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Neuropeptides are conserved metazoan signaling molecules, and represent useful markers for comparative investigations on the morphology and function of the nervous system. However, little is known about the variation of neuropeptide expression patterns across closely related species in invertebrate groups other than insects. In this study, we compare the immunoreactivity patterns of 14 neuropeptides in three closely related microscopic dinophilid annelids (Dinophilus gyrociliatus, D. taeniatus and Trilobodrilus axi). The brains of all three species were found to consist of around 700 somata, surrounding a central neuropil with 3-5 ventral and 2-5 dorsal commissures. Neuropeptide immunoreactivity was detected in the brain, the ventral cords, stomatogastric nervous system, and additional nerves. Different neuropeptides are expressed in specific, non-overlapping cells in the brain in all three species. FMRFamide, MLD/pedal peptide, allatotropin, RNamide, excitatory peptide, and FVRIamide showed a broad localization within the brain, while calcitonin, SIFamide, vasotocin, RGWamide, DLamide, FLamide, FVamide, MIP, and serotonin were present in fewer cells in demarcated regions. The different markers did not reveal ganglionic subdivisions or physical compartmentalization in any of these microscopic brains. The non-overlapping expression of different neuropeptides may indicate that the regionalization in these uniform, small brains is realized by individual cells, rather than cell clusters, representing an alternative to the lobular organization observed in several macroscopic annelids. Furthermore, despite the similar gross brain morphology, we found an unexpectedly high variation in the expression patterns of neuropeptides across species. This suggests that neuropeptide expression evolves faster than morphology, representing a possible mechanism for the evolutionary divergence of behaviors., (© 2017 Wiley Periodicals, Inc.)

The anterior sensory anatomy (not including amphids) of the nematode Aphelenchus avenae (Tylenchomorpha) has been three-dimensionally reconstructed from serial, transmission electron microscopy thin sections. Models, showing detailed morphology and spatial relationships of cuticular sensilla and internal sensory receptors, are the first computerized reconstruction of sensory structures of a Tylenchomorpha nematode. Results are analyzed with respect to similarly detailed reconstructions of Rhabditida outgroup nematodes, Acrobeles complexus (Cephalobomorpha) and Caenorhabditis elegans (Rhabditomorpha). Homologies identified in A. avenae demonstrate the general conservation of the anterior sensory system between freeliving nematodes and the largely plant parasitic Tylenchomorpha. A higher degree of similarity is shown between A. avenae and A. complexus, with common features including: the presence of a second, internal outer labial dendrite (OL1); a second cephalic dendrite in the female (CEP2/CEM); an accessory process loop of inner labial dendrite 1; and terminus morphology and epidermal associations of internal sensory receptors BAG and URX. Unique to A. avenae is a pair of peripheral, lateral neurons of unknown homology but with axial positions and intercellular relationships nearly identical to the "posterior branches" of URX in A. complexus. Knowledge of homologies and connectivity of anterior sensory structures provides a basis for expansion of the experimental behavioral model of C. elegans to the economically important nematodes of Tylenchomorpha.

Periviscerokinins (PVKs) and pyrokinins (PKs) are neuropeptides known in several arthropod species. Sequence homology of these peptides with the molluscan small cardioactive peptides reveals that the occurrence of PVKs and PKs is not restricted to arthropods. Our study focuses on the biochemical and immunocytochemical identification of neuropeptides with sequence homology to PVKs and PKs in the central and peripheral nervous system of the earthworm Eisenia fetida. By means of affinity chromatography, nanoflow liquid chromatography, and high accuracy mass spectrometry, six peptides, SPFPR(L/I)amide, APFPR(L/I)amide, SPLPR(L/I)amide, SFVR(L/I)amide, AFVR(L/I)amide, and SPAFVR(L/I)amide, were identified in the central nervous system with the common -XR(L/I)amide C-terminal sequence. The exact anatomical position of 13 labeled XR(I/L)amide expressing neuron groups and numerous peptide-containing fibers were determined by means of immunocytochemistry and confocal laser scanning microscopy in whole-mount preparations of ventral nerve cord ganglia. The majority of the stained neurons were interneurons with processes joining the distinct fine-fibered polysegmental tracts in the central neuropil. Some stained fibers were seen running in each segmental nerve that innervated metanephridia and body wall. Distinct groups of neurosecretory cells characterized by small round soma and short processes were also identified. Based on immunoelectron microscopy six different types of labeled cells were described showing morphological heterogeneity of earthworm peptides containing elements. Our findings confirm that the sequence of the identified earthworm neuropeptides homologous to the insect PVKs and PKs suggesting that these peptides are phylogenetically conservative molecules and are expressed in sister-groups of animals such as annelids, mollusks, and insects.

We used transmission electron microscopic montages to examine the composition of nerve bundles serving the infrared pit organs of two species of crotaline snakes, Agkistrodon blomhoffii and A. brevicaudus. In the three main bundles, the myelinated fibers totaled 2,200-3,700, and unmyelinated fibers 2,400. We also discovered for the first time two accessory bundles composed almost entirely of unmyelinated fibers running alongside the main bundles, containing an average total of 3,300 unmyelinated fibers vs. an average of 10 myelinated fibers. Thus, the average total of unmyelinated fibers was nearly twice that of myelinated fibers. To study the nature of the unmyelinated fibers, we did double staining immunohistochemistry with antibodies for substance P (SP) and vasoactive intestinal polypeptide (VIP) in combination with and without capsaicin pretreatment. SP and VIP immunoreactive varicose fibers ran straight toward the center of the pit membrane in parallel with arterioles and venules, and also formed a dense network around the periphery of the membrane. There were three types of fibers: fibers containing only SP, fibers containing only VIP, and fibers containing both peptides. SP-only fibers were distributed singly throughout the pit membrane and in small bundles around the periphery. SP+VIP fibers were distributed sparsely in the pit membrane and around its periphery. VIP-only fibers were distributed throughout the pit membrane and were of smaller diameter than SP and SP+VIP fibers. After treatment with capsaicin, most of the three types of varicose fibers disappeared from the central part of the pit membrane, but those around the periphery remained unaffected. The capsaicin-sensitive fibers may be unmyelinated sensory types, and the unaffected ones may be autonomic nerve fibers., (Copyright 2002 Wiley-Liss, Inc.)

The antennules of decapod crustaceans are covered with thousands of chemosensilla that mediate odor discrimination and orientation behaviors. Most studies on chemoreception in decapods have focused on the prominent aesthetasc sensilla. However, previous behavioral studies on lobsters following selective sensillar ablation have revealed that input from nonaesthetasc antennular chemosensilla is sufficient for many odor-mediated behaviors. Our earlier examination of the setal types on the antennules of the Caribbean spiny lobster Panulirus argus revealed three types of nonaesthetasc chemosensilla. The most abundant and widely distributed of these is the hooded sensillum. The present study describes the detailed ultrastructure of antennular hooded sensilla and the physiological response properties of their receptor neurons. Light and scanning and transmission electron microscopy were used to examine structural characteristics, and electrophysiology was used to examine single-unit responses elicited by focal chemical and mechanical stimulation of antennular hooded sensilla. Hooded sensilla have a porous cuticle and are innervated by 9-10 chemosensory and 3 mechanosensory neurons whose dendrites project to the distal end of the sensillum. Hooded sensillar chemosensory neurons responded to waterborne chemicals, were responsive to only one of the six tested single compounds, and had different specificities. Hooded sensillar mechanosensory neurons were not spontaneously active. They had low sensitivity in that they responded to tactile but not waterborne vibrations, and they responded to sensillar deflection with phasic bursts of activity. These results support the idea that hooded sensilla are bimodal chemo-mechanosensilla and are receptors in an antennular chemosensory pathway that parallels the well-described aesthetasc chemosensory pathway., (Copyright 2002 Wiley-Liss, Inc.)

The nervous system of temnocephalid flatworms consists of the brain and three pairs of longitudinal connectives extending into the trunk and tail. The connectives are crosslinked by an invariant number of regularly spaced commissures. Branches of the connectives innervate the tentacles of the head and the sucker organ in the tail. A set of nerve rings encircling the pharynx and connected to the brain and connectives constitute the pharyngeal nervous system. The nervous system is formed during early embryogenesis when the embryo represents a multilayered mesenchymal mass of cells. Gastrulation and the formation of separate epithelial germ layers that characterize most other animal groups are absent. The brain arises as a bilaterally symmetric condensation of postmitotic cells in the deep layers of the anterior region of the embryonic mesenchyme. The pattern of axon outgrowth, visualized by labeling with anti-acetylated tubulin (acTub) antibody, shows marked differences from the pattern observed in other flatworm taxa in regard to the number of neurons that express the acTub epitope. Acetylated tubulin is only expressed in neurons that form long axon tracts. In other flatworm species, such as the typhloplanoid Mesostoma and the polyclad Imogine, which were investigated by us with the acTub antibody (Hartenstein and Ehlers [2000] Dev. Genes Evol. 210:399-415; Younossi-Hartenstein and Hartenstein [2000] Dev. Genes Evol. 210:383-398), only a small number of "pioneer neurons" become acTub positive during the embryonic period. By contrast, in temnocephalids, most, if not all, neurons express acTub and form long, large-diameter axons. Initially, the brain commissure, pharyngeal nerve ring, and the connectives are laid down. Commissural tracts and tentacle nerves branching off the connectives appear later. We speculate that the precocious differentiation of the nervous system may be related to the fact that temnocephalids move by muscle action, and possess a massive and complex muscular system when they hatch. In addition, they have muscular specializations such as the anterior tentacles and the posterior sucker that are used as soon as they hatch. By contrast, juveniles of Mesostoma and larvae of polyclads move predominantly by ciliary action that may not require a complex neural circuitry for coordination., (Copyright 2001 Wiley-Liss, Inc.)

Morphologic and functional parameters altered during neuronal death were investigated in chick ciliary ganglion (CG) neurons in vitro isolated from embryonic day 8 (E8, stage 34). Neurons were separated from nonneuronal cells to investigate their inherent cell death program and were cultured with or without trophic support (choroid, iris, pigment epithelium) from their appropriate target tissue. The cell death process was characterized with investigations focused on the earliest events at the onset of commitment to cell death at 11 hours after plating. Initial morphologic changes in the process of cell death were cytoplasmic; swelling, dendritic retraction, blebbing, vacuolization, which are all characteristics of necrosis. Later, nuclear chromatin condensation occurred, a characteristic of apoptosis. An increase in membrane permeability was measured earlier at 8 hours. During these alterations (associated with the initiation of cell death) single cell analysis was performed to evaluate mobile Ca(2+) changes in the same trophically deprived neurons during the course of the death process; Ca(2+) levels remained at 50 nM. Transient Ca(2+) entry was buffered in control and deprived cells at 13 hours but with different parameters. During the execution stage of death mobile Ca(2+) levels were variable. In this final stage of cell death, neurons demonstrated nuclear damage, cytosol disintegration, or morphology sharing both characteristics. These observations define embryonic CG cell death in vitro as neither purely apoptotic nor necrotic but a form of that exhibits features of both. These results also demonstrate a disassociation of changes in cytosolic Ca(2+) from both CG neuronal survival and trophically deprived cell death in vitro., (Copyright 2000 Wiley-Liss, Inc.)

When infective larvae of Haemonchus contortus (a highly pathogenic, economically important, gastric parasite of ruminants) are ingested by grazing hosts, they are exposed to environmental changes in the rumen, which stimulate resumption of development. Presumably, resumption is controlled by sensory neurons in sensilla known as amphids. Neuronal function can be determined by ablation of specifically recognized neurons in hatchling larvae (L1) in which neuronal cell bodies are easily visualized using differential interference microscopy. Using three-dimensional reconstructions from electron micrographs of serial transverse sections, amphidial structure of the L1 is described. Each amphid of H. contortus is innervated by 12 neurons. The ciliated dendritic processes of 10 neurons lie in the amphidial channel. Three of these end in double processes, resulting in 13 sensory cilia in the channel. One process, that of the so-called finger cell, ends in a number of digitiform projections. Another specialized dendrite enters the amphidial channel, but leaves it to end within the sheath cell, a hollow, flask-shaped cell that forms the base of the amphidial channel. Although not flattened, this process is otherwise similar to the wing cells in Caenorhabditis elegans; we consider it AWC of this group. Two other neurons, ASA and ADB, appear to be homologs of wing cells AWA and AWB in C. elegans, although they end as ciliated processes in the amphidial channel, rather than as flattened endings seen in C. elegans. Each of the 12 amphidial neurons was traced to its cell body in the lateral ganglion, posterior to the worm's nerve ring. The positions of these bodies were similar to their counterparts in C. elegans; they were named accordingly. A map for identifying the amphidial cell bodies in the living L1 was prepared, so that laser microbeam ablation studies can be conducted. These will determine which neurons are involved in the infective process, as well as others important in establishing the host-parasite relationship.

Early embryonic development of the nervous system of a lamprey, Lampetra japonica, was studied by using immunohistochemical techniques and by scanning electron microscopy. The earliest appearance of axons was detected at Tahara's stage 21-, when dorsolateral and ventral longitudinal fasciculi were present in the hindbrain and spinal cord regions. The branchiomeric nerve roots began to appear at stage 22; the fibers were joined to the dorsolateral fasciculus proximally and also extended distally into each pharyngeal arch. The anterior neural tube was divided into several neuromeres: the mid-hindbrain sulcus became apparent first, then the portion rostral to this sulcus was subdivided into two portions by the syn-parencephalic boundary. In the hindbrain around stage 23, rhombomeres developed transiently, of which, rhombomere 4 was the most distinctive. Putative crest cells forming the octavofacial nerve root anlage were selectively adhering to rhombomere 4, whereas no crest cells were found on rhombomere 3. The assignment of the crest-derived nerve anlage to rhombomeres is conserved between gnathostomes and L. japonica. The neuromerical scheme of the neural tube of L. japonica is also mostly in accordance with that in gnathostomes, sharing the basic developmental patterning of axon bundles at early developmental stages. The most distinct difference between these two groups is the topographical relationships between the hindbrain neuraxis and pharyngeal arches, as well as the otic placode.

The impact of increased levels of skin-derived nerve growth factor (NGF) neurotrophin on sensory and sympathetic innervation to the mouse mystacial pad and postero-orbital vibrissae was determined. Consistent with an approximate doubling of neuron number in trigeminal and superior cervical ganglia, many components of the sensory and sympathetic innervation were substantially enhanced. Although the increased number of neurons raised the possibility that all types of innervation were increased, immunohistochemical analysis indicated that enhanced NGF production had a differential effect upon sensory innervation, primarily increasing unmyelinated innervation. This increased innervation occurred in specific locations known to be innervated by small, unmyelinated fibers, suggesting that NGF modulated sensory innervation density, but not targeting. In contrast, sympathetic innervation was not only increased but also was distributed to some aberrant locations. In the intervibrissal fur of the mystacial pad, both the number of sensory axons and branches appeared increased, whereas in vibrissal follicle sinus complexes, only branching increased. In some areas, sensory ending density was lower than expected based upon the size of the source nerve bundles suggesting that many axons and branches were surviving but failing to form functional endings. Furthermore, the immunochemical profile of innervation was altered in some sensory populations as demonstrated by the coexistence of RT-97 neurofilament labeling in calcitonin gene-related peptide (CGRP) positive axons, by the loss of substance P colocalization in some CGRP axons, and by an absence of neuropeptide Y labeling in tyrosine hydroxylase positive sympathetic axons. Collectively, these results indicate that the NGF mediated increase in neuron number may be selective for particular sets of innervation and that increases among some populations may result from phenotypic switching.

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and subsequent immunoblotting of neuronal membrane proteins derived from thoracic ganglia of adult Locusta and Schistocerca reveal that a polyclonal antiserum raised against the Locusta nicotinic acetylcholine receptor (nAChR), binds strongly to an identical polypeptide band corresponding to 65 kDa in both locust species. This polyclonal antiserum was used to analyze the distribution of antigenic sites within the developing thoracic central nervous system of Schistocerca embryos. Axonal outgrowths from the earliest differentiated neurons are first labeled between 30% and 35% development. By 40% to 45% development, labeled granules appear in the cytoplasm of neuronal cell bodies. When the developing neuropil is first enclosed at approximately 45% to 50% development, it appears uniformly labeled, but by 55% development, unlabeled areas appear that represent the sites of future tracts and commissures. By 75%, an adult pattern of neuropil immunogenicity is established in which synaptic regions are stained but tracts and commissures are not. This suggests that during the early development of the thoracic nervous system nAChR-like antigenic sites are evenly distributed, but later become concentrated in the developing synaptic areas.

Crustacean cardioactive peptide-immunoreactive neurons have been mapped in whole-mount preparations and sections of the ventral nervous system of the crayfish Astacus astacus and Orconectes limosus. Based on their morphology, projection patterns, and staining characteristics, two types of contralaterally projecting neurons are individually identifiable. In both species, these neurons occur in all neuromers as apparent serial homologs. In adult specimens, one type of cell has a small, densely stained dorsal lateral perikaryon, and a descending axon, and appears to be an interneuron. Each neuromer contains a single pair of these cells. Only in maxillary ganglia, these cells may have an additional ascending projection. The other type, a neurosecretory cell, has a larger, weakly stained perikaryon and a projection to the segmental third root of the next anterior neuromer. All neuromers contain a single pair of these neurons adjacent to the interneurons except for the abdominal neuromers, which contain two pairs of the neurosecretory cells. Central arborizations and varicose processes toward the surface of the third roots and within the perineural sheath of the ventral nerve cord arise from these neurons. Electron microscopy of granule-containing terminals substantiated that these newly discovered extensive neurohemal areas are release sites for the peptide. In young immature specimens, the perikarya of both neuron types do not differ in size. Additional weakly stained small perikarya occur in all neuromers of Astacus astacus. These two types of crayfish neurons and other comparable aminergic and peptidergic neurons of crayfish and lobster are differentially distributed in the ventral cord. Furthermore, comparison of similar neuron types in crab, locust, meal worm, and moth species indicates intra- and interphyletic structural homologies.

Neural connections between the mushroom body (MB) and other protocerebral areas of the honeybee's brain were studied with the help of cobalt chloride and Golgi staining methods. Focal injections of cobalt ions into the alpha-lobe neuropil of the MB reveal seven clusters of somata located in the protocerebrum and deutocerebrum of each brain hemisphere. These neurons connect the mushroom body neuropil with protocerebral areas and number approximately 400. They contact the layered organization of the alpha-lobe at different locations. Some project not only into the alpha-lobe, but also into the beta-lobe and pedunculus neuropils. Fifteen cell types which form intraprotocerebral circuits are morphologically described. They can be divided into three categories: 1) unilateral neurons, with projection fields restricted to the ipsilateral protocerebrum; these neurons connect the alpha-lobe with areas in the protocerebral lobe and ramify with densely layered arborisations arranged perpendicularly to the longitudinal axis of the alpha-lobe; 2) recurrent neurons, which interconnect subcompartments of the MB, forming loops at different levels of the neuropil; their arborisations are mainly restricted to the alpha-lobe, beta-lobe, pedunculus, and calyces of the ipsilateral MB; they also ramify sparsely around the neuropil of the alpha-lobe; and 3) bilateral neurons, which either interconnect both alpha-lobes or connect the ipsilateral alpha-lobe and protocerebral lobe with the dorsolateral protocerebral lobe of the contralateral hemisphere. The connections of different compartments of the MB with other parts of the protocerebrum as revealed in this study are discussed in the context of hypotheses about the functional role of MBs in the honeybee brain.

The size and shape of vesicles at junctional appositions of the rectifying electrical synapses between the medial giant fibre and motor giant neurone of the crayfish were measured during the first 2 months after hatching. Summed data over this period reveal a bimodal distribution in vesicle diameter. From the day of hatching until about 7 days of age, small vesicles (circa 25 nm diameter) predominate. From day 7 onwards, larger vesicles (circa 55 nm diameter) occur in increasing numbers, until at day 56 they constitute about 85% of the population at any one junctional apposition. At intermediate ages (day 7-28) individual junctional appositions may show the same bimodal distribution in size as does the age group as a whole, indicating that large and small vesicles occur together at the same junction. The larger vesicles are mainly circular, while the small vesicles are pleomorphic, with shapes ranging from almost circular down to a shape factor of about 0.6.

The phenomenon of postlarval cell addition to the peripheral nervous system of fish has been reported for some sensory systems, but has yet to be characterized for the gustatory system. Many fishes, such as catfish, possess taste buds scattered across their body surface, and presumably, the number of taste buds increases during growth of the animal. The present study was undertaken in order to examine the process of growth in the peripheral gustatory system and to determine whether the degree of convergence of receptors onto primary sensory afferents changes during growth. The recurrent facial nerve of channel catfish was used for these studies since this nerve contains no general cutaneous components and innervates taste buds along the fish's body surface. Electron micrographs were made of cross sections of this nerve taken from individuals ranging in size from 5.1 to 39.5 cm standard length. In addition, estimates were made of the number of taste buds innervated by this nerve by determining taste bud density along selected regions of the flank and fins in large and small fish. As catfish get larger, the number of both myelinated and unmyelinated axon profiles in the recurrent facial nerve increases, but at a slower rate than the number of taste buds innervated by this nerve. Thus, on average, the number of taste buds innervated by each fiber increases as the fish enlarges; on average there are 2 taste buds per axons profile in small fish and nearly 14 taste buds per axon profile in large fish. The rate of addition of new axon profiles to the nerve is estimated at roughly 70 per day over the range of sizes studied. Although generation of new ganglion cells and axons may contribute to this increase, several lines of evidence indicate that axonal branching occurs. In addition, the mean axon diameter for both myelinated and unmyelinated axons increases during postlarval growth. The finest myelinated fibers (0.2 micron) in small animals were significantly smaller than the finest myelinated fibers (0.7 micron) in larger animals.

Lateral-to-motor and medial-to-motor synapses in crayfish nerve cords are composed of an electrical and a chemical component. The presynaptic terminals showed localized clusters of synaptic vesicles, electron-dense areas, coated pits, and coated vesicles. In thin sections, active zones were defined by electron-dense regions where synaptic vesicles attached and, in freeze-fracture replicas, by clusters of intramembrane particles localized in bands with vesicle openings on the sides of these bands. The cytoplasmic surface of the postsynaptic membrane opposite the active zones was coated with electron-dense material that in freeze-fracture replicas was seen as an increase in intramembrane particles located in the external leaflet (EF-face). This specialization of the postsynaptic membrane may correspond to the neurochemical receptor. Also, pre- and postsynaptic membranes were separated by a wider extracellular gap than those of adjacent nonsynaptic regions and electrical synapses or gap junctions. Synaptic vesicles were located exclusively at the synaptic regions by means of a cytoskeleton that was different for the electrical and the chemical components. The vesicles associated with the electrical component were anchored to a cytoskeleton composed of a beaded layer of densities located parallel to the membrane. This cytoskeleton maintained the synaptic vesicles separated from the presynaptic membrane by a distance of 13 +/- 2 nm. The synaptic vesicles associated with the chemical component were anchored to electron-dense regions formed by filaments arranged in bundles, anchored to the presynaptic membrane. Vesicles lined both sides positioned to discharge their contents into the extracellular space and to replace the discharged vesicles.

Cytophotometric analyses of Feulgen-stained nuclei present in homogenates of vertical and subesophageal lobes of octopus brain have shown that the latter region contains larger nuclei with up to several times the amount of DNA present in vertical nuclei. No obvious relationship was found between DNA content and nuclear size. Except for a rather small minority, nuclei of the vertical lobe have a uniform size and the expected diploid amount of DNA. These parameters are not substantially dependent on body weight. In contrast, the DNA content of subesophageal nuclei increases progressively with body weight. The amount of DNA found in subesophageal nuclei does not seem to be a simple multiple of the diploid or haploid value.

Neuroanatomical and ultrastructural studies of the paired lateral giant nerve fibers (LGFs) in posterior segments of the tubificid worm, Branchiura sowerbyi, demonstrate that the fibers are 1) segmental in origin (two cell bodies per segment), 2) joined longitudinally and transversely to form an intersegmental syncytial network, and 3) surrounded by a myelinlike sheath. The LGFs are unique among paired giant fiber systems because of their extreme asymmetry, the diameter of the left fiber being several times greater than that of the right. Electrophysiological studies demonstrate that the small, right fiber has a high input resistance and, during mechanosensory stimulation, functions as the locus for LGF spike initiation. The larger, left fiber contributes by enhancing the speed of LGF spike conduction along the animal. One physiological benefit of this asymmetric arrangement may be optimization of escape reflex sensitivity to mechanosensory inputs.

To determine the organizational principles underlying the peripheral electrosensory nervous system of weakly electric gymnotiform teleosts we labelled each of the four anterior lateral line nerve branches with HRP. We determined the position of labelled cell bodies within the ganglion and followed anterogradely filled fibers to their termination sites in one of the four somatotopic maps in the electroreceptive lateral line lobe (ELL). Within the ganglion, cell bodies exhibit a loose somatotopy based on nerve branch position: trunk electroreceptors have their cell bodies located in the caudal ganglion; cell bodies to the head receptors are rostral. Cell bodies to the head exhibit a rough dorsoventral polarity, supraorbital cells tend to be located dorsally, infraorbital cells centrally, and mandibular cells ventrally. Despite this general somatotopy there is substantial overlap (up to 30%) of cell bodies among regions. There appears to be no rostrocaudal topography within nerve branch regions. Iontophoretic WGA-HRP injected into the medial segment of the ELL retrogradely labelled cell bodies that innervate ampullary organs. These cell bodies were dispersed throughout the ganglion, indicating that cell bodies do not cluster by receptor type. Peripherally directed axons from the ganglion appear to undergo an active reorganization in order to form the nerve branches. Within nerve branches, axons to a particular area of skin do not cluster together. Centrally from the ganglion, axons retain the position of their cell body until they reach the ELL border. Once in the ELL, fibers become sorted in the deep fiber layer according to receptor type and the map they terminate in. This reorganization involves rearrangement of fascicles and axons within fascicles. In toto, proceeding from peripheral to central, the electrosensory periphery loses at least a portion of its receptor topography in the distal nerve and ganglion and then acquires both a functional and somatotopic organization after reaching the ELL; conceptually it is torn down and rebuilt again. From an ontogenetic perspective, axonal growth occurs from the ganglion outward; the fact that ganglion cell bodies are not highly organized while the receptors they innervate and their central processes are suggests that active axonal guidance mechanisms are involved.

Central distribution of afferent and efferent components of the pudendal nerve was examined in the cat by the HRP method after applying HRP to the central cut end of the pudendal nerve. Retrogradely labeled neuronal cell bodies were located primarily in the feline homologue of the Onuf's X nucleus, constituting a slender longitudinal cell column in the ventral horn of the S1 and S2 cord segments. The Onuf's nucleus was present constantly from middle S1 to high S2 cord segments, and occasionally extended rostrally to high S1 or low L7, and caudally to middle S2, low S2, or high S3 cord segments. No sex differences were observed in the distribution pattern, number, and soma size of labeled neurons in the Onuf's nucleus. Transganglionically labeled dorsal root fibers were found to terminate ipsilaterally in the lamina I of the dorsal horn at levels of lower lumbar, sacral, and higher coccygeal cord segments and the gracile nucleus, and bilaterally with an ipsilateral predominance in the dorsal commissural gray and laminae III, IV, V, and VI of the dorsal horn at levels of lower lumbar, sacral, and higher coccygeal cord segments. Some labeled dorsal root fibers appeared to end ipsilaterally in the regions where the sacral parasympathetic preganglionic neurons have been shown to be located.

The distribution of FMRFamidelike peptides was studied in the nervous system of the lobster Homarus americanus by using immunocytochemical and radioimmunological techniques. By radioimmunoassay FMRFamidelike immunoreactivity (FLI) was found in low levels (ca. 1 pmol/mg protein) throughout the ventral nerve cord and in much higher amounts (60-100 pmol/mg protein) in the neurosecretory pericardial organs. Immunocytochemical studies showed FLI in approximately 300-350 cell bodies, and in distinct neuropil regions, neuronal fiber tracts, and varicose endings. Specificity of the immunostaining was tested by preabsorbing the antiserum with FMRFamide, with peptides having similar carboxyl termini to FMRFamide (Met-enkephalin-Arg-Phe, Phe-Met-Arg-Tyr-amide), with several amidated peptides (alpha-melanocyte-stimulating hormone, substance P, oxytocin), and with proctolin, a peptide found widely distributed in the lobster nervous system. Of these substances, only FMRFamide blocked the staining. In addition to the pericardial organs, significant levels of FLI were found in neurosecretory regions associated with thoracic second roots and in the connective tissue sheath that surrounds the ventral nerve cord. In all three regions, immunocytochemical studies showed the FLI to be localized to fine fibers and associated terminal varicosities lying close to the surface of the tissue, with no obvious target in their immediate vicinity. When examined at the ultrastructural level, the immunoreactive varicosities of the thoracic second roots and of the ventral nerve cord sheaths were found a few microns from the surface of the tissue and contained electron-dense granules. In the immunoreactive nerve cord sheath endings, in addition to the large, dense granules, small, clear vesicles were found. The appearance and location of these terminals suggest a neurohormonal role for FMRFamidelike peptides in lobsters. The observation that low levels of FLI are found in the hemolymph supports this suggestion. In addition, the localization of FLI to particular neuronal somata, fiber tracts, and neuropil regions suggests possible functional roles for these peptides in (1) integration of visual and olfactory information, (2) function of the anterior and posterior gut, and (3) the control of exoskeletal muscles.

The pentapeptide proctolin (H-Arg-Tyr-Leu-Pro-Thr-OH) is a well-studied bioactive substance in insects. With an antiserum against proctolin we have mapped proctolinlike-immunoreactive (PLI) neurons in the nervous system of the blowfly Calliphora erythrocephala. In the brain, including the suboesophageal ganglia, 80-90 neurons were found to be PLI. A further 200-250 PLI neurons innervate the lobula of the optic lobe. The thoracic ganglia contain 100-130, and the abdominal ca. 60 PLI neurons. In the brain and ventral ganglia the immunoreactive neurons are of different types: interneurons, efferents (possibly some motorneurons), and neurosecretory cells. Some of these neurons are individually identifiable; others can be identified collectively as clusters. Identifiable neurons innervate protocerebral neuropil associated with the pars intercerebralis and the beta-lobes of the mushroom bodies as well as tritocerebral neuropil. Some of the prominent clusters innervate the central body of the protocerebrum, tritocerebrum, and possibly leg motor neurons. One abdominal cluster is of special interest because it consist of efferent neurons with processes in the lateral abdominal nerves. Some of these processes are located in the neural sheath in neurohaemal regions, and electron microscopy demonstrates that their terminals are outside the blood-brain barrier. The PLI processes in the protocerebrum contain large granular vesicles and form chemical synapses with different kinds of nonimmunoreactive neural elements. Thus, in Calliphora the proctolinlike substance may be used as a central transmitter/modulator, a neuromuscular transmitter, and a neurohormone released into the circulation.

As demonstrated with immunocytochemistry, specific cells and axons in the nervous system of female Aedes aegypti contain antigens immunologically related to FMRFamide (phenylalanine-methionine-arginine-phenylalanine-amide) and locust adipokinetic hormone I (AKH). In the supra-esophageal ganglion, including some medial neurosecretory cells, and in all ganglia of the ventral nerve cord, there are 100-120 cells immunoreactive to a FMRFamide antiserum. The same cells cross-react with a bovine pancreatic polypeptide antiserum, but when the latter antiserum is preabsorbed with FMRFamide, immunoreactivity is lost. However, immunoreactivity is maintained when FMRFamide antiserum is preabsorbed with pancreatic polypeptide, suggesting that the immunoreactive peptide is more closely related to FMRFamide. There are 6-12 cells in the supra- and subesophageal ganglia immunoreactive to an AKH antiserum, and some of the same cells are reactive to the FMRFamide antiserum. As well, unpaired cells in each of the abdominal ganglia are positive for both AKH and FMRFamide. Although the function of the FMRFamide- and AKH-like peptides in mosquitoes is unknown, this study, combined with previous reports on the localization of FMRFamide-like peptides in midgut endocrine cells, supports the concept of a brain-midgut neuroendocrine axis in this insect.

The ganglion of Cephalodiscus gracilis M'Intosh 1882 is entirely intraepithelial and located in the dorsal epidermis immediately behind the tentacular apparatus that is formed by the mesosome (collar). A characteristic feature of the ganglion is a well-developed neuropile in which different types of nerve fibres can be discerned, many of which contain small granules with electron-dense contents. There are no glia-like cells in association with these fibres. Only slender basal processes of epidermal epithelial cells traverse the neuropile. In the depth of the epithelium the neuropile borders the epidermal basal lamina; apically it is covered by a layer of cell bodies, the majority of which belong to what appear to be ordinary ciliated epidermal cells. Besides these epidermal cells the perikarya of two additional types of cells, which are considered to be neurons, can be discerned. One type is characterised by many rough endoplasmic reticulum cisterns and mitochondria, the other by abundant small, electron-dense granules. The nuclei of these cells are comparatively pale and contain a prominent nucleolus. The neuron cell bodies do not form a distinct layer; but they are loosely distributed somewhat deeper than those of the ordinary epidermal cells. They probably send off an apical process to the epidermal surface and a basally directed one into the neuropile. The ganglion has been compared to the nervous systems in cnidarians, some spiralians, and especially other hemichordates, echinoderms, and chordates; it is found to be of primitive rather than degenerate nature. Furthermore, the possible functional significance of its close connection to the food-capturing tentacular apparatus is discussed.

All neuronal cells in the dwarf male of the dimorphic polychaete species Dinophilus gyrociliatus were individually identified by means of serial ultrathin sections. Altogether 68 neural cells--including 40 sensory neurons and 2 glial cells--constitute a small but complex nervous system. Fifty-three neural cells are located in three pairs of ganglia and connected by paired nerve cords. The prominent frontal ganglia, each consisting of a well-developed neuropile and surrounded by 20 or 21 neural cells, represent the animal's brain. The ventral ganglia contain only 2 neurons each. The penis ganglia--four cells each--are associated with the copulatory organ. A conspicuous circumpenial fiber mass surrounds the basal part of the penis. The effector cells--22 multiciliated epidermal cells, 34 muscle cells, and different gland cells (?)--were also reconstructed and their innervation was partly elucidated. Sensory-motor neurons were unambiguously identified. They are discussed in regard to the small body size of the animal. The male's nervous organization resembles a very simple rope ladder and may represent a reduced derivative of a nervous system in normal-sized males of monomorphic species. Similarities, however, also occur with the developing nervous system of a planktotrophic metatrochophore. The neuronal organization, with its two centers (frontal ganglia and ventral ganglia vs. penis ganglia and circumpenial fiber mass), accords well with the bipartite behavioral pattern, which is entirely devoted to locomotion and copulation, respectively.

The structure and distribution of synapses on nonspiking local interneurones in the metathoracic ganglion of the locust was revealed by electron microscopy following intracellular injection of horseradish peroxidase (HRP). Before staining, each interneurone was characterized physiologically as nonspiking and its output effects on motor neurones innervating muscles in a hindleg were investigated. Three nonspiking interneurones of different morphologies, each typical of a previously described population, were selected for detailed study. The first has a dorsal soma and ipsilateral neuropilar branches, the second a ventral soma and ipsilateral branches, and the third a ventral soma and contralateral branches. The somata have few trophospongial invaginations, and most of their volume is occupied by the nucleus. The initial parts of the primary neurites are either wrapped in glia or isolated in tracts from the neuropile and thus do not participate in synaptic interactions. Some of the larger secondary neurites are also wrapped in glia, but others both make and receive synaptic contacts. Output synapses have an array of some 500-1,600 round, agranular vesicles (diameter 47.0 +/- 5.7 nm; mean +/- S.D., n = 97) associated with a bar-shaped presynaptic density up to 0.3 micron long. Two postsynaptic processes, whose diameter can vary greatly, are usually associated with each presynaptic density. Processes making input synapses onto nonspiking local interneurones typically contain round, agranular vesicles and often make several contacts within a few microns. Serial reconstructions from one of the interneurones revealed input and output synapses intermingled on the larger processes with outputs dominating by a factor of 3:1, whereas on some of the thinner processes only input synapses are present. In the other two interneurones, however, both input and output synapses are present on the fine branches. No feature of the structure or distribution of synapses observed here on the nonspiking local interneurones distinguishes them from spiking neurones in the same ganglia.