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.     

Morphology of the dorsal vessel in the abdomen of the blood-feeding insect Rhodnius prolixus

The dorsal vessel (DV) in the abdomen of the blood-feeding insect Rhodnius prolixus was divided functionally into two regions, the heart, into which haemolymph entered the DV through four pairs of ostia located in abdominal segment VII, and the aorta, along which the haemolymph was propelled from abdominal segment VI to the thorax. Osmium-fixed whole mounts revealed the DV to consist of spirally arranged striated muscle fibers and to possess two rows of ventrally attached longitudinal fibers extending the length of the abdomen. Seven pairs of alary muscles were found attached to the DV in the posterior abdominal segments. Contractions of the alary muscles attached to the ventral surface of abdominal segments VII and VIII served to expand the heart. Electron microscopy revealed the DV to consist of a thin layer of contractile elements surrounded by an inner (intima) and outer (adventitia) connective tissue layer. Embedded in the intima along each lateral side of the DV were two large groups of endocardial cells extending the length of the DV. A small group of pericardial cells was embedded in the adventitia along the mid-ventral side of the DV, and clusters of pericardial cells were found attached to the alary muscles. Nerve terminals were found only on the heart: they contained agranular synaptic vesicles approximately 30 nm in diameter and densely stained granules approximately 100-120 nm in diameter. These structural components are discussed in relation to the role of the DV in circulation.     

Excitatory Transmission in Insect Neuromuscular Systems

Many, but not all, visceral muscles in insects are innervatedby neurosecretory axons. The neurosecretory junctions with theheart muscle of the American cockroach, Periplaneta americana,show ultrastructural and electrophysiological evidence of chemicallytransmitting synapses, and cytochemical evidence for the presenceof monoamines. Electron microscopy of nerve terminals showsthat synaptic vesicles may be formed directly from electron-dense"neurosecretory" granules Neurotomy of motor axons to skeletal muscles in insects leadsto aggregation and clumping of synaptic vesicles after 48 hours.Treatment of in vitro nerve-muscle preparations with variousrespiratory poisons caused aggregation similar to that developedin neurotomized animals. This suggested that vesicle aggregationin both cases may have resulted from a decrease in availableadenosine triphosphate in the nerve terminal with subsequentalteration in the normal charge density which supports a repulsiveforce between the vesicles.     

Innervation of heart and alary muscles in Sphinx ligustri L. (Lepidoptera)

Summary The origin and orientation of the heart nerves in Sphinx ligustri and Ephestia kuehniella were investigated by scanning electron microscopy using a special technique which involved pinning the dissected specimens on a stabilizing metal pad. The heart and alary muscles in Sphinx particularly their caudal extremity were also examined by transmission electron microscopy. The alary muscles form an incomplete sheath around the heart with a mainly longitudinal fibre orientation, e.i. antagonistically to the fibres of the heart itself. The heart and alary muscles are multiterminally innervated by branches of the transverse segmental nerves. All branches contain a single electron lucent axon; the thickest branches also possess several neurosecretory axons. Swellings of the segmental nerves may indicate the position of nerve cell bodies. There are no lateral heart nerves. Only one type of neuromuscular junction is abundant in the alary muscles but less frequently found in the heart. The terminals originate from the central axon only. They are capped by glial cells, which interdigitate with the muscle cells. They penetrate into the T-system toward the Z-discs and form a complex intercellular space system. Exocytosis of dense-cored vesicles into this perisynaptic reticulum seems likely. Sites of neurohaemal release are distributed along the nerve branches and special nerve endings occur at the level of the ostia. The possible nervous influence upon heart activity is discussed.The transmission electron microscopic part of this investigation was supported by a research scholarship from the Deutsche Forschungsgemeinschaft     

The hypothalamo-hypophysial system of the wild carp,Cyprinus carpio L.

Summary The posterior neurohypophysis (PNH)-pars intermedia complex of the wild and pond carp, Cyprinus carpio L., has been studied by light, fluorescence and electron microscopy. Gomori-positive neurosecretory fibres are abundant in the main trunk of the neurohypophysis as well as its roots penetrating the pars intermedia. Terminals of these fibres are in contact with capillaries of the general circulation and with glandular cells of the pars intermedia. Monoaminergic fibres with a weak green fluorescence, somewhat increasing after injection of nialamide into the pond carp, have largely the same distribution. Three types of neurosecretory fibres and their terminals have been recognized in the PNH-pars intermedia complex. Types-A₁ and -A₂ fibres, containing granules of 140–180 nm and 100–160 nm in diameter respectively, are peptidergic Gomori-positive. Type-A₂ fibres predominate in the PNH. The least frequent monoaminergic type-B fibres have granules of 60–100 nm in diameter. Numerous peptidergic and few monoaminergic neurosecretory terminals make contact with the capillaries located within the roots of the PNH as well as at the border between them and the pars intermedia. Both peptidergic and monoaminergic terminals make direct synaptoid contacts with the gland cells or end close to connective tissue septa, basal lamina or pituicytes. The PAS-positive gland cells and to a lesser degree the leadhaematoxylin-positive gland cells show these relationships with neurosecretory terminals. The question concerning the mode of interaction between peptidergic and monoaminergic structures in the dual control of the gland cells of the pars intermedia of teleosts is discussed.     

Effect of sympathomimetic substances and DOPA on the ultrastructure of the nervous apparatus of the heart]

After a single administration of norepinephrine or DOPA to albino rats there occurred an incorporation of norepinephrine into the adrenergic axons of the heart and its deposition in the form of granules in small synaptic vesicles, about 300 A in diameter. The adrenergic and cholinergic axons can be thus differentiated. The amount of cholinergic axons in the auricles is greater than that of the adrenergic ones. The adrenergic terminals came into the most intimate contact with the cholinergic terminals and with the endothelial cells of the blood capillaries and the myocardial muscle cells. It is supposed that adrenergic fibers can act upon the heart muscle in three ways: by means of presynaptic inhibition through the cholinergic axons, by humoral route, and directly on the myocardial muscle cells.     

The hypothalamo-hypophysial system of the frog Rana temporaria

Summary The median eminence (ME) of the adult frog, Rana temporaria, was studied by means of electron microscopy including quantitative electron-microscopic autoradiography. In frogs captured in May and June numerous peptidergic neurosecretory fibres extending via the internal zone to the pars nervosa display large swellings containing few granules, mitochondria, neurotubules and cisternae of the smooth endoplasmic reticulum. In addition, few secretory globules up to 1.5 m in diameter occur in these varicosities. In animals collected during the autumn period many of these neurosecretory swellings filled with neurosecretory granules and polymorphic inclusions resemble Herring bodies. Three types of granule-containing neurosecretory fibres were observed in the external zone (EZ) of the ME of adult R. temporaria. Peptidergic A₁- and A₂-type fibres are characterized by granules 150–220 nm and 100–160 nm in diameter, respectively. Monoaminergic fibres of type B with granules approximately 100 nm in diameter represent 50% of all neurosecretory elements in the EZ of the frog ME; 12% of the total number of granule-bearing axons in the EZ actively taking up radiolabelled 5-hydroxytryptophan are thought to be serotoninergic terminals. Neurosecretory terminals of all types and glial vascular endfeet establish direct contacts with the perivascular space of the primary portal capillaries. Some neurosecretory terminals are separated from the lumen of the third ventricle by a thin cytoplasmic lamella of tanycytes. The possible physiological significance of this structural pattern is discussed.     

Vasoactive intestinal polypeptide (VIP)- and neuropeptide Y (NPY)-containing nerve fibres in the penile cavernous tissue of green monkeys (Cercopithecus aethiops)

Summary The cavernous body of green monkeys contains many unmyelinated and few myelinated axons. The unmyelinated axons form terminals in the adventitia of the arteries, between trabecular muscle cells, in the interstitium, and close to endothelium cells of the sinuses. All terminals displayed predominantly small clear vesicles and very few large granular vesicles; small granular vesicles were not seen. However, in rabbit penises, terminals with many large granular vesicles are prominent. Immunohistochemistry (PAP technique) showed a dense network of VIP- and NPY-reactive fibres around the arteries and around trabecular muscles. The density of nerve fibres was particularly high around the subendothelial cushions of the helicine arteries. Double staining for NPY and VIP revealed that both peptides were colocalized. Immunocytochemistry (preembedding PAP technique) showed VIP- and NPY-reactivity in terminals with small clear vesicles; the reaction product was bound to the cytoplasmic face of different membrane types. Although the intracellular localization of the reaction product is probably due to artefactual displacement during preparation, the uniformity of the terminals questions the view that large and small granular vesicles in all species characterize peptidergic and noradrenergic terminals, respectively. The essential findings can be summarized as (1) a high degree of uniformity of nerve terminals, (2) colocalization of VIP and NPY, (3) heavy innervation of the subendothelial cushions of the helicine arteries, and (4) possible innervation of endothelial cells.     

Ultrastructural and immunocytochemical studies of neuromuscular junctions in oviduct of Locusta migratoria

The ultrastructure of nerve endings in the oviduct visceral muscles of Locusta migratoria was studied by electron microscopy and by immunogold labeling for two kinds of neuromodulators, the pentapeptide proctolin and FMRFamide-related peptides. Nerve endings contained electron-lucent round vesicles and two kinds of granules (round and avoid), and formed two types of synapses or release sites with the muscle. The morphologically distinct nerve endings were classified into three different categories based on the composition of synaptic vesicles and granules. Type-I nerve endings were dominated by electron-lucent round vesicles and contained only a few round electron-dense granules. Type-II nerve endings contained mostly electron-dense round granules and electron-lucent round vesicles. A few electron-dense ovoid granules were also present. Electron-dense ovoid granules dominated the type-III nerve endings, which usually contained less electron-lucent vesicles than either type-I or II nerve endings. Both proctolin and FMRFamide-like immunoreactivity was associated with electron-dense round granules. However, FMRFamide-like immunoreactivity was only found in the type-II nerve endings, while proctolin immunoreactivity was found within type-I nerve endings as well as in some type-II nerve endings. Immunological results therefore allow us to further divide type-II nerve endings into type-IIa (immunonegative for proctolin) and type-IIb (immunopositive for proctolin). Type-III nerve endings show no immunolabeling to either proctolin or FMRFamide.     

Two types of neuromuscular junctions in the pharyngeal retractor muscle ofHelix lucorum

The pharyngeal retractor muscle of the snailHelix lucorum is innervated by a pair of nerves containing axons of two types, for which there are two corresponding types of myoneural junctions with the muscle cells. The junctions of type I correspond to the thick axons. The terminals of these axons, which contain numerous spherical transparent vesicles (41±5 nm) and fewer vesicles of the dense-core type (67±3 nm), make contact mainly with noncontracting sarcoplasmic projections of the muscle cells. Junctions of type II correspond to thin axons, containing many granules. The terminals of these axons make contact with contractile parts of the muscle cells and they contain a heterogeneous population of vesicles: small spherical clear vesicles (44±2 nm), granules with fine-grained contents (135±5 nm), and a few spherical dense-core vesicles. The distance between the muscle cells is usually great — over 50 nm, but in the region of the sarcoplasmic processes the surface membranes come together to form a gap which in some areas does not exceed 10 nm.N. K. Kol'tsov Institute of Developmental Biology, Academy of Sciences of the USSR, Kiev. Translated from Neirofiziologiya, Vol. 9, No. 5, pp. 539–542, September–October, 1977.     

Fine structure of pars neuro-intermedia of the loach,Misgurnus fossilis L.

Summary Two types of glandular cells are present in the pars intermedia of the loach, Misgurnus fossilis. Basophils are characterized by the presence in their cytoplasm of numerous secretory granules containing electron-dense and homogeneous material and by scarce endoplasmic reticulum. Weak acidophils contain in their cytoplasm abundant endoplasmic reticulum and numerous granules of different electron densities.The distal part of the neurohypophysis is composed of several types of neurosecretory axons, strongly branched pituicytes, numerous capillaries, and connective tissue elements. The axon terminals form the neuroglandular, neurovascular and neurointerstitial contacts. Some axon terminals are closely apposed to the basement membrane separating neurohypophysis from meta-adenohypophysis. At points of absence of continuity of this membrane, some neurosecretory axons become directly contiguous with cytoplasmic membranes of the intermedia cells.The investigation was partly supported by a research grant from the Zoological Committee of the Polish Academy of Sciences.     

Ultrastructural changes in the gracile nucleus of the spontaneously diabetic BB rat.

The present study describes the structural changes in the gracile nucleus of the spontaneously diabetic BB rat. At 3-7 days post-diabetes, axons, axon terminals and dendrites showed electron-dense degeneration. Degenerating axons were characterized by swollen mitochondria, vacuolation, accumulation of glycogen granules, tubulovesicular elements, neurofilaments and dense lamellar bodies. Degenerating axon terminals consisted of an electron-dense cytoplasm containing swollen mitochondria, vacuoles and clustering of synaptic vesicles. These axon terminals made synaptic contacts with cell somata, dendrites and other axon terminals. Degenerating dendrites were postsynaptic to normal as well as degenerating axon terminals. At 1-3 months post-diabetes, degenerating electron-dense axons, axon terminals and dendrites were widely scattered in the neuropil. Macrophages containing degenerating electron-dense debris were also present. At 6 months post-diabetes, the freshly degenerating neuronal elements encountered were similar to those observed at 3-7 days. However, there were more degenerating profiles at 6 months post-diabetes compared to the earlier time intervals. Terminally degenerating axons were vacuolated and their axoplasm appeared amorphous. It is concluded that degenerative changes occur in the gracile nucleus of the spontaneously diabetic BB rat.     

Electron microscopic and pharmacological studies on the rat median eminence

Summary The rat median eminence contains at least three kinds of granules or vesicles: 1. large electron-dense granules (perhaps carriers of neurohypophysial hormones), 2. small electron-dense granules with or without haloes (perhaps carriers of catecholamines) and 3. synaptic vesicle-like structures (perhaps carriers of acetylcholine). The former two electrondense granules exist in separate axons but they coexist with the latter vesicles in the same axons.The pars nervosa shows basically a similar structure to the median eminence. However, the axons containing the small electron-dense granules are very few. In the pars tuberalis, there are at least two types of cells: the cells of one type contain much cytoplasm with large round nuclei and those of the other type contain a small amount of cytoplasm with polymorphic nuclei. The cells of the former include multivesicular bodies and secretory granules, but those of the latter do not. Some of capillaries of the primary plexus are surrounded by the cells of the pars tuberalis on one side and by neurosecretory axon endings on the other side.The median eminence contains high concentration of acetylcholine or an acetylcholine-like substance and shows neurohypophysial hormone activity.Aided by Grant A-3678 from the United States National Institute of Arthritis and Metabolic Diseases. The authors are indebted to Dr. Welsh, Harvard University, for the kind gift of Mytolon.     

Electron-microscopic study of innervation of smooth muscle cells surrounding collecting tubules of the fish kidney

Summary The fine structure of the collecting tubules of the trout and killifish kidney was studied. These tubules are surrounded by layers of smooth muscle cells which are commonly innervated. The nerve terminals contain synaptic vesicles and, occasionally, a few dense-cored granules as well. Capillaries occur in the connective tissue space between these smooth muscle cells and the collecting tubule. Epithelial cells of the collecting tubules contain abundant mitochondria and a well developed membrane system displaying parallel arrays, and were considered to be actively involved in the transport of materials. In the trout, the collecting tubules contain peculiar cells in addition to regular tubule cells. The fine structure of these peculiar cells is highly reminiscent of that of gill chloride cells. The significance of these findings may be summarized as follows: If the smooth muscles around the collecting tubule contract under neural influence, intratubular pressure may be increased and, thus affect glomerular filtration rate. The contraction of these muscles may also cause the collapse of peritubular capillaries, affecting the transport activity of tubule cells.     

Immuno-electron microscopy of sorting and release of neuropeptides in Lymnaea stagnalis

Summary The cerebral caudodorsal cells of the pulmonate snail Lymnaea stagnalis control egg laying and egglaying behavior by releasing various peptides derived from two precursors. The biosynthesis, storage, intracellular breakdown and release of three caudodorsal cell peptides were studied by means of immuno-electron microscopy using antisera raised to fragments of these peptides: (1) Caudodorsal Cell Hormone-I (CDCH-I; derived from precursor I), (2) Caudodorsal Cell Hormone-II (CDCH-II; from precursor II), and (3) -Caudodorsal Cell Peptide ( CDCP; from both precursors). After affinity purification of the antisera, the specificity of the sera was confirmed with dotting immunobinding assays. From the ultrastructural immunocytochemical data it has been concluded that the precursor molecules are cleaved at the level of the Golgi apparatus after which the C-terminal parts (containing CDCP) and N-terminal parts (containing CDCH-I or CDCH-II) are sorted and preferentially packaged into large electron-dense granules (MD 150 nm), respectively. Very probably, the content of the large electron-dense granules is degraded within the cell body. The immunoreactivity of the secretory granules increases during discharge from the Golgi apparatus, indicating further processing. At least a portion of the secretory granules contains all three peptides, as shown by double and triple immunopositive stainings whereas other granules appear to contain only one or two of these peptides. The caudodorsal cells release multiple peptides via exocytosis from neurohemal axon terminals into the hemolymph and from blindly ending axon collaterals into the intercellular space of the cerebral commissure (nonsynaptic release).     

Electron microscopic and histochemical observations on different types of nerve endings in the extraocular muscles of the rat

Summary Nerve endings in the extraocular muscles of the rat were submitted to histochemical tests for formalin-induced fluorescence and carboxylic esterases. Acetylthiocholine, butyrylthiocholine and -naphthyl acetate were used as substrates and iso-OMPA, 284C51, eserine and E-600 as inhibitors. The ultrastructure of the endings was studied with the electron microscope.Both single and multiple nerve terminals were observed in all six extraocular muscles. The single terminals of myelinated axons were comparable in their light and electron microscopic structure with the typical motor end plates of other striated muscles, and like these they exhibit acetylcholinesterase (AChE), non-specific cholinesterase (ns. ChE) and non-specific esterase (ns. E) activity. These endings were apposed to twitch-type muscle fibres.The multiple terminals were classified with the light microscope into two types. The larger type was 1/3 of the size of the motor end plate; 2–5 endings innervated the same muscle fibre; subneural infoldings were weakly developed and possessed only slight AChE and ns. ChE and probably no ns. E activity. No subneural lamellae were visible under the light microscope in the smaller type, which also possessed AChE and ns. ChE and was composed of 10–20 small dots dispersed along a single muscle fibre. The Schwann cells along nerve fibres leading to these two types of multiple endings exhibited ns. ChE but not AChE and ns. E activity.The ultrastructure of the two types of multiple endings was principally similar. The main difference, compared with the motor end plate, was that these endings were derived from unmyelinated axons which either make synaptic contacts along their course with the muscle fibre at variable distances (smaller-type) or these terminals were grouped closely together (larger-type).A few dense-core vesicles were observed in these unmyelinated nerves and in their terminals which were considerably smaller than those in the motor end plate. They were not always separated from each other by sarcoplasm and teloglia (larger-type) and contained also empty vesicles. The secondary synaptic clefts were often sparse and irregular or even absent, but the typical myoneural postsynaptic electron density was always observed. These multiple endings, in contrast to the motor end plate, were apposed only to muscle fibres with slow contraction.No catecholamine containing nerve endings were observed in the extraocular muscles. These observations indicate that the rat extraocular muscles have a double cholinergic innervation.The author wishes to express his gratitude to Professor Antti Telkkä, M. D., Head of the Electron Microscope Laboratory, University of Helsinki, for permission to avail himself of the electron microscope facilities.     

The pedal muscle of the land snail Megalobulimus oblongus (Gastropoda, Pulmonata): an ultrastructure approach

M. Cristina Faccioni-Heuser, Denise M. Zancan, Christiane Q. Lopes and Matilde Achaval. 1999. The pedal muscle of the land snail Megalobulimus oblongus (Gastropoda, Pulmonata): an ultrastructure approach. — Acta Zoologica (Stockholm) 80: 325–337
The ultrastructure of the pedal muscle of the Megalobulimus oblongus is described. This muscle consists of transverse, longitudinal and oblique bundles ensheathed in collagenous tissue. Each muscle cell is also ensheathed by collagen. The smooth muscle cells contain thin and thick filaments; the thin filaments are attached to dense bodies. These cells contain a simple system of sarcoplasmic reticulum, subsarcolemmal caveolae and mitochondria with dense granules in the matrix, and glycogen. Three types of muscle cells were identified. Type A cells exhibited densely packed myofilaments, abundant glycogen rosettes, numerous mitochondria and sarcoplasmic reticulum profiles. Type B cells exhibited scanty glycogen and mitochondria, few cisternae of sarcoplasmic reticulum and large intermyofibrillar spaces. Type C cells exhibited intermediate characteristics between type A and type B cells. Neither nexus nor desmosomes were observed between the muscle cell membranes. The muscle contains well developed connective tissue and blood vessels. These structures and the distribution of muscle cells are probably involved in the muscular-hydrostat system. The muscle is richly innervated, having neuromuscular junctions with clear and electron-dense synaptic vesicles. The clear vesicles probably contain acetylcholine because the axons to which they are connected arise from acetylcholinesterase positive neurones of the pedal ganglion. The other vesicles may secrete monoamines such as serotonin and/or neuropeptides such as substance P.     

Localization and function of an FMRFamide-like substance in the aorta of Helix aspersa

Summary With an antiserum (aFM) against the molluscan cardio-active FMRFamide (Phe-Met-Arg-Phe-NH₂) numerous immunoreactive axons were found in the outer, longitudinal, muscle layer of the anterior aorta of Helix aspersa. Immunoreactive axons were rare in the inner, circular, muscle layer. At the ultrastructural level four types of axons could be distinguished. The granules containing the immunoreactive substance (mean diameter ca. 100 nm) are present in type-2 axons. The effect of synthetic FMRF-amide was tested in vitro on preparations of ring- and tubule-shaped pieces of the anterior aorta. Physiological doses (3 × 10^-7 M) provoked contractions of the circular muscle fibres, but had no effect on the longitudinal muscle cells. Apparently in vivo the FMRF-like substance diffuses from the richly innervated longitudinal muscle layer to the circular muscle layer, where it exerts its effect. This conclusion is sustained by the observation that the contents of the aFM-immunoreactive granules in type-2 axons are released by exocytosis in a non-synaptic fashion.     

The efferent innervation of the genital chamber by an identified serotonergic neuron in the female cricket Acheta domestica

Summary The serotonergic innervation of the genital chamber of the female cricket, Acheta domestica, has been investigated applying anti-serotonin (5-HT) immunocyto-chemistry at both light- and electron-microscopic levels as well as using conventional electron microscopy. Whole mount and pre-embedding chopper techniques of immuno-cytochemistry reveal a dense 5-HT-immunoreactive network of varicose fibers in the musculature of the genital chamber. All of these immunoreactive fibers originate from the efferent serotonergic neuron projecting through the nerve 8v to the genital chamber (Hustert and Topel 1986; Elekes et al. 1987). At the electron-microscopic level, 5-HT-immunoreactive nerve terminals, which contain small (50–60 nm) and large ( 100 nm) agranular vesicles as well as granular vesicles (100nm), contact the muscle fibers or the sarcoplasmic processes without establishing specialized neuromuscular connections. In addition to the 5-HT-immunoreactive axons, two types of immunonegative axons can also be found in the musculature. By use of conventional electron microscopy, three ultrastructurally distinct types of axon processes can be observed, one of which resembles 5-HT-immunoreactive axons. While the majority of the varicosities do not synapse on the muscle fibers, terminals containing small (50–60 nm) agranular vesicles occasionally form specialized neuromuscular contacts. It is suggested that the 5-HTergic innervation plays a non-synaptic modulatory role in the regulation circular musculature in the genital chamber of the cricket, while the musculature as a whole may be influenced by both synaptic and modulatory mechanisms.Fellow of the Alexander von Humboldt-Stiftung     

The neurosecretory cells of the brain and suboesophageal ganglion of Chironomus riparius

The neurosecretory cells of the supra- and suboesophageal ganglia of young, unmated, adult male midges, Chironomus riparius, have been examined by both light and electron microscopy. The 5 cell types recognized have been placed in three major categories on the basis of their ultrastructural characteristics:—α₁ cells, of which there are 8 in each medial neurosecretory cell (MNC) group and 3 in each group of ventral neurosecretory cells (VNC), contain electron-dense granules, 150 to 200 nm in diameter; α₂ cells containing irregular, electron-dense granules, 70 to 120 nm in diameter comprise the remaining 3 cells in each VNC group and the 2 or 3 cells in each outer neurosecretory cell (ONC) group; α₃ cells, of which there are 1 or 2 on each side of the midline in the ventral cortex of the sub-oesophageal ganglion (SNC₂), contain electron-lucent, spherical granules, 70 to 120 nm in diameter. The β cells contain spherical or ellipsoidal, electron-lucent granules, 80 to 100 nm in diameter, and make up the lateral neurosecretory cell (LNC) groups, each of three or four cells. The γ cells contain both spherical and flattened, electron-dense granules, 130 to 160 nm in diameter and 150 to 250 by 70 to 150 nm in size respectively, only 1 cell of this category being found in each half of the suboesophageal ganglion in the dorsal cortex (SNC₁). Axons from the MNC and VNC form the nervi corporis cardiaci I (NCCI) and those of the LNC and ONC, the nervi corporis cardiaci II (NCCII). Those of the SNC₁ appear to enter the wall of the stomodaeum but axons of the SNC₂ could not be traced.