Outer Nerve Ring Research Articles

Unusual lophophore innervation in ctenostome Flustrellidra hispida (Bryozoa).

Since ctenostomes are traditionally regarded as an ancestral clade to some other bryozoan groups, the study of additional species may help to clarify questions on bryozoan evolution and phylogeny. One of these questions is the bryozoan lophophore evolution: whether it occurred through simplification or complication. The morphology and innervation of the ctenostome Flustrellidra hispida (Fabricius, 1780) lophophore have been studied with electron microscopy and immunocytochemistry with confocal laser scanning microscopy. Lophophore nervous system of F. hispida consists of several main nerve elements: cerebral ganglion, circumoral nerve ring, and the outer nerve ring. Serotonin-like immunoreactive perikarya, which connect with the circumoral nerve ring, bear the cilium that directs to the abfrontal side of the lophophore and extends between tentacle bases. The circumoral nerve ring gives rise to the intertentacular and frontal tentacle nerves. The outer nerve ring gives rise to the abfrontal neurites, which connect to the outer groups of perikarya and contribute to the formation of the abfrontal tentacle nerve. The outer nerve ring has been described before in other bryozoans, but it never contributes to the innervation of tentacles. The presence of the outer nerve ring participating in the innervation of tentacles makes the F. hispida lophophore nervous system particularly similar to the lophophore nervous system of phoronids. This similarity allows to suggest that organization of the F. hispida lophophore nervous system may reflect the ancestral state for all bryozoans. The possible scenario of evolutionary transformation of the lophophore nervous system within bryozoans is suggested.

Detailed morphology of tentacular apparatus and central nervous system in Owenia borealis (Annelida, Oweniidae)

The Oweniidae are marine annelids with many unusual features of organ system, development, morphology, and ultrastructure. Together with magelonids, oweniids have been placed within the Palaeoannelida, a sister group to all remaining annelids. The study of this group may increase our understanding of the early evolution of annelids (including their radiation and diversification). In the current research, the morphology and ulta-anatomy of the head region of Owenia borealis is studied by scanning electron microscopy (SEM), 3D reconstructions, transmission electron microscopy (TEM), and whole-mount immunostaining with confocal laser scanning microscopy. According to SEM, the tentacle apparatus consists of 8–14 branched arms, which are covered by monociliary cells that form a ciliary groove extending along the oral side of the arm base. Each tentacle contains a coelomic cavity with a network of blood capillaries. Monociliary myoepithelial cells of the tentacle coelomic cavity form both the longitudinal and the transverse muscles. The structure of this myoepithelium is intermediate between a simple and pseudo-stratified myoepithelium. Overall, tentacles lack prominent zonality, i.e., co-localization of ciliary zones, neurite bundles, and muscles. This organization, which indicates a non-specialized tentacle crown in O. borealis and other oweniids with tentacles, may be ancestral for annelids. TEM, light, and confocal laser scanning microscopy revealed that the head region contains the anterior nerve center comprising of outer and inner (=circumoral) nerve rings. Both nerve rings are organized as concentrated nerve plexus, which contains perikarya and neurites extending between basal projections of epithelial cells (radial glia). The outer nerve ring gives rise to several thick neurite bundles, which branch and extend along aboral side of each tentacle. Accordingly to their immunoreactivity, both rings of the anterior nerve center could be homologized with the dorsal roots of circumesophageal connectives of the typical annelids. Accordingly to its ultrastructure, the outer nerve ring of O. borealis and so-called brain of other oweniids can not be regarded as a typical brain, i.e. the most anterior ganglion, because it lacks ganglionic structure.

Peculiarities of Tentacle Innervation of Flustrellidra hispida and Evolution of Lophophore in Bryozoa.

The study of the lophophore organization is of great importance for the reconstruction of lophophorate phylogeny and for understanding the evolutionary transformation in each phylum of Lophophorata. The innervation of the lophophore in ctenostome bryozoan Flustrellidra hispida was studied using immunocytochemistry and confocal laser scanning microscopy. It has been demonstrated that this species has an outer nerve ring giving rise to the tentacle nerves. The outer nerve ring was earlier described in some ctenostomates and cyclostomates, but not as connected with nerves. The discovered feature of lophophore innervation in F. hispida suggests the evolutionary transformation from a hypothetical phoronida-like ancestor lophophore bearing a prominent outer nerve ring with numerous tentacle nerves emanating from it, to the complex bell-shaped lophophore of F. hispida with a well-pronounced outer nervous ring bearing a few tentacle nerves. The next one in this hypothetical row is the lophophore of the other ctenostomates and some cyclostomates with no ring-nerve connection and cheilostomates lophophore with no outer nerve ring at all.

The nervous system in the cyclostome bryozoan Crisia eburnea as revealed by transmission electron and confocal laser scanning microscopy

IntroductionAmong bryozoans, cyclostome anatomy is the least studied by modern methods. New data on the nervous system fill the gap in our knowledge and make morphological analysis much more fruitful to resolve some questions of bryozoan evolution and phylogeny.ResultsThe nervous system of cyclostome Crisia eburnea was studied by transmission electron microscopy and confocal laser scanning microscopy. The cerebral ganglion has an upper concavity and a small inner cavity filled with cilia and microvilli, thus exhibiting features of neuroepithelium. The cerebral ganglion is associated with the circumoral nerve ring, the circumpharyngeal nerve ring, and the outer nerve ring. Each tentacle has six longitudinal neurite bundles. The body wall is innervated by thick paired longitudinal nerves. Circular nerves are associated with atrial sphincter. A membranous sac, cardia, and caecum all have nervous plexus.ConclusionThe nervous system of the cyclostome C. eburnea combines phylactolaemate and gymnolaemate features. Innervation of tentacles by six neurite bundles is similar of that in Phylactolaemata. The presence of circumpharyngeal nerve ring and outer nerve ring is characteristic of both, Cyclostomata and Gymnolaemata. The structure of the cerebral ganglion may be regarded as a result of transformation of hypothetical ancestral neuroepithelium. Primitive cerebral ganglion and combination of nerve plexus and cords in the nervous system of C. eburnea allows to suggest that the nerve system topography of C. eburnea may represent an ancestral state of nervous system organization in Bryozoa. Several scenarios describing evolution of the cerebral ganglion in different bryozoan groups are proposed.

Innervation of the lophophore suggests that the phoronid Phoronis ovalis is a link between phoronids and bryozoans

The validity of the Lophophorata as a monophyletic group remains controversial. New data on the innervation of the lophophore, which is a unique feature of the lophophorates, may help clarify the status of the Lophophorata and provide new information on the early evolution of the group. In this paper, the organization of the nervous system of the lophophore is described in adults of the minute phoronid Phoronis ovalis. The lophophore nervous system includes a dorsal ganglion, a tentacular nerve ring, an inner ganglion, an inner nerve ring, and six nerves in each tentacle. The inner ganglion and inner nerve ring, which is associated with sensory cells, are described for the first time in adult phoronids. The general plan of the nervous system of the lophophore and tentacles is similar in P. ovalis and bryozoans. These new results suggest the presence of two nerve centers and two nerve rings in the last common ancestor of phoronids and bryozoans. During evolution, bryozoans may have lost the outer nerve center and outer nerve ring, whereas phoronids may have lost the inner nerve center and inner nerve ring. These morphological results evidence the lophophorates are monophyletic.

The nervous system of the lophophore in the ctenostome Amathia gracilis provides insight into the morphology of ancestral ectoprocts and the monophyly of the lophophorates.

BackgroundThe Bryozoa (=Ectoprocta) is a large group of bilaterians that exhibit great variability in the innervation of tentacles and in the organization of the cerebral ganglion. Investigations of bryozoans from different groups may contribute to the reconstruction of the bryozoan nervous system bauplan. A detailed investigation of the polypide nervous system of the ctenostome bryozoan Amathia gracilis is reported here.ResultsThe cerebral ganglion displays prominent zonality and has at least three zones: proximal, central, and distal. The proximal zone is the most developed and contains two large perikarya giving rise to the tentacle sheath nerves. The neuroepithelial organization of the cerebral ganglion is revealed. The tiny lumen of the cerebral ganglion is represented by narrow spaces between the apical projections of the perikarya of the central zone. The cerebral ganglion gives rise to five groups of main neurite bundles of the lophophore and the tentacle sheath: the circum-oral nerve ring, the lophophoral dorso-lateral nerves, the pharyngeal and visceral neurite bundles, the outer nerve ring, and the tentacle sheath nerves. Serotonin-like immunoreactive nerve system of polypide includes eight large perikarya located between tentacles bases. There are two analmost and six oralmost perikarya with prominent serotonergic “gap” between them. Based on the characteristics of their innervations, the tentacles can be subdivided into two groups: four that are near the anus and six that are near the mouth. Two longitudinal neurite bundles - medio-frontal and abfrontal - extend along each tentacle.ConclusionThe zonality of the cerebral ganglion, the presence of three commissures, and location of the main nerves emanating from each zone might have caused by directive innervation of the various parts of the body: the tentacles sheath, the lophohpore, and the digestive tract. Two alternative scenarios of bryozoan lophophore evolution are discussed. The arrangement of large serotonin-like immunoreactive perikarya differs from the pattern previously described in ctenostome bryozoans. In accordance with its position relative to the same organs (tentacles, anus, and mouth), the lophophore outer nerve ring corresponds to the brachiopod lower brachial nerve and to the phoronid tentacular nerve ring. The presence of the outer nerve ring makes the lophophore innervation within the group (clade) of lophophorates similar and provides additional morphological evidence of the lophophore homology and monophyly of the lophophorates.Electronic supplementary materialThe online version of this article (doi:10.1186/s12862-016-0744-7) contains supplementary material, which is available to authorized users.

Action potential shape change in an electrically coupled network during propagation: a computer simulation

We applied compartmental computer modeling to test a model of spike shape change in the jellyfish, Polyorchis penicillatus, to determine whether adaptive spike shortening can be attributed to the inactivation properties of a potassium channel. We modeled the jellyfish outer nerve-ring as a continuous linear segment, using ion channel and membrane properties derived in earlier studies. The model supported action potentials that shortened as they propagated away from the site of initiation and this was found to be largely independent of potassium channel inactivation. Spike broadening near the site of initiation was found to be due to a depolarization plateau that collapsed as two spikes spread from the point of initiation. The lifetime of this plateau was found to depend critically on the inward current flux and the space constant of the membrane. These data suggest that the spike shape changes may be due not only to potassium channel inactivation, but also to the passive properties of the membrane.

Nitric oxide regulates swimming in the jellyfish Aglantha digitale.

The cnidarian nervous system is considered by many to represent neuronal organization in its earliest and simplest form. Here we demonstrate, for the first time in the Cnidaria, the neuronal localization of nitric oxide synthase (NOS) in the hydromedusa Aglantha digitale (Trachylina). Expression of specific, fixative-resistant NADPH-diaphorase (NADPH-d) activity, characteristic of NOS, was observed in neurites running in the outer nerve ring at the base of the animal and in putative sensory cells in the ectoderm covering its tentacles. At both sites, diphenyleneiodonium (10(-4) M) abolished staining. Capillary electrophoresis confirmed that the NO breakdown products NO2- and NO3- were present at high levels in the tentacles, but were not detectable in NADPH-d-negative areas. The NADPH-d-reactive neurons in the tentacles send processes to regions adjacent to the inner nerve ring where swimming pacemaker cells are located. Free-moving animals and semi-intact preparations were used to test whether NO is involved in regulating the swimming program. NO (30-50 nM) and its precursor L-arginine (1 mM) stimulated swimming, and the effect was mimicked by 8-Br-cGMP (50-100 microM). The NO scavenger PTIO (10-100 microM) and a competitive inhibitor of NOS, L-nitroarginine methyl ester (L-NAME, 200 microM), significantly decreased the swimming frequency in free-moving animals, while its less-active stereoisomer D-nitroarginine methyl ester (D-NAME, 200 microM) had no such effect. 1H-[1,2,4]oxadiazolo[4,3-a]quinoxaline-1-one (ODQ, 5-20 microM), a selective inhibitor of soluble guanylyl cyclase, suppressed spontaneous swimming and prevented NO-induced activation of the swimming program. We suggest that an NO/cGMP signaling pathway modulates the rhythmic swimming associated with feeding in Aglantha, possibly by means of putative nitrergic sensory neurons in its tentacles.

Central circuitry in the jellyfish Aglantha. I: The relay system

1. The relay system is an interneuronal pathway in the margin of the jellyfish Aglantha digitale. It excites a second interneuronal pathway, the carrier system, and is itself excited by pacemaker neurones concerned with slow swimming. It also excites a slow conduction pathway in the tentacles causing graded, tonic contractions of all the tentacles during slow swimming. 2. The pacemakers, the carrier system and the relay system all contribute to the production of excitatory postsynaptic potentials (EPSPs) in a giant axon that runs in the outer nerve ring (ring giant axon). These EPSPs may cause the latter to spike during slow swimming. If it does so, it will fire tentacle giant axons, producing twitch contractions of the tentacles. Such contractions probably help to contract the tentacles rapidly at the start of slow swimming. This is an unusual case of a giant axon that normally mediates escape behaviour being appropriated for use during a non-escape activity. 3. The relay system can conduct impulses on its own but their conduction velocity is greatly increased when preceded by either pacemaker or ring giant spikes. This phenomenon, termed the 'piggyback effect', may be due to extracellular field effects rather than to actions mediated by chemical or electrical synapses. 4. Recordings from the epithelial cells that ensheath the ring giant and outer nerve ring neurones show miniature synaptic potentials and other events that seem to reflect events in the nervous system, but no functions can be assigned to them. 5. There is no obvious counterpart to the relay system in medusae lacking escape circuitry.

Central circuitry in the jellyfish Aglantha. II: The ring giant and carrier systems

1. The ring giant axon in the outer nerve ring of the jellyfish Aglantha digitale is a multinucleate syncytium 85 % of which is occupied by an electron-dense fluid-filled vacuole apparently in a Gibbs­Donnan equilibrium with the surrounding band of cytoplasmic cortex. Micropipette recordings show small (-15 to -25 mV) and large (-62 to -66 mV) resting potentials. Low values, obtained with a high proportion of the micropipette penetrations, are assumed to be from the central vacuole; high values from the cytoplasmic cortex. Background electrical activity includes rhythmic oscillations and synaptic potentials representing hair cell input caused by vibration. 2. After the ring giant axon has been cut, propagating action potentials evoked by stimulation are conducted past the cut and re-enter the axon on the far side. The system responsible (the carrier system) through-conducts at a velocity approximately 25 % of that of the ring giant axon and is probably composed of small neurones running in parallel with it. Numerous small neurones are seen by electron microscopy, some making one-way and some two-way synapses with the ring giant. 3. Despite their different conduction velocities, the two systems normally appear to fire in synchrony and at the velocity of the ring giant axon. We suggest that, once initiated, ring giant spikes propagate rapidly around the margin, firing the carrier neurones through serial synapses and giving them, in effect, the same high conduction velocity. Initiation of ring giant spikes can, however, require input from the carrier system. The spikes are frequently seen to be mounted on slow positive potentials representing summed carrier postsynaptic potentials. 4. The carrier system fires one-for-one with the giant axons of the tentacles and may mediate impulse traffic between the latter and the ring giant axon. We suggest that the carrier system may also provide the pathways from the ring giant to the motor giant axons used in escape swimming. 5. The findings show that the ring giant axon functions in close collaboration with the carrier system, increasing the latter's effective conduction velocity, and that interactions with other neuronal sub-systems are probably mediated exclusively by the carrier system.

Synaptic potentials and threshold currents underlying spike production in motor giant axons of Aglantha digitale.

1. Motor giant axons that excite swimming muscles in the jelly-fish Aglantha digitale interface with units of the inner and outer nerve rings in the margin at the base of the bell. External recording electrodes were used to monitor electrical activity at different sites within the nerve ring while events in the motor giant axon were recorded with intracellular micropipettes placed within 100 microns of the synaptic area. In some experiments, 4- to 6-micron-diam patch pipettes were used to record in situ from ion channel clusters at different locations along the axon. 2. Independently propagating calcium and sodium spikes in the motor giant axon were found to arise from different excitatory postsynaptic potentials (EPSPs). Two separate inputs were identified; one EPSP class represented an input from the pacemaker system in the inner nerve ring, whereas another represented an input from the giant axon in the outer nerve ring. EPSPs from the two nerve rings had significantly different time courses and amplitudes. EPSPs from the ring giant axon reached a peak in little more than 1 ms, whereas EPSPs from the pacemaker system reached a maximum in approximately 7 ms. These slower EPSPs may be compound events composed of postsynaptic potentials from multiple synapses excited in series by the passage of the pacemaker neuron signal. 3. The threshold for the production of calcium spikes by the slow EPSPs of the pacemaker system (-51 +/- 2.2 mV, mean +/- SD; n = 5) corresponded well with the voltage at which a net inward "T"-type calcium current first appeared in recordings from axon membrane patches (-55 to -50 mV); the threshold for the initiation of the sodium spike by the fast EPSPs of the ring giant system (-32 +/- 1.2 mV, mean +/- SD; n = 6) corresponded well with the voltage at which a net inward sodium current first appeared (-35 to -30 mV). 4. Inward currents were rarely observed in membrane patches formed using pipettes with tips of < 1 micron OD. Even with 4-micron pipettes, patches of membrane were sometimes obtained with a channel population consisting exclusively of potassium channels; calcium and sodium currents were found in highly discrete areas ("hot spots"). Preliminary findings on the undersurface of the axon, which makes synaptic contact with the myoepithelium, are consistent with a similar distribution. 5. The pathway by which the ring giant excites the motor giant axon is not definitely known. The synaptic delay between the peak of the ring giant action potential (monitored externally) and the initial rise of the fast EPSP (1.64 +/- 0.15 ms, mean +/- SD; n = 21) would allow for transmission at two synapses, because single synaptic delays at neuromuscular junctions in Aglantha are approximately 0.7 ms at 12 degrees C. The mean synaptic delay at the slow EPSP synapse was 0.88 +/- 0.09 (SD) ms (n = 12). 6. The delay between the impulse in the ring giant axon and the subsequent excitation of the motor giant axon may permit the animal to withdraw its tentacles and so lower the drag that would otherwise reduce the effectiveness of any escape swim and might induce tentacle autotomy.

Hair Cell Mechanoreception in the Jellyfish Aglantha Digitale

ABSTRACT The jellyfish Aglantha digitale is equipped with clusters of hair cells on the velum and on the tentacle bases. The cells have a central non-motile cilium surrounded by a collar of microvilli. The microvilli are graded in length, from long on one side to short on the other, giving the collar a marked polarity. The hair cells are set in specific orientations in all regions where they occur, as shown by this polarity. Small mechanical displacements of the velum or tentacles in the vicinity of the hair cells evoke bursts of potentials which can be recorded extracellularly from the outer nerve ring. Intracellular recordings from the ring giant axon, which is implicated in escape locomotion, show patterns of facilitating EPSPs correlated with these potentials. Hair cell input may be important in feeding as well as in locomotory behaviour. Three classes of hair cells were studied : those of the tentacle bases (T cells) and those of the velum (C cells and F cells). Direct stimulation of the tentacle bases shows that the T cells respond to slight mechanical displacements. To distinguish the functions of the C cells and F cells, which lie close together, a laser was used to ablate selectively one or other kind. It was found that the C cells respond to small mechanical displacements of the velum but no response could be assigned to the F cells.

Distribution of Nerve Elements Showing FMRFamide‐like Immunoreactivity in Hydromedusae

AbstractImmunofluorescence tests with antisera to various peptides reveal the presence of a FMRFamide‐like peptide in neurons of hydromedusae belonging to three orders. An avian pancreatic polypeptide‐like peptide may also be present in certain neurons. Distribution of FMRFamide‐like immunoreactivity (Fa‐IR) has been studied in Proboscidactyla (O. Limnomedusae), Phialidium and Aequorea (O. Leptomedusae) and Aglantha (O. Trachymedusae). These findings are compared with results obtained with Polyorchis (O. Anthomedusae) by Grimmelikhuijzen and Spencer [J. comp. Neurol. 230, 361 (1984)]. All species show Fa‐IR neurons in the tentacles linked by interconnecting neurites running in the outer marginal nerve ring. A Fa‐IR plexus is present in the manubrium and, except in Aglantha, this system merges with a plexus associated with the subumbrellar radial muscles. As in Polyorchis, the swimming motor neurons are unstained. In contrast to this species, only a small percentage of the neurites composing the nerve rings are stained. The giant axons of Aglantha show no Fa‐IR. The cnidothylaces of Proboscidactyla contain neurons reacting with antisera to FMRFamide and APP. Present evidence suggests that in hydromedusae Fa‐IR is confined to distinct subsets of neurons. These appear to be either sensory units or units supplying smooth muscles, but they are not involved in the innervation of striated muscles.

Putative extraocellar photoreceptors in the outer nerve ring of Polyorchis penicillatus

AbstractPutative sensory cells have been described in the outer nerve ring of the hydrozoan jellyfish Polyorchis penicillatus that resemble ocellar photoreceptors characteristic of this animal group. Each receptor cell bears an apical structure consisting of verticillate lamellae representing evaginations of ciliary membranes (9 + 2 axonemes). Verticils are grouped in clusters of three or more and are distributed throughout the outer nerve ring. Individual lamellae of each verticil are up to 0.2 μM wide and are frequently reflexly folded to form a tightly packed whorl. Multiple axonemes contribute to each verticil. Each receptor cell body contributes a single axon (up to 1.75 μm in diameter) to the neuronal tracts of the outer nerve ring. The structural similarity between the described cells and hydrozoan photoreceptors, and the close agreement between the location of these cells and the localized photoresponse in Polyorchis and related medusae suggest that the putative receptor cells function as photoreceptors.

FMRFamide immunoreactivity in the nervous system of the medusa Polyorchis penicillatus.

Three different antisera to the molluscan neuropeptide Phe-Met-Arg-Phe-amide (FMRFamide) and two different antisera to the fragment RFamide were used to stain sections or whole mounts of the hydrozoan medusa Polyorchis penicillatus. All antisera stained the same neuronal structures. Strong immunoreactivity was found in neurons of the ectodermal nerve nets of the manubrium and tentacles, in neurons of the sensory epithelium, and in neurons at the periphery of the sphincter muscle. Strong immunoreactivity was also present in processes and perikarya of the whole outer nerve ring, in the ocellar nerves, and in nerve cells lying at the periphery of the ocellus. The inner nerve ring contained a moderate number of immunoreactive processes and perikarya, which were distinct from the swimming motor neurons. In contrast to the situation in the hydrozoan polyp Hydra attenuata, no immunoreactivity was found with several antisera to oxytocin/vasopressin and bombesin/gastrin-releasing peptide. The morphology and location of most FMRFamide-immunoreactive neurons in Polyorchis coincides with two identified neuronal systems, which have been recently discovered from neurophysiological studies.

Microanatomy of the subumbrellar motor innervation in Aglantha digitale (Hydromedusae: Trachylina).

The hydrozoan medusa Aglantha digitale (Müller 1776) has eight syncytical giant motor axons, up to 40 micrometers in diameter, running from the margin, up the inside of the bell towards the apex. Giant motor axons injected with Lucifer Yellow CH are connected with lateral neurons running circumferentially across the subumbrellar muscle. These processes fill with the dye. Bundles of 20 to 50 small dye-coupled neurons extend circumferentially along the margin for up to 0.85 mm. Giant motor axons injected with horseradish peroxidase divided into a few short branches on entering the inner nerve ring. Here the giant motor axon forms both chemical synapses and gap junctions with neurons that also send their axons into the inner nerve ring. In this region the inner and outer nerve ring are connected by axons passing through openings in the intervening mesoglea.

Fine structure studies of the ocelli of Polyorchis penicillatus (Hydrozoa, Anthomedusae) and their connection with the nerve ring

The fine structure of the small compact ocelli (50–100 μm in diameter) of Polyorchis penicillatus is described. The ocellar cup is formed of pigment cells and receptor cells. The pigment cells occur in approximately a 2:1 ratio to the receptor cells. Each pigment cell has a process that may pass through the presumed photosensory region. Pigment cells are connected to adjacent receptor cell processes by septate junctions. The sensory cells are bipolar with the apical part forming the receptor process and the basal part forming an axon 8–15 μm long and 1–2 μm in diameter. Each receptor cell axon forms a synapse with a single second order neuron but the sensory cells are also connected to the second order neurons postsynaptically. There are also synapses between adjacent second order neurons. The second order neurons lie outside the ocellar cup, next to the tentacular mesogloea. Each second order neuron forms an axon of about 1 μm thickness. The axons on each side group together to form an optic nerve having 30–40 axons that travel around the tentacle base on either side and enter the outer nerve ring independently.

The giant axon escape system of a hydrozoan medusa, Aglantha digitale.

ABSTRACT Aglantha digitale (Hydrozoa) has a ring giant axon, up to 35 μm in diameter, which runs all round the margin of the bell in the outer nerve-ring. Running from the margin, up the inside of the bell towards the apex are eight motor giant axons, up to 40 μm in diameter. These synapse with the sub-umbrellar myoepithelium and are therefore motoneurones. The myoepithelial cells line the inside of the bell and have striated muscular tails running circumferentially. Contraction of this circular musculature forces water out of the bell and propels the medusa through the water. A large axon (up to 7 μm in diameter) runs on the aboral side of each tentacle. The tentacles retract during swimming and contain longitudinally aligned striated muscle tails. Intra- and extracellular recordings from the giant axons indicate that they are involved in the rapid escape swimming response of Aglantha. Stimuli which evoke escape swimming lead to a brief burst of two to six impulses (2–3 ms in duration) in the ring giant axon. These propagate round the margin (at up to 2·6 m s−1). The large tentacle axons fire one-to-one with the ring giant axon and the tentacles contract. The motor giant axons are excited at chemical synapses (blocked by divalent cations and with a synaptic delay of 1·6 ± 1 ms), where large, facilitating epsps evoke an impulse which then propagates up the bell at up to 4 m s−1. The overshooting motor giant axon impulses are Na+ dependent, 2–3 ms in duration, and excite the myoepithelium at synapses with a 1·6 ± 1 ms synaptic delay. A regenerative muscle impulse is evoked. It is of long duration (15–70 ms), Ca2+ dependent, overshoots zero, and propagates through the myoepithelium at 0·22 to 0·29 ms−1. A nearly synchronous contraction of the whole subumbrellar circular musculature is evoked. Isolated strips of muscle can reach peak tension in 40 ms. In the absence of sarcoplasmic reticulum, Ca2+ for excitationcontraction coupling probably enters the muscle during the impulse. This preparation has allowed a clear understanding of how the two types of axon co-ordinate escape behaviour. It should also be valuable for the study of synaptic transmission, previously very inaccessible in Cnidarians.

Neurobiology of Polyorchis. II. Structure of effector systems.

The gross and fine morphology of the major effector systems in the anthomedusan, Polyorchis penicillatus, is described and discussed in relation to the known physiological and behavioral properties of these systems. Swimming is controlled by an anastomosing network of giant neurons within the inner nerve ring and radial nerves. Although these neurons may be coupled by gap junctions it is likely that they form a syncytium. The photosensitivity of the "giants" is attributed to reflexive membranes within the cytoplasm. Giant neurons act as both the pre- and postsynaptic cell when forming synapses with other neurons of the inner nerve ring. Neuromuscular synapses between "giants" and the striated swimming muscle are found around the margin and along the radii. Swimming muscle cells are connected laterally by gap junctions and end-to-end by desmosomes which are sometimes elaborated with extra-thick filaments. Unstriated sphincter and radial muscles, the major muscles associated with crumpling, are both greatly folded over mesogloeal ridges and have processes that cross the mesogloea to contact the ring and radial canals, respectively. Synapses or other sites that might be responsible for exciting these muscles during crumpling have not been found. The ability of the endodermal lamella and canals to propagate action potentials can be accounted for by the numerous gap junctions that are seen in these tissues. The precise location where excitation is transferred to the nervous system to initiate crumpling is not known but epithelial bridges crossing the mesogloea are likely routes. Synapses between neurons originating in the outer nerve ring and tentacle longitudinal muscle can account for the control of tentacle length. Neurons of the outer nerve ring also synapse onto velar, radial fibers and the sphincter muscle. The inner and outer nerve rings have nervous connections. The organisation of the outer nerve ring and the arrangement of nerves within the endodermal plexus is described. A diagram showing the major connections and interactions of components of the effector systems is presented.

Locomotion and neuromuscular system of Aglantha digitale.

Aglantha digitale swims in two ways: a slow rhythmical swim typical of hydromedusae in general and a sudden rapid movement that appears to be an escape response. The swimming musculature is an extremely well developed striated circular muscle layer that possesses a sarcoplasmic reticulum. The nervous system of this species can be divided into three units: an inner nerve ring and an outer nerve ring, which are joined by unusually large transmesogleal pathways, a group of giant axons that extends over the surface of the swimming muscle, and the radial canal. Well developed ciliated sensory cells are located on the exumbrellar surface of the margin. Consideration of these properties of the organisation of this species suggests that normal slow swimming is controlled by a mechanism similar to that found in other medusae, while the escape response is the result of the action of the giant axons.