Circumoesophageal Connectives Research Articles

May the palps be with you \u2013 new insights into the evolutionary origin of anterior appendages in Terebelliformia (Annelida)

BackgroundHead appendages in Annelida contribute significantly to the immense morphological diversity in this spiralian taxon. Nevertheless, the evolutionary origin of annelid antennae, palps, cirri and tentacles are part of vast theories and debates that took place over decades. One of these heavily discussed groups are the Terebelliformia, which bear numerous anterior tentacles originating from different regions of the head. The question, whether these tentacles are homologous to feeding palps in other annelids or if these structures evolved convergently in terebellids and the remaining taxa, has been highly debated in the past.ResultsBy using morphological methods including immunohistochemistry, confocal microscopy, Azan-stained serial sections and 3D-visualisation, we are able to shed new light and a fresh look on the old question of the evolutionary origin of the buccal tentacles and their associated head structures in Terebelliformia. Our investigations show that the brains of the ampharetid Hypania invalida and the aulophora larvae of Lanice conchilega (Terebellidae) consist of a dorsal, more prominent and a more slender, ventral brain region. Neurite bundles innervating the buccal tentacles split off from the ventral and dorsal root within the ventral brain region and thus originate from the dorsal and ventral root of the circumoesophageal connectives. Hence, the observed neurite bundles fulfil the morphological criteria for the innervating neurite bundles of feeding palps known from Paleoannelida.ConclusionsWe disagree with former conclusions that buccal tentacles are part of the alimentary canal. Based on the presented data, the buccal tentacles of terebelliform taxa are innervated by neurite bundles and can be homologized with peristomial feeding palps of other Annelida.Our comparative investigations reveal important insights into morphological changes during the evolution of anterior head appendages in Terebelliformia and Annelida in general. Nevertheless, our analyses also illustrate the gaps in knowledge and that more investigations throughout the annelid tree are necessary to explain and understand the huge diversity of annelid anterior appendages.

Within-family plasticity of nervous system architecture in Syllidae (Annelida, Errantia)

BackgroundThe ground pattern underlying the nervous system of the last common ancestor in annelids was long thought to be settled, consisting of a dorsal brain, circumoesophageal connectives and a subepithelial, ladder-like ventral nerve cord with segmental ganglia connected by paired connectives. With the advent of immunocytochemical stainings and confocal laser scanning microscopy, it becomes evident that its architecture is extremely diverse, which makes the reconstruction of a ground pattern in annelida challenging. Whereas the nervous systems of many different families has already been described, only very few studies looked at the diversity of nervous systems within such clades to give a closer estimate on how plastic the annelid nervous system really is. So far, little is known on syllid nervous system architecture, one of the largest and most diverse groups of marine annelids.ResultsThe position of the brain, the circumoesophageal connectives, the stomatogastric nervous system, the longitudinal nerves that traverse each segment and the innervation of appendages are relatively uniform within the clade. Both the number of connectives within the ventral nerve cord and the number of segmental nerves, which in earlier studies were used to infer phylogenetic relationships and to reconstruct an annelid ground pattern, are highly diverse and differ between genera or even within a given genus. Differences in the distribution of somata of the brain, the nuchal innervation and its associated cell bodies were found between Syllinae and Exogoninae and may be subfamily-specific.ConclusionsThe nervous system morphology of syllids very likely depends on the taxon-specific ecological requirements. Thus, it is not surprising that in a clade, which occupies such diverse niches as the Annelida, we find similar patterns in phylogenetically widely separated species in similar niches and a high degree of modularity within a family. Only standardized protocols and staining methods can lead to comparable results, but so far different approaches have been taken to describe annelid nervous systems, making homologization of certain structures difficult. This study provides the first thorough description of the nervous system in the family Syllidae, allowing more detailed comparisons between annelid families in the future.

Pigmented eyes, photoreceptor‐like sense organs and central nervous system in the polychaete Scoloplos armiger (Orbiniidae, Annelida) and their phylogenetic importance

The phylogenetic position of Orbiniidae within Annelida is unresolved. Conflicting hypotheses place them either in a basal taxon Scolecida, close to Spionida, or in a basal position in Aciculata. Because Aciculata have a specific type of eye, the photoreceptive organs in the orbiniid Scoloplos armiger were investigated to test these phylogenetic hypotheses. Two different types of prostomial photoreceptor-like sense organs were found in juveniles and one additional in subadults. In juveniles there are four ciliary photoreceptor-like phaosomes with unbranched cilia and two pigmented eyes. The paired pigmented eyes lie beside the brain above the circumoesophageal connectives. Each consists of one pigmented cell, one unpigmented supportive cell and three everse rhabdomeric sensory cells with vestigial cilia. During development the number of phaosomes increases considerably and numerous unpigmented sense organs appear consisting of one rhabdomeric photoreceptor cell and one supportive cell. The development and morphology of the pigmented eyes of S. armiger suggest that they represent miniaturized eyes of the phyllodocidan type of adult eye rather than persisting larval eyes resulting in small inverse eyes typical of Scolecida. Moreover, the structure of the brain indicates a loss of the palps. Hence, a closer relationship of Orbiniidae to Phyllodocida is indicated. Due to a still extensive lack of ultrastructural data among polychaetes this conclusion cannot be corroborated by considering the structure of the unpigmented ciliary and rhabdomeric photoreceptor-like sense organs.

Immunocytochemical studies on the naupliar nervous system of Balanus improvisus (Crustacea, Cirripedia, Thecostraca)

The nervous system of nauplii of the crustacean taxon Cirripedia was analysed in the species Balanus improvisus Darwin, 1854 using for the first time immunocytochemical staining against serotonin, RFamide and α-tubulin in combination with confocal laser scanning microscopy. This approach revealed a circumoesophageal neuropil ring with nerves extending to the first and second antennae and to the mandibles, all features typical for Crustacea. In addition, RFamidergic structures are present in the region of the thoraco-abdomen. A pair of posterior nerves and a pair of lateral nerves run in anterior-posterior direction and are connected by a thoracic nerve ring and a more posteriorly situated commissure. A median nerve is situated along the ventral side of the thoraco-abdomen. The innervation of frontolateral horns and the frontal filaments are α-tubulin-positive. Several pairs of large neurons in the protocerebrum, along the circumoesophageal connectives and in the mandibular ganglion stain only for serotonin. Due to the almost complete absence of comparable data on the neuroanatomy of early (naupliar) stages in other Crustacea, we include immunocytochemical data on the larvae of the branchiopod, Artemia franciscana Kellogg, 1906 in our analysis. We describe several characteristic neurons in the brains of the nauplius larvae of both species which are also found in decapod larvae and in adult brains of other crustaceans. Furthermore, our data reveal that the naupliar brain of cirripedes is more complex than the adult brain. It is concluded that this ontogenetic brain reduction is related to the sessile life style of adult Cirripedia.

Three-dimensional reconstruction of the central nervous system of Macrobiotus hufelandi (Eutardigrada, Parachela): implications for the phylogenetic position of Tardigrada

The morphology of the central nervous system of the tardigrade species Macrobiotus hufelandi was analysed with anti α-tubulin immunostaining in combination with confocal-laser-scanning-microscopy and computer aided three-dimensional reconstruction. The brain anatomy is unexpectedly complex with distinct tracts and highly intermingled nerve fibres. In contrast to older descriptions, we could not detect a suboesophageal ganglion. Furthermore, we found no evidence for a tripartite/three-segmented brain organisation. The median part of the brain is directly connected to the first pair of trunk ganglia via circumoesophageal connectives. Surprisingly, the four paired ventral ganglia do not show segmental commissures typical for the ladder-like nervous system of arthropods. Our findings constrain the phylogenetic position of Tardigrada. The much simpler organisation of the central nervous system of Tardigrada compared to that of Onychophora and Euarthropoda and some similarities to the nervous system of Cycloneuralia support a phylogenetic position of Tardigrada outside an Onychophora/Euarthropoda clade. This means that Tardigrada might be either the sister group to all other Arthropoda or they are more closely related to Cycloneuralia.

The nervous system of Leptodora kindtii (Branchiopoda, Cladocera) surveyed with Confocal Scanning Microscopy (CLSM), including general remarks on the branchiopod neuromorphological ground pattern

The present study describes the nervous system of Leptodora kindtii Focke, 1844 (Branchiopoda, Cladocera) and compares it with that of other branchiopods, with the aim of determining the characters of the branchiopod neuromorphological ground pattern. L. kindtii shares certain correspondences with all studied branchiopods, including a diffuse deutocerebrum, a separated tritocerebrum and a ventral nerve cord with two commissures in each segment. We document a central complex consisting of central body, a protocerebral bridge and lateral lobes which is homologous to that recently described for the anostracan Artemia salina Linnaeus, 1758 and also known for malacostracans. Other characters in L. kindtii, such as the fused compound eye, are shared with other cladocerans, and some, such as optic fibres, the elongated circumoesophageal connectives and the short ventral nerve cord without ganglia in the thoracal part, are unique to Leptodora. The innervation of the “lateral lobes” of the “lower lip” indicates a correspondence between these lobes and the maxillules. We found evidence that the ganglia of the maxillula and maxilla segments have not been incorporated in other ganglia, as suggested earlier.

Sense organs and central nervous system in an enigmatic terrestrial polychaete, Hrabeiella perighndulata (Annelida)-implications for annelid evolution

Abstract. The terrestrial polychaete Hrabeiella periglandulata has many features in common with the Clitellata and the polychaete taxon Parergodrilidae. An ultrastructural investigation of the central nervous system and the sense organs of H. periglandulata individuals was undertaken to look for structural similarities with these taxa as well as to elucidate whether these structures might exhibit adaptive characters typical of terrestrial annelids in general. The central nervous system of H. periglandulata is subepidermal and consists of a brain situated in the first achaetigerous segment. The circumoesophageal connectives are without dorsal and ventral roots, and the ventral nerve cord has closely associated connectives and ill-defined ganglia. In contrast to clitellates and the terrestrial parergodrilid Parergodrilus heideri, nuchal organs are present. They are internal and highly modified compared with those of marine polychaetes but are similar to those of the intertidal parergodrilid Stygocapitella subterranea. A pair of ciliary sense organs is present inside the brain, resembling similar structures in many microdrile oligochaetes. These observations indicate that there are, in fact, structural similarities between the nervous system and the sense organs of clitellates, parergodrilids, and Hrabeiella individuals. These similarities may very likely be the result of convergent evolution in adaptation to the terrestrial environment.

Effects of neck and circumoesophageal connective lesions on posture and locomotion in the cockroach

Few studies in arthropods have documented to what extent local control centers in the thorax can support locomotion in absence of inputs from head ganglia. Posture, walking, and leg motor activity was examined in cockroaches with lesions of neck or circumoesophageal connectives. Early in recovery, cockroaches with neck lesions had hyper-extended postures and did not walk. After recovery, posture was less hyper-extended and animals initiated slow leg movements for multiple cycles. Neck lesioned individuals showed an increase in walking after injection of either octopamine or pilocarpine. The phase of leg movement between segments was reduced in neck lesioned cockroaches from that seen in intact animals, while phases in the same segment remained constant. Neither octopamine nor pilocarpine initiated changes in coordination between segments in neck lesioned individuals. Animals with lesions of the circumoesophageal connectives had postures similar to intact individuals but walked in a tripod gait for extended periods of time. Changes in activity of slow tibial extensor and coxal depressor motor neurons and concomitant changes in leg joint angles were present after the lesions. This suggests that thoracic circuits are sufficient to produce leg movements but coordinated walking with normal motor patterns requires descending input from head ganglia.

Morphology and ultrastructure of the anterior end of Diplocirrus longisetosus Marenzeller, 1890 (Flabelligeridae, Polychaeta, Annelida)

The morphology and ultrastructure of the sedentary polychaete Diplocirrus longisetosus Marenzeller, 1890, collected from the White Sea, were studied using dissection, histological methods, light microscopy, and both scanning and transmission electron microscopy. The prostomium and peristomium carry a pair of palps, eight branchiae, a pair of nuchal organs and two nephridiopores, ciliated folds and the mouth. The prostomium, peristomium and the first chaetigerous segment with all appendages comprise the so-called siphon complex. The mouth leads to a pharyngeal organ that is closed ventrally and composed of a ventral muscle bulb adjoined dorsally by two folds projecting into the pharyngeal lumen. These parts are connected and enveloped by the longitudinal investing muscle. No tongue-like organ is present. The nervous system of the siphonal part comprises the brain, the circum-oesophageal connectives and the ganglia of the peristomium and first chaetigerous segment.

Putative neurohemal release zones in the stomatogastric nervous system of decapod crustaceans.

The stomatogastric nervous system (STNS) of decapod crustaceans has long been used to study the modulation of small neural circuits. Profiles in the sheath of the nerves and ganglia of the STNS, which contain only dense-core vesicles, have been described in electron microscopical studies (Friend [1976] Cell Tissue Res. 175:369-380; Kilman and Marder [1997] Soc Neurosci Abstr. 23:477; Skiebe and Ganeshina [2000] J Comp Neurol 420:373-397). These profiles resemble those found in neurohemal organs and suggest the presence of neurohemal release zones in the STNS. To map these putative neurohemal release zones, a combination of two antibodies was used in the present study. A synapsin antibody recognizing vesicle proteins of clear vesicles was combined with a synaptotagmin antibody recognizing vesicle proteins of clear and dense-core vesicles. Exclusive synaptotagmin-like staining, therefore, indicated the regions with only dense-core vesicles. Such a staining was found in a mesh in the perineural sheath of nerves in the STNS of all three species investigated. In the crayfish Cherax destructor and the lobster Homarus americanus, the stained mesh was located in the sheath of nerves connecting all four ganglia of the STNS, whereas in the crab Cancer pagurus it was found on different nerves, which are more directly exposed to the hemolymph in this species. Exclusive synaptotagmin-like staining was also found in a putative neurohemal release zone in the sheath of the circumoesophageal connectives and the postoesophageal commissure in C. destructor. These data suggest that an important source of modulation of the networks and the muscles of the stomach is a compartmentalized release of neurohormones from zones in the STNS.

Analysis of the Central Nervous System and Sense Organs in Potamodrilus fluviatilis (Annelida: Potamodrilidae)

The Potamodrilidae are small limnetic annelids of apparently simple organization and uncertain phylogenetic position. This group, which was regarded either as a subtaxon of the Aeolosomatidae or united with the Aeolosomatidae as Aphanoneura, has been placed in various positions within the Annelida: (1) at the base of the Clitellata, (2) as a highly derived clitellate taxon closely related to the Naididae, or (3) excluded from the Clitellata because of fundamental differences in the reproductive system and the lack of a clitellum. Moreover, a sister-group relationship between the Aeolosomatidae and Potamodrilidae has been questioned as well. The results of an immunohistochemical (cLSM) and ultrastructural analysis of the central nervous system in Potamodrilus fluviatilis support an exclusion from the Clitellata. The circumoesophageal connectives enter the brain through dorsal and ventral roots, a situation unknown in Clitellata but generally occurring in polychaetes. The ciliated pits, located in front of the brain, are not nuchal organs as previously thought but different sensory structures. However, modified nuchal organs are present behind the brain close to the ventral roots of the circumoesophageal connectives in a position corresponding to that of the nuchal organs found in Aeolosoma spp. Such organs are characteristic for polychaetes and absent in clitellates without exception. These morphological results clearly support the removal of the Potamodrilidae from the Clitellata. Furthermore, the characters present in the nervous system and sense organs show a general correspondence between the Potamodrilidae and the Aeolosomatidae.

Anatomy and in vivo activity of neurons connecting the crustacean stomatogastric nervous system to the brain.

In decapod crustaceans, the inferior ventricular nerve connects the cerebral ganglia (brain) with the stomatogastric nervous system (STNS). In the ivn of the crayfish, eight axons with diameters between 3.5 microm and 10 microm were found in close proximity to the oesophageal ganglion. Two of these axons terminate with their cell body within the ivn. The projections of the other six axons spread inside many neuropiles of the brain, mainly within the protocerebrum and the neuropils of the first and second antennae. Several fibers also send neurites via the circumoesophageal connectives toward the paired commissural ganglia and further down to the ventral nerve cord. The activity of motoneurons within the STNS and of axons in the ivn was recorded with implanted electrodes before, during and after times of feeding. At the beginning of feeding all tonically active ivn neurons accelerated their discharge rate and initially silent neurons also started to fire. Spike frequency was correlated with the quantity of food consumed. The ivn response was accompanied by a corresponding increase in pyloric frequency and an initiation of a gastric rhythm. The two motor rhythms showed a strong phasic interaction, but there was no phase coupling to the ivn activity.

On the anatomy of the central nervous system and the morphological value of the anterior end appendages of Ampharetidae, Pectinariidae and Terebellidae (Polychaeta)

AbstractEarlier papers dealing with the anatomy of the central nervous system of ampharetids, pectinariids and terebellids were studied. On this basis a re‐investigation appeared necessary: statements in the literature about the structure of the brain and the innervation of the appendages of the anterior end were incomplete and contradictory. In the present paper, the brain, the circum‐oesophageal connectives and the innervation of, inter alia, the tentacular membrane (including the dorsal ridge), the buccal tentacles, the alimentary canal, the nuchal organs and the branchiae (when present) of Amphicteis gunneri, Anobothrus gracilis, Melinna cf. cristata, Pectinaria auricoma, P. belgica, P. koreni, Petta pusilla, Pista cristata, Eupolymnia nebulosa, Thelepus cincinnatus and Polycirrus medusa are described. The results are summarized in schematic diagrams and compared with each other and with the central nervous system of other polychaetes. It is concluded that the ampharetids, the pectinariids and the terebellids bear no antennae and no palps and that their buccal tentacles belong to the alimentary canal. It is emphasized that all attempts to range their cephalic nervous system into previously proposed common and general schemes of ‘the polychaete nervous system’ seem totally fruitless.

Immunohistochemical (cLSM) and ultrastructural analysis of the central nervous system and sense organs in Aeolosoma hemprichi (Annelida, Aeolosomatidae)

The Aeolosomatidae are very small limnetic or terrestrial annelids of apparently simple organisation and uncertain phylogenetic position. They have been placed either at the base of the Clitellata, as a highly derived taxon within the Clitellata closely related to the Naididae, or as their sister group within the „Polychaeta”. A combined immunohistochemical (cLSM) and ultrastructural investigation of the central nervous system and the sense organs in Aeolosoma hemprichi was undertaken to look for characters which might support one of these theories. The position of the brain within the prostomium and the organisation of the ventral nerve cord, with its intraepithelial paired longitudinal nerves lying far apart from each other and the presence of a median longitudinal nerve, are atypical for the Clitellata and clearly differ from the situation found in Naididae. Moreover, the circumoesophageal connectives are bifurcated and enter the brain as dorsal and ventral roots; this arrangement is unknown in Clitellata, in which these connectives are unbranched. An ultrastructural analysis of the ciliated pits located laterally in the furrow between prostomium and peristomium in A. hemprichi and other Aeolosomatidae show that they are in fact nuchal organs. Such presumed chemosensory organs are typical of „Polychaeta” and absent in all Clitellata. Two pairs of ciliary sense organs are present in the prostomium in front of the brain of A. hemprichi. Although similarly organised sensory structures occur in many species of the Clitellata, they differ in position and certain ultrastructural features and are known from other Annelida as well. These results clearly support the exclusion of the Aeolosomatidae from the Clitellata and do not provide any evidence for a sister-group relationship between these two taxa.

Sulfakinin neuropeptides in a crustacean. Isolation, identification andtissue localization in the tiger prawn Penaeus monodon.

The sulfakinin (SK) family of neuropeptides are characterized by a C-terminal octapeptide sequence that begins with two acidic residues (most commonly DD), and ends with YGHMRF-NH2, usually with the tyrosyl residue sulfated. So far, sulfakinins have only been identified in insects and the present study was initiated to investigate if the family is more widely distributed within the arthropods. Purification of an extract of the central nervous system of the giant tiger prawn Penaeus monodon has revealed three novel members of the sulfakinin peptide family. One of the peptides, Pem SKI, has the sequence <QFDEY(SO3H)GHMRF-NH2, where <Q denotes a pyroglutamic acid residue, and is for all criteria typical of insect sulfakinins, several of which also have an N-terminal pyroglutamic acid. Tyrosyl O-sulfation was verified by mass spectrometry. The two other peptides have a hitherto unknown L/M variation at position three from the C-terminus. One of these, Pem SKII, has a particularly glycine-rich N-terminus, AGGSGGVGGEYDDYGHLRF-NH2. The other, Pem SKIII, is a truncated form of Pem SKII, with the sequence VGGEYDDYGHLRF-NH2. Mass spectrometry of the latter two peptides indicated that only one of the two tyrosyl residues is sulfated. By analogy, it is suggested that the sulfation is located at the residue in position six from the C-terminus. A small amount of a nonsulfated variant of Pem SKII was also present in the extract. Immunocytochemical studies with sulfakinin antisera show a sparse neuronal distribution pattern, similar to that of insects. A prominent pair of large (approximately 25 micrometer) cells and 6-8 pairs of smaller (approximately 10 micrometer) cells are present in the protocerebrum. The larger cells have prominent neurites that give rise to varicosities in the centre of the brain. Their axons exit the brain via the circumoesophageal connectives and continue along the intersegmental connectives. Each of the thoracic and abdominal ganglia has sulfakinin-immunoreactive arborizations as a result of branching from the intersegmental nerves. This distribution pattern strongly suggests a role in neurotransmission or neuromodulation, although it remains to be elucidated what the exact role(s) is. However, on account of the conservation of peptide structure during the evolutionary period spanning the insect/crustacean lineage, especially between Pem SKI and insect sulfakinins, it may be assumed that the sulfakinins have a significant physiological role.

Dopaminergic modulation of spontaneous activity in the brain of the crayfishCherax destructor(Decapoda, Crustacea)

The role of dopamine in the crayfish (Cherax destructor) brain was studied in an isolated head preparation by means of extracellular recordings from unidentified neurons located in the circumoesophageal connectives. Perfusion of dopamine and the dopamine receptor agonist (±)-2-amino-6,7-dihydroxy-1,2,3,4-tetrahydronaphthalene hydrobromide into the brain induced excitatory as well as inhibitory modulation of spontaneous activity. The physiological effects of both drugs were reversible and showed concentration dependency in the concentration range 10−7to 10−3 M. Two vertebrate-derived dopamine receptor antagonists, chlorpromazine and fluphenazine, reversibly blocked the action of dopamine (10−5 M). The threshold for antagonistic blockade by chlorpromazine and fluphenazine occurred at a relatively low concentration (10−7 M) and was concentration dependent. These data collectively suggest that dopamine plays a physiological role in the crayfish brain, stimulating putative dopamine receptor(s) to alter neuronal activity.

Structure of the prostomial appendages and the central nervous system in the Protodrilida (Polychaeta)

The prostomial appendages and the central nervous system have been investigated by electron microscopy in Protodriloides chaetifer, P. symbioticus, Protodrilus haurakiensis, P. oculifer, P. ciliatus, P. helgolandicus, P. adhaerens, Saccocirrus krusadensis and S. papillocereus. The tentacles are highly developed, mobile sensory structures and consist of cuticle, epidermis, a different number of intraepithelial nerves, a small blind-ending blood vessel and a bundle of longitudinal muscle fibres. An internal canal is only present in Protodrilus and Saccocirrus species. On the tentacles seven types of sensory cells have been found including different multiciliated and uniciliated sensory cells with cilia penetrating the cuticle, sensory cells with non-penetrative cilia, phaosomes and basal ciliated sensory cells. The latter are described for the first time in polychaetes. From the specific pattern of innervation by up to five nerves originating close to the brain from the dorsal and ventral roots of the circumoesophageal connectives it is evident that the prostomial appendages represent palps. In the palps the nerve fibres form neuroneuronal, myoneuronal and epithelioneuronal synapses. The brain also gives rise to the stomatogastric nerves and various dorsal nerves. The palp canals are separated from the surrounding tissue by a prominent extracellular matrix. The wall is formed by muscle cells. The centre is usually completely filled with the cell bodies of these muscle fibres and large coelenchyme-like cells. These cells move freely in the canals and they are very likely the structural basis for the hydroskeletal function of the canals. The canals are completely separated from other body cavities and fluid is probably driven into the canals from the blood vascular system via podocytes located in a specific zone in the prostomium. In particular, the structure of the central nervous system with its nerves, the pattern of innervation of the palps and the palp canal system are compared with those of other polychaetes with special emphasis to the Spionida, the taxon presumed to include the sister group of the Protodrilida.

Suboesophageal DUM neurons innervate the principal neuropiles of the locust brain

The morphology of the dorsal unpaired median (DUM ) neurons of the suboesophageal ganglion (SOG) of the migratory locust, Locusta migratoria , were studied by using intracellular staining. The SOG lacks segmental DUM neurons with peripheral axons. All DUM neurons are either intersegmentally projecting (towards the brain or the thoracic nerve cord) or they are local. In addition to previously described DUM neurons with axons in peripheral nerves of the brain (Bräunig 1990), the SOG contains DUM neurons which, in the brain, innervate principal neuropile areas such as the antennal lobes, the pedunculi and calyces of the mushroom body, and the central complex. The number and location of DUM cell bodies stained with intracellular fills is compared with those obtained with either backfilling cervical or circumoesophageal connectives, or octopamine-immunocytochemistry. Additional experiments show that the locust brain, like the SOG, lacks both segmental DUM neurons with peripheral axons, and axons descending into the ventral nerve cord.

Antennal circulatory organs in Onychophora, Myriapoda and Hexapoda: Functional morphology and evolutionary implications

A comparative investigation of the antennal circulatory organs in representatives of the Onychophora, all subtaxa of the Myriapoda and numerous taxa of the Hexapoda (comprising a total of 54 species) revealed an unexpected diversity in structure and function. In the Onychophora, antennal vessels exist which are connected to the enlarged anterior end of the aorta dorsal to the brain. In the Chilopoda, Diplopoda and Symphyla, antennal vessels exist which originate from the dorsal vessel caudal to the brain. They extend under the optic lobes, lateral to the circumoesophageal connectives, into the antennae. In the Hexapoda, the investigations include representatives of all higher taxa, apart from the Paraneoptera and the Holometabola. Generally, antennal vessels exist. In the Diplura, they originate from the anterior end of the aorta in front of the brain. In all other insects the antennal vessels are separate from the dorsal vessel. Their proximal ends form ampullary enlargements which are attached to the frontal cuticle near the antenna bases. They communicate via valved ostia with the haemolymph sinus in front of the brain. In the Archaeognatha, Zygentoma, Odonata, certain Plecoptera and the Notoptera, no muscles are connected to these organs. In all other groups the ampullae are pulsatile as a result of associated muscles (“antennal hearts”). These muscles diverge widely in their attachments and act either as compressors (Dermaptera) or dilators of the ampullae (Embioptera, Blattopteroidea, Orthopteroidea, and some Plecoptera). In the Collembola and Ephemeroptera, special antennal circulatory organs are lacking. In some forms the anatomical arrangement of the inner organs, in conjunction with short diaphragms at the antenna bases, apparently leads to a channelling of haemolymph flow. This condition may be explained by the very short antennae of these insects and is considered as a convergent and apomorphic state in these taxa. The antennal vessels are supposed to be homologous within the Tracheata and to represent the lateral arteries of the antenna segment. An origin from the dorsal vessel is considered an ancestral state, which was lost in the stem lineage of the Ectognatha. Specific space constraints within the cephalic capsule are discussed as the possible reason for this loss. The evolution of pulsatile antennal circulatory organs in the Neoptera is the result of the association of muscles with the proximal ampullary ends of the antennal vessels. The attachments and innervation of these muscles indicate a derivation from precerebral pharyngeal dilators.

Alkaline phosphatase activity in the brain of the American cockroachPeriplaneta americana L.

The supra- and suboesophageal ganglia of the American cockroach contain material which catalyses the alkaline hydrolysis (pH 9.5) of 5-bromo-4-chloro-3-indolyl phosphate in the presence of Nitro blue tetrazolium. Histochemical studies on unfixed cryostat sections indicate that this type of alkaline phosphatase is restricted to discrete regions in the cockroach brain. Highest enzyme activity is encountered in the mushroom bodies, central body, antennal glomeruli and specific parts of some distinct neural connections including the optic nerve, antennal nerve, circumoesophageal connectives and nerves leaving the suboesophageal ganglion. Tissue fixation by use of formaldehyde-type fixatives, as well as routine paraffin-embedding, completely destroy all histochemically detectable enzyme activity. Native polyacrylamide gradient electrophoresis suggests that the alkaline phosphatase activity is present as multiple isozymic forms, which show up in the 120-130 kD range of standard proteins. Enzyme activity becomes undetectable after fixation (trichloroacetic acid, formaldehyde containing fixatives) of electrophoretically separated native proteins, as well as after electrophoresis in denaturing conditions (SDS and beta-mercapto-ethanol, boiling). However, the enzyme activity remains virtually unaffected after storage of the sample for prolonged periods at -20 to -80 degrees C.