Structure and development of the insect antennodeutocerebral system

The structure and postembryonic development of antennae and deutocerebrum in various insect orders are reviewed. First, the number and/or size of system components, i.e. antennal sensilla, neuroreceptors, deutocerebral neurons and synaptic complexes (glomeruli), are compared in adult insects. Second, the neuronal organization of the system is examined. Evidence of projection of the neuroreceptors from the 2 basal antennal segments (scape and pedicel) in the antennal mechanosensory and motor center and from the distal segments (flagellum) in the antennal lobe is discussed. Third, the types of cerebral neurons found in the antennal lobe are described. Fourth, all synaptic contacts between neurons in the antennal lobe take place in discrete glomeruli whose ultrastructure and neurotransmitters are examined. Evidence of individual identifiability of glomeruli is given. Fifth, various sexual dimorphisms present in the antenna and antennal lobe, related or not to sex pheromone perception, are described at sensillar, neuronal, and glomerular levels. Sixth, the postembryonic development of neurons and glomeruli is analyzed in holo- and hemimetabolous insects. In conclusion, the columnar organization of the system is emphasized, alternative models of antennal-neuron projection into glomeruli are considered, and the functional significance of identified glomeruli is discussed.     

A neuroanatomical study on the organization of the central antennal pathways in insects

Summary Receptor cell axons from the antennal flagellum terminate in the glomeruli of the ipsilateral deutocerebrum in Periplaneta americana and Locusta migratoria. Processes from several groups of deutocerebral neurons also enter the glomeruli and terminate in characteristic branching patterns. There, they contact the antennal axons. Connections are both convergent and divergent. Not only do single central neurons collect the inputs from many receptor cells, but receptor axons were often observed to branch and terminate at more than one deutocerebral neuron. The axons from a portion of the neurons go to form the deutocerebral bundle of the tractus olfactorioglobularis. These axons of the bundle terminate in the ipsilateral calyx of the corpus pedunculatum and in the lateral lobus protocerebri. The processes of the majority of the deutocerebral neurons stay within the deutocerebrum itself and may serve as local interneurons. Part of some antennal fibers terminate in the lobus dorsalis. The lobus glomeratus receives inputs from the maxillary palps and also from processes of deutocerebral neurons.Electron microscopy of synaptic connections and anatomical experiments reveal a complicated pattern of connections between receptor axons and higher order neurons as well as between higher order neurons themselves within the glomeruli.The ratio of the number of antennal fibers to that of relay fibers could easily lead to the interpretation, that the deutocerebrum merely serves as a device for reducing the number of transmission channels. However, coupled with physiological data, anatomical details such as conand divergence of input and interconnections between input channels suggest rather a filtering system and a highly complicated integrative network.     

Aspects of the embryology and neural development of the American lobster.

It is feasible to study the anatomical, physiological, and biochemical properties of identifiable neurons in lobster embryos. To exploit fully the advantages of this preparation and to lay the foundation for single-cell studies, our recent goals have been to 1) establish a quantitative staging system for embryos, 2) document in detail the lobster's embryonic development, 3) determine when uniquely identifiable neurons first acquire their transmitter phenotypes, and 4) identify particular neurons that may serve developmental functions. Behavioral, anatomical, morphometric, and immunocytochemical studies have led to a detailed characterization of the growth and maturation of lobster embryos and to the adoption of a percent-staging system based upon the eye index of Perkins (Fish. Bull., 70:95-99, 1972). It is clear from these studies that the lobster nauplius molts at approximately 12% embryonic development (E12%) into a metanauplius, which subsequently undergoes a complete molt cycle within the egg. This molt cycle climaxes with the emergence of the first-stage larva shortly after hatching. Serotonin and proctolin, neurohormones widely distributed in the lobster nervous system, appear at different times in development. Serotonin immunoreactive neurons begin to appear at approximately E10%, with the adult complement being established by E50%. In contrast, proctolin immunoreactive neurons appear later and attain their full complement over a protracted period including larval and juvenile stages. The development of serotonergic deutocerebral neurons and their targets, the olfactory and accessory lobes in the brain, are also examined. The olfactory lobes are forming by E10% and have acquired their glomerular organization by E50%, whereas the formation of the accessory lobes is delayed; the early rudiments of the accessory lobes are seen by E50%, and glomeruli do not form until the second larval stage.     

A neuroanatomical study on the organization of the central antennal pathways in insects

1. Single unimodal (olfactory) or multimodal (olfactory and mechanosensory) neurons in the antennal lobe of the deutocerebrum of the American cockroach were characterized functionally by microelectrode recording, and their morphological types and positions in the brain were established by dye injection. Thus individual, physiologically identified neurons of known shape could be mapped in reference to the areas of soma groups, glomeruli, tracts and their projection regions in the brain. 2. All of these neurons send processes to deutocerebral glomeruli, i.e., the regions in which the axons of antennal sensory cells terminate. Output neurons have axons that leave the deutocerebrum whereas local interneurons are anaxonic. 3. An output neuron innervates only one glomerulus, sending its axon into the calyces of the corpora pedunculata (CP) in the protocerebrum, where by multiple branching they reach many CP neurons. The same axons send collaterals into the lateral lobe of the protocerebrum. Because of this arrangement, each deutocerebral glomerulus is represented individually and separately in the two projection regions. The fine structure of the endings of the deutocerebral axons in the protocerebrum is described. In the CP calyces they form microglomeruli with typical divergent connectivity. 4. A local interneuron innervates many glomeruli without sending processes to other parts of the brain. 5. Unimodal olfactory and multimodal neurons can be either output neurons or local interneurons; multimodal information is sent to the protocerebrum directly, in parallel with the unimodal information. 6. At least one glomerulus--the macroglomerulus of the male deutocerebrum--is specialized so as to provide an exclusive topographic representation of certain olfactory stimuli not represented elsewhere (female sexual pheromone).     

Antennal sensory system of Periplaneta americana L.

The morphology of the antennal hair-sensilla of Periplaneta americana, their distribution and frequency on the antennal flagellum have been examined by transmission- and scanning-electron microscopy. The types of sensilla were distinguished with respect to physiologically relevant criteria such as wall structure and number of sensory cells. Among the sensilla of the antenna of the adult male, long, single-walled sensilla with four sensory cells (types sw B), Probably responsible for reception of sexual pheromones, are most frequent, representing about 54% of the antennal sensilla. About half of these sensilla are newly-formed at the imaginal ecdysis; the other half are derived from the shorter type sw B sensilla of the nymphal antenna. Short type sw B sensilla are present in all larval stages of both sexes and in adult females as well. During the imaginal ecdysis of males, however, the length of these sensilla increases to double that found in nymphs. Dendritic branches also increase in number. During postembryonic development, the number of sensory fibers in the antennal flagellum increases nearly 20-fold, from 14,000 in the first larval instar to about 270,000 in the adult male. The greatest increase, approximately 90%, occurs during the last developmental stage.     

Leucokinin and callitachykinin immunoreactive neurons during postembryonic development of calliphora vomitoria (L.) (DIPTERA : CALLIPHORIDAE)

Neurons containing 2 types of myotropic neuropeptides were investigated by immunocytochemistry during postembryonic development of the brain and ventral nerve cord of the blowfly Calliphora vomitoria (Diptera : Calliphoridae). Antisera raised against the insect neuropeptides Callitachykinin II (CavTK II), Locustatachykinin I (LomTK I), and Leucokinin I (LK I) were used. Callitachykinin immunoreactive (CavTK–IR) neurons were detected from the 1st-instar larva throughout development to adult. The number of CavTK–IR cell bodies in the brain was 4–16 in larval stages, 10–84 in pupal stages, and over 140 neurons in the newly emerged fly. With the CavTK antiserum, the fibers of only 4 descending neurons were detected in thoracico–abdominal ganglia throughout development. The antiserum to LomTK displayed the same neurons as that to CavTK II as well as a small number of additional neurons. Notably, there were seen about 14–20 locustatachykinin-like immunoreactive (LomTK-LI) cell bodies in the thoracico–abdominal ganglia throughout development. Leucokinin-like immunoreactive (LK-LI) neurons were labeled throughout postembryonic development. In the brain, 2–4 LK-LI cell bodies were labeled from 1st-instar larva to 8-day-old pupa, and 6 LK-LI cell bodies were labeled in the adult brain. In the abdominal ganglia, 7 pairs of LK-LI cell bodies were labeled from 1st-instar larva to 96-h-old pupa, 8 pairs in 8-day-old pupa, and 9 pairs in newly emerged fly, respectively. The CavTK containing neurons in the brain displayed a drastic increase in numbers from larval stages to adult, which indicates an addition of functional roles for this type of peptide. During earlier pupal stages, the number of CavTK–IR neurons decreased. The LK-LI neurons, however, were strongly immunoreactive throughout postembryonic development. Only one additional pair of cells appeared in the brain and 2 additional pair of cells appeared in the abdominal ganglia of the adult as compared with larvae. The continuous high expression of LK-LI material may suggest a functional role for this type of peptide during development.     

中华蜜蜂嗅叶胚后发育过程中的神经胶质

本研究通过形态解剖、免疫组织化学等技术,对中华蜜蜂Apis cerana cerana Fabricius工蜂嗅叶胚后发育中神经胶质的模式进行了比较研究。结果表明:神经胶质在蜜蜂嗅叶的发育过程中起着重要的作用,它们在嗅叶发育早期就划定了神经纤维网的边界;在嗅觉神经轴突进入嗅叶之前,对神经轴突进行"分选",并引导它们进入嗅叶特定的区域,形成神经纤维球;它们不仅规定了神经纤维网的边界和区域,还为神经纤维网提供内部的分隔。中华蜜蜂神经胶质增殖的高峰期集中在幼虫发育末期和预蛹期。     

Phenotypic plasticity in number of glomeruli and sensory innervation of the antennal lobe in leaf‐cutting ant workers (A. vollenweideri)

In the leaf‐cutting ant Atta vollenweideri, the worker caste exhibits a pronounced size‐polymorphism, and division of labor is dependent on worker size (alloethism). Behavior is largely guided by olfaction, and the olfactory system is highly developed. In a recent study, two different phenotypes of the antennal lobe of Atta vollenweideri workers were found: MG‐ and RG‐phenotype (with/without a macroglomerulus). Here we ask whether the glomerular numbers are related to worker size. We found that the antennal lobes of small workers contain ～390 glomeruli (low‐number; LN‐phenotype), and in large workers we found a substantially higher number of ～440 glomeruli (high‐number; HN‐phenotype). All LN‐phenotype workers and some small HN‐phenotype workers do not possess an MG (LN‐RG‐phenotype and HN‐RG‐phenotype), and the remaining majority of HN‐phenotype workers do possess an MG (HN‐MG‐phenotype). Using mass‐staining of antennal olfactory receptor neurons we found that the sensory tracts divide the antennal lobe into six clusters of glomeruli (T1–T6). In LN‐phenotype workers, ～50 glomeruli are missing in the T4‐cluster. Selective staining of single sensilla and their associated receptor neurons revealed that T4‐glomeruli are innervated by receptor neurons from the main type of olfactory sensilla, the Sensilla trichodea curvata. The other type of olfactory sensilla (Sensilla basiconica) exclusively innervates T6‐glomeruli. Quantitative analyses of differently sized workers revealed that the volume of T6 glomeruli scales with the power of 2.54 to the number of Sensilla basiconica. The results suggest that developmental plasticity leading to antennal‐lobe phenotypes promotes differences in olfactory‐guided behavior and may underlie task specialization within ant colonies. © 2009 Wiley Periodicals, Inc. Develop Neurobiol 70: 222–234, 2010.     

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.     

Sexual dimorphism in the antennal lobe of the ant Camponotus japonicus

The carpenter ant, a social hymenopteran, has a highly elaborated antennal chemosensory system that is used for chemical communication in social life. The glomeruli in the antennal lobe are the first relay stations where sensory neurons synapse onto interneurons. The system is functionally and structurally similar to the olfactory bulbs of vertebrates. Using three-dimensional reconstruction of glomeruli and subsequent morphometric analyses, we found sexual dimorphism of the antennal lobe glomeruli in carpenter ants, Camponotus japonicus. Female workers and unmated queens had about 430 glomeruli, the highest number reported so far in ants. Males had a sexually dimorphic macroglomerulus and about 215 ordinary glomeruli. This appeared to result from a greatly reduced number of glomeruli in the postero-medial region of the antennal lobe compared with that in females. On the other hand, sexually isomorphic glomeruli were identifiable in the dorsal region of the antennal lobe. For example, large, uniquely shaped glomeruli located at the dorso-central margin of the antennal lobe were detected in all society members. The great sexual dimorphism seen in the ordinary glomeruli of the antennal lobe may reflect gender-specific tasks in chemical communications rather than different reproductive roles.     

Metamorphic development of locusta-tachykinin immunoreactive neurons of the antennal lobes of the beetle Tenebrio molitor and the effect of fenvalerate application

The effects of the widely used neurotoxic pyrethroid insecticides on neuronal development or plasticity are unclear. To expand knowledge about the influence of the pyrethroid fenvalerate on neuronal development, metamorphic remodelling of the primary olfactory neuropil of the beetle Tenebrio molitor has been studied. The antennal lobe is subdivided into distinct glomeruli before metamorphosis. This is in contrast to that which occurs in other well-studied holometabolous insects such as the moth Manduca sexta and the honeybee. As an indicator of antennal lobe interneurons, locusta-tachykinin immunoreactive neurons have been used. They project into the antennal lobes and form tufted arbors in larval and adult stages within glomeruli throughout the neuropil. These glomerular structures are invaded by glomerular sensory afferent axons and are surrounded by processes of glia cells. With pupation, the glomerulization is lost and no locusta-tachykinin or substance P immunoreactivity is visible in the antennal lobe. The immunoreactivity reappears during metamorphosis, starting with diffusely branched arbors that later become tufted. Application of the neurotoxic insecticide fenvalerate at pupation in sublethal concentrations resulted in a loss or reduction of glomerular pattern formation by neurons and glia cells during metamorphosis. Labelling of antennal sensory axons revealed that the olfactory neuropil was not deafferented, and also that the sensory axons were not organized into a normal glomerular pattern. In addition to the morphological differences, fenvalerate treatment caused locusta-tachykinin immunoreactivity to reappear prematurely during metamorphosis. Possible reasons for fenvalerate-induced alterations in antennal lobe development and their implications for normal development are discussed.     

Caste and sex specific olfactory glomerular organization and brain architecture in two sympatric ant species Camponotus sericeus and Camponotus compressus (Fabricius, 1798)

We use monoclonal antibodies against synaptic proteins and anterograde tracing with neurobiotin to describe the architecture of the antennal lobes in different castes of two ant species – Camponotus sericeus and Camponotus compressus. The reproductives and worker classes are readily categorized based on size and external morphology. The overall organization of brain neuropile is comparable between castes with differences only in the visual ganglia. Males have a larger fraction of neuropile occupied by the medulla and lobula than females. In the diurnal species, C. sericeus these regions are more highly represented, than in the nocturnal species C. compressus. The most striking differences are in the antennal lobe where males possess a macroglomerulus, which is about ten times larger in volume than the other glomeruli; such a specialization is absent in females. Minor workers possess a significantly larger number of glomeruli than the majors despite the smaller overall volume of the lobe. These caste-specific differences occur mainly within glomerular clusters that receive input from sensory neurons that project in tracts – T4 and T5 – within the antennal nerve. The comparative anatomy of different castes of ants provides an entry point into a future systematic analysis of how divergent brain architectures can arise within a single species.     

The developmental expression of serotonin-immunoreactivity in the brain of the pupal honeybee

The biogenic amine serotonin is a neurotransmitter and modulator in both vertebrates and invertebrates. In the CNS of insects, serotonin is expressed by identifiable subsets of neurons. In this paper, we characterize the onset of expression in the brain and suboesophageal ganglion of the honeybee during pupal development. Several identified serotonin-immunoreactive neurons are present in the three neuromeres of the suboesophageal ganglion the dorsal protocerebrum, and the deutocerebrum at pupal ecdysis. Further immunoreactive neurons are incorporated into the developing pupal brain in two characteristic developmental phases. During the first phase, 5 days after pupal ecdysis, serotonin immunoreactivity is formed in the protocerebral central body, the lamina and lobula, and the deutocerebral antennal lobe. During the second phase, 2 days later, immunoreactivity appears in neurons of the protocerebral noduli of the central complex, the medulla, and the pedunculi and lobes of the mushroom bodies. Three novel serotonin-immunoreactive neurons that innervate the central complex and the mushroom bodies can be individually identified.     

Development of an identified serotonergic neuron in the antennal lobe of the moth and effects of reduction in serotonin during construction of olfactory glomeruli

Each olfactory (antennal) lobe of the moth Manduca sexta contains a single serotonin (5-HT) immunoreactive neuron whose processes form tufted arbors in the olfactory glomeruli. To extend our present understanding of the intercellular interactions involved in glomerulus development to the level of an individual, identified antennal lobe neuron, we first studied the morphological development of the 5-HT neuron in the presence and absence of receptor axons. Development of the neuron's glomerular tufts depends, as it does in the case of other multiglomerular neurons, on the presence of receptor axons. Processes of the 5-HT neuron are excluded from the region in which the initial steps of glomerulus construction occur and thus cannot provide a physical scaffolding on which the array of glomeruli is organized. Because the neuron's processes are present in the antennal lobe neuropil throughout postembryonic development, 5-HT could provide signals that influence the pattern of development in the lobe. By surgically producing 5-HT-depleted antennal lobes, we also tested the importance of 5-HT in the construction of olfactory glomeruli. Even in the apparent absence of 5-HT, the glomerular array initiated by the receptor axons was histologically normal, glial cells migrated to form glomerular borders, and receptor axons formed terminal branches in their normal region within each glomerulus. In some cases, 5-HT-immunoreactive processes from abnormal sources entered the lobe and formed the tufted intraglomerular branches typical of most antennal lobe neurons, suggesting that local cues strongly influence the branching patterns of developing antennal lobe neurons. © 1995 John Wiley & Sons, Inc.     

Morphogenesis and cellular proliferation pattern in the developing antennal lobe of Drosophila melanogaster

The adult antennal lobe of Drosophila melanogaster emerges from a precursor, the larval antennal lobe. Pulse and pulse-chase labelling of dividing cells in larvae and pupae with bromodeoxyuridine confirmed previous data that some of the interneurons of the adult antennal lobe derive from a lateral neuroblast which starts to divide early in the first larval instar. However, the majority of these interneurons originate from neuroblasts that initiate mitosis at later stages, with a peak of about 10–12 pairs of dividing neuroblasts in the late third larval instar. No clustering of adult antennal lobe neurons according to their birthdates was observed. In contrast to neurons, terminal divisions of glia in the antennal lobe reach their maximum only 12 h after puparium formation.     

The tracheal system of the developing primary olfactory pathway of Manduca sexta: tracheae do not play a guidance or targeting role for ingrowing receptor axons

Axons navigate to their targets by detecting signals within the environment through which they are growing. The surfaces of tracheae, which are prominent features of the insect body plan, could be detected as favorable pathways for sensory axons growing toward the brain. The pattern of the tracheal investment of the adult antennal lobe of the moth Manduca sexta suggested two specific possibilities for interaction between tracheae and axons during development: that tracheae might be involved in guiding olfactory receptor axons to their target region of the brain, the antennal lobe; and that tracheae could provide an address system within the lobe that defines the sites of glomeruli, which are olfactory-axon target areas within the lobe. To determine whether tracheae contribute to development of the primary olfactory pathway, the distribution of tracheae in the adult and developing antennal lobes was examined with both confocal and electron microscopes. During the major stages in which axons are growing into the antennal lobe and in which glomeruli are forming, the tracheal investment of the nerve and lobe was found to be minimal. Tracheae thus cannot serve as axon guides or as local address sites for newly forming glomeruli during the initial targeting of receptors onto the antennal lobe.     

Distribution of tachykinin-related neuropeptide in the developing central nervous system of the moth Spodoptera litura

Neuropeptides with similarities to vertebrate tachykinins, designated tachykinin-related peptides (TRPs), have been identified in several insect species. In this investigation we have utilized an antiserum raised to one of the locust TRPs, locustatachykinin-I (LomTK-I), to determine the distribution pattern of LomTK-like immunoreactive (LTKLI) neurons in the developing nervous system of the moth Spodoptera litura. A number of LTKLI neurons could be followed from the larval to the adult nervous system: a set of median neurosecretory cells (MNCs) in the brain, a pair of brain descending neurons and a few sets on neurons in the ventral nerve cord. The distribution of LTKLI neurons in the adult brain is very similar to that seen in other insect species with prominent arborizations in the central body, antennal lobes, mushroom body calyces, optic lobe neuropils and other distinct neuropil areas in the protocerebrum and tritocerebrum. A new finding is the presence of LTKLI neurosecretory cells with axon terminals in the anterior aorta and corpora cardiaca, suggesting for the first time a neurohormonal role of tachykinin-related peptide(s) in insects. During postembryonic development the number of LTKLI neurons in the ventral nerve cord decreases somewhat, whereas the number increases in the brain. Thus the functional roles of TRPs may change to some extent during development.     

Sensory Neuron-Derived Eph Regulates Glomerular Arbors and Modulatory Function of a Central Serotonergic Neuron

Olfactory sensory neurons connect to the antennal lobe of the fly to create the primary units for processing odor cues, the glomeruli. Unique amongst antennal-lobe neurons is an identified wide-field serotonergic neuron, the contralaterally-projecting, serotonin-immunoreactive deutocerebral neuron (CSDn). The CSDn spreads its termini all over the contralateral antennal lobe, suggesting a diffuse neuromodulatory role. A closer examination, however, reveals a restricted pattern of the CSDn arborization in some glomeruli. We show that sensory neuron-derived Eph interacts with Ephrin in the CSDn, to regulate these arborizations. Behavioural analysis of animals with altered Eph-ephrin signaling and with consequent arborization defects suggests that neuromodulation requires local glomerular-specific patterning of the CSDn termini. Our results show the importance of developmental regulation of terminal arborization of even the diffuse modulatory neurons to allow them to route sensory-inputs according to the behavioural contexts.     

Structure and metamorphic changes in the brain of the lemon butterfly Papilio demoleus L. (Lepidoptera: Papilionidae)

The morphology of the larval and adult brain of Papilio demoleus, and changes in the cell population and neuropile morphology during the pupal period have been described. The larval brain has more simple fibre areas than that of the adult. Dividing neuroblasts have been found which form the adult neurones. The larval brain contains the three neuromeres (proto-, deuto-, and tritocerebrum). The protocerebrum has well developed corpora pedunculata, a central body, a pons cerebralis and developing optic centres. The corpora ventralia are joined with each other by paired ventral commissures (single in adult). The deutocerebrum is simple and small, the antennal centres are small and simple (ef. adult). The glomerular tritocerebrum is posteroventral to the deutocerebrum, and fibres from the former travel to the crura cerebri. The cortex of the brain consists of four types of glial cells and of association cells, and large and medium sized motor neurones. The number of mitoses is greatest in the larval and prepupal stages; in the pupa it decreases gradually and in late stages it does not occur. Histolysis and pyknosis begin in the prepupa and decrease considerably in the late pupa. The entire neural lamella is broken down in the early pupa. Numerous haemocytes penetrate the laminae of the neural lambella and envelop the entire brain. In the adult, behind the well-developed central body is an ellipsoid body. The medulla interna is divided into two smaller lobes and the deutocerebral lobes are differentiated into cortical and medullary zones. Chiasmata between optic centres are also formed during the pupal period.