A subpopulation of mushroom body intrinsic neurons is generated by protocerebral neuroblasts in the tobacco hornworm moth, Manduca sexta (Sphingidae, Lepidoptera)

Subpopulations of Kenyon cells, the intrinsic neurons of the insect mushroom bodies, are typically sequentially generated by dedicated neuroblasts that begin proliferating during embryogenesis. When present, Class III Kenyon cells are thought to be the first born population of neurons by virtue of the location of their cell somata, farthest from the position of the mushroom body neuroblasts. In the adult tobacco hornworm moth Manduca sexta, the axons of Class III Kenyon cells form a separate Y tract and dorsal and ventral lobelet; surprisingly, these distinctive structures are absent from the larval Manduca mushroom bodies. BrdU labeling and immunohistochemical staining reveal that Class III Kenyon cells are in fact born in the mid-larval through adult stages. The peripheral position of their cell bodies is due to their genesis from two previously undescribed protocerebral neuroblasts distinct from the mushroom body neuroblasts that generate the other Kenyon cell types. These findings challenge the notion that all Kenyon cells are produced solely by the mushroom body neuroblasts, and may explain why Class III Kenyon cells are found sporadically across the insects, suggesting that when present, they may arise through de novo recruitment of neuroblasts outside of the mushroom bodies. In addition, lifelong neurogenesis by both the Class III neuroblasts and the mushroom body neuroblasts was observed, raising the possibility that adult neurogenesis may play a role in mushroom body function in Manduca.     

Developmental organization of the mushroom bodies of Thermobia domestica (Zygentoma, Lepismatidae): insights into mushroom body evolution from a basal insect

The mushroom bodies of the insect brain are sensory integration centers best studied for their role in learning and memory. Studies of mushroom body structure and development in neopteran insects have revealed conserved morphogenetic mechanisms. The sequential production of morphologically distinct intrinsic neuron (Kenyon cell) subpopulations by mushroom body neuroblasts and the integration of newborn neurons via a discrete ingrowth tract results in an age-based organization of modular subunits in the primary output neuropil of the mushroom bodies, the lobes. To determine whether these may represent ancestral characteristics, the present account assesses mushroom body organization and development in the basal wingless insect Thermobia domestica. In this insect, a single calyx supplied by the progeny of two neuroblast clusters, and three perpendicularly oriented lobes are readily identifiable. The lobes are subdivided into 15 globular subdivisions (Trauben). Lifelong neurogenesis is observed, with axons of newborn Kenyon cells entering the lobes via an ingrowth core. The Trauben do not appear progressively during development, indicating that they do not represent the ramifications of sequentially produced subpopulations of Kenyon cells. Instead, a single Kenyon cell population produces highly branched axons that supply all lobe subdivisions. This suggests that although the ground plan for neopteran mushroom bodies existed in early insects, the organization of modular subunits composed of separate Kenyon cell subpopulations is a later innovation. Similarities between the calyx of Thermobia and the highly derived fruit fly Drosophila melanogaster also suggest a correlation between calyx morphology and Kenyon cell number.     

Functional division of intrinsic neurons in the mushroom bodies of male Spodoptera littoralis revealed by antibodies against aspartate, taurine, FMRF-amide, Mas-allatotropin and DC0

The aim of this study was to further reveal the organization of Kenyon cells in the mushroom body calyx and lobes of the male moth Spodoptera littoralis, by using immunocytochemical labeling. Subdivisions of the mushroom bodies were identified employing antisera raised against the amino acids taurine and aspartate, the neuropeptides FMRF-amide and Mas-allatotropin, and against the protein kinase A catalytic subunit DC0. These antisera have previously been shown to label subsets of Kenyon cells in other species. The present results show that the organization of the mushroom body lobes into discrete divisions, described from standard neuroanatomical methods, is confirmed by immunocytology and shown to be further elaborated. Anti-taurine labels the accessory Y-tract, the gamma division of the lobes, and a thin subdivision of the most posterior component of the lobes. Aspartate antiserum labels the entire mushroom body. FMRF-amide-like immunolabeling is pronounced in the gamma division and in the anterior perimeter of the alpha/beta and alpha'/beta' divisions. Mas-allatotropin-like immunolabeling shows the opposite of FMRF-amide-like and taurine-like immunolabeling: the gamma division and the accessory Y-system is immunonegative whereas strong labeling is seen in both the alpha/beta and alpha'/beta' divisions. The present results agree with findings from other insects that mushroom bodies are anatomically divided into discrete parallel units. Functional and developmental implications of this organization are discussed.     

Fate of neuroblast progeny during postembryonic development of mushroom bodies in the house cricket,Acheta domesticus

Mushroom bodies represent the main sensory integrative center of the insect brain and probably play a major role in the adaptation of behavioral responses to the environment. Taking into account the continuous neurogenesis of cricket mushroom bodies, we investigated ontogenesis of this brain structure. Using BrdU labeling, we examined the fate of neuroblast progeny during the postembryonic development. Preimaginal Kenyon cells survived throughout larval and imaginal moults and persisted during adulthood. Our results indicate that the location of labelled Kenyon cells in the cortex of the adult cricket mainly depends upon the period when they were produced during development. The present data demonstrate that cricket mushroom bodies grow from the inside out and that, at any developmental stage, the center of the cortex contains the youngest Kenyon cells. This study also allowed us to observe the occurrence of quiescent neuroblasts. Kenyon cell death during postembryonic and adult life seems to be reduced. Although preimaginal Kenyon cells largely contribute to adult mushroom body structure, a permanent remodeling of the mushroom body occurs throughout the whole insect life due to the persistence of neurogenesis in the house cricket. Further studies are needed to understand the functional significance of these findings.     

Separate distribution of deutocerebral projection neurons in the mushroom bodies of the cricket brain

Deutocerebral projection neurones in the brain of the cricket (Gryllus bimaculatus) have been investigated by experimental dextran staining, viewed by light and electron microscopy. These neurones of two separate somata clusters innervate two separate primary glomerular neuropils of the deutocerebral segment, either the antennal lobe receiving only antennal nerve sensory input, or the glomerular lobe, receiving input from sensory neurones of lower segmental origin, including chemosensory fibres from mouth parts. Projection neurones of the antennal lobe only invade the anterior calyx of the mushroom body neuropil via the inner antenno glomerular tract, while glomerular relay neurones of the glomerular lobe innervate only the posterior calyx via the tritocerebral tract. All types of projection neurones give rise to presynaptic boutons. forming the central core of microglomeruli with patterned distribution. These projection neurons are cholinergic. The results are discussed in view of maintained segregated modal information, first processed in the separated primary deutocerebral neuropiles and further on in the second order input neuropils of the mushroom bodies. The large posterior calyces are proposed as a compartment for gustatory information.     

Development and evolution of the insect mushroom bodies: towards the understanding of conserved developmental mechanisms in a higher brain center

The insect mushroom bodies are prominent higher order neuropils consisting of thousands of approximately parallel projecting intrinsic neurons arising from the minute basophilic perikarya of globuli cells. Early studies described these structures as centers for intelligence and other higher functions; at present, the mushroom bodies are regarded as important models for the neural basis of learning and memory. The insect mushroom bodies share a similar general morphology, and the same basic sequence of developmental events is observed across a wide range of insect taxa. Globuli cell progenitors arise in the embryo and proliferate throughout the greater part of juvenile development. Discrete morphological and functional subpopulations of globuli cells (or Kenyon cells, as they are called in insects) are sequentially produced at distinct periods of development. Kenyon cell somata are arranged by age around the center of proliferation, as are their processes in the mushroom body neuropil. Other aspects of mushroom body development are more variable from species to species, such as the origin of specific Kenyon cell populations and neuropil substructures, as well as the timing and pace of the general developmental sequence.     

Evolution of insect mushroom bodies: old clues,new insights

The mushroom bodies are a morphologically diverse sensory integration and learning and memory center in the brains of various invertebrate species, of which those of insects are the best described. Insect mushroom bodies are composed of numerous tiny intrinsic neurons (Kenyon cells) that form calyces with their dendrites and a pedunculus and lobes with their axons. The identities of conserved Kenyon cell subpopulations and the correlations between morphological and functional specializations of the mushroom bodies are just beginning to be elucidated, providing insight into mechanisms of mushroom body evolution. Comparisons of mushroom body organization in different insect lineages reveal trends in the evolution of subcompartments correlated with the elaboration, reduction, acquisition or loss of Kenyon cell subpopulations. Furthermore, these changes often appear correlated with variation in type and strength of afferent input and in behavioral ecology. These and other features of mushroom body organization suggest a striking convergence with mammalian cortex, with Kenyon cell subpopulations displaying evolutionary modularity in a manner reminiscent of cortical areas.     

Mushroom bodies in the brain of carrion beetles (Coleoptera, Silphidae)

Mushroom bodies are developed similarly in all the carrion beetles studied, and the transition from necrophagy to predation in some species does not affect considerably their level of development. The Kenyon cells form two small groups in each brain hemisphere and are subdivided into central and peripheral cells. Two closely located inputs of the Kenyon cell processes enter a single, pillow-shaped calyx region. The pedunculus comprises two shafts along a considerable part of its length, which fuse in the lobes. In the degree of development, the mushroom bodies of carrion beetles resemble those of basal lamellicorn and longhorned beetles.     

Taurine in the insect central nervous system

1. Taurine is one of the most abundant free amino acids found in the tissues of insect nervous systems. A brief survey of its immunocytochemical distribution is provided for the brain of worker honeybees.2. The protocerebral mushroom bodies are prominent neuropiles of the insect brain. Immunoreactivity for taurine was compared in the mushroom body intrinsic Kenyon cells of Apis, Drosophila, and Locusta.3. In all three species Kenyon cells expressed immunoreactivity.4. The intensity of the immunoreactivity was, however, graded, depending on the species.5. Recent technical advances in the primary culture of the Kenyon cells of honeybees in a defined taurine-free medium provide the opportunity to investigate the action of taurine in a controlled environment.6. Taurine-like immunoreactivity has been described in the photoreceptor cells of insect and mammalian visual systems. Physiological evidence for similar functions of taurine in mammalian and insect nervous systems is reviewed.     

General structure of the mushroom body calyx in brachycera orthorrhapha flies (Diptera)

The mushroom body calyx in Brachycera Orthorrhapha flies is extremely diverse in the degree of development. In general, the calyx has the anterior, posterior, and dorsal lobes, as well as “sleeves” of glomerular neuropil surrounding Kenyon cell fibers. The anterior lobe of the calyx is found in all species studied. The most complex structure of the calyx is characteristic of the families Empididae and especially Bombyliidae, in which it has all the parts listed above. Brachycera Orthorrhapha flies have three fiber bundles of Kenyon cells, in contrast to four bundles in Drosophila. It is assumed that each mushroom body in Brachycera Orthorrhapha flies is formed by descendants of three single neuroblasts.     

Some cetoniinae (Coleoptera,Scarabaeidae) have structurally different medial and lateral calyces of mushroom bodies

Structural differences between the medial and lateral calyces of mushroom bodies in insects are described for the first time. In two cetoniine scarab beetles, Cetonia aurata and Oxythyrea funesta, the lateral calyces are subdivided into two portions showing a different neuropil structure. This feature is not reflected in the structure of the pedunculus and lobes, as well as in the relative neuropil volume occupied by transformed lateral calyx as compared with unmodified lateral calyx of related scarab beetles. The lateral calyx modification is considered to be related to changes in dendritic arborizations of central Kenyon cells. The subdivision of lateral calyx occurs only in adults and was not observed in larvae.     

Structure of the mushroom bodies in Scarabaeoidea (Insecta: Coleoptera): 1. Basal families and coprophagous scarabaeidae

Mushroom bodies are in general similarly developed in most taxons studied. The calyx region appears as a single structure, and its dual nature is not yet realized. An anterio-posterior asymmetry of the calyx region with Kenyon cell processes running mostly behind the glomerular neuropil of the calyx is characteristic of all the species studied. In this respect, the calyx region of basal Scarabaeoidea resembles greatly the calyx of many dipterans. Lobe compartmentalization occurs at the initial stage. The passalid beetles represent an exception, as their mushroom bodies are much more developed than in related families. This may be connected with the complicated social behavior of Passalidae.     

On structural-functional organization of dragonfly mushroom bodies and some general considerations about purpose of these formations

Anatomy as well as (for the first time) the fine structure have been studied of the mushroom bodies located in protocerebrum of the supraesophageal ganglion of dragonflies—the most ancient flying insects on Earth. Used in the work are larvae of the last age (prior to winging), in which the mushroom body structure has already been completely formed and corresponds to that in imago. The total organization of the dragonfly mushroom bodies has been established to be more primitive than that of other insects studied so far. This involves both the number of interneurons (Kenyon cells) present in the mushroom bodies and the character of anaptic connections formed by these cells. There is confirmed the absence in dragonflies of the mushroom body calyces that in opinion of some authors are obligatory input gates into these structures. Peculiarities of the neuropil structure in the area of the absent calyces are studied in detail. For the first time in insects there are revealed the direct (without additional synaptic switching) pathways forming the afferent input from optic lobes into the mushroom body calyx area. Also detected are the direct pathways going from the mushroom bodies to the abdominal chain (efferent output). A possible functional significance of these findings as well as the general role of mushroom bodied in control of some forms of insect behavior are discussed.Translated from Zhurnal Evolyutsionnoi Biokhimii i Fiziologii, Vol. 40, No. 6, 2004, pp. 495–507     

中华蜜蜂蕈形体的发育

中华蜜蜂(Apis cerana cerana)的脑由前脑、中脑和后脑三部分构成,蕈形体位于前脑的背侧,是其重要的学习及其他复杂行为的整合中心。通过对中华蜜蜂工蜂的幼虫、蛹及成虫的蕈形体形态发育的观察研究,发现中华蜜蜂的蕈形体包含约1000个成神经细胞,它们最终形成了蕈形体的所有Kenyon细胞。这些成神经细胞来自于在新孵化的幼虫脑中已存在的四丛成神经细胞,每一丛细胞的数量不多于45个。蕈形体柄区的出现约在3龄幼虫,而α叶和β叶在5龄幼虫已可明显辨认。冠区出现较晚,大约在蛹期的第二天以后。由于社会性昆虫复杂的学习、记忆和认知需求,其蕈形体的体积和复杂程度都优于其他昆虫。     

General morphology of the brain of the blind cave beetle,Neaphaenops tellkampfii Erichson (Coleoptera : Carabidae)

Neaphaenops tellkampfii (Coleoptera : Carabidae) was collected from Cartmill Cave located in Hart County, Kentucky, U.S.A. This is a cave insect with complete absence of external evidence of eyes or ocelli. The brain of N. tellkampfii has been studied at the light microscope level using Rowell's (1963) silver staining method. Particular attention has been paid to the protocerebrum. One of the notable features of the brain is the dominance of the corpora pedunculata. The corpora pedunculata consists of a calyx, with 3 groups of fibers originating from the Kenyon cells. The stalk is arranged into 2 distinct layers with alpha and beta lobes. The central complex is located anterio-medially beneath the pons cerebralis. It consists of central and ellipsoid bodies and a single ventral tubercle. The ellipsoid body is connected to the beta lobes by a unique chiasmatic fiber tract. The pons cerebralis appears to be formed by 3 distinct groups of globuli cells sending fibers into the pons. The accessory lobes are situated posterio-laterally. The antennal lobes are located posterio-ventrally. Two tubercles were observed lateral to the protocerebrum which may be vestigial optic tubercles. There is no evidence of typical optic lobes or associated fiber tracts. Fiber connections were observed between the calyx and pons cerebralis, the calyx and central body, and also between the calyx and antennal lobe. Two fiber tracts not previously described were observed extending obliquely from the accessory lobe to the beta lobe and protocerebrum.     

Behavioral functions of the insect mushroom bodies

New methods of intervention in Drosophila and other insect species reveal that the mushroom bodies are involved in a diverse set of behavioral functions. The intrinsic Kenyon cells (those neurons with projections within the mushroom bodies) house part of the short-term memory trace for odors and are required for courtship conditioning memory. A pair of extrinsic mushroom body neurons (neurons with projections both inside and outside the mushroom bodies) provides a neuropeptide important for 1-hour olfactory memory. In addition, the mushroom bodies are necessary for context generalization in visual learning and for regulating the transition from walking to rest.     

Neurons with dopamine-like immunoreactivity target mushroom body Kenyon cell somata in the brain of some hymenopteran insects

The mushroom bodies of the insect brain are centers for olfactory and multimodal information processing and they are involved in associative olfactory learning. They are comprised of numerous (340,000 in the bee brain), small (3–8 μm soma diameter) local interneurons, the Kenyon cells. In the brain of honeybees (Apis mellifera) of all castes (worker bees, drones and queens), wasps (Vespula germanica) and hornets (Vespa crabro) immunostaining revealed fibers with dopamine-like immunoreactivity projecting from the pedunculus and the lip neuropil of the mushroom bodies into the Kenyon cell perikaryal layer. These fibers terminate with numerous varicosities, mainly around the border between medial and lateral Kenyon cell soma groups. Visualization of immunostained terminals in the transmission electron microscope showed that they directly contact the somata of the Kenyon cells and contain presynaptic elements. The somata of the Kenyon cells are clearly non-immunoreactive. Synaptic contacts at the somata are unusual for the central nervous systems of insects and other arthropods. This finding suggests that the somata of the Kenyon cells of Hymenoptera may serve an integrative role, and not merely a supportive function.     

Metamorphosis and adult development of the mushroom bodies of the red flour beetle, Tribolium castaneum

The insect mushroom bodies play important roles in a number of higher processing functions such as sensory integration, higher level olfactory processing, and spatial and associative learning and memory. These functions have been established through studies in a handful of tractable model systems, of which only the fruit fly Drosophila melanogaster has been readily amenable to genetic manipulations. The red flour beetle Tribolium castaneum has a sequenced genome and has been subject to the development of molecular tools for the ready manipulation of gene expression; however, little is known about the development and organization of the mushroom bodies of this insect. The present account bridges this gap by demonstrating that the organization of the Tribolium mushroom bodies is strikingly like that of the fruit fly, with the significant exception that the timeline of neurogenesis is shifted so that the last population of Kenyon cells is born entirely after adult eclosion. Tribolium Kenyon cells are generated by two large neuroblasts per hemisphere and segregate into an early-born delta lobe subpopulation followed by clear homologs of the Drosophila gamma, alpha'/beta' and alpha/beta lobe subpopulations, with the larval-born cohorts undergoing dendritic reorganization during metamorphosis. BrdU labeling and immunohistochemical staining also reveal that a proportion of individual Tribolium have variable numbers of mushroom body neuroblasts. If heritable, this variation represents a unique opportunity for further studies of the genetic control of brain region size through the control of neuroblast number and cell cycle dynamics.     

Mushroom Body Neuroblasts of the Lepidopteran Brain (Insecta: Lepidoptera)

A study of the brain of 47 species from 15 lepidopteran families has revealed that only one neuroblast corresponds to each calyx cup of the mushroom body and that mushroom body neuroblasts have been found in the imagoes of 13 out of 25 species caught in the field. It is considered that the proliferative centers consisting of several neuroblasts are not characteristic of lepidopteran mushroom bodies, whereras Kenyon cell neurogenesis in the imago appears to be a widespread phenomenon.