maturity (Richard, 1967, 1971), and in the octopus Eledone, starvation is held to induce maturation (Mangold and Boucher-Rodoni, 1973). The matter is further complicated by the possibility that the optic gland has other functions (Mangold and Froesch, 1977; Froesch and Mangold, 1976a; Froesch et al., 1978; Froesch, 1979). The subject has been extensively reviewed for octopods (Wells and Wells, 1977a,b; Wells, 1978) and for decapods (Arnold and Williams-Arnold, 1977).

Fig. 4 The various neurovenous tissues of Octopus vulgaris. The diagram shows the buccal mass, parts of the brain and optic lobes, and an eye, all displaced from their normal position in order to display the neurovenous organs. [From Young (1970).]

Another, clearly neurosecretory system is found in the anterior vena cava. A large proportion of the venous blood returns to the heart through a single, ventral, anterior vena cava. This vessel divides into two lateral vena cavae, which run to the branchial hearts which distribute blood to the gill of each side (Smith and Boyle, 1983). Confirming the results of earlier authors it was noted by Alexandrowicz (1964, 1965) that the anterior vena cava of Eledone and Sepia, respectively, was intensively innervated from the palliovisceral lobe of the brain, and he suggested this indicated a neurosecretory role. The fine structure (Martin, 1968) was consistent with this suggestion, and subsequently Berry and Cottrell (1970) demonstrated the excitatory effect of extracts of the vena cava on the heart of Eledone. The active factor was not 5-HT, noradrenaline, or adrenaline, and electrical stimulation of the nerves to the anterior vena cava could not regularly produce the same effect as the injection of tissue extract into the bloodstream.

Cells thought to be neurosecretory on structural evidence have been described in the brain of Illex and Ommastrephes (Martin, 1966) as well as Octopus (Bonichon, 1967, 1968; Martin, 1968; Froesch, 1974). The subpedunculate lobe (Young, 1970), which gives rise to the nerve to the optic gland, also supplies the pharyngo-ophthalmic vein (POV) (octopods, not sepiids) which drains the suborbital sinus. The termination of the nerve in the POV also appears to be neurosecretory (Froesch, 1974), and acceleratory effects of the extract on the systemic heart have been demonstrated (Froesch and Mangold, 1976b). The high levels of pharmacologically active substances already noted in the posterior salivary glands are associated with the presence of the same substances in the innervating superior buccal lobe of the brain (Martin and Barlow, 1972;Matus, 1971), and this may constitute another neurosecretory system (Ducros, 1972a,b; Barlow et al., 1974; Arluison and Ducros, 1976).

The paravertical tissue of the brain (Young, 1970), the juxtaganglionic tissue partly investing the inferior buccal ganglion (Young, 1965c), and the juxtaganglionic tissue of the gastric ganglion (Young, 1967 b) are also potential neurosecretory tissues but may be non-nervous.

Two other tissues associated with the nervous system and once suspected of a neurosecretory function are now known to be sensory: the parolfactory vesicles of squids, pigmented sacs close to the optic tract; and the epistellar body of octopods, a small pigmented body lying on the stellate ganglion and particularly conspicuous in Eledone. Structural, ultrastructural, and biochemical studies have shown unequivocally that these structures are photoreceptors (Nishioka et al., 1962, 1966a,b; Messenger, 1967c; Baumann et al., 1970; Perrelet and Mauro, 1972). Moreover, depolarizing generator potentials have been recorded from both the epistellar body (Mauro and Baumann, 1968) and parolfactory vesicles (Mauro and Sten-Knudsen, 1972; Sperling et al., 1973).

III Intrinsic Movements

Separated from the main sources of their innervation, some muscle systems exhibit considerable activity in vitro. This level of intrinsic activity complicates the interpretation of their control by the nervous system.

A Heart

The main circulatory pump in cephalopods, the systemic heart, consists of a single, median ventricle supplied with blood by a pair of lateral auricles. Output from the ventricle is distributed through an anteriorly running dorsal aorta, abdominal and gonadial arteries, respectively. Isolated from the body, the ventricle continues to beat rhythmically and steadily so long as muscle tonus is maintained. The physiological properties of this preparation have been extensively investigated in Eledone (Smith, 1979).

With the gonadial and abdominal arteries sealed off and output back pressure in the dorsal aorta held constant, the ventricular beat frequency and stroke volume are both sensitive to variations in the input pressure mediated via one of the auricles (Smith, 1981b). Beat frequency increases with input pressure.

A consistent and complex electrocardiogram can be recorded from the beating ventricle in vitro (Smith, 1981a), the form of which depends on the precise position of the recording electrodes. This implies a rather controlled sequence for the depolarization of ventricular muscle and indicates that some form of nodal pacemaker may be present (Smith, 1981a). A potential anatomical location, apparently consisting of atypical ventricular muscle, has been described by Smith (1979). If such a pacemaker area is present, then it contrasts with the general diffuse myogenicity thought to control most molluscan hearts (Hill and Welsh, 1966). This feature would also further complicate the present picture of the coordination of contractile components of the circulatory system (Section IV).

The isolated beating ventricle responds directly to the application of certain drugs. When perfused in seawater into the beating heart of Eledone, 5-hydroxytryptamine, adrenaline, and noradrenaline were found to cause transient increases in the frequency and amplitude (Berry and Cottrell, 1970). Epinephrine produces variable effects in Eledone (Smith, 1979), but in Octopus it causes an increase in ventricular frequency in vitro (Fänge and Östland, 1954) or an inhibitory effect in vivo (Johansen and Huston, 1962).

Extracts of some cephalopod tissues are known to have a direct effect on the activity of the cephalopod ventricle. Introduction of anterior vena cava extract into the perfusing seawater causes a prolonged cardioexcitatory effect (Berry and Cottrell, 1970). On immunocytochemical evidence, enkephalin-related peptides have been identified in the neurosecretory region of the anterior vena cava of Octopus (Martin et al., 1979, 1980). Few of these peptides have much effect on the heart at reasonable concentrations, but the molluscan cardioexcitatory peptide Phe-Met-Arg-Phe-NH2 causes a strong excitatory response (Voigt et al., 1981). Extracts of putative neurosecretory tissue in the pharyngo-ophthalmic vein of Eledone have been also reported to have excitatory effects on isolated heartbeat frequency and amplitude (Froesch and Mangold, 1976b), but the concentrations required in Octopus make it unlikely that release of substances from this source could be effective in the intact animal (Wells and Mangold, 1980).

The auricles show remarkably little contractile activity in the experimental conditions used by most authors (Johansen and Martin, 1962; Wells, 1978, p. 43; Smith, 1979), and it is doubtful whether they contribute much to ventricular filling.

B Branchial Hearts

Sited at the base of the gill on either side is a large, pigmented, contractile body, the branchial heart. The branchial heart has a muscular wall and its lumen is traversed by muscular trabeculae. The lumen is partly occluded by the presence of quantities of a loose matrix of apparently secretory cells (Schipp and Schäfer, 1969a,b).

In isolation, the branchial heart beats rhythmically. Location of the source of this activity is difficult to ascertain because the cardiac ganglion is closely attached to the muscle surface. In Eledone extirpation of the ganglion by freezing stops the contractile activity (Smith, 1979), but this does not always occur in Octopus (Wells, 1978, p. 43; Wells, 1980). In octopods, but not decapods, the ganglion itself is visibly pulsatile, due to the movement of a small intraganglionic body with its own muscle fibers (Alexandrowicz, 1963). The branchial hearts each receive venous blood via the lateral vena cava of each side. Their function is commonly held to be to boost the flow of blood out through the afferent branchial vessel to the gill (Wells, 1980), although the evidence for effective pumping has been doubted (Smith, 1982). Blood returns from the gill through the efferent branchial vessel to the auricle of each side.

C Blood Vessels

Most of the other blood vessels also exhibit intrinsic contractile activity. Such peristaltic activity is widely thought to be significant in maintaining the circulatory blood flow in cephalopods (Johansen and Martin, 1962; L. S. Smith, 1962; Wells, 1980; P. J. S. Smith, 1981b). The contractions of the lateral venae cavae in particular may well contribute more to the blood flow to the gills than the branchial hearts themselves (Smith, 1982).

The anatomy and innervation of blood vessels and central circulatory organs have been described by several authors in recent years (Smith, 1963; Barber and Graziadei, 1965; Smith and Boyle, 1983; Andrews and Tansey, 1983a).

D Gut

The smooth muscle of the esophagus and crop is innervated via the sympathetic nerves from the inferior buccal ganglion, but also much of the innervation to the gut arises from the visceral nerves directly and through the gastric ganglion which is located on the wall of the cecum (Young, 1967b). Isolated sections of the gut of Octopus vulgaris, e.g., esophagus, crop, stomach, caecum, intestine, and rectum, show spontaneous contractile activity when maintained in an organ bath (Andrews and Tansey, 1983b).

In the crop and stomach, acetylcholine reduces the level of spontaneous activity and the resting tonus of the muscle. Noradrenaline, in contrast, stimulates crop activity, leading Andrews and Tansey (1983b) to suggest that these two agents may normally control gut activity by their antagonistic actions; 5-HT was found to be more effective in stimulating the intestine and rectum. The caecum was relatively unaffected by drugs.

In Loligo pealei, where sections of isolated gut are also intrinsically active, acetylcholine is inhibitory, adrenaline and 5-HT excitatory (Bacq, 1934; Wood, 1969). Since the gut tissues contain a very extensive intramural nerve plexus, it is difficult to separate the effects of nervous action from intrinsic myogenicity. The view of those authors who have worked on squids (Wood, 1969) and octopuses (Andrews and Tansey, 1983b) is that the activity of the gut is basically myogenic and only modulated by nerves. The fact that ACh antagonists have an excitatory effect and that the presence of nerve-blocking drugs (benzoquinonium chloride, d-tubocurarine) abolished the effect of ACh inhibition also suggests that this is a nerve-mediated influence (Wood, 1969).

E Chromatophores

In isolated pieces of skin, detached from the surface of the animal, chromatophores continue to be active individually or as part of coordinated waves of chromatophore expansion.

The chromatophore organs are unique to cephalopods. Located in the subdermal layers of the skin, overlying the somatic musculature, each one consists of an elastic pigment sac on which radiating muscle fibers are inserted. When the muscles contract, the pigment sac expands, spreading the area of contained pigment; when the muscles relax, the elasticity of the pigment sac causes it to retract, the pigment area is reduced to a small spot, and the white background of the somatic musculature shows through. These muscular chromatophores are the basic elements of the complex pattern and color change abilities of the modern cephalopods (coleoids only; the pigmentation of Nautilus is not actively variable).

There is a significant amount of early literature on the distribution and activity of the chromatophores. Interest remains high in the physiology of individual chromatophores and their coordination into large-scale groups generating the patterned appearance of octopuses and squids. Not only are these organs fascinating from the point of view of cephalopod biology, but they also provide a preparation in which the results of the activity of individual muscle fibers can be readily seen and recorded as movement. Cloney and Florey (1968) and Florey (1969) give thorough general accounts of the structure and function of cephalopod chromatophores. Mirow (1972a,b) has described the morphology of the squid chromatophores and also the structural reflecting elements of the skin, the leucophores and iridophores (see also Denton and Land, 1971; Brocco, 1977; Brocco and Cloney, 1980).

The innervation of the chromatophore muscles originates in the chromatophore lobes of the brain, but the organization and coordination of patterned chromatophore activity with other aspects of overt behavior are influenced by several of the higher centers. The lateral basal lobes and the optic lobes, at least, are important parts of the controlling system (Young, 1971). Individual chromatophore muscles are each innervated directly with a motor innervation (Florey, 1966). No evidence has been obtained for inhibitory innervation (Florey and Kriebel, 1969), but multiple innervation of individual muscle fibers is known (Weber, 1968; Dubas, 1982).

Florey and Kriebel (1969) used glass microelectrodes placed within the muscle fibers of the large chromatophores of Loligo opalescens. cetylcholine caused a conspicuous increase in the frequency of spontaneous miniature potentials in the muscle fiber but had no measurable effect on the overall resting potential of the cell membrane. Applied superficially, acetylcholine at concentrations above 10–7 g/ml caused tonic contraction of the muscle fibers (chromatophore expansion) and pulsations (Florey, 1966). The 5-HT appears to act antagonistically to acetylcholine in causing the relaxation of chromatophore muscle after spontaneous or ACh-induced contraction (Florey, 1966; Florey and Kriebel, 1969).

The source of the pulsating activity of individual chromatophores and their coordinated waves of contraction in detached skin is unknown. This intrinsic activity of chromatophores is noted to be much more common in aged skin preparations, even several days old, when the innervation would be expected to be degenerate (Florey and Kriebel, 1969; Dubas, 1982). In Loligo, rhythmic depolarizations, so-called pacemaker potentials, develop in individual fibers at frequencies of about 1/sec. They result in the synchronous activity of all the muscle fibers of the individual chromatophore. Florey and Kriebel (1969) also showed that this coordination was probably achieved by the electrical coupling of neighboring muscle fibers via low-resistance pathways.

The peripheral coordination of neighboring chromatophores in the absence of innervation is another dimension of the control of chromatophore activity. In Eledone cirrhosa these wandering clouds of chromatophore expansion travel across the skin at speeds of about 3–5 mm/sec. (Dubas, 1982). The wave generally travels in one direction, rather than radiating from the point of origin. Several waves may originate from the same point at intervals of about 6–8 per minute, the interval gradually lengthening until that part of the skin ceases to be spontaneously active. Dubas (1982) observed that if a superficial cut is made in the skin in the path of a moving wave, the path of the wave is interrupted to detour around the cut. The same area of skin electrically stimulated through its nervous supply resulted in normal chromatophore expansion on both sides of the cut, demonstrating that the effect of the cut in stopping transmission of the wave of chromatophore expansion was not due to interruption of the innervation by the cut. This result supports the conclusion of Florey and Kriebel (1969), who found that the intrinsic activity of chromatophore muscles was not abolished by tetrodotoxin, which should block neuromuscular transmission. Generally, pulsating activity is more common in the large squid chromatophores, and wandering clouds are more frequent in octopuses. Structural evidence has been produced to argue that neighboring chromatophores in Octopus can be directly connected mechanically or electrically by common muscle fibers (Froesch-Gaetzi and Froesch, 1977). In Eledone such common fibers are possibly, but rarely, present (Dubas, 1982,) although mechanical or electrical coupling could occur at the frequent points where chromatophore muscles from different chromatophores cross each other and are in close apposition.

Despite the recent efforts in this field, the peripheral origin (in the absence of CNS and major ganglia) and means of propagation of intrinsic activity in chromatophores remain a mystery. To some extent these uncertainties complicate the interpretation of chromatophore patterns generated by nerve stimulation and during normal behavior (Section VII,C).

IV Local Organization of Movement, Lower and Intermediate Motor Centers

Those areas of the nervous system known to contain final motor neurons together with the centers of the subesophageal brain which are associated directly with the effectors of a particular region are reviewed in this section.

A Arms

Uniquely equipped with discoid suckers and with 40–70% of total body tissue mass as arms, an octopus over 2–3 kg in weight can exert sufficient strength and adhesion to defy human efforts to dislodge it from a hard surface. As predators, they grip and handle with ease food items ranging in size up to about their own weight. Dilly et al. (1964) investigated the pull exerted by Octopus on a crab tied to nylon monofilament line before it tore free or was released (Fig. 5). The maximum pulling force increased at the rate of 2 kg/0.1 kg body weight up to about 0.5 kg, after which it increased much more slowly. Maintained pulls never exceeded 50% of the value obtained for maximum pulls in the same animal. Using an isometric myograph attached to a line sewn through the head cartilage, Trueman and Packard (1968) found that Octopus vulgaris with five arms holding to the side of the tank can exert a holding tension of up to 100 times its body weight. Squids may have suckers which are also toothed around the rim, and all the decapods (squids, sepiolids, and cuttlefish) have, in addition, a pair of tentacles. The tentacles are used exclusively for prey capture and only have suckers on the terminal portion. They remain tightly packed into two ventral pouches until extended rapidly forward to strike and grip a prey. Extension is due to the contraction of transverse and circular muscle fibers (Kier, 1982). Comparative details of sucker surfaces in relation to prey capture are given by Nixon and Dilly (1977). Nautilus does not have suckers on its numerous prehensile arms.

Fig. 5 ) of Octopus vulgaris ). [From Dilly et al. (1964).]

The manipulative abilities of the cephalopod arms are unparalleled among molluscs. At full extension the arm tips are very fine and mobile and actively explore the surroundings. The arms and suckers manipulate prey, and they are effective in disarticulating crustacea, extracting shelled molluscs, etc., and passing food to the mouth. In Octopus they are equally able to gather together stones and shells to form a home or lair on the bottom. Arms and suckers in a resting octopus are frequently active, moving over the body surface, apparently sloughing off epidermal layers which then can be clearly seen in the water. At sexual maturity one of the arms assumes responsibility as the male intromittent organ. The tip of one arm (third right in octopods) becomes morphologically modified and, as the hectocotylus, or hectocotylized arm, transfers spermatophores from the male duct to the female.

Collectively the arms are estimated to contain more nerve cells than the rest of the nervous system altogether (Young, 1963a). The anatomy of their complex arrangements of muscle and nerve has been extensively studied by early authors such as Guérin (1908) and ten Cate (1928). More recently, the anatomy was further investigated by Rossi and Graziedei (1954, 1956a,b, 1958) and has been summarized by Graziadei (1971). Anatomical detail is also available for some squids (Graziadei, 1960a,b) and sepioids (Graziadei, 1959; Martoja and May, 1956). The basic arrangement of a thick axial nerve cord is shown in Figure 2 taken from Graziadei (1965a). Nerve cells are present all along the axial cord but accumulate as swellings opposite the base of each sucker to form the brachial ganglionic chain of sucker ganglia. Separated from the axial cord and sited just at the base of each sucker is a further ganglion, the acetabular ganglion. The final motor neurons innervating the muscles responsible for all movements of the arms and suckers lie in these cords and ganglia.

The mobility of amputated arms of Octopus excited the curiosity of early workers and was the object of considerable investigations by von Uexküll (1894) and ten Cate (1928). More recently Rowell (1963) stimulated the axial nervous system and classified the results and in a later series of experiments (Rowell, 1966) studied electrical activity in the arms of intact octopuses. Altman (1968) made a detailed study of the nervous control of arm movements in Octopus organized at the level of the arm nerve cords and at higher levels.

Table II, taken from Altman (1968), shows her classification of the reflexes of the isolated arm in response to three levels of mechanical stimulation. Tactile stimulation of the sucker disk results in a grip reflex, a raising of the sucker rim which will then grip the stimulating probe or cause adherence to a flat surface. More intense stimulation causes protraction of adjacent suckers toward the source of stimulation. Such recruitment of the suckers is proportional to the strength of the stimulus and begins in those on the side distal to the level of stimulation. If the disk of the protracted sucker does not make contact with the probe, then searching occurs in which the sucker disk moves in semicircles relative to the peduncle. Several suckers contacting the probe result in grasping and are assisted by slight bending of the arm. If the peduncle of a sucker already attached to the substrate is touched, the peduncle contracts, causing retraction of the disk. A serial stepping of suckers was sometimes caused by a strong retraction reflex or strong stimulation of the dorsal or lateral skin of the arms. Since each sucker touched the peduncle of the adjacent sucker as it was protracted, it is possible that stepping can be coordinated by local stimulation of the suckers rather than by neural integration at the level of the arm cord. The coordinated pattern of sucker movement in the isolated arm is also described in accept and reject reflexes whereby an item (e.g., food morsel) is passed proximally or distally along the sucker row (Altman, 1971). The reflex behavior of the arm of Octopus denervated in situ (chronic arm preparation) was found to be similar to that of the amputated arm, but it was altogether less active, with lowered muscle tone and no sucker grip. It is possible that an excitatory input to the arm cords from the central ganglia is required to maintain these features which, in the acute preparation, is duplicated by injury discharges resulting from the amputation.

TABLE II

Strength of the Mechanical Stimuli Required to Elicit Various Reflexes from the Isolated Arm of Octoous vulearisa,b

aSimplified from Altman (1968). I am grateful to Dr. J. Altman for permisson to reproduce this table from her unpublished work.

bStimulus strengths, + light (bristle); ++ medium (seeker); +++ strong (pinch with forceps); – no response to any stimulus.

The suckers of Octopus, Sepia, and Todarodes (Ommastrephes) which have been investigated are liberally supplied with tapered and multipolar nerve cells which appear to be the primary tactile receptors. A category of peripheral cell bodies identified as encapsulated cells is also present below the epithelium. Through short, stumpy dendritic processes and longer, branching collaterals, these encapsulated cells appear to be interneurons (Fig. 6) linking the receptors of the sucker surface to the associated ganglion of the arm cord (Graziadei, 1965b). Axodendritic contacts between