Cerebral Neurosecretory Cells Research Articles

Neuroendocrine control of cocoon production in native earthworm Perionyx ceylanensis subjected to seasonal variation

Aim: The aim of the present study was to observe the role of cerebral ganglionic neurosecretory cells (NSCs) during cocoon production in native earthworm species Perionyx ceylanensis subjected to amputation and seasonal variations. Methodology: Histological studies (using Aldehyde Fuchsin and Chrome Alum Haematoxylin Phloxin stain) were carried out on brain NSCs in the two groups of earthworms (Group I and Group II) maintained in earthen culture pots (2L) with cowdung as food. Group I comprised of brain amputed earthworms was subjected to observe the role of brain NSCs in production of cocoon and Group II earthworms subjected to seasonal changes in the cerebral NSCs during cocoon production. Five replications were kept for Group I (1 individual per pot) and Group II (1 pair per pot). Results: Group I debrained earthworms started to lay cocoons from the 31st day following regeneration of cerebral ganglionic type A NSCs. In group II worms the highest neurosecretory activity was registered in the cerebral type A cells, especially during monsoon coinciding with the hike of cocoon generation. Interpretation: Appearance of type A NSCs in regenerated brain and peak of type A neurosecretory cell activity during peak reproductive period of earthworm species (as indicated by peak of cocoon production) indicates the possible role of cerebral type A NSCs in cocoon laying.

Effects of dehydration and subsequent hydration stress on the cerebral neurosecretory cells of Eudrilus eugeniae (Kinberg, 1867)

Effects of dehydration and subsequent hydration stress on the cerebral neurosecretory cells of Eudrilus eugeniae (Kinberg, 1867)

Effects of dehydration and subsequent hydration stress on the cerebral neurosecretory cells of Eudrilus eugeniae (Kinberg, 1867)

Moisture is a limiting factor for the survival and distribution of earthworms. During rainy season earthworms come out from their water-filled burrows and during summer move deeper into the soil to prevent acute desiccational water loss. Moisture loving worms like Eudrilus eugeniae when subjected to the dry soil and then subsequently hydrated, both the types of neurosecretory cells (A and B cells) show contrasting neurosecretory changes. During dehydration stress (2, 4 & 6 hours), both A and B cells deplete, lose their cellular conformity with extensive vacuolation. But in subsequent hydration of desiccated worms for 4 hrs, there is accumulation of neurosecretory materials in both types of cells, chiefly in A cells. The result indicates involvement of both A and B neurosecretory cells during osmotic stress from the perspective of histomorphic variations and secretory dynamics.

Ultrastructural studies on the neurosecretory cells in the pars intercerebralis of larvae of tasar silkworm, Antheraea mylitta (D) eco-race Bhandara (Lepidoptera: Saturniidae)

The cerebral neurosecretory cells (NSC) constitute four paired groups, medial (MNC), lateral (LNC-I, LNC-II) and posterior (PNC) in the brain of larvae of tasar silkworm Antheraea mylitta (D) Eco-race Bhandara. The MNC is the largest groups of peptidergic neurosecretory cells and are located in the pars intercerebralis region. The transmission electron microscopic (TEM), ultrastructure of the NSC confirmed the presence of mitochondria, endoplasmic reticulum, Golgi bodies, lysosomes and neurosecretory granules. The median neurosecretory cells shows secretory activity and release of secretory products, the neurosecretory granules (NSG) in the axons of NSC as well as the blood sinus.

Partial Synchronization Dynamics of Coupled Ultradian Oscillators Comprising an Insect Neurosecretory Cell System

An insulin-related peptide, bombyxin, in the silkmoth Bombyx mori is secreted by four pairs of cerebral neurosecretory cells that form a weakly coupled oscillator system to produce a pulsatile pattern of hormone secretion. The activity of individual bombyxin-producing (BP) cells oscillated with different periods (20-70 min). The population of BP cells exhibited complex phase dynamics, including spontaneous synchronization and desynchronization of different combinations of cells. Statistical cross-correlation analyses of oscillation patterns between BP cells revealed that one cell usually correlated closely with a few particular cells of similar periodicity. Close investigation of the phase differences between individual active phases of the related cell pairs revealed that an inphase synchronous state was usually maintained for many cycles, whereas an antiphase state was transient, lasting for a few cycles. In contrast, antiphase synchronous states often occurred between several cell pairs when the brain containing the cerebral neurosecretory cell system was disconnected from the ventral nerve cord containing the neuronal mechanism that induced periodic heartbeat reversals at intervals of 80-110 min and exerted a periodic suppressive or phase-resetting effect on individual BP cells. These results suggest that the internal coupling mechanism in the BP cell system is not sufficient to maintain an in-phase synchronous state in the heterogeneous cell population, and that the external phase resetting mechanism may assist in-phase synchronization of many neurosecretory cells to generate an overall pulsatile pattern of bombyxin secretion.

Co-ordination of firing activity of neurosecretory cells with cardiac activity in the silkmoth Bombyx mori

Electrocardiograms and electrical action potentials of cerebral neurosecretory cells producing bombyxin (an insulin-related neuropeptide) were simultaneously recorded from male pupae of the silkmoth Bombyx mori. A pupa showed alternations in the flow of haemolymph due to a rhythmic heartbeat reversal: a train of retrograde heartbeat with a slow pulse rate followed a train of anterograde heartbeat with a higher pulse rate. Intervals of heartbeat reversals changed throughout the pupal period. At any stage of the pupal period, firing activity of a population of bombyxin-producing (BP) cells rapidly declined after the start of anterograde heartbeat and an inactive state of cells continued during an anterograde heartbeat period. Analyses of ultradian bursting rhythmicity of a single BP cell revealed that a bursting phase of the cell often delayed at a time when the anterograde cardiac activity occurred at the preceding inter-burst period of firing rhythm. The results support the postulation that firing (secretory) activity of an insect neurosecretory cell system may be co-ordinated with circulation of haemolymph for rapid and pulsatile delivery of the peptides released to target organs.

Mutual coupling among insect neurosecretory cells with an ultradian firing rhythm

Firing correlations were examined among four pairs of cerebral neurosecretory cells with an ultradian bursting rhythm in the pupal silkmoth Bombyx mori. Cross-correlation analyses of spiking activities between pairs of simultaneously active cells usually revealed a significant correlation of firings: 20–50% of spikes of a cell fired within 0.2 s before or after the other cell's spikes fired. The mutual, excitatory communication among neurosecretory cells make the set of cells a weakly coupled multi-oscillator system that may generates a pulsatile release pattern of their product, an insulin-related neuropeptide (bombyxin).

A Neurosecretory Phenomenon in Some Cerebral Nervous Cells of the Land Snail Helix aspersa maxima (Mollusca, Gastropoda), and Its Relationship to Circadian Locomotor and Food-Intake Activities

Some cerebral neurosecretory cells of the land snail Helix aspersa maxima have been studied using light microscopy combined with image analysis. The amount of neurosecretion varies during the course of the day, displaying a peak at 22:00 GMT. This peak is in phasis with the maxima of the locomotor and food-intake activities recorded by means of a video apparatus.

Experimental aspergillosis in the German cockroach Blattella germanica: a histopathological study.

Inoculation of a spore suspension of Aspergillus flavus in the haemocoel of the German cockroach (Blatella germanica) resulted in large-scale invasion of various internal organs as seen in tissue sections stained with a fungal stain. The organs affected were the alimentary canal, fat bodies, muscle fibres, malpighian tubules and the cerebral neurosecretory cells. No invasion of the trachea and tracheoles was noted.

Neuroendocrine control of insect ecdysis by eclosion hormone

The peptides eclosion hormone (EH) and ecdysis-triggering hormone (ETH) trigger ecdysis behaviors and other physiological changes which occur at the end of the molt. EH is synthesized in a discrete set of cerebral neurosecretory cells, and is released into the central nervous system and into the hemolymph in response to the declining ecdysteroid titers. The onset of behavioral sensitivity to EH action is similarly controlled by ecdysteroids. EH is released from the ventral nerve cord for larval and pupal ecdyses, and from the corpora cardiaca for adult ecdysis. This transition is mediated by the restructuring of the EH cells during adult development, so that they elaborate an axon which extends to the CC for storage and release of EH just prior to adult ecdysis. ETH is released from the Inka cells, a component of the segmentally paired epitracheal glands, and acts directly on the central nervous system (CNS) to trigger preecdysis and ecdysis behavior. Although EH is best understood in lepidopteran insects, accumulating evidence suggests that EH is present in all insects and may provide a general hormonal mechanism to regulate ecdysis.

Control of cell proliferation in the corpora allata during the reproductive cycle of the cockroachDiploptera punctata

Unlike in Blettelta gcrmanica and Supella longipalpa, the corpora allata (CAI of Diploptera punctafa exhibited cyclic changes in cell number during the reproductive cycle. In mated females, a wave of DNA synthesis followed by mitosis resulted in a significant increase in CA cell number from about 9,000 cells on day 0 to 12,000 cells at ovulation on day 8. Subsequently, the number of cells per CA underwent a gradual decline to about 10,000 cells by day 64. During this long period of gestation, mitotic activity was undetectable (by colchicine arrest) and pycnotic nuclei were frequently observed by transmission electron microscopy. Just before parturition on day 72 another mitotic wave was detected and CA cell number increased again. The early wave of CA cell proliferation could be postponed by delaying mating or abolished by maintaining females as virgins. Neural disconnection of the CA from the brain mimicked the effect of mating, suggesting that enhanced cell proliferation is permitted by the removal of inhibitory signals from cerebral neurosecretory cells. The proliferative activities after mating were neither abolished by ovariectomy, which suppressed the normal increase in JH synthesis, nor elevated by unilateral allatectomy, which doubled the rates of JH synthesis in the remaining CA. These data corroborate previous results (Szibbo and Tobe, 1981 a; Tobe et al., 1984; Johnson et at., 1993) and suggest that waves of cell proliferation and JH synthesis, though simultaneous, are regulated independently by inhibitory signals from cerebral neurosecretory cells. D 1996 Witey-Liss, Inc.

Control of cell proliferation in the corpora allata during the reproductive cycle of the cockroach Diploptera punctata

Unlike in Blettella germanica and Supella longipalpa, the corpora allata (CA) of Diploptera punctata exhibited cyclic changes in cell number during the reproductive cycle. In mated females, a wave of DNA synthesis followed by mitosis resulted in a significant increase in CA cell number from about 9,000 cells on day 0 to 12,000 cells at ovulation on day 8. Subsequently, the number of cells per CA underwent a gradual decline to about 10,000 cells by day 64. During this long period of gestation, mitotic activity was undetectable (by colchicine arrest) and pycnotic nuclei were frequently observed by transmission electron microscopy. Just before parturition on day 72 another mitotic wave was detected and CA cell number increased again. The early wave of CA cell proliferation could be postponed by delaying mating or abolished by maintaining females as virgins. Neural disconnection of the CA from the brain mimicked the effect of mating, suggesting that enhanced cell proliferation is permitted by the removal of inhibitory signals from cerebral neurosecretory cells. The proliferative activities after mating were neither abolished by ovariectomy, which suppressed the normal increase in JH synthesis, nor elevated by unilateral allalectomy, which doubled the rates of JH synthesis in the remaining CA. These data corroborate previous results (Szibbo and Tobe, 1981a; Tobe et al., 1984; Johnson et al., 1993) and suggest that waves of cell proliferation and JH synthesis, though simultaneous, are regulated independently by inhibitory signals from cerebral neurosecretory cells. © 1996 Wiley-Liss, Inc.

Developmental expression of the prothoracicotropic hormone in the CNS of the Tobacco Hornworm Manduca sexta

The prothoracicotropic hormone is an insect neuropeptide released into the hemolymph to signal molting and metamorphosis through its stimulation of steroidogenesis. The only known source of the prothoracicotropic hormone in the tobacco hornworm, Manduca sexta, has been a group of lateral cerebral neurosecretory cells, the L-NSC III. In this study, the developmental and spatial distribution of the prothoracicotropic hormone was examined throughout the life cycle of Manduca. In common with many vertebrates and invertebrates in which neuropeptides are located in several regions within the central nervous system (CNS), the prothoracicotropic hormone phenotype in Manduca is expressed by CNS neurons in addition to the L-NSC III. These neurons are located in the brain, frontal ganglion, and subesophageal ganglion. One cerebral neurosecretory cell group, the ventromedial neurons, expresses the prothoracicotropic hormone phenotype and the behavioral neurohormone, eclosion hormone. Whereas the L-NSC III and the ventromedial neurons express the peptide phenotype throughout the life cycle, the other neurons express the peptide only during the embryonic and larval stages. This precise spatial and temporal expression of the prothoracicotropic hormone by different groups of neurosecretory cells raises the possibility that in Manduca the peptide may, in addition to its known neuroendocrine function, play other physiological roles in different ways at different stages of the life cycle.

Three‐dimensional architecture of identified cerebral neurosecretory cells in an insect

The organization of identified neurosecretory cell groups in the larval brain of the tobacco hornworm, Manduca sexta, was investigated immunocytologically. Computer-assisted three-dimensional reconstruction was used to examine the architecture of the neurosecretory cell groups. The group III lateral neurosecretory cells (L-NSC-III) which produce the prothoracicotropic hormone are located dorsolaterally in the protocerebrum and extend axons medially that decussate to the contralateral lobe prior to exiting the brain through the nervi corporis cardiaci I + II. The group IIa2 medial neurosecretory cells (M-NSC IIa2) are located anteriorly in the medial dorsal protocerebrum. The axons of these cells also exit the brain via the contralateral nervi corporis cardiaci I + II. However, their axons traverse a different pathway through the brain from that of the L-NSC III axons. Each of the cell groups possesses elaborate dendrites with terminal varicosities. The dendrites can be classified into specific fields based upon their location and projection pattern within the brain. The dendrites for these two neurosecretory cell groups overlap in specific regions of the protocerebral neuropil. After the axons of these neurosecretory cells exit the brain through the retrocerebral nerve, they innervate the corpus allatum where they arborize to form neurohemal terminals in strikingly different patterns. The L-NSC III penetrate throughout the glandular structure and the M-NSC IIa2 terminals are restricted to the external sheath. A third group of cerebral neurosecretory cells, the ventromedial neurons (VM) which stain with the monoclonal antibody to prothoracicotropic hormone in Manduca, are located anteriorly in the medial region of the brain. The axons of these cells do not exit the brain to the retrocerebral complex, but rather pass through the circumesophageal connectives and ventral nerve cord. These neurons appear to be the same VM neurons that produce eclosion hormone. One dendritic field of the L-NSC III terminates in close apposition to the VM neurons. The distinct morphologies of these neurosecretory cell groups in relation to other cell groups and the distribution of neuropeptides within the neurons suggest that insect neurosecretory cells, like their vertebrate counterparts, may have multiple regulatory roles.

Peptide‐immunocytochemistry of neurosecretory cells in the brain and retrocerebral complex of the sphinx moth Manduca sexta

Antisera against a variety of vertebrate and invertebrate neuropeptides were used to map cerebral neurosecretory cells in the sphinx moth Manduca sexta. Intense immunoreactive staining of distinct populations of neurosecretory cells was obtained with antisera against locust adipokinetic hormone, bovine pancreatic polypeptide, FMRFamide, molluscan small cardioactive peptide (SCPB), leucine-enkephalin, gastrin/cholecystokinin, and crustacean beta-pigment dispersing hormone (beta PDH). Other antisera revealed moderate to weak staining. Each type of neurosecretory cell is immunoreactive with at least one of the antisera tested, and most of these neurons can be identified anatomically. The staining patterns provide additional information on the organization of cerebral neurosecretory cells in M. sexta. Based upon anatomical and immunocytochemical characteristics, 11 types of neurosecretory cells have been recognized in the brain, one type in the suboesophageal ganglion, and one in the corpus cardiacum. Extensive colocalization experiments show that many neurosecretory cells are immunoreactive with several different antisera. This raises the possibility that these cells may release mixtures of neuropeptides into the hemolymph, as has been demonstrated in certain other systems. The immunocytochemical data should be helpful in efforts to identify additional peptide neurohormones released from the brain of this and other insects.

Extirpation and transplantation of the brain of the snail Helix aspersa: a study of the survival of the animal and the implant

Dormant, hibernating snails whose brains (supraoesophageal cerebral ganglia and associated dorsal bodies) were removed by surgery continued to live retracted in their shells. The remaining ganglia of the circumoesophageal ring did not regenerate in the 12 months following the removal of the brain. The transplanted brains in the haemocoel (auto- or allo-graft) were not rejected and many of their cerebral neurosecretory cells and the endocrine dorsal bodies appeared normal structurally and functionally, as evidenced by ultrastructural and immunocytochemical studies. The data from these experiments demonstrate the possibility of studying by in vivo culture methods the roles of cerebral ganglia and endocrine dorsal bodies on the control of reproduction under different temperature regimes.

Regeneration of the neurohemal terminals for identified cerebral neurosecretory cells in an insect

The axons of specific neurosecretory cells, L-NSC III, in the brain of the tobacco hornworm, Manduca sexta, were transected during larval-pupal development to study the effects of this type of lesion on these peptidergic neurons and to begin to identify factors that may regulate their regeneration and growth. The two somata of these bilaterally paired neurons produce the prothoracicotropic hormone and are located in the pars intercerebralis. Their axons exit from the contralateral brain lobe via a retrocerebral nerve and pass through the corpus cardiacum before terminating at the glandular corpus allatum. At the corpus allatum, the L-NSC III axons arborize to form the terminal neurohemal organ for prothoracicotropic hormone release. The retrocerebral nerve was severed either in vitro followed by brain transplantation or in situ; in either protocol, the distal axon segments and corpus allatum were removed. The ability of the injured L-NSC III axons to regenerate was assessed immunocytologically by using a monoclonal antibody against the prothoracicotropic hormone. In both treatments, the proximal axon stumps exhibited regenerative growth as early as 1 day after axotomy, and, by the third day, neurites had extended. By the fifth day, the regenerating axons had branched to form terminal varicosities similar to those of a normal neurohemal organ. The regenerated neurohemal structure appeared to be functional, because larvae that had been bilaterally axotomized were able to metamorphose to pupae, a process requiring temporally precise periods of prothoracicotropic hormone release. In addition to the regeneration of the terminal axon structures, several other responses to axotomy and retrocerebral organ excision occurred. These included an apparent accumulation of prothoracicotropic hormone in the axons and regenerating neurohemal-like structure, sprouting of ectopic neurites from the axotomized somata, and a change in shape of the cell bodies from spherical to avoid.

Production of antibodies to Manduca sexta cerebral neurosecretory cells

AbstractUnderstanding the neuroendocrine regulation of insect development depends upon having antibody probes to the neurohormones involved, which are usually present at trace levels, making antibody generation difficult. This report describes a simple method for producing antibodies specific to cerebral neurosecretory cells (NSC) of the tobacco hornworm, Manduca sexta, and to the neurohormone(s) they produce. The method involves the isolation of only a few hundred NSC somata (∼ 0.3 μg of protein) that serve as the immunogen. The cerebral NSC used were the L‐NSC III, the prothoracicotropes, that produce the prothoracicotropic hormone (PTTH), the principal neuroendocrine effector of insect molting and metamorphosis. A PTTH‐containing extract of microsurgically isolated somata of the L‐NSC III was injected intraperitoneally into a Balb/c mouse. The antiserum produced specifically immunostained the L‐NSC III in wholemounts of brains from different developmental stages. This antiserum also contained antibodies directed against PTTH, as shown by its ability to inhibit the neurohormone's biological activity in an in vitro prothoracic gland bioassay. Such antiserum can be used to investigate the ontogeny and phylogeny of NSCs. With the hybridoma technique, monoclonal antibodies to individual NSC proteins (PTTH) could be obtained, circumventing the need to purify them for antibody production. This method should be applicable to comparable neurosecretory cell systems in other insect species.

Identification of the cerebral neurosecretory cells that contain eclosion hormone in the moth Manduca sexta

Eclosion hormone (EH) is an insect neuropeptide that is released at the end of metamorphosis from the CNS and triggers the stereotyped motor program of adult emergence. Using three distinct experimental approaches, we have identified a discrete set of neurosecretory cells in the brain of the moth Manduca sexta that contains and releases EH. By isolating the neurosecretory somata and testing them with a sensitive behavioral bioassay, we identified a cluster of ipsilaterally projecting cells (Group Ia) that contain EH. Intracellular stimulation of individual cells within this group induced the release of bioactive EH into the hemolymph surrounding the neurohemal organs of the brain, whereas stimulation of cells in the other cerebral neurosecretory clusters did not. We also developed a polyclonal antiserum against purified EH that precipitated all bioactive material from samples containing the peptide. This antiserum selectively stained 5 of the Group Ia cells on either side of the brain, as well as their central and terminal processes. Preincubation of the serum with EH dramatically reduced its ability to bind the peptide subsequently. The combined application of these physiological and immunological techniques has led to the unequivocal identification of the EH neurons in the moth brain.

Control of neurosecretion in the moth Manduca sexta: physiological regulation of the eclosion hormone cells.

Metamorphosis in the moth Manduca sexta culminates with the secretion of the peptide eclosion hormone (EH), which triggers the stereotyped behavior of adult emergence (eclosion) from the pupal cuticle. In restrained but spontaneously behaving animals, the release of EH occurred shortly before the onset of subjective night (Fig. 3) and coincided with a depletion of EH from the neurohemal organs of the brain, the corpora cardiaca-corpora allata complex (CC-CA; Fig. 4). EH is produced by neurons within a bilaterally paired group of brain neurosecretory cells (Group Ia) which project to the CC-CA via the nervi corporis cardiaci- 1 + 2 (NCC-1 + 2; Fig. 1). Electrical stimulation of the NCC-1 + 2 caused a marked increase in the levels of EH secreted from isolated CC-CA (Fig. 2), while stimulation of the other nerves innervating the neurohemal organs did not. Electrical activity in the NCC-1 + 2 paralleled that of the cerebral neurosecretory cells (Fig. 1). Chronic extracellular recordings revealed a sudden increase in the tonic firing of several units within this nerve approximately 2 to 3 h before normal eclosion (Fig. 5), coincident with the release of EH bioactivity from the CC-CA (Fig. 6). The Group Ia neurons were electrically inactive on the day before eclosion (Day-1), but on the day of eclosion (Day 0) a subgroup of these cells exhibited both enhanced synaptic input and elevated rates of tonic firing during the normal time of EH release (Fig. 7). No significant differences in resting membrane potential or spike waveform characteristics were detected among the various subsets of Group Ia cells on either Day-1 or Day 0, while a significant increase in the resting input resistance was seen in the active subgroup on Day 0 (Fig. 8). This increase may be due to the regulatory effects of the steroid 20-hydroxyecdysone, which inhibits the release of EH and may act by preventing the synaptic activation of the EH neurons until the final day of adult development.