Sarah M. Farris

Ground plan of the insect mushroom body: Functional and evolutionary implications

In most insects with olfactory glomeruli, each side of the brain possesses a mushroom body equipped with calyces supplied by olfactory projection neurons. Kenyon cells providing dendrites to the calyces supply a pedunculus and lobes divided into subdivisions supplying outputs to other brain areas. It is with reference to these components that most functional studies are interpreted. However, mushroom body structures are diverse, adapted to different ecologies, and likely to serve various functions. In insects whose derived life styles preclude the detection of airborne odorants, there is a loss of the antennal lobes and attenuation or loss of the calyces. Such taxa retain mushroom body lobes that are as elaborate as those of mushroom bodies equipped with calyces. Antennal lobe loss and calycal regression also typify taxa with short nonfeeding adults, in which olfaction is redundant. Examples are cicadas and mayflies, the latter representing the most basal lineage of winged insects. Mushroom bodies of another basal taxon, the Odonata, possess a remnant calyx that may reflect the visual ecology of this group. That mushroom bodies persist in brains of secondarily anosmic insects suggests that they play roles in higher functions other than olfaction. Mushroom bodies are not ubiquitous: the most basal living insects, the wingless Archaeognatha, possess glomerular antennal lobes but lack mushroom bodies, suggesting that the ability to process airborne odorants preceded the acquisition of mushroom bodies. Archaeognathan brains are like those of higher malacostracans, which lack mushroom bodies but have elaborate olfactory centers laterally in the brain. J. Comp. Neurol. 513:265–291, 2009.

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.

Parasitoidism, not sociality, is associated with the evolution of elaborate mushroom bodies in the brains of hymenopteran insects

The social brain hypothesis posits that the cognitive demands of social behaviour have driven evolutionary expansions in brain size in some vertebrate lineages. In insects, higher brain centres called mushroom bodies are enlarged and morphologically elaborate (having doubled, invaginated and subcompartmentalized calyces that receive visual input) in social species such as the ants, bees and wasps of the aculeate Hymenoptera, suggesting that the social brain hypothesis may also apply to invertebrate animals. In a quantitative and qualitative survey of mushroom body morphology across the Hymenoptera, we demonstrate that large, elaborate mushroom bodies arose concurrent with the acquisition of a parasitoid mode of life at the base of the Euhymenopteran (Orussioidea + Apocrita) lineage, approximately 90 Myr before the evolution of sociality in the Aculeata. Thus, sociality could not have driven mushroom body elaboration in the Hymenoptera. Rather, we propose that the cognitive demands of host-finding behaviour in parasitoids, particularly the capacity for associative and spatial learning, drove the acquisition of this evolutionarily novel mushroom body architecture. These neurobehavioural modifications may have served as pre-adaptations for central place foraging, a spatial learning-intensive behaviour that is widespread across the Aculeata and may have contributed to the multiple acquisitions of sociality in this taxon.

Taurine-, aspartate- and glutamate-like immunoreactivity identifies chemically distinct subdivisions of Kenyon cells in the cockroach mushroom body.

The lobes of the mushroom bodies of the cockroach Periplaneta americana consist of longitudinal modules called laminae. These comprise repeating arrangements of Kenyon cell axons, which like their dendrites and perikarya have an affinity to one of three antisera: to taurine, aspartate, or glutamate. Taurine‐immunopositive laminae alternate with immunonegative ones. Aspartate‐immunopositive Kenyon cell axons are distributed across the lobes. However, smaller leaf‐like ensembles of axons that reveal particularly high affinities to anti‐aspartate are embedded within taurine‐positive laminae and occur in the immunonegative laminae between them. Together, these arrangements reveal a complex architecture of repeating subunits whose different levels of immunoreactivity correspond to broader immunoreactive layers identified by sera against the neuromodulator FMRFamide. Throughout development and in the adult, the most posterior lamina is glutamate immunopositive. Its axons arise from the most recently born Kenyon cells that in the adult retain their juvenile character, sending a dense system of collaterals to the front of the lobes. Glutamate‐positive processes intersect aspartate‐ and taurine‐immunopositive laminae and are disposed such that they might play important roles in synaptogenesis or synapse modification. Glutamate immunoreactivity is not seen in older, mature axons, indicating that Kenyon cells show plasticity of neurotransmitter phenotype during development. Aspartate may be a universal transmitter substance throughout the lobes. High levels of taurine immunoreactivity occur in broad laminae containing the high concentrations of synaptic vesicles. J. Comp. Neurol. 439:352–367, 2001.

Development of laminar organization in the mushroom bodies of the cockroach: Kenyon cell proliferation, outgrowth, and maturation

The mushroom bodies of the insect brain are lobed integration centers made up of tens of thousands of parallel‐projecting axons of intrinsic (Kenyon) cells. Most of the axons in the medial and vertical lobes of adult cockroach mushroom bodies derive from class I Kenyon cells and are organized into regular, alternating pairs (doublets) of pale and dark laminae. Organization of Kenyon cell axons into the adult pattern of laminae occurs gradually over the course of nymphal development. Newly hatched nymphs possess tiny mushroom bodies with lobes containing a posterior lamina of ingrowing axons, followed by a single doublet, which is flanked anteriorly by a γ layer composed of class II Kenyon cells. Golgi impregnations show that throughout nymphal development, regardless of the number of doublets present, the most posterior lamina serves as the “ingrowth lamina” for axons of newborn Kenyon cells. Axons of the ingrowth lamina are taurine‐ and synaptotagmin‐immunonegative. They produce fine growth cone tipped filaments and long perpendicularly oriented collaterals along their length. The maturation of these Kenyon cells and the formation of a new lamina are marked by the loss of filaments and collaterals, as well as the onset of taurine and synaptotagmin expression. Class I Kenyon cells thus show plasticity in both morphology and transmitter expression during development. In a hemimetabolous insect such as the cockroach, juvenile stages are morphologically and behaviorally similar to the adult. The mushroom bodies of these insects must be functional from hatching onward, while thousands of new neurons are added to the existing structure. The observed developmental plasticity may serve as a mechanism by which extensive postembryonic development of the mushroom bodies can occur without disrupting function. This contrasts with the more evolutionarily derived holometabolous insects, such as the honey bee and the fruit fly, in which nervous system development is accomplished in a behaviorally simple larval stage and a quiescent pupal stage. J. Comp. Neurol. 430:331–351, 2001.

Development and morphology of Class II Kenyon cells in the mushroom bodies of the honey bee, Apis mellifera

Class II Kenyon cells, defined by their early birthdate and unique dendritic arborizations, have been observed in the mushroom bodies of evolutionarily divergent insects. In the fruit fly Drosophila melanogaster, Class II (also called clawed) Kenyon cells are well known for their extensive reorganization that occurs during metamorphosis. The present account reports for the first time the occurrence of mushroom body reorganization during metamorphosis in holometabolous insect species outside of the Diptera. In the honey bee, Apis mellifera, Class II Kenyon cells show signs of degeneration and undergo a subtle reshaping of their axons during metamorphosis. Unlike in Drosophila, reorganization of Class II Kenyon cells in the honey bee does not involve the loss of axon branches. In contrast, the mushroom bodies of closely related hymenopteran species, the polistine wasps, undergo a much more dramatic restructuring near the end of metamorphosis. Immunohistochemistry, dextran fills, and Golgi impregnations illuminate the heterogeneous nature of Class II Kenyon cells in the developing and adult honey bee brain, with subpopulations differing in the location of dendritic arbors within the calyx, and branching pattern in the lobes. Furthermore, polyclonal antibodies against the catalytic subunit of Drosophila protein kinase A (anti‐DC0) label an unusual and previously undescribed trajectory for these neurons. The observed variations in morphology indicate that subpopulations of Class II Kenyon cells in the honey bee can likely be further defined by significant differences in their specific connections and functions within the mushroom bodies. J. Comp. Neurol. 474:325–339, 2004.

Miniaturization of Nervous Systems and Neurons

Miniaturized species have evolved in many animal lineages, including insects and vertebrates. Consequently, their nervous systems are constrained to fit within tiny volumes. These miniaturized nervous systems face two major challenges for information processing: noise and energy consumption. Fewer or smaller neurons with fewer molecular components will increase noise, affecting information processing and transmission. Smaller, more densely-packed neural processes will increase the density of energy consumption whilst reducing the space available for mitochondria, which supply energy. Although miniaturized nervous systems benefit from smaller distances between neurons, thus saving time, space and energy, they have also increased the space available for neural processing by expanding and contorting their nervous systems to fill any available space, sometimes at the expense of other structures. Other adaptations, such as multifunctional neurons or matched filters, may further alleviate the pressures on space within miniaturized nervous systems. Despite these adaptations, we argue that limitations on information processing are likely to affect the behaviour generated by miniaturized nervous systems.

A unique mushroom body substructure common to basal cockroaches and to termites.

The mushroom bodies of the cockroach Periplaneta americana are made up of intrinsic neurons (class I and class II Kenyon cells) with dendrites in a dorsal calyx and axons that bifurcate into medial and vertical lobes. Here, we describe a substructure of the cockroach mushroom bodies composed of a previously unrecognized class of Kenyon cells with distinct morphologies. The embryonically produced class III Kenyon cells form a separate accessory calyx below the calyx proper. The medial branches of class III Kenyon cell axons form the previously described “γ bulb,” whereas the vertical branches leave the vertical lobe to form a toroidal “lobelet” around the posterior surface. Taking advantage of the morphologically and immunochemically distinct nature of the lobelet, we have attempted to determine the distribution of this unique structure in other insects of the taxon Dictyoptera (cockroaches, mantises, and termites). Our data indicate that the lobelet is present only in basal cockroaches and in termites, supporting existing theories of a close phylogenetic relationship between these groups. Higher termites possess a duplicated lobe structure due to immense elaboration of the processes of class III Kenyon cells. The degree of complexity in the mushroom body lobes of termites agrees with current taxonomic arrangements of the Isoptera based on non‐neural morphological and DNA sequence analyses. It thus appears that the evolution of the Dictyoptera has been accompanied by increasing complexity of the mushroom bodies, achieved in part through the further specialization and elaboration of a subset of Kenyon cells. J. Comp. Neurol. 456:305–320, 2003.

Structural, functional and developmental convergence of the insect mushroom bodies with higher brain centers of vertebrates.

Convergence of higher processing centers has been proposed for insects and vertebrates, but the extent of these similarities remains controversial. The present study demonstrates that one higher brain center of insects, the mushroom bodies, displays a number of similarities with mammalian higher brain centers that are arguably the products of adaptation to common behavioral ecologies, despite their deeply divergent origins. Quantitative neuroanatomy, immunohistochemistry, fluorescent tract tracing and BrdU labeling are employed to investigate the relationships among behavioral ecology and mushroom body size, sensory input and mode of development in one taxon, the scarab beetles (Coleoptera: Scarabaeidae). Comparisons are extended to a taxon in which similar mushroom body architectures have arisen independently, the cockroaches (Dictyoptera), and to published accounts of vertebrate brain evolution. This study demonstrates that evolutionary increases in higher brain center size and intrinsic neuron number are associated with flexibility in food acquisition behaviors in both vertebrates and insects. These evolutionarily expanded higher brain centers are divided into novel structural subcompartments that acquire novel processing functions. Increased numbers of neurons comprising enlarged higher brain centers are generated by expanded neural precursor pools, and the time for development of these brain centers is protracted. Taken together, these findings extend our understanding of how evolutionarily constrained neural substrates might converge under shared adaptive landscapes, even after 600 million years of divergence, and even at the level of higher brain centers that generate complex behaviors.

Evolutionary Convergence of Higher Brain Centers Spanning the Protostome-Deuterostome Boundary

Currently available evidence supports a single origin for the centralized nervous system of bilaterally symmetrical animals. Beneath the staggering diversity of protostome and deuterostome nervous systems lies a fundamental groundplan consisting of a tripartite brain and a nerve cord divided into distinct antero-posterior and medio-lateral zones. As divergent lineages have taken independent paths towards increased encephalization, complex brain centers have arisen that serve multiple levels of sensory processing and advanced behavioral coordination and execution. Many questions arise as one surveys the distribution of these brain centers across the bilaterian phylogenetic trees. What environments did these lineages encounter that promoted the acquisition of energetically expensive brain centers composed of thousands, millions or even trillions of neurons? What novel behavioral capabilities did these brain centers in turn give rise to? Comparative studies within vertebrate clades have revealed instances of parallelism and convergence that have been instructive in associating evolutionary changes in brain structure and function with specific behavioral ecologies. The present account reviews these findings and extends them to invertebrate animals that have independently evolved higher brain centers. By expanding the scope of comparative studies across phyla, it will be possible to uncover structural and functional constraints imposed by deep homology, and to better understand the environmental pressures that have given rise to brain and behavioral complexity.