The neural control of spermathecal contractions in the locust,Locusta migratoria

The innervation of the spermatheca and demonstration of neural control of spermathecal contractions in Locusta migratoria was illustrated using anterograde and retrograde fills, combined with electrophysiological stimulation and recording. The anterior portion of the spermatheca receives innervation via the receptaculum seminis nerve (N2B2) from two large ventral neurons and one dorsal neuron. All were bilaterally paired and situated in the VIIIth abdominal ganglion. Three ventral bilaterally paired neurons situated in the VIIIth abdominal ganglion also provide innervation to the posterior portion of the spermatheca via the ductus seminalis aperture nerve (N2B3). Six DUM neurons, located in the VIIIth abdominal ganglion, in addition to two centroposteriorly situated DUM neurons in the VIIth abdominal ganglion, are also associated with these two nerves. N2B4 also provides innervation to the posterior portion of the spermatheca. N2B6b is associated with sensory cells identified in the anterior lateral regions of the genital chamber. The spermatheca contracts spontaneously, with peristaltic contractions beginning at the spermathecal sac and continuing along the length of the spermathecal duct. However electrical stimulation of the ventral ovipositor nerve (VON or N2B), receptaculum seminis nerve (N2B2) and the ductus seminalis aperture nerve (N2B3) indicates that contractions are also under neural control. In particular contractions of the spermathecal sac, coil duct and anterior straight duct are initiated via motor projections from the receptaculum seminis nerve (N2B2) and posterior straight duct contractions are controlled by motor input from the ductus seminalis aperture nerve (N2B3). The results suggest that spermathecal contractions of the anterior and posterior portions of the spermatheca are under separate neural control.     

The fate of the normal-anucleated spermatozoa in inseminated females of the silkworm Bombyx mori

Both typical (haploid) and atypical (anucleated) spermatozoa reach the receptaculum seminis of inseminated females of Bombyx mori intermingled. However, only typical spermatozoa both leave the receptaculum and fertilize the eggs. Atypical spermatozoa, which are in fact anucleated flagellar apparatuses, probably function in transporting typical fertilizing spermatozoa to the receptaculum seminis. In the male ejaculatory duct both kinds of spermatozoa are wrapped with extra-cellular sleeves that presumably protect them on their way to the receptaculum. Typical spermatozoa “hatch” from the sleeves before leaving the receptaculum to fertilize the eggs. The presence of a centriole in the extra-testicular spermatozoa of this species supports the generalization that insect spermatozoa do have a centriole at the base of the flagellum.     

Evidence of a neural loop involved in controlling spermathecal contractions in Locusta migratoria

The control of spermathecal contractions in Locusta migratoria via a neural loop was demonstrated using mechanical stimulation and electrophysiological recordings. Extracellular electrophysiological recordings from the receptaculum seminis nerve (N2B2), which innervates the spermathecal sac, were conducted during mechanical stimulation of the genital chamber sensory cells. Activation of the genital chamber sensory cells, using a glass probe approximating the shape and size of an egg, was found to increase the action potential frequency and initiate bursts of action potentials if a tonic frequency of action potentials was present prior to stimulation. If the motor pattern initially consisted of bursts of action potentials, then mechanical stimulation of the genital chamber sensory cells resulted in an increase in firing frequency, in most preparations, with the bursting remaining. Removal of the probe from the genital chamber always returned the motor activity to that noted prior to sensory cell stimulation. Simultaneous electrophysiological recordings from both the left and right receptaculum seminis nerves (N2B2) revealed that the bursts of action potentials were coordinated, although individual action potentials were not coupled one to one. Activation of the genital chamber sensory cells also resulted in increases in spermathecal contraction frequency, an effect which was coordinated with the changes in motor activity. It is proposed that an egg in the genital chamber activates the sensory cells resulting in increases in spermathecal contraction frequency and the subsequent release of spermatozoa onto the micropyle of the egg for fertilization.     

Ultrastructure of receptaculum seminis in Haemaphysalis longicornis (Acari: Ixodidae) in relation to inserted endospermatophore

The receptaculum seminis, opening into the female genital tract, is found only in the metastriate ixodid ticks. An endospermatophore that has been inserted into the female genital aperture at copulation is first stored in the receptaculum seminis, where spermiogenesis is completed before the sperm ascend the oviducts. The receptaculum seminis consists of a simple cuticularized epithelium. Epithelial cells in sexually matured females develop during feeding and become active in secretion. Secretions discharged from epithelial cells are released into the lumen of this organ through the cuticle and may act on the wall of the inserted endospermatophore. The fact that resumption of spermiogenesis (spermateleosis) has already occurred before destruction of the endospermatophore just after copulation suggests that secretions from epithelial cells of the receptaculum seminis are not the trigger of spermateleosis, but a destructive agent of the endospermatophore wall. J Morphol 231:143–147, 1997. © 1997 Wiley-Liss, Inc.     

Evidence for the association of FMRFamide-related peptides with the spermatheca of Locusta migratoria

The association of FMRFamide-related peptides (FaRPs) with the spermatheca of Locusta migratoria was demonstrated using radioimmunoassay and immunohistochemical techniques. The physiological effects of various FaRPs on the neurally evoked contractions of the spermatheca were also examined. FMRFamide-like immunoreactivity (FLI) was demonstrated in processes and cell bodies situated in the VIIIth (terminal) abdominal ganglion. These included an anterior, central and posterior pair of ventral cell bodies positioned near the midline of the ganglion, in addition to two bilaterally paired dorsal cell bodies in the posterior region of the VIIIth abdominal ganglion. Two axons displaying FLI proceed down the ventral ovipositor nerve (VON) and into the receptaculum seminis nerve which innervates the anterior regions of the spermatheca. FLI was also noted in processes on the spermathecal muscle with the highest density occurring on the spermathecal sac and coil duct. FaRPs applied to the spermathecal muscle included GQERNFLRFamide, NFIRFamide, ADDRNFIRFamide, YGGFMRFamide, FMRFamide, ADVGHVFLRFamide and SchistoFLRFamide (PDVDHVFLRFamide). Dose-dependent physiological effects were only noted for FMRFamide, ADVGHVFLRFamide and SchistoFLRFamide. FMRFamide led to a dose-dependent increase in the amplitude of neurally evoked contractions with a threshold of approximately 5 x 10(-7) M. SchistoFLRFamide, and ADVGHVFLRFamide, had an inhibitory effect, decreasing the amplitude of neurally evoked spermathecal contractions.     

Spermiogenesis and sperm ultrastructure of Cyathocephalus truncatus (Pallas, 1781) Kessler, 1868 (Cestoda: Spathebothriidea)

Spermiogenesis and sperm ultrastructure of adult Cyathocephalus truncatus, a member of presumably basal group of "true" cestodes (Eucestoda), have been examined for the first time by using transmission electron microscopy. The process of sperm formation corresponds in basic pattern to that of the Pseudophyllidea. In addition, the 2 pairs of electron-dense attachment zones are present in median cytoplasmic process of C. truncatus. However, mature spermatozoa of C. truncatus differ significantly from those of the pseudophyllideans, especially in the morphology of the proximal and distal spermatozoon extremities. The proximal extremity of the mature spermatozoon lacks a crested body, which is present in more derived cestodes and some pseudophyllideans. The distal end of the mature spermatozoa exhibits different morphology in the gametes from testes and those from receptaculum seminis. New for the Eucestoda is a finding that a lateral cytoplasmic extension creates the distal end of the spermatozoa from testes, resembling sperm of some Monogenea and Digenea. In contrast, the distal extremity of the spermatozoa from receptaculum seminis contains only a nucleus. Despite the above-mentioned peculiarities, the ultrastructural data on sperm/spermiogenesis suggest close relationships of the Spathebothriidea and Pseudophyllidea.     

Different effects of the biogenic amines dopamine,serotonin and octopamine on the thoracic and abdominal portions of the escape circuit in the cockroach

1. The escape behavior of the cockroach, Periplaneta americana, is known to be modulated under various behavioral conditions (Camhi and Volman 1978; Camhi and Nolen 1981; Camhi 1988). Some of these modulatory effects occur in the last abdominal ganglion (Daley and Delcomyn 1981a, b; Libersat et al. 1989) and others in the thoracic ganglia (Camhi 1988). Neuromodulator substances are known to underlie behavioral modulation in various animals. Therefore, we have sought to determine whether topical application of putative neuromodulators of the escape circuit enhance or depress this circuit, and whether these effects differ in the last abdominal vs. the thoracic ganglia. 2. Topical application of the biogenic amines serotonin and dopamine to the metathoracic ganglion modulates the escape circuitry within this ganglion; serotonin decreases and dopamine enhances the response of leg motoneurons to activation of interneurons in the abdominal nerve cord by electrical or wind stimulation. 3. The neuropil of the thoracic ganglia contains many catecholamine-histofluorescent processes bearing varicosities, providing a possible anatomical substrate for dopamine release sites. 4. Topical application of octopamine to the terminal abdominal ganglion enhances the response of abdominal interneurons to wind stimulation of the cerci. In contrast, serotonin and dopamine have no effect at this site. 5. It is proposed that release of these biogenic amines may contribute to the known modulation of the cockroach escape response.     

The terminal abdominal ganglion of the wood cricket Nemobius sylvestris

The abdominal cerci of the wood cricket, Nemobius sylvestris, are covered by a variety of hair‐like sensilla that differ in length, thickness, and articulation. Fillings from the cercal nerves with cobalt chloride and fluorescent dyes revealed the projection of sensory axons into the terminal abdominal ganglion of the ventral nerve chain. Two projection areas on each side of the terminal abdominal ganglion midline could be identified: a posterior cercal glomerulus and an anterior bristle neuropil. Axons from some cercal sensilla ascend through the connectives to reach the metathoracic ganglionic mass. As their axons pass through each segmental abdominal ganglion, they project medial arborization. Cross‐sections of the terminal abdominal ganglion and retrograde fills with cobalt chloride and fluorescent dyes from connectives revealed several small cells and seven pairs of giant ascending interneurons organized symmetrically. Giant somata are located contralateral to their axons (diameters between 20 and 45 μm). The cercal projections overlap extensively with the dendritic fields of the giant interneurons. In the terminal abdominal ganglion, we identified nine longitudinal tracts, two major tracts, and seven smaller ones. The functional implications of the neuranatomical organization of the system are discussed on a comparative basis. J. Morphol., 2008. © 2008 Wiley‐Liss, Inc.     

绿盲蝽腹神经节的解剖结构

【目的】揭示绿盲蝽Apolygus lucorum腹神经节的组成结构。【方法】采用免疫组织化学染色方法,利用突触蛋白抗体对绿盲蝽成虫的腹神经节进行免疫标记,激光共聚焦扫描显微镜扫描照相获得原始数据,用图像分析软件进行标记,构建三维结构模型。【结果】绿盲蝽成虫腹神经节位于腹神经索的末端,与其前方的后胸神经节和中胸神经节紧密融合,形成后部神经节。与脑和胸神经节类似,腹神经节由周围的细胞体和内部的神经髓构成。腹神经节的神经纤维束主要包括位于腹侧的两条纵向神经连索和向两侧发出的9束神经纤维。9束神经纤维连接着9个神经原节,即富含突触联系的神经髓。这些神经原节紧密融合,无明显的边界,最后两节形成膨大的末端腹神经节。两侧的神经原节由横向的神经连锁连接起来。腹神经节外周的细胞体数量较多,排列紧密,大小一致,仅在前端背侧中间和后端腹侧中间位置分别有2个和5个体积较大的细胞体。【结论】本研究结果明确了绿盲蝽腹神经节的结构,为进一步研究昆虫的行为调控及神经系统发育和演化奠定一定的形态学基础。     

Release mechanism of sex pheromone in the female gypsy moth <Emphasis Type="Italic">Lymantria dispar</Emphasis>: a morpho-functional approach

A morpho-functional investigation of the sex pheromone-producing area was correlated with the pheromone release mechanism in the female gypsy moth Lymantria dispar. As assessed by male electroantennograms (EAG) and morphological observations, the pheromone gland consists of a single-layered epithelium both in the dorsal and ventral halves of the intersegmental membrane between the 8th and 9th abdominal segments. By using the male EAG as a biosensor of real-time release of sex pheromone from whole calling females, we found this process time coupled with extension movements of the ovipositor. Nevertheless, in females in which normal calling behavior was prevented, pheromone release was detected neither in absence nor in presence of electrical stimulation of the ventral nerve cord/terminal abdominal ganglion (TAG) complex. Tetramethylrhodamine-conjugated dextran amine stainings also confirm the lack of any innervation of the gland from nerves IV to VI emerging from the TAG. These findings indicate that the release of sex pheromone from the glands in female gypsy moths is independent of any neural control exerted by the TAG on the glands, at least by way of its three most caudally located pairs of nerves, and appears as a consequence of a squeezing mechanism in the pheromone-producing area.     

Ultrastructure of the reproductive system of the house dust mitesDermatophagoides farinae andD. pteronyssinus (Acari,Pyroglyphidae) with remarks on spermatogenesis and oogenesis

Several parts of the reproductive system of both sexes ofDermatophagoides farinae andD. pteronyssinus are investigated and compared by light-, scanning electron-, and transmission electron microscopy. New techniques have been employed for scanning of the internal structures of these mites. The male reproductive system consists of unpaired testis, paired vasa deferentia, an accessory gland, ejaculatory duct, and copulatory organ. The female reproductive system consists of bursa copulatrix, ductus bursae, receptaculum seminis, paired ducti receptaculi, ovaries, oviducts, one chorion gland, ovipositor, and oviporus. Testis as well as ovaries are characterized by syncytia of nutritional function. The specifics of spermatogenesis are discussed in connection with sperm transfer. Similarities between the construction of the ovaries and oogenesis in astigmatic mites and telotrophic meroistic ovaries in insects are discussed.     

Anatomy and ultrastructure of the reproductive systems ofAcarus siro (Acari: Acaridae)

Anatomy and ultrastructure of the female and male reproductive system inAcarus siro L. were investigated by light and electron microscopy. The female system consists of paired ovaries of nutrimentary type in which oogonia and oocytes are connected by bridges with a large central cell. The oviducts empty into the uterus, which passes into preoviporal duct lined bycuticle, and opening as a longitudinal slit (oviporus). An elongated accessory gland composed of one type of secretory cell is located along each oviduct. The copulatory opening occurs at the posterior margin of the body and leads, via the inseminatory canal, to the receptaculum seminis, consisting of the basal and saccular part. Both inseminatory canal and basal part of receptaculum seminis are lined by cuticle, whereas the wall of the sac is formed by cells covered only by long, numerous microvilli. The basal part of the receptaculum seminis joins the ovaries via two lumenless transitory cones.The male reproductive system contains paired testes, in which spermatogonia tightly surround the central cell. The proximal part of the paired vasa deferentia serves as a sperm reservoir, while the distal one has a glandular character. An unpaired, cuticle-lined ejaculatory duct opens into the apex of the aedeagus. The single accessory gland is located asymmetrically at the level of, or slightly posterior to, coxae IV.The structure of the genital papillae, which are topographically related to the genital opening in both sexes, is also briefly described.     

The neurophysiology of larval firefly luminescence: Direct activation through four bifurcating (DUM) neurons

Summary The paired lanterns of the larval fireflyPhoturis versicolor are bilaterally innervated by four dorsal unpaired median (DUM) neurons the somata of which are found in the terminal abdominal ganglion (A8) and which stain with Neutral Red (Fig. 1A). Both intra- and extracellularly recorded activity in these neurons is always associated with a bilateral glow response, or BGR (Figs. 3 and 4). Luminescence cannot be initiated or maintained in the absence of DUM neuron excitation. Furthermore, there is a linear causative relationship between the frequency of DUM neuron activity and the amplitude of the resultant BGR (Figs. 6 and 7).Due to the intrinsic bilateral morphology, firefly DUM neurons may be antidromically activated through either lantern nerve, resulting in the initiation of luminescence in the contralateral lantern (Figs. 8 and 9). This activation is unaffected by high Mg⁺⁺ saline indicating that the DUM neurons provide a direct pathway for conduction through the ganglion (Fig. 9). The DUM neurons receive synaptic input from axons descending through both anterior connectives, however, stimulation of only one connective results in a BGR since excitation is carried to both sides of the periphery through the bilateral axons.Firefly DUM neurons exhibit physiological qualities typical of neurosecretory cells: spikes are characterized by a slow time course and a long and deep afterhyperpolarization (Fig. 10). This is consistent with the observation that spontaneous firing rates are usually below 3 Hz, but nevertheless elicit a strong BGR (Figs. 3 and 5). The physiological evidence presented in this study correlates well with the morphological, pharmacological and biochemical evidence compiled from previous studies, which indicates that the four DUM neurons represent the sole photomotor output from the central nervous system to the larval lanterns. Evidence is discussed which indicates that these effects are mediated throught the release of octopamine, long presumed to be the lantern neurotransmitter. These results, therefore, describe a novel and unexpected role for DUM neurons in regulating an unusual invertebrate effector tissue and further expands the growing list of functions for octopamine in neural control mechanisms.Abbreviations A1-A7 first through seventh abdominal ganglia - A8 terminal abdominal ganglion - DUM dorsal unpaired median - BGR bilateral glow response     

Structure of allotransplanted ganglia and regenerated neuromuscular connections in crayfish

In adult crayfish, Procambarus clarkii, motoneurons to a denervated abdominal superficial flexor muscle regenerate long-lasting and highly specific synaptic connections as seen from recordings of excitatory postsynaptic potentials, even when they arise from the ganglion of another crayfish. To confirm the morphological origins of these physiological connections we examined the fine structure of the allotransplanted tissue that consisted of the third abdominal ganglion and the nerve to the superficial flexor muscle (the fourth ganglion and the connecting ventral nerve cord were also included). Although there is considerable degeneration, the allotransplanted ganglia display intact areas of axon tracts, neuropil, and somata. Thus in both short (6–8 weeks) and long (24–30 weeks) term transplants approximately 20 healthy somata are present and this is more than the five axons regenerated to the host muscle. The principal neurite and dendrites of these somata receive both excitatory and inhibitory synaptic inputs, and these types of synaptic contacts also occur among the dendritic profiles of the neuropil. Axon tracts in the allotransplanted ganglia and ventral nerve cord consist largely of small diameter axons; most of the large axons including the medial and lateral giant axons are lost. The transplanted ganglia have many blood vessels and blood lacunae ensuring long-term survival. The transplanted superficial flexor nerve regenerates from the ventral to the dorsal surface of the muscle where it has five axons, each consisting of many profiles rather than a single profile. This indicates sprouting of the individual axons and accounts for the enlarged size of the regenerated nerve. The regenerated axons give rise to normal-looking synaptic terminals with well-defined synaptic contacts and presynaptic dense bars or active zones. Some of these synaptic terminals lie in close proximity to degenerating terminals, suggesting that they may inhabit old sites and in this way ensure target specificity. The presence of intact somata, neuropil, and axon tracts are factors that would contribute to the spontaneous firing of the transplanted motoneurons. © 1996 John Wiley & Sons, Inc.     

Cardiac reflexes and their neural pathways in lepidopterous insects

Previously described salines for lepidoptera did not maintain a constant heart rate for a very long. We have been successful in maintaining a normal heartbeat for many hours in a newly designed saline. This saline was also suitable for maintaining normal neuromuscular junctional potentials. The cardiac reflexes studied in larvae of Bombyx and Agrius were five types of cardiac responses induced by mechanical stimuli to sensillar setae. The cardiac responses were caused by electrical stimulation of nerves in the reflex pathways. The antidromic heartbeat was triggered even in larvae before the 5th instar by stimulation of axons in the visceral nerve arising from the frontal ganglion and terminating at the aorta, while spontaneous heartbeat reversal started to occur in wandering larvae. Other axons in the visceral nerve terminate at the rear end of the heart. Electrical stimuli to the nerves caused cardiac inhibition of the orthodromic heartbeat. Nerves extending from the visceral nerve to the alary muscles of the 2nd abdominal segment contain axons to increase the tone of the muscles. Nerves extending from the 7th abdominal ganglion to the most posterior alary muscles also contain axons to increase the tone of the muscles, and were responsible for acceleration of the antidromic and orthodromic heartbeat, respectively.     

Neural activation of the sex-pheromone gland in the moth Manduca sexta: real-time measurement of pheromone release

Abstract. Investigations of various species of moths have suggested that the biosynthesis of sex pheromone in the abdominal pheromone glands of females may be at least partly regulated by neuroendocrine mechanisms. Few studies, however, have explored the mechanisms underlying the release of sex pheromone. In experiments on the sphinx moth Manduca sexta (L.) (Lepidoptera: Sphingidae), we have monitored the time course of sex-pheromone release in scotophase females with the aid of an electroantennogram bioassay based on the highly sensitive and selective sex-pheromone receptor neurones of the male antenna. Pheromone release was evoked by orthodromic stimulation of the ventral nerve cord. Neurally stimulated release occurred with a subsecond latency and did not depend on bioactive factors in the haemolymph or on movement of the abdomen or the ovipositor. Severing the most medial pair of nerves posterior to the terminal abdominal ganglion (the terminal nerves) eliminated pheromone release, but not abdominal contractions. Release was also inhibited reversibly if the descending Ca²⁺-dependent synaptic input to the terminal ganglion was blocked by exposure to elevated concentrations of Mg²⁺. These findings indicate that the release of sex pheromone from the pheromone gland in female M. sexta is a true neuroeffector response and that the gland appears to be controlled by neurones that project to it from the terminal abdominal ganglion.     

Anatomy and physiology of neurons composing the commissural ring nerve of the cricket, Acheta domesticus

The commissural ring nerve (RN) of the cricket Acheta domesticus links together the two cercal motor nerves of the terminal abdominal ganglion. It contains the axons of almost 100 neurons including two bilateral clusters of eight to 13 ventrolateral neurons and approximately 75 neurons with midline somata within the terminal abdominal ganglion. The ventrolateral neurons have an ipsilateral dendritic arborization within the dorsal neuropil of the ganglion and their axons use the RN as a commissure in order to enter the contralateral nerves of the tenth ganglionic neuromere. In contrast, most midline neurons have bifurcating axons projecting bilaterally into the neuropil of the ganglion as well as into the RN where they often branch extensively before entering the contralateral tenth nerves. Most RN neurons have small, non-spiking somata with spike initiation zones distant from the soma. Many midline neurons also produce double-peaked spikes in their somata, indicative of multiple spike initiation zones. Spontaneous neuronal activity recorded extracellularly from the RN reveals several units, some with variable firing patterns, but none responding to sensory stimuli. The RN is primarily composed of small (50 nm diameter) axon profiles with a few large (0.5-1 microm diameter) profiles. Occasionally, profiles of nerve terminals containing primarily small clear vesicles and a few large dense vesicles are observed. These vesicles can sometimes be clustered about an active zone. We conclude that the primary function of the RN is to serve as a peripheral nerve commissure and that its role as a neurohemal organ is negligible. J. Exp. Zool. 286:350-366, 2000.Copyright 2000 Wiley-Liss, Inc.     

Commissural ring nerve: A female-specific neurosecretory tract supplied by bifurcating median neurons in the cockroach Periplaneta americana (L.) and the cricket Teleogryllus commodus (Walker)

Summary In the American cockroach, Periplaneta americana, and the Australian field cricket, Teleogryllus commodus, the two nerves supplying the bases of the cerci are joined by a branch that crosses behind the last abdominal ganglion. This commissural ring nerve is restricted to females, and it contains many axons filled with granular and agranular vesicles. The axons stem from somata located within the ganglion. There are one (Periplaneta) or two (Teleogryllus) groups of median neurons with bilaterally symmetrical bifurcations, and a group of postero-ventral neurons on each side. In T. commodus, these neurons are distinct from others associated with the cerci. In the two species, the ring nerve neurons contribute to a neuropile near the root of each cereal nerve. The bifurcating median neurons arborize on both sides before entering the ring nerve, while the postero-ventral ones branch more extensively ipsilateral to their somata. The possibilities are discussed that the bifurcating neurons may be homologous to dorsal unpaired median neurons, and that the ring nerve may be a neurohemal area.     

The distribution of pheromone-biosynthesis-activating neuropeptide (PBAN) immunoreactivity in the central nervous system of the corn earworm moth,Helicoverpa zea

Summary Production of sex pheromone in several species of moths has been shown to be under the control of a neuropeptide termed pheromone-biosynthesis-activating neuropeptide (PBAN). We have produced an antiserum to PBAN from Helicoverpa zea (Lepidoptera: Noctuidae) and used it to investigate the distribution of immunoreactive peptide in the brain-suboesophageal ganglion complex and its associated neurohemal structures, and the segmental ganglia of the ventral nerve cord. Immunocytochemical methods reveal three clusters of cells along the ventral midline in the suboesophageal ganglion (SOG), one cluster each in the presumptive mandibular (4 cells), maxillary (12–14 cells), and labial neuromeres (4 cells). The proximal neurites of these cells are similar in their dorsal and lateral patterns of projection, indicating a serial homology among the three clusters. Members of the mandibular and maxillary clusters have axons projecting into the maxillary nerve, while two additional pairs of axons from the maxillary cluster project into the ventral nerve cord. Members of the labial cluster project to the retrocerebral complex (corpora cardiaca and cephalic aorta) via the nervus corpus cardiaci III (NCC III). The axons projecting into the ventral nerve cord appear to arborize principally in the dorsolateral region of each segmental ganglion; the terminal abdominal ganglion is distinct in containing an additional ventromedial arborization in the posterior third of the ganglion. Quantification of the extractable immunoreactive peptide in the retrocerebral complex by ELISA indicates that PBAN is gradually depleted during the scotophase, then restored to maximal levels in the photophase. Taken together, our findings provide anatomical evidence for both neurohormonal release of PBAN as well as axonal transport via the ventral nerve cord to release sites within the segmental ganglia.Abbreviations A aorta - Br-SOG brain-suboesophageal ganglion complex - CC corpus cardiacum - PBS phosphate-buffered saline - PLI PBAN-like immunoreactivity - TAG terminal abdominal ganglion - VNC ventral nerve cord     

Secretion of axonally transported neural peptides from the nervous system of Aplysia

The possibility that proteins reaching the abdominal ganglion of Aplysia by axonal transport from the circumesophageal ganglia might be subject to secretion in that structure was examined. Transported labeled protein was found to be released from the abdominal ganglion; such release was enhanced by exposure to a high K⁺ medium and by electrical stimulation of the transporting axons. Stimulation of release was inhibited by lowering the Ca²⁺/Mg²⁺ ratio of the medium. The released material is predominantly of 1–2000 daltons in molecular weight and appears to have been derived from a group of transported peptides of about the same size. The possibility is raised that these data may reflect the existence of a peptidergic second-order neurosecretory pathway in this nervous system.