Morphology of neurones associated with the antennal heart of Periplaneta americana (Blattodea,Insecta)

Summary Innervation of the antennal heart, an independent accessory circulatory motor in the head of insects, was investigated in the cockroach Periplaneta americana by use of axonal cobalt filling and transmission electron microscopy. The muscles associated with this organ are innervated by neurones located in a part of the suboesophageal ganglion, generally considered to be formed by the mandibular neuromere. Dorsal unpaired median (DUM) and paired contralateral neurones were stained. The axons of all these neurones run along the circumoesophageal connectives and through the paired nervus corporis cardiaci III into the corpora cardiaca. They pass through these organs forming fine arborizations there and exit anteriorly as a small pair of nerves which terminate at the antennal heart-dilator muscles. Numerous branches of these nerves extend beyond the lateral borders of the large transverse dilator muscle and terminate in the ampullar walls of the antennal heart. These neurosecretory fibres form neurohaemal areas which obviously release their products into the haemolymph, which is pumped into the antennae. The possible functions of the neurones associated with the antennal heart are discussed with respect to both, their role as a modulatory input for the circulatory motor and as a neurohormonal release site.     

The diverticulated crop of adult Phormia regina

The crop of adult Phormia regina consists of a duct that diverges from the esophagus, just in front of the cardia, and extends ventrally and posteriorly into the thorax and abdomen where it forms a bilobed sac. Flattened epithelial cells produce the cuticular lining of the crop. When empty, or partially full, the epithelial cells and cuticular lining form folds extending into the lumen, thus providing for expansion as the crop sac fills. Covering the sac on the hemolymph side is a layer of anastomosed, intrinsic muscles connected to one another by intercellular cytoplasmic bridges. Mitochondria are located at the periphery of the sarcomere. Also inside the sarcomere are glycogen, sarcoplasmic reticula, and transverse tubular systems (T-system). I, A, and Z-bands are present and the Z-bands are not in register making the muscle-type supercontractile. Important structures, not previously researched and associated with the crop muscles, are the crop nerves. Coming off the corpora cardiaca, and running down each side of the crop duct, is a pair of nerves, each housing several axons. These nerves extend to and branch over the crop sac. Here they penetrate the muscle mass and form neuromuscular junctions where electron-dense droplets of neurosecretion are released. Based on the literature, and research in our laboratory, it has now been shown that these nerves carry adipokinetic hormone, Drosophila insulin-like peptide, and a dromyosuppressin-like neuropeptide.     

Helicostatins: brain-gut peptides of the moth, Helicoverpa armigera (Lepidoptera: Noctuidae)

Gene expression and immunolocalisation studies have determined that the helicostatins are brain-gut peptides in larvae of the lepidopteran, Helicoverpa armigera. Mapping of the distribution of these peptides in the nervous system and alimentary canal has provided evidence for multifunctional regulatory roles. In situ hybridisation studies have shown that the helicostatin precursor gene is expressed in neurones of the central and stomatogastric nervous systems, and endocrine cells of the midgut demonstrating that the helicostatins are true brain-gut peptides. Antisera raised against Leu-callatostatin 3 (ANRYGFGL-NH(2)), a peptide isolated from the blowfly, Calliphora vomitoria was used to map the distribution of allatostatin-like immunoreactive (Ast-ir) material in H. armigera to elucidate possible functions of the helicostatins. In situ hybridisation studies verified that the helicostatin precursor gene is expressed in neurones shown to contain Ast-ir, providing strong evidence that the Ast-ir material is helicostatins. Extensive immunoreactive axonal projections into complex regions of neuropile indicate that the helicostatins may have a neuromodulatory role in the brain and segmental ganglia of the ventral nerve cord. The presence of large amounts of immunoreactive material in axons within the corpora cardiaca (CC) and transverse nerves of the perisympathetic nervous system, two known neurohaemal organs, provides evidence for a neurohormonal role. The corpora allata (CA) were innervated only sparsely by Ast-ir axons suggesting that the CA are not a neurohaemal release site or a target. Thus, it is unlikely that the helicostatins regulate juvenile hormone (JH) biosynthesis or release. Ast-ir axons extended from the frontal ganglion through the recurrent nerve and many branches were closely associated with muscles of the foregut, stomodeal valve, and anterior midgut, implicating helicostatins in regulation of foregut motility. Ast-ir material was also present in nerves associated with muscles of the pyloric valve and rectum, and in endocrine cells of the midgut.     

Innervation of chromatophore muscle fibres in the octopus Eledone cirrhosa

Summary Cephalopod chromatophores are made of a central pigment cell surrounded by 10 to 20 radially arranged muscle fibres under direct nervous control. Innervation of these muscle fibres was studied with anterograde cobalt fills of peripheral nerve bundles and light and electron microscopy. Individual axons branch repeatedly to innervate the muscles of chromatophores scattered over several millimeters. Axons contained in several dermal nerves converge to innervate the same chromatophores. Among the chromaophores, axons were found running either singly or in small bundles, often accompanied by sheath cells. Single chromatophore muscles were innervated by at least one axon running across or along its length. Since nerves terminating on chromatophore muscles are very rare, neuromuscular contact seems to be made en passant. Varicosities of the axons apposed to the muscles are thought to be presynaptic sites. However, morphological differentiations of the pre-or post-synaptic membranes were not visible. Two types of innervating processes were found containing either electron-clear or a mixture of electron-clear and dark-core synaptic vesicles.Supported by a postgraduate award from the University of Aberdeen (GB)     

Stomatogastric nervous system of Galleria mellonella L. (Lepidoptera : Pyralidae): Changes during metamorphosis with special reference to FMRFamide neurons

Scanning electron microscopy and immunohistochemical staining for FMRFamide-like peptides revealed that the stomatogastric nervous system of Galleria mellonella (Lepidoptera : Pyralidae) includes 5 ganglia: the frontal ganglion with 4, the hypocerebral ganglion with 2, the ingluvial ganglion with 2–4, and each of the paired proventricular ganglia with 6–8 immunoreactive perikarya. Immunoreactivity was also found in axons to and within the corpora cardiaca, in the nerves connecting stomatogastric ganglia, as well as in 8 gastric nerves that extend along longitudinal midgut muscles. Adhesion of corpora cardiaca to the hypocerebral ganglion and partial merging and shortening of gastric nerves were the only conspicuous changes of the stomatogastric system that occurred during metamorphosis.     

The effect of age on motor neurone death following axotomy in the mouse.

The ability of mouse motor neurones to survive axotomy during the first month of life was studied. The motor neurones that lie in the dorsolateral columns of spinal segments C7 and C8 and supply the flexor muscles of the forepaw were axotomized by cutting and removing part of the median and ulnar nerves above the elbow. The number and position of cell bodies with axons in these nerves were confirmed by retrograde labelling of the cut axons with horseradish peroxidase. The ability of these neurones to survive axotomy varies with the age of the animal at the time of axotomy. When the axons are sectioned within the first four postnatal days, 80-90% of the cell bodies will die, more than half of this death occurring in less than one week after axotomy. If the animals are one week old at the time the nerves are cut, a significantly smaller number (50%) die (P = 0.013), and the time-course of death is different, with eight to ten days elapsing before half the death has occurred. 40% of the neurones will die if sectioned at two weeks of age, and it is not until four weeks of age that more than 90% of the cells can survive axotomy. We conclude, therefore, that the kinetics of motor neurone death, as well as the final extent of neuronal loss, are affected by the age at which the animal is axotomized.     

Abdominal stretch reception in Dipetalogaster maximus (Hemiptera: Reduviidae)

Each half abdominal segment in 5th-instar larvae of the giant bloodsucking reduviid, Dipetalogaster maximus, contains 3 stretch receptor neurones, one associated with the tergosternal muscles, one with the ventral intersegmental muscles and one with the dorsal intersegmental muscles. Each of the three receptors respond phasically to the onset of stretch in its respective muscle group, but none show persistent activity upon prolonged stretch. By contrast, stretch of the main abdominal nerves (which run between the thoracic ganglion and the ventral intersegmental muscles of each abdominal segment) is accompanied by a prolonged and sustained pattern of discharge by an as yet unidentified neurone, the rate of discharge being proportional to the degree of stretch. In life, the abdominal nerves become stretched to about 145% of their resting length when the larva takes a bloodmeal. Thus it appears that in Dipetalogaster stretch of the abdominal nerves themselves is the only mechanism for stretch reception after a blood meal.     

Organization of motoneurones in the prothoracic ganglion of the cockroach Periplaneta americana (L.).

The location within the prothoracic ganglion of neurone somata with axons in identified peripheral nerves is examined by the cobalt iontophoresis technique. Axons are filled with cobalt by diffusion through their cut ends and the cobalt is then precipitated as the black sulphide inside the neurone. It is assumed that neurones with axons in peripheral nerves and somata in central ganglia are either motor or neuro-secretory. Fifteen nerves are examined and maps of the location of somata with axons in each nerve are presented. The axon distribution in peripheral nerves of three common inhibitory neurones is described. Dendritic morphology of one common inhibitory neurone and two coxal depressor motoneurones is illustrated. It is proposed that some individual neurones can be reliably identified from their soma dimensions and location within the ganglion. The number of motoneurones with somata in the prothoracic ganglion and their homology with cells in the other thoracic ganglia are discussed.     

Triple co-localisation of two types of allatostatin and an allatotropin in the frontal ganglion of the lepidopteran Lacanobia oleracea (Noctuidae): innervation and action on the foregut

The triple co-localisation of peptidergic material immunoreactive to antisera raised against allatostatins of the Y/FXFGL-NH2 type, Manduca sexta allatostatin (Mas-AS), and allatotropin has been demonstrated in a single pair of anterodorsal neurones in the frontal ganglion of the tomato moth, Lacanobia oleracea (Noctuidae). Another pair of posterior neurones contain only Y/FXFGL-NH2-type allatostatin immunoreactivity. The neurites of all four cells trifurcate, and axons project to the brain in the frontal connectives and to the foregut in the recurrent nerve. Axons from the anterior neurones, within the recurrent nerve, have prominent lateral branches supplying muscles of the crop, and axons from both anterior and posterior cells show profuse branching and terminal arborisations in the region of the stomodeal valve. The brain contributes Y/FXFGL-NH2-immunoreactive material, but not allatotropin or Mas-AS, to the recurrent nerve via NCC 1+2 and NCC 3. All three peptides have a reversible effect on the spontaneous (peristaltic) contractions of the foregut (crop) in vitro. Thus, both types of allatostatin are inhibitory at 10(-12) to 10(-7) M, whereas allatotropin is strongly myostimulatory at 10(-14) M. This is the first demonstration of the gut myoinhibitory effects of Mas-AS and, taken together with the effects of Y/FXFGL-NH2-type allatostatins and allatotropin, reveals a different functional aspect to that normally attributed to these three peptides, i.e. control of juvenile hormone synthesis by the corpus allatum.     

Muscles and nerves of the posterior abdomen and genitalia of male Periplaneta americana (L.) (Dictyoptera : Blattidae)

Segmental and intersegmental muscles of abdominal segments 7–10 are described for adult, male Periplaneta americana (L.) (Dictyoptera : Blattidae). Locations of extrinsic and intrinsic genitalic muscles are documented, and the actions of those associated with the right phallomere are hypothesized. Muscles of the 5 abdominal segments are innervated by branches from 5 pairs of segmental nerves and 3 pairs of transverse nerves. These stem from a terminal synganglion, formed during embryogenesis by fusion of neuromeres of abdominal segments 7–11. One pair of segmental nerves issues from each of the 5 neuromeres, and one pair of transverse nerves arises from neuromeres of abdominal segments 7–9. The nerves are traced to the muscles, integument, and reproductive glands, and their peripheral unions are characterized. Serial homologies of the nerves and muscles are proposed, and comparisons are made with neuromusculature of the female.     

Muscles and innervation of the genital and postgenital abdominal segments of the female american cockroach Periplaneta Americana (L.) (Dictyoptera : Blattidae)

The locations of skeletal muscles in abdominal segments 7–10 of female Periplaneta americana (Dictyoptera : Blattidae) are described, and the action of some of those attached to the gonapophyses is hypothesized. The muscles are innervated from a terminal synganglion, a composite formed by fusion of embryonic neuromeres of abdominal segments 7–11. Pathways of the 5 pairs of segmental nerves and 3 pairs of transverse nerves that issue from the synganglion are documented. In addition to supplying skeletal muscles, certain of the nerves can be traced to the oviducts, spermatheca, colleterial glands, ventral glands, hindgut, and apparently, to sensilla of the cerci, gonapophyses, paraprocts, tergum, pleura, and sternum. Homologous relationships of the nerves are proposed, and the observations are compared to those reported by others who have, for the most part, examined males only. Putative neurosecretory somata occur in abdominal segments 7 and 8, at the junction of the transverse and segmental nerves. The association may be comparable to the “link nerve” complex described by others.     

Structural features associated with movement and ‘catch’ of sea-urchin spines

Spine catch ligaments of a sea urchin Arbacia punctulata were extended under constant load. Ligaments from an undisturbed animal may show any extension rate from zero (catch state) to rapid extension to failure. Replacing the preparation bath with Ca²⁺- and Mg²⁺-free sea water reversibly abolishes the catch state. The fine structure of the outer muscle layer and inner ligament cone associated with the spine base is described. The unstriated paramyosin muscles bear thin flanges and form compact interlocking rows. Subsurface cisternae are associated with the plasma membrane. The muscles are innervated by glia-free axons ending in bulbous terminals containing lucent synaptic vesicles. The ligament comprises cylindrical bundles of collagen fibrils: one or more minute muscle fibers (paramyosin) lie parallel with and closely adjoining each bundle. The mean diameter of these muscles is 0.3 μg and they occupy 2–3 % of the ligament's cross-sectional area. Axons containing electronopaque secretory droplets accompany the muscles between the collagen bundles: the cell bodies of these neurones generally lie on the outer surface of the ligament. When an urchin points a spine, the ligament on the side of the contracting spine muscle shortens but does not buckle. A function of the intraligamental muscles is to effect this non-buckled shortening. The catch mechanism (which resides entirely within the ligament) may be due either to the intraligamental muscles and/or to a locked polymer mechanism in which matrix molecules between collagen fibrils are reversibly crosslinked by divalent cations.     

Fine structure of the alary muscles of the American cockroach

The alary muscles of the cockroach, Periplaneta americana, are striated with an A-band of 3·0 to 3·5 μm long. Each muscle fibre was 10 to 12 μm in diameter and Z-lines appeared as small discrete units staggered throughout the sarcoplasm. Mitochondria were conspicuously located near the Z-line areas and were absent from the middle portion of the sarcomere. A transverse membrane system was present which formed dyad structures with a relatively sparse sarcoplasmic reticulum. Cockroach alary muscles were innervated by axons containing electron-dense granules of near 100 nm in diameter. These are thought to be typical of ‘neurosecretory’ axons based on their ultrastructural appearance.     

The distribution and colocalization of neuropeptides and catecholamines in nerves supplying the gall bladder of the toad,Bufo marinus

The indirect immunofluorescence technique was used to determine the distribution of peptide-containing axons in the gall bladder of the cane toad, Bufo marinus. In addition, the adrenergic innervation of the gall bladder was examined by use of immunoreactivity to the catecholamine-synthesizing enzyme, tyrosine hydroxylase, and glyoxylic acid-induced fluorescence. On the basis of peptide coexistence, two intrinsic populations of neurones and their projecting fibres could be distinguished substance P neurones and vasoactive intestine peptide neurones. Neither of these two types of neurones contained any other colocalized neuropeptides. Four populations of nerve fibres arising from cell bodies outside the gall bladder were identified: nerves containing colocalized galanin, somatostatin and vasoactive intestinal peptide; nerves containing colocalized calcitonin gene-related peptide and substance P; adrenergic nerves containing neuropeptide Y; and nerves containing only adrenaline.     

The Sense Organs and Ventral Ganglion of Sagitta (Chaetognatha)

This paper describes some features of the chaetognath nervous system from ultrastructural observations and observations on material stained with specific techniques for nervous tissue, and from records of the activity of the locomotor muscles and ventral ganglion. Sensory cells grouped on the ventral surface of the head bear ciliary processes (some with multiple tubules), and are probably in connexion with the central nervous system by their own axons, unlike the sensory cells of the hair fan vibration receptors of head and body. The ventral ganglion is motor to the locomotor muscles of the body, and controls the rhythmic locomotor activity of the animal. Electrical events associated with contraction of these muscles are compound non-overshooting spike-like potentials. The ventral ganglion contains several large nerve fibres constant in position and connexions in different individuals. Some of these arise from cells in the ganglia of the head, and pass to the ventral ganglion, others from cells within the ventral ganglion, and probably supply the ciliary hair fan receptors of the body, whilst the motor axons issuing from the ventral ganglion are smaller in diameter. The ganglion is arranged on a ladder-like plan, and axons of the lateral cell bodies cross the central neuropil transversely before they contribute to the longitudinal tracts or pass out in the radial nerves. Synapses in the neuropil contain 30–40 nm electron lucent vesicles; the transmitter is unknown, but is unlikely to be either acetylcholine or l -glutamate. Occasional larger electron dense vesicles up to 70 nm in diameter are also found within nerve fibres of the neuropil. It is concluded that the arrangement of the peripheral nervous system is unlike that of several groups which have been suggested as related to chaetognaths.     

Novel peripheral motor neurons in the posterior tentacles of the snail responsible for local tentacle movements

Three flexor muscles of the posterior tentacles of the snail Helix pomatia have recently been described. Here, we identify their local motor neurons by following the retrograde transport of neurobiotin injected into these muscles. The mostly unipolar motor neurons (15–35 µm) are confined to the tentacle digits and send motor axons to the M2 and M3 muscles. Electron microscopy revealed small dark neurons (5–7 µm diameter) and light neurons with 12–18 (T1 type) and 18–30 µm diameters (T2 type) in the digits. The diameters of the neurobiotin-labeled neurons corresponded to the T1 type light neurons. The neuronal processes of T1 type motor neurons arborize extensively in the neuropil area of the digits and receive synaptic inputs from local neuronal elements involved in peripheral olfactory information processing. These findings support the existence of a peripheral stimulus–response pathway, consisting of olfactory stimulus—local motor neuron—motor response components, to generate local lateral movements of the tentacle tip (“quiver”). In addition, physiological results showed that each flexor muscle receives distinct central motor commands via different peritentacular nerves and common central motor commands via tentacle digits, respectively. The distal axonal segments of the common pathway can receive inputs from local interneurons in the digits modulating the motor axon activity peripherally without soma excitation. These elements constitute a local microcircuit consisting of olfactory stimulus—distal segments of central motor axons—motor response components, to induce patterned contraction movements of the tentacle. The two local microcircuits described above provide a comprehensive neuroanatomical basis of tentacle movements without the involvement of the CNS.     

Internal and external morphology of the deutonymph of Caloglyphus boharti (Arachnida: Acari)

A band of flexible cuticle encircles the deutonymph, separating the dorsal and ventral plates. The coxae are large, flat and fused with one another to form most of the ventor. Individual coxal margins are redefined as sternites, epimerites or simply apodemes according to which margins fuse with which others. A given area of cuticle may have patches of dark or light cuticle not corresponding to particular structures or cuticular contours; this is a source of confusion to taxonomists. Each leg has a dicondylic coxal-trochantal (adduction-abduction) and trochantal-femoral (promotion-remotion) joint with opposing muscles. The three more distal monocondylic joints (flexion-extension) have only flexor muscles; extension is by increased haemolymph pressure. The five apodemes of the sucker plate provide rigidity; the four suckers attach by a flexible cuticular ring to a solid flange or socket in the sucker plate. The sucker muscles attach to the center of each sucker. The flat, external face of the sucker plate apodemes may complement sucker action by adhesion. Coxal discs and sucker plate discs are identical, contain birefringent cuticular elements, and are considered modified setae. Functional mouthparts and a pharynx are lacking, but a cheliceral anlage is present. The esophagus, midgut and caecae, and malpighian tubules are lumenless and the cells small. The hindgut has a lumen, larger cells and opens externally via the anus. Whereas the digestive tract is regressed, the reproductive system is yet incompletely developed. In older deutonymphs anlagen of ducts, accessory glands and gonads are discernible. The nature of the haemocoel and peritoneum remains nuclear. The central nerve mass is conspicuously large for the size of the deutonymph. The supraesophageal ganglion gives rise to the cheliceral nerves; all other nerves arise from the subesophageal ganglion. Most major nerves were traced to the effector organs. The muscles are divided into leg, dorso-ventral (derived from coxal muscles), dorsal, sucker, and anogenital muscles. The trochantal adductor muscles originate on an endosternite, which is supported by muscles running to the dorsal hysterosoma. The dorso-ventral and propodosomal retractor muscles affect haemolymph pressure. The massive sucker retractor muscles are unique to this instar. Anogenital muscles are not well developed.     

Structure of the intramural nerves of the rat bladder

The bladder of adult female rats receives ～16,000 axons (i.e., is the target of that many ganglion neurons) of which at least half are sensory. In nerves containing between 40 and 1200 axons cross-sectional area is proportional to number of axons; >99% of axons are unmyelinated. A capsule forms a seal around nerves and ends abruptly where nerves, after branching, contain ～10 axons. A single blood vessel is present in many of the large nerves but never in nerves of <600 axons. The number of glial cells was estimated through the number of their nuclei. There is a glial nucleus profile every 76 axonal profiles. Each glial cell is associated with many axons and collectively covers ～1,000 μm of axonal length. In all nerves a few axonal profiles contain large clusters of vesicles independent of microtubules. The axons do not branch; they alter their relative position along the nerve; they vary in size along their length; none has a circular profile. All the axons are fully wrapped by glial cells and never contact each other. The volume of axons is larger than that of glial cells (55%–45%), while the surface of glial cell is twice as extensive as that of axons; there are ～2.27 m² of axolemma and ～4.60 m² of glial cell membrane per gram of nerve. Of the mitochondria of a nerve ～3/4 are in axons and ～1/4 in glial cells.     

The ultrastructure of the cardiac ganglion of the desert scorpion,Paruroctonus mesaensis (Scorpionida: Vaejovidae)

Light and electron microscopy of the pacemaker ganglion of the scorpion heart indicate that it is about 15 mm long and 50 μm in diameter and extends along the dorsal midline of the heart. The largest cell bodies (30–45 μm in diameter) occur in clusters along the length of the ganglion. The ganglion appears to be innervated with fibers from the subesophageal and first three abdominal ganglia. The cardiac ganglion is surrounded by a neurilemma and a membranous sheath. The latter is apparently derived from connective tissue cells seen outside the ganglion. Nerve fibers other than those in the neuropil areas are usually surrounded by membrane and cytoplasm of glial cells. Often there are several layers of glial membrane, forming a loose myelin. The cardiac nerves to the heart muscle are also surrounded by a neurilemma, and the axons are surrounded by glia. The motor nerves contain lucent vesicles 60–100 nm and opaque granules 120–180 nm in diameter. In the cardiac ganglion, some nerve cell bodies have complex invaginations of glial processes forming a peripheral trophospongium. In the neuropil areas, nerve cell processes are often in close apposition. The septilaminar configuration typical of gap junctions is common, with gap distances of 1–4 nm. In tissues stained with lanthanum phosphate during fixation, we found gaps with unstained connections (1–2 nm diameter) between nerve-nerve and glial-nerve cell processes. Annular or double-membrane vesicles in various stages of formation were also seen in some nerve fibers in ganglia stained with lanthanum phosphate. Nerve endings with electron-lucent vesicles 40–60 nm in diameter are abundant in the cardiac ganglion, suggesting that these contain the excitatory transmitter of intrinsic neurons of the ganglion. Less abundant are fibers with membrane-limited opaque granules, circular or oblong in shape and as much as 330 nm in their longest dimension. Also seen were some nerve endings with both vesicles and granules.