Serotonin-like immunoreactivity in the optic lobes of three insect species

The cellular localization of 5-HT in the optic lobes of three insect species was assayed with the use of antibodies raised against 5-HT. In Schistocerca, Periplaneta, and Calliphora all neuropil regions of the optic lobe, the lamina, medulla and lobula, contain 5-HT-immunoreactive varicose fibres in different patterns, like columns and layers. Such fibres also connect the lobula to neuropil in the lateral protocerebrum. In Calliphora also 5-HT-positive fibres of the medulla and lobula plate have projections to the lateral protocerebrum, whereas the origin of the lamina fibres is not certain. In all species the processes displaying 5-HT-like immunoreactivity appear to be derived from a relatively small number of cell bodies, each neuron thus having processes over a large volume of the neuropil of the optic lobe in different layers.     

Immunocytochemical characterization of the accessory medulla in the cockroach Leucophaea maderae

Several lines of evidence suggest that pigment-dispersing hormone-immunoreactive neurons with ramifications in the accessory medulla are involved in the circadian system of insects. The present study provides a detailed analysis of the anatomical and neurochemical organization of the accessory medulla in the brain of the cockroach Leucophaea maderae. We show that the accessory medulla is compartmentalized into central dense nodular neuropil surrounded by a shell of coarse fibers. It is innervated by neurons immunoreactive to antisera against serotonin and the neuropeptides allatostatin 7, allatotropin, corazonin, gastrin/cholecystokinin, FMRFamide, leucokinin I, and pigment-dispersing hormone. Some of the immunostained neurons appear to be local neurons of the accessory medulla, whereas others connect this neuropil to various brain areas, including the lamina, the contralateral optic lobe, the posterior optic tubercles, and the superior protocerebrum. Double-label experiments show the colocalization of immunoreactivity against pigment-dispersing hormone with compounds related to FMRFamide, serotonin, and leucokinin I. The neuronal and neurochemical organization of the accessory medulla is consistent with the current hypothesis for a role of this brain area as a circadian pacemaking center in the insect brain.     

Comparative anatomy of pigment-dispersing hormone-immunoreactive neurons in the brain of orthopteroid insects

Summary In a comparative study, the anatomy of neurons immunoreactive with an antiserum against the crustacean -pigment-dispersing hormone was investigated in the brain of several orthopteroid insects including locusts, crickets, a cockroach, and a phasmid. In all species studied, three groups of neurons with somata in the optic lobes show pigment-dispersing hormone-like immunoreactivity. Additionally, in most species, the tritocerebrum exhibits weak immunoreactive staining originating from ascending fibers, tritocerebral cells, or neurons in the inferior protocerebrum. Two of the three cell groups in the optic lobe have somata at the dorsal and ventral posterior edge of the lamina. These neurons have dense ramifications in the lamina with processes extending into the first optic chiasma and into distal layers of the medulla. Pigment-dispersing hormone-immunoreactive neurons of the third group have somata near the anterior proximal margin of the medulla. These neurons were reconstructed in Schistocerca gregaria, Locusta migratoria, Teleogryllus commodus, Periplaneta americana, and Extatosoma tiaratum. The neurons have wide and divergent arborizations in the medulla, in the lamina, and in several regions of the midbrain, including the superior and inferior lateral protocerebrum and areas between the pedunculi and -lobes of the mushroom bodies. Species-specific differences were found in this third cell group with regard to the number of immunoreactive cells, midbrain arborizations, and contralateral projections, which are especially prominent in the cockroach and virtually absent in crickets. The unusual branching patterns and the special neurochemical phenotype suggest a particular physiological role of these neurons. Their possible function as circadian pacemakers is discussed.     

Distribution of PER protein,pigment-dispersing hormone,prothoracicotropic hormone,and eclosion hormone in the cephalic nervous system of insects

Investigations performed on adult insects revealed that putative components of the central pacemaker, the protein Period (PER) and the pigment-dispersing hormone (PDH), are immunocytochemically detectable in discrete sets of brain neurons throughout the class of Insecta, represented by a bristletail, mayfly, damselfly, 2 locust species, stonefly, 2 bug species, goldsmith beetle, caddisfly, honeybee, and 2 blowfly species. The PER-positive cells are localized in the frontal protocerebrum and in most species also in the optic lobes, which are their only location in damselfly and goldsmith beetle. Additional PER-positive cells occur in a few species either in the deuto- and tritocerebrum or in the suboesophageal ganglion. The PER staining was always confined to the cytoplasm. The PDH immunoreactivity consistently occurs in a cluster of perikarya located frontoventrally at the proximal edge of the medulla. The mayfly and both locust species possess additional PDH neurons in 2 posterior cell clusters at the proximal edge of the medulla, and mayfly, waterstrider, and 1 of the blowfly species in the central brain. PDH-positive fibers form a fanlike arrangement over the frontal side of the medulla. Two or just 1 bundle of PDH-positive fibers run from the optic lobe to the protocerebrum, with collaterals passing over to the contralateral optic lobe. Antisera to the prothoracicotropic (PTTH) and the eclosion (EH) hormones, which in some insects regulate the molting and ecdysis rhythms, respectively, typically react with a few neurons in the frontal protocerebrum. However, the PTTH-positive neurons of the mayfly and the damselfly and the EH-positive neurons of the caddisfly are located in the suboesophageal ganglion. No PTTH-like antigen was detected in locusts, and no EH-like antigens were detected in the damselfly, stonefly, locusts, and the honeybee. There are no signs of co-localization of the PER-, PDH-, PTTH-, and EH-like antigens in identical neurons.     

Distribution of corazonin and pigment-dispersing factor in the cephalic ganglia of termites

Distribution of neurones detectable with antisera to the corazonin (Crz) and the pigment-dispersing factor (PDF) was mapped in the workers or pseudergates of 10 species representing six out of seven termite families. All species contained two triads of Crz-immunoreactive (Crz-ir) neurones in the protocerebrum. Their fibres were linked to the opposite hemisphere, formed a network in the fronto-lateral protocerebrum, and projected to the corpora cardiaca (CC); in most species the fibres also supplied the deuto- and tritocerebrum and the frontal ganglion. Some species possessed additional Crz-ir perikarya in the protocerebrum and the suboesophageal ganglion (SOG). The PDF-ir somata were primarily located in the optic lobe (OL) and SOG. OL harboured a group (3 groups in Coptotermes) of 2-6 PDF-ir cells with processes extending to the medulla, connecting to the contralateral OL, forming 1-2 networks in the protocerebrum, and in most species running also to CC. Such a PDF-ir system associated with the OL was missing in Reticulitermes. Except for Mastotermes, the termites contained 1-2 PDF-ir cell pairs in the SOG and two species had additional perikarya in the protocerebrum. The results are consistent with the view of a monophyletic termite origin and demonstrate how the Crz-ir and PDF-ir systems diversified in the course of termite phylogeny.     

Connections between wide-field monocular and binocular movement detectors in the brain of a hawk moth

Summary Recordings were made in the brain of Sphinx ligustri of pairs of directionally selective movement detectors, and the spike trains analysed with a computer for possible synaptic connections between two classes of movement detector. (1) Neurones with large binocular fields which arise in the medial protocerebrum and project to the medulla or lobula of one optic lobe, or to the ventral nerve cord. (2) Movement detectors which project from the lobula complex of one optic lobe to the opposite medial protocerebrum. The majority of the second group had back-to-front preferred directions over the ipsilateral eye, and of these many were weakly sensitive to stimuli to the opposite eye. The ipsilateral receptive field covered most of the eye.Optic lobe output cells with the appropriate preferred direction provide a powerful excitatory input to the binocular movement detectors centrifugal to the medulla. Each centrifugal movement detector probably receives excitatory inputs from no more than two optic lobe output cells with back-to-front preferred direction. The same set of optic lobe output neurones probably feeds several cells projecting to the medulla and lobula of both optic lobes, and, possibly, to the ventral nerve cord.Evidence was obtained that the optic lobe output cells themselves receive few excitatory inputs, and that therefore the receptive fields of their input cells are large.Two moving stimuli were presented in different areas of the receptive field. Movement through the null direction in one area inhibited the response to movement in the preferred direction in another area. This suppression was stronger in optic lobe output cells with front-to-back preferred direction than in units with back-to-front preferred direction. Thus the optic lobe output cells, or wide-field units feeding them, receive inhibitory inputs from wide-field units with the opposite preferred direction.Similar tests in which moving stimuli were presented to both eyes gave results indicating that the binocular centrifugal movement detectors may receive inhibitory inputs from movement detectors with back-to-front preferred direction. The possible functional significance of these inhibitory inputs is discussed.I am very greatful to F. A. Miles for helpful discussion and criticism. Financial support came from the U. K. Science Research Council.     

Postembryonic changes in circadian photo-responsiveness rhythms of optic lobe interneurons in the cricket Gryllus bimaculatus

The photo-responsiveness of 2 groups of interneurons responding to light in the protocerebrum was investigated at 2 developmental stages, the last instar nymphs and adults, in the cricket Gryllus bimaculatus. The cricket is diurnally active during the nymphal stage but becomes nocturnal as an adult. In both adults and nymphs, light-induced responses of optic lobe light-responding interneurons that conduct light information from the optic medulla to the lobula and the cerebral lobe showed a circadian rhythm peaking during the subjective night. Amplitudes of the rhythms were not significantly different between adults and nymphs, but adults showed more stable day and night states than did nymphs. The medulla bilateral neurons that interconnect the bilateral medulla areas of the optic lobe also showed circadian rhythms in their light-induced responses in both adults and nymphs. The rhythm had a clear peak and a trough in adults, and its amplitude was significantly greater than that of nymphs. These results suggest that the 2 classes of interneurons are differentially controlled by the circadian clock. The difference might be related to their functional roles in the animal's circadian behavioral organization.     

蟋蟀脑的解剖结构与生理机能

蟋蟀脑由前脑、中脑和后脑三部分组成。前脑由1对蕈形体、中央复合体和视叶构成;每个蕈形体由2个冠、柄及与柄相连的α叶和β叶组成,是信息联络整合部位;中央复合体由中央体和脑桥组成,主要参与感觉信息的加工过程;视叶由神经节层、外髓和内髓组成,是视觉系统的中心。中脑由主要组成成分为嗅觉纤维球的嗅叶组成,是嗅觉系统的中心。后脑向后与食道下神经节相连。     

Postembryonic development of serotonin-immunoreactive neurons in the central nervous system of the blowfly,Calliphora erythrocephala

Summary The postembryonic development of serotonin-immunoreactive (5-HTi) neurons was studied in the optic lobe of the blowfly. In the adult fly there are 24 5-HTi neurons invading each optic lobe. The perikarya of two of these neurons are situated in the dorso-caudal part of the protocerebrum (LBO-5HT neurons; large bilateral optic lobe 5-HTi neurons). The cell bodies of the remaining 22 neurons are located anteriorly at the medial base of the medulla (2 innervating the lobula, LO-5HT neurons; and 20 neurons innervating the medulla, ME-5HT neurons). The two central neurons (LBO-5HT neurons) are derived from metamorphosing larval neurons, while the ME- and LO-5HT neurons are imaginai optic lobe neurons differentiating during pupal development.The 5-HTi neurons of the optic lobe seem to have different ancestors. The LBO-5HT neurons are probably derived from segmental protocerebral neuroblasts, whereas the ME-and LO-5HT neurons are most likely derived from the inner optic anlage. The first 5-HTi fibers to reach the imaginal optic lobes are seen in the late third instar larva and are derived from the LBO-5HT neurons. The first ME- and LO-5HT neurons become immunoreactive at 24 h (10%) pupal development. At about 96 h (40%) of pupal development all the 5-HTi neurons of the optic lobes have differentiated and attained their basic adult morphology. The further development mainly entails increase in volume of arborizations and number of finer processes. The differentiation and outgrowth of 5-HTi processes follows that of, e.g., columnar neurons in the optic lobe neuropils. Hence, 5-HTi processes invade neuropil relatively late in the differentiation of the optic lobe.     

Physiology and morphology of olfactory neurons associating with the protocerebral lobe of the honeybee brain

Physiology and morphology of olfactory neurons associated with the protocerebral lobe around the alpha-lobe of the mushroom body were studied in the brain of the honeybee Apis mellifera using intracellular recording and staining techniques. The responses of neurons to behaviorally relevant odorants (a blend, and components of the Nasonov pheromone, and some other non-pheromonal odors) were recorded. Different response patterns were observed within different neurons, and often within the same neuron, in response to different stimuli. All the neurons stained had innervations in the protocerebral lobe. The cell profiles varied from cells connecting the antennal lobe with both the protocerebral and lateral protocerebral lobes (projection neurons), cells linking the pedunculus of the mushroom body with both the protocerebral and lateral protocerebral lobes (PE1 neurons), cells linking the alpha-lobe and protocerebral lobe with the calyces of the mushroom body (feedback neurons), and cells linking the alpha-lobe and protocerebral lobe with the antennal lobe (recurrent neurons), to cells connecting the protocerebral lobe with the contralateral protocerebrum (bilateral neurons). These findings suggest that the protocerebral lobe acts as an olfactory center associating with other centers, and provides multi-layered recurrent networks within the protocerebrum and between the deutocerebrum and the protocerebrum in honeybee olfactory pathways.     

Morphological and physiological identification of medulla interneurons in the visual system of the tiger beetle larva

The morphology of visual interneurons in the tiger beetle larva was identified after recording their responses. Stained neurons were designated as either medulla or protocerebral neurons according to the location of their cell bodies. Medulla neurons were further subdivided into three groups. Afferent medulla neurons extended processes distally in the medulla neuropil and a single axon to the brain through the optic nerve. They received their main input from stemmata on the ipsilateral side. Two distance-sensitive neurons, near-by sensitive and far-sensitive neurons, were also identified. Atypical medulla neurons extended their neurites distally in the medulla and proximally to the brain, as afferent medulla neurons, but their input patterns and the shapes of their spikes differed from afferent neurons. Protocerebral neurons sent a single axon to the medulla neuropil. They spread collateral branches in the posterior region of the protocerebrum on its way to the medulla neuropil. They received main input from stemmata on the contralateral side. Medulla intrinsic neurons did not extend an axon to the brain, and received either bilateral or contralateral stemmata input only. The input patterns and discharge patterns of medulla neurons are discussed with reference to their morphology.     

Distribution and morphology of descending brain neurons in the cricket Gryllus bimaculatus

The number and distribution of descending brain neurons have been investigated in the cricket. The results are based on retrograde labeling of these cells with either Lucifer yellow or Neurobiotin via whole or small split portions of the cervical connectives. Various groups of cells and single neurons have been identified, and the morphology of more than 40 cells is described. Nearly 200 descending brain neurons can be stained via one cervical connective. Their perikarya are concentrated in clusters that occur ipsi- and contralateral to the filled connective and that lie dorsal and ventral in the brain. Descending cells only arborize in the nonglomerular neuropils of the brain and never branch in the optic lobe. Cells descending ipsilaterally never arborize in the contralateral hemisphere, whereas contralateral descending neurons often branch in both hemispheres. Irrespective of soma position, cells can arborize in the ventral and/or dorsal neuropils of the brain. Neurons with somata in the protocerebrum often have branches in the deutocerebrum and vice versa. The main arborizations of the cells from the prominent ventral i5 group are found in the same part of the protocerebrum. In contrast, various cells arborize in the ventral posterior deutocerebrum, but their somata are not located in different clusters. Thus, neurons from the same cluster may, but need not necessarily, arborize in the same brain area.     

DPKQDFMRFamide expression is similar in two distantly related Drosophila species

Drosophila melanogaster DPKQDFMRFamide was isolated and its expression reported. Distribution of DPKQDFMRFamide immunoreactivity is now described in Drosophila virilis. DPKQDFMRFamide antibody stained a cell in the subesophageal ganglion in embryo. DPKQDFMRFamide antibody stained cells in the superior protocerebrum, subesophageal ganglion, thoracic ganglia, and an abdominal ganglion in larva, pupa, and adult. DPKQDFMRFamide antibody stained an additional pair of cells in the optic lobe and a cell in the lateral protocerebrum in adult. Structure identity and similar distribution of DPKQDFMRFamide in D. virilis and D. melanogaster, two distantly related Drosophila species, suggests an important and conserved activity for the peptide.     

Postembryonic changes in the optic primordia and optic bud in the flesh fly Sarcophaga ruficornis fabr. (Diptera: Sarcophagidae)

Differentiation of the optic lobe anlagen begin in the brain of second instar. Each is an elongated disc of cortical cells placed on the dorsolateral border of each protocerebrum. In the late second instar the disc elongates and its two ends bend inwards which gradually separate from the central region, thus giving three imaginal discs. The protocerebral neuropile extends into these discs and medulla interna and externa are formed. The rudiments of compound eyes (cephalic complex) appear in the early laid larva. These are attached with the brain and pharyngeal wall separately. The posterior portion of cephalic complex (optic bud), after establishing a nervous association with the central optic lobe anlage (lamina ganglionaris), forms the compound eye. Ech optic bud is attached to the brain by a non-nervous stalk. The epiblast cells of the optic bud do not migrate into the brain and the lamina is formed by the proliferation of the central imaginal disc. The reorientation of the optic lobe anlagen starts in the late third instar and the medulla interna divides into two unequal lobes. In 2 day pupa the nerve fibres from the lamina travel into the optic stalk and the optic nerve is formed. The epiblast cells of the optic bud differentiate to form a peripheral epithelial layer which becomes pigmented and gets apposed to the lateral boundary of the brain. The central epiblast cells of the optic bud form several ommatidia. The optic nerve degenerates gradually and various components of the compound eye are formed by the epiblast cells. Chiasm internum is present but chiasm externum is absent.     

The optic lobes of Lepidoptera

Variants of the Golgi-Colonnier (1964) selective silver procedure have been used to show up neurons in insect brains. Neural elements are particularly clearly impregnated in the optic lobes. Three classes of nerve cells can be distinguished; perpendicular (class I), tangential (class II) and amacrine cells (class III). There are many types of neurons in each class which together have a very wide variety of form. Their components are related to specific strata in the optic lobe regions. Short visual cells from the retina terminate in the lamina in discrete groups of endings (optic cartridges). Pairs of long visual fibres from ommatidia pass through the lamina and end in the medulla. Class I cells link these two regions in parallel with the long visual fibres and groups of these elements define columns in the medulla. These in turn give rise to small-field fibres that project to the lobula complex. Tangential processes intersect the parallel arrays of class I cells at characteristic levels. Some are complex in form and may invade up to three regions. Another type provides a direct link between the ipsi- and contralateral optic lobe. Amacrine cells are intrinsic to single lobe regions and have processes situated at the same levels as those of classes I and II cells. A fifth optic lobe region, the optic tubercle, is connected to the medulla and lobula and also receives a set of processes from the mid-brain. There are at least six separate types of small-field relays which could represent the retina mosaic arrangement in the lobula.     

Serotonin- and two putative serotonin receptors-like immunohistochemical reactivities in the ground crickets Dianemobius nigrofasciatus and Allonemobius allardi

Serotonin (5-hydroxytryptamine; 5-HT)- and two putative serotonin receptors, 5-HT1A- and 5-HT1B-like, immunohistochemical reactivities were investigated in the cephalic ganglia of two ground crickets, Dianemobius nigrofasciatus and Allonemobius allardi. 5-HT-ir was strongly expressed in the central body, accessory medulla region of the optic lobe, frontal ganglion, posterior cortex of the protocerebrum, dorsolateral region of the protocerebrum, and the suboesphageal ganglion (SOG) in both crickets. However, 5-HT1A-ir and 5-HT1B-ir showed quite mutually distinct patterns that were also distinct from 5-HT-ir. 5-HT1A-ir was located in the pars intercerebralis, dorsolateral region of the protocerebrum, optic tract, optic lobe, and the midline of the SOG in both crickets. 5-HT1B-ir was located in the pars intercerebralis and dorsolateral region of the protocerebrum, and detected weakly in the optic lobe, tritocerebrum, and the midline of the SOG in both crickets. Interspecific differences were observed with 5-HT1A-ir. 5-HT1A-ir was expressed weakly in two neurons in the mandibular neuromere of the SOG in D. nigrofasciatus, while it was expressed strongly in the tritocerebrum, mandibular neuromere, and maxillary neuromere of the SOG in A. allardi and co-localized with CLOCK-ir (CLK-ir). 5HT-1B-ir was co-localized with CLK-ir in the tritocerebrum, mandibular neuromere, and maxillary neuromere of the SOG when double-labeling was conducted in both crickets. These results indicated that 5-HT and both types of 5-HT receptors may regulate circadian photo-entrainment or photoperiodism in A. allardi, while only 5-HT1B may be involved in circadian photo-entrainment or photoperiodism in D. nigrofasciatus.     

Immunocytochemical distribution of pigment-dispersing hormone in the cephalic ganglia of polyneopteran insects

Material detectable with antisera to the pigment-dispersing hormone (PDH) is regarded as a component of the circadian clock residing in some insects in the optic lobe. This paper demonstrates that the position of the PDH-positive neurones and the course of their processes are similar in all representatives of the insect cohort Polyneoptera. A basic morphological pattern, which includes the proximal frontoventral (Pfv), distal posteriodorsal (Dpd) and posterioventral (Dpv) clusters of PDH-positive neurones, was found in the examined species of locusts, crickets, walking sticks, cockroaches, earwigs and termites. The Pfv cluster is located close to the accessory medulla and usually consists of a set of smaller and a set of larger perikarya. The Dpd and Dpv clusters occupy a dorsal and a ventral position, respectively, at the distal edge of the medulla. These clusters are lacking in stonefly and praying mantid species. The fan-like arrangement of PDH-positive fibres within the frontal medulla face (the locusts and the praying mantid have an additional, smaller fan on the posterior medulla face) is another characteristic feature of Polyneoptera. One (two in the locusts and the praying mantid) nerve bundle runs from the optic lobe to the lateral protocerebrum where it ramifies. One branch gives rise to a fibre network frontally encircling brain neuropile in the area of mushroom bodies. One thin fibre in the crickets and the earwig, and several thicker and anastomosing fibres in the other insects, connect the brain hemispheres. The arrangement of other PDH-positive structures specifies taxa within Polyneoptera. Specific features comprise the presence of PDH-positive perikarya in protocerebrum (walking stick and termite), deutocerebrum (cricket, walking stick, and one cockroach species), tritocerebrum (another cockroach species), and the suboesophageal ganglion (cricket, walking stick and termite). In the walking stick and the termite, PDH-positive fibres pass from the cephalic to the frontal ganglion and from there via the recurrent nerve to the corpora cardiaca where they make varicosities indicative of peptide release into the haemolymph.     

Octopaminergic neurons in the locust brain: morphological,biochemical and electrophysiological characterisation of potential modulators of the visual system

The two Protocerebral-Medulla 4 neurons (PM4a and b) in the locust brain have adjacent cell bodies in the medial deutocerebrum. They project through the posterior protocerebrum, forming limited arborisations en route, and enter the lobula and medulla of the ipsilateral optic lobe, where they form extensive, overlapping arborisations. The PM4a and b neurons are octopamine immunoreactive. Their octopamine content (approximately 25 pg per cell) is confirmed by gas chromatography-mass spectrometry; each cell contains approximately 25 pg p-octopamine. Simultaneous intracellular recording from exposed PM4a and b cell bodies reveals that the two cells are physiologically indistinguishable. They receive multimodal sensory inputs. Tactile/mechanosensory stimuli to much of the animal's body and head, acoustic stimuli, and simple visual stimuli all give rise to e.p.s.p.s and action potentials in the PM4 cell body. Simultaneous recording from the cell body in the deutocerebrum and the axon in the lobula demonstrates that action potentials are predominantly initiated in the deutocerebrum and propagate centrifugally, towards the optic lobe. Occasionally, bright light flashes will initiate an action potential in the axon in the optic stalk, which probably propagates bidirectionally: centripetally to the cell body, and centrifugally into the optic lobe. The extensive arborisations in the lobula and medulla are therefore likely to be sites of octopamine release. Because PM4 neurons are octopaminergic, project to the optic lobe, and receive modalities of sensory input known to dishabituate the Descending Contralateral Movement Detector (DCMD) visual interneuron, it is proposed that PM4 neurons are neuromodulatory — mediating dishabituation or arousal of the visual system.     

Central connections of Limulus ventral photoreceptors revealed by intracellular staining

We stained the central terminations of Limulus ventral photoreceptors by intracellular injection of cobalt chloride into the cell bodies. Axons of these photoreceptors enter the protocerebrum via the ventral optic nerve and pass to the medulla. As they reach the surface of the medullar neuropil they branch profusely in fine processes with intermittent varicosities. Each axonal arborization covers about 0.01-0.02 mm2 of this surface immediately adjacent to the medullar ganglion cell layer. Each point on the surface of the medullar neuropil receives, on the average, input from about 6 ventral photoreceptor axons.