Postembryonic Brain Development Research Articles

Developmental Accumulation of Gene Body and Transposon Non-CpG Methylation in the Zebrafish Brain.

DNA methylation predominantly occurs at CG dinucleotides in vertebrate genomes; however, non-CG methylation (mCH) is also detectable in vertebrate tissues, most notably in the nervous system. In mammals it is well established that mCH is targeted to CAC trinucleotides by DNMT3A during nervous system development where it is enriched in gene bodies and associated with transcriptional repression. Nevertheless, the conservation of developmental mCH accumulation and its deposition by DNMT3A is largely unexplored and has yet to be functionally demonstrated in other vertebrates. In this study, by analyzing DNA methylomes and transcriptomes of zebrafish brains, we identified enrichment of mCH at CAC trinucleotides (mCAC) at defined transposon motifs as well as in developmentally downregulated genes associated with developmental and neural functions. We further generated and analyzed DNA methylomes and transcriptomes of developing zebrafish larvae and demonstrated that, like in mammals, mCH accumulates during post-embryonic brain development. Finally, by employing CRISPR/Cas9 technology, we unraveled a conserved role for Dnmt3a enzymes in developmental mCAC deposition. Overall, this work demonstrates the evolutionary conservation of developmental mCH dynamics and highlights the potential of zebrafish as a model to study mCH regulation and function during normal and perturbed development.

Visual Experience Facilitates BDNF-Dependent Adaptive Recruitment of New Neurons in the Postembryonic Optic Tectum.

Postembryonic brain development is sensitive to environmental input and sensory experience, but the mechanisms underlying healthy adaptive brain growth are poorly understood. Here, we tested the importance of visual experience on larval zebrafish (Danio rerio) postembryonic development of the optic tectum (OT), a midbrain structure involved in visually guided behavior. We first characterized postembryonic neurogenic growth in OT, in which new neurons are generated along the caudal tectal surface and contribute appositionally to anatomical growth. Restricting visual experience during development by rearing larvae in dim light impaired OT anatomical and neurogenic growth, specifically by reducing the survival of new neurons in the medial periventricular gray zone. Neuronal survival in the OT was reduced only when visual experience was restricted for the first 5 d following new neuron generation, suggesting that tectal neurons exhibit an early sensitive period in which visual experience protects these cells from subsequent neuronal loss. The effect of dim rearing on neuronal survival was mimicked by treatment with an NMDA receptor antagonist early, but not later, in a new neuron's life. Both dim rearing and antagonist treatment reduced BDNF production in the OT, and supplementing larvae with exogenous BDNF during dim rearing prevented neuronal loss, suggesting that visual experience protects new tectal neurons through neural activity-dependent BDNF expression. Collectively, we present evidence for a sensitive period of neurogenic adaptive growth in the larval zebrafish OT that relies on visual experience-dependent mechanisms.SIGNIFICANCE STATEMENT Early brain development is shaped by environmental factors via sensory input; however, this form of experience-dependent neuroplasticity is traditionally studied as structural and functional changes within preexisting neurons. Here, we found that restricting visual experience affects development of the larval zebrafish optic tectum, a midbrain structure involved in visually guided behavior, by limiting the survival of newly generated neurons. We found that new tectal neurons exhibit a sensitive period soon after cell birth in which adequate visual experience, likely mediated by neuronal activity driving BDNF production within the tectum, would protect them from subsequent neuronal loss over the following week. Collectively, we present evidence for neurogenic adaptive tectal growth under different environmental lighting conditions.

The labial gene is required to terminate proliferation of identified neuroblasts in postembryonic development of the Drosophila brain

The developing brain of Drosophila has become a useful model for studying the molecular genetic mechanisms that give rise to the complex neuronal arrays that characterize higher brains in other animals including mammals. Brain development in Drosophila begins during embryogenesis and continues during a subsequent postembryonic phase. During embryogenesis, the Hox gene labial is expressed in the developing tritocerebrum, and labial loss-of-function has been shown to be associated with a loss of regional neuronal identity and severe patterning defects in this part of the brain. However, nothing is known about the expression and function of labial, or any other Hox gene, during the postembryonic phase of brain development, when the majority of the neurons in the adult brain are generated. Here we report the first analysis of Hox gene action during postembryonic brain development in Drosophila. We show that labial is expressed initially in six larval brain neuroblasts, of which only four give rise to the labial expressing neuroblast lineages present in the late larval brain. Although MARCM-based clonal mutation of labial in these four neuroblast lineages does not result in an obvious phenotype, a striking and unexpected effect of clonal labial loss-of-function does occur during postembryonic brain development, namely the formation of two ectopic neuroblast lineages that are not present in wildtype brains. The same two ectopic neuroblast lineages are also observed following cell death blockage and, significantly, in this case the resulting ectopic lineages are Labial-positive. These findings imply that labial is required in two specific neuroblast lineages of the wildtype brain for the appropriate termination of proliferation through programmed cell death. Our analysis of labial function reveals a novel cell autonomous role of this Hox gene in shaping the lineage architecture of the brain during postembryonic development.

A Mechanism to Enhance Cellular Responsivity to Hormone Action: Krüppel-Like Factor 9 Promotes Thyroid Hormone Receptor-β Autoinduction During Postembryonic Brain Development.

Thyroid hormone (TH) receptor (TR)-β (trb) is induced by TH (autoinduced) in Xenopus tadpoles during metamorphosis. We previously showed that Krüppel-like factor 9 (Klf9) is rapidly induced by TH in the tadpole brain, associates in chromatin with the trb upstream region in a developmental stage and TH-dependent manner, and forced expression of Klf9 in the Xenopus laevis cell line XTC-2 accelerates and enhances trb autoinduction. Here we investigated whether Klf9 can promote trb autoinduction in tadpole brain in vivo. Using electroporation-mediated gene transfer, we transfected plasmids into premetamorphic tadpole brain to express wild-type or mutant forms of Klf9. Forced expression of Klf9 increased baseline trb mRNA levels in thyroid-intact but not in goitrogen-treated tadpoles, supporting that Klf9 enhances liganded TR action. As in XTC-2 cells, forced expression of Klf9 enhanced trb autoinduction in tadpole brain in vivo and also increased TH-dependent induction of the TR target genes klf9 and thbzip. Consistent with our previous mutagenesis experiments conducted in XTC-2 cells, the actions of Klf9 in vivo required an intact N-terminal region but not a functional DNA binding domain. Forced expression of TRβ in tadpole brain by electroporation-mediated gene transfer increased baseline and TH-induced TR target gene transcription, supporting a role for trb autoinduction during metamorphosis. Our findings support that Klf9 acts as an accessory transcription factor for TR at the trb locus during tadpole metamorphosis, enhancing trb autoinduction and transcription of other TR target genes, which increases cellular responsivity to further TH action on developmental gene regulation programs.

Orthodenticle is required for the development of olfactory projection neurons and local interneurons in Drosophila.

ABSTRACTThe accurate wiring of nervous systems involves precise control over cellular processes like cell division, cell fate specification, and targeting of neurons. The nervous system of Drosophila melanogaster is an excellent model to understand these processes. Drosophila neurons are generated by stem cell like precursors called neuroblasts that are formed and specified in a highly stereotypical manner along the neuroectoderm. This stereotypy has been attributed, in part, to the expression and function of transcription factors that act as intrinsic cell fate determinants in the neuroblasts and their progeny during embryogenesis. Here we focus on the lateral neuroblast lineage, ALl1, of the antennal lobe and show that the transcription factor-encoding cephalic gap gene orthodenticle is required in this lineage during postembryonic brain development. We use immunolabelling to demonstrate that Otd is expressed in the neuroblast of this lineage during postembryonic larval stages. Subsequently, we use MARCM clonal mutational methods to show that the majority of the postembryonic neuronal progeny in the ALl1 lineage undergoes apoptosis in the absence of orthodenticle. Moreover, we demonstrate that the neurons that survive in the orthodenticle loss-of-function condition display severe targeting defects in both the proximal (dendritic) and distal (axonal) neurites. These findings indicate that the cephalic gap gene orthodenticle acts as an important intrinsic determinant in the ALl1 neuroblast lineage and, hence, could be a member of a putative combinatorial code involved in specifying the fate and identity of cells in this lineage.

A multipotent transit-amplifying neuroblast lineage in the central brain gives rise to optic lobe glial cells in Drosophila

The neurons and glial cells of the Drosophila brain are generated by neural stem cell-like progenitors during two developmental phases, one short embryonic phase and one more prolonged postembryonic phase. Like the bulk of the adult-specific neurons, most of glial cells found in the adult central brain are generated postembryonically. Five of the neural stem cell-like progenitors that give rise to glial cells during postembryonic brain development have been identified as type II neuroglioblasts that generate neural and glial progeny through transient amplifying INPs. Here we identify DL1 as a novel multipotent neuroglial progenitor in the central brain and show that this type II neuroblast not only gives rise to neurons that innervate the central complex but also to glial cells that contribute exclusively to the optic lobe. Immediately following their generation in the central brain during the second half of larval development, these DL1 lineage-derived glia migrate into the developing optic lobe, where they differentiate into three identified types of optic lobe glial cells, inner chiasm glia, outer chiasm glia and cortex glia. Taken together, these findings reveal an unexpected central brain origin of optic lobe glial cells and central complex interneurons from one and the same type II neuroglioblast.

The labial gene is required to terminate proliferation of identified neuroblasts in postembryonic development of the Drosophila brain

SummaryThe developing brain of Drosophila has become a useful model for studying the molecular genetic mechanisms that give rise to the complex neuronal arrays that characterize higher brains in other animals including mammals. Brain development in Drosophila begins during embryogenesis and continues during a subsequent postembryonic phase. During embryogenesis, the Hox gene labial is expressed in the developing tritocerebrum, and labial loss-of-function has been shown to be associated with a loss of regional neuronal identity and severe patterning defects in this part of the brain. However, nothing is known about the expression and function of labial, or any other Hox gene, during the postembryonic phase of brain development, when the majority of the neurons in the adult brain are generated. Here we report the first analysis of Hox gene action during postembryonic brain development in Drosophila. We show that labial is expressed initially in six larval brain neuroblasts, of which only four give rise to the labial expressing neuroblast lineages present in the late larval brain. Although MARCM-based clonal mutation of labial in these four neuroblast lineages does not result in an obvious phenotype, a striking and unexpected effect of clonal labial loss-of-function does occur during postembryonic brain development, namely the formation of two ectopic neuroblast lineages that are not present in wildtype brains. The same two ectopic neuroblast lineages are also observed following cell death blockage and, significantly, in this case the resulting ectopic lineages are Labial-positive. These findings imply that labial is required in two specific neuroblast lineages of the wildtype brain for the appropriate termination of proliferation through programmed cell death. Our analysis of labial function reveals a novel cell autonomous role of this Hox gene in shaping the lineage architecture of the brain during postembryonic development.

Multipotent neural stem cells generate glial cells of the central complex through transit amplifying intermediate progenitors in Drosophila brain development

The neural stem cells that give rise to the neural lineages of the brain can generate their progeny directly or through transit amplifying intermediate neural progenitor cells (INPs). The INP-producing neural stem cells in Drosophila are called type II neuroblasts, and their neural progeny innervate the central complex, a prominent integrative brain center. Here we use genetic lineage tracing and clonal analysis to show that the INPs of these type II neuroblast lineages give rise to glial cells as well as neurons during postembryonic brain development. Our data indicate that two main types of INP lineages are generated, namely mixed neuronal/glial lineages and neuronal lineages. Genetic loss-of-function and gain-of-function experiments show that the gcm gene is necessary and sufficient for gliogenesis in these lineages. The INP-derived glial cells, like the INP-derived neuronal cells, make major contributions to the central complex. In postembryonic development, these INP-derived glial cells surround the entire developing central complex neuropile, and once the major compartments of the central complex are formed, they also delimit each of these compartments. During this process, the number of these glial cells in the central complex is increased markedly through local proliferation based on glial cell mitosis. Taken together, these findings uncover a novel and complex form of neurogliogenesis in Drosophila involving transit amplifying intermediate progenitors. Moreover, they indicate that type II neuroblasts are remarkably multipotent neural stem cells that can generate both the neuronal and the glial progeny that make major contributions to one and the same complex brain structure.

A Transient Expression of Prospero Promotes Cell Cycle Exit of Drosophila Postembryonic Neurons through the Regulation of Dacapo

Cell proliferation, specification and terminal differentiation must be precisely coordinated during brain development to ensure the correct production of different neuronal populations. Most Drosophila neuroblasts (NBs) divide asymmetrically to generate a new NB and an intermediate progenitor called ganglion mother cell (GMC) which divides only once to generate two postmitotic cells called ganglion cells (GCs) that subsequently differentiate into neurons. During the asymmetric division of NBs, the homeodomain transcription factor PROSPERO is segregated into the GMC where it plays a key role as cell fate determinant. Previous work on embryonic neurogenesis has shown that PROSPERO is not expressed in postmitotic neuronal progeny. Thus, PROSPERO is thought to function in the GMC by repressing genes required for cell-cycle progression and activating genes involved in terminal differentiation. Here we focus on postembryonic neurogenesis and show that the expression of PROSPERO is transiently upregulated in the newly born neuronal progeny generated by most of the larval NBs of the OL and CB. Moreover, we provide evidence that this expression of PROSPERO in GCs inhibits their cell cycle progression by activating the expression of the cyclin-dependent kinase inhibitor (CKI) DACAPO. These findings imply that PROSPERO, in addition to its known role as cell fate determinant in GMCs, provides a transient signal to ensure a precise timing for cell cycle exit of prospective neurons, and hence may link the mechanisms that regulate neurogenesis and those that control cell cycle progression in postembryonic brain development.

DEVELOPMENTAL AND THYROID HORMONE-INDUCED EXPRESSION OF DNA METHYLTRANSFERASES AND METHYL-CPG BINDING PROTEINS IN XENOPUS TADPOLE BRAIN DURING METAMORPHOSIS

Event Abstract Back to Event DEVELOPMENTAL AND THYROID HORMONE-INDUCED EXPRESSION OF DNA METHYLTRANSFERASES AND METHYL-CPG BINDING PROTEINS IN XENOPUS TADPOLE BRAIN DURING METAMORPHOSIS Yasuhiro Kyono1, 2 and Robert J. Denver1* 1 University of Michigan, Molecular, Cellular and Developmental Biology, United States 2 University of Michigan, Neuroscience Graduate Program, United States Postembryonic brain development is critically dependent on thyroid hormone (T3), which influences neural cell proliferation, migration, differentiation, morphology and function. Thyroid hormone is known to promote neurogenesis, and recent findings showed that proteins that methylate the genome, DNA methyltransferases (DNMTs), and proteins that bind to methylated DNA, methyl-CpG binding proteins (MBDs), also play critical roles in cell lineage specification and neurogenesis in the developing brain. We hypothesized that T3 promotes neurogenesis through its regulation of DNMT and MBD expression. To test this we investigated the developmental expression and T3 regulation of DNMT (dnmt1 and dnmt3a) and MBD (mecp2, mbd1-4 and kaiso) genes in the brain of tadpoles of Xenopus laevis. We microdissected tadpole brains and analyzed mRNAs in the preoptic area/diencephalon by RTqPCR. We found that mRNAs for dnmt1, dnmt3a, mbd3 and kaiso were upregulated during spontaneous metamorphosis. Treatment of premetamorphic tadpoles with T3 (5nM) caused a time-dependent upregulation of each of these genes. Dnmt3a and mbd3 mRNAs were upregulated by 12 hr; whereas, dnmt1 and kaiso mRNAs were upregulated by 24 hr after T3 treatment. Using in situ hybridization histochemistry, we confirmed the developmental and T3-dependent increase in dnmt3a and mbd3 mRNAs in the tadpole brain, and we investigated their regional expression. Dnmt3a mRNA is widely expressed throughout the tadpole brain in regions where cells are undergoing migration and differentiation, but is excluded from the ventricular and subventricular zones (VZ/SVZ) where cell proliferation occurs. By contrast, the expression of mbd3 mRNA is restricted to the periphery of the VZ/SVZ throughout the tadpole brain. Our findings support that T3 may control neurogenesis through regulation of expression of DNMTs and MBDs. Acknowledgements Supported by NSF grants IBN 0235401 and IOS 0922583 to RJD Keywords: Brain, DNA Methylation, epigenetics, metamorphosis, Thyroid hormone Conference: NASCE 2011: The inaugural meeting of the North American Society for Comparative Endocrinology, Ann Arbor, United States, 13 Jul - 16 Jul, 2011. Presentation Type: Poster Topic: Developmental endocrinology Citation: Kyono Y and Denver RJ (2011). DEVELOPMENTAL AND THYROID HORMONE-INDUCED EXPRESSION OF DNA METHYLTRANSFERASES AND METHYL-CPG BINDING PROTEINS IN XENOPUS TADPOLE BRAIN DURING METAMORPHOSIS. Front. Endocrinol. Conference Abstract: NASCE 2011: The inaugural meeting of the North American Society for Comparative Endocrinology. doi: 10.3389/conf.fendo.2011.04.00028 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 18 Jul 2011; Published Online: 09 Aug 2011. * Correspondence: Prof. Robert J Denver, University of Michigan, Molecular, Cellular and Developmental Biology, Ann Arbor, United States, Rdenver@umich.edu Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Yasuhiro Kyono Robert J Denver Google Yasuhiro Kyono Robert J Denver Google Scholar Yasuhiro Kyono Robert J Denver PubMed Yasuhiro Kyono Robert J Denver Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

Segregation of postembryonic neuronal and glial lineages inferred from a mosaic analysis of the Drosophila larval brain

Due to its intermediate complexity and its sophisticated genetic tools, the larval brain of Drosophila is a useful experimental system to study the mechanisms that control the generation of cell diversity in the CNS. In order to gain insight into the neuronal and glial lineage specificity of neural progenitor cells during postembryonic brain development, we have carried an extensive mosaic analysis throughout larval brain development. In contrast to embryonic CNS development, we have found that most postembryonic neurons and glial cells of the optic lobe and central brain originate from segregated progenitors. Our analysis also provides relevant information about the origin and proliferation patterns of several postembryonic lineages such as the superficial glia and the medial-anterior Medulla neuropile glia. Additionally, we have studied the spatio-temporal relationship between gcm expression and gliogenesis. We found that gcm expression is restricted to the post-mitotic cells of a few neuronal and glial lineages and it is mostly absent from postembryonic progenitors. Thus, in contrast to its major gliogenic role in the embryo, the function of gcm during postembryonic brain development seems to have evolved to the specification and differentiation of certain neuronal and glial lineages.

Thebrain tumorgene negatively regulates neural progenitor cell proliferation in the larval central brain ofDrosophila

Brain development in Drosophila is characterized by two neurogenic periods, one during embryogenesis and a second during larval life. Although much is known about embryonic neurogenesis, little is known about the genetic control of postembryonic brain development. Here we use mosaic analysis with a repressible cell marker (MARCM) to study the role of the brain tumor (brat) gene in neural proliferation control and tumour suppression in postembryonic brain development of Drosophila. Our findings indicate that overproliferation in brat mutants is due to loss of proliferation control in the larval central brain and not in the optic lobe. Clonal analysis indicates that the brat mutation affects cell proliferation in a cell-autonomous manner and cell cycle marker expression shows that cells of brat mutant clones show uncontrolled proliferation, which persists into adulthood. Analysis of the expression of molecular markers, which characterize cell types in wild-type neural lineages, indicates that brat mutant clones comprise an excessive number of cells, which have molecular features of undifferentiated progenitor cells that lack nuclear Prospero (Pros). pros mutant clones phenocopy brat mutant clones in the larval central brain, and targeted expression of wild-type pros in brat mutant clones promotes cell cycle exit and differentiation of brat mutant cells, thereby abrogating brain tumour formation. Taken together, our results provide evidence that the tumour suppressor brat negatively regulates cell proliferation during larval central brain development of Drosophila, and suggest that Prospero acts as a key downstream effector of brat in cell fate specification and proliferation control.

The Drosophila castor gene is involved in postembryonic brain development.

castor (cas) encodes a zink finger protein expressed in a subset of Drosophila embryonic neuroglioblasts where it controls neuronal differentiation. We show here that cas is expressed at larval and pupal stages in brain cell clusters where it participates in the elaboration of the adult structures. In particular using the MARCM system (mosaic analysis with a repressible cell marker), we show that cas is required postembryonically for correct axon pathfinding of the central complex (CX) and mushroom body (MB) neurons. The linotte (lio) gene encodes a transmembrane protein expressed at larval/pupal stage in a glial structure, the TIFR, and interacts with the no-bridge (nob) gene. We show that cas interacts genetically with lio and nob. These interactions do not involve direct transcription regulation but probably cellular communication processes.

Drosophila Pax-6/eyeless is essential for normal adult brain structure and function

A role for the Pax-6 homologue eyeless in adult Drosophila brain development and function is described. eyeless expression is detected in neurons, but not glial cells, of the mushroom bodies, the medullar cortex, the lateral horn, and the pars intercerebralis. Furthermore, severe defects in adult brain structures essential for vision, olfaction, and for the coordination of locomotion are provoked by two newly isolated mutations of Pax-6/eyeless that result in truncated proteins. Consistent with the morphological lesions, we observe defective walking behavior for these eyeless mutants. The implications of these data for understanding postembryonic brain development and function in Drosophila are discussed.

Central brain postembryonic development indrosophila: Implication of genes expressed at the interhemispheric junction

Postembryonic brain development of Drosophila has become recently a subject of intense investigations. In particular, the linotte (lio) mutants display strong structural defects in the mushroom bodies and the central complex. The Lio kinase is expressed in a glial structure at the interhemispheric junction of late larval and young pupal brain. With the aim of identifying new genes involved in the formation of adult central brain structures, 821 enhancer-trap Gal4 lines were generated and screened for late larval expression. We identified 167 lines showing expression at or near the interhemispheric junction of third-instar larval brain, an area from which the central complex differentiates. Adult brains from 104 of these 167 lines were analyzed through paraffin sections. This secondary screen allowed the recovery of five central brain mutants. Of 89 control lines showing various patterns of expression excluding the interhemispheric junction, only one anatomical mutant was isolated. These six mutations, which have been thoroughly characterized, affect the midline area of the adult brain with phenotypes of split central complex structures and/or fused mushroom body lobes. This work opens the way for further analysis of the molecular and cellular events involved in central brain reorganization during metamorphosis.

GABA and serotonin immunoreactivity during postembryonic brain development in the beetle Tenebrio molitor.

Analysis of the serotonin immunoreactive neurons in the central brain of the beetle Tenebrio molitor during postembryonic development shows that the basic structural characteristics of larval brain resemble those of the adult. Most, if not all, serotonin immunoreactive central brain neurons persist with metamorphosis. Their fate can be followed during development. GABA immunoreactivity occurs in about 360 neurons assembled in ten different clusters of somata in the larval midbrain. During metamorphosis no additional clusters are formed. However, the number of immunoreactive neurons increases to 450. Their morphological analysis is restricted to location of the somata and the distribution of arborizations within neuropil areas. Metamorphic transition of glomerular sub-units in the antennal lobes as well as ellipsoid body development can be followed by GABA immunohistochemistry. Furthermore, the study of these transitions proved useful in displaying changes during metamorphic pattern formation induced by sublethal application of the pyrethroid insecticide fenvalerate.

A protein related to p21-activated kinase (PAK) that is involved in neurogenesis in the Drosophila adult central nervous system

Brains are organized by the developmental processes generating them. The embryonic neurogenic phase of Drosophila melanogaster has been studied in detail at the genetic, cellular and molecular level [1–3]. In contrast, much of what is known of postembryonic brain development has been gathered by neuroanatomical and gene expression studies. The molecular mechanisms underlying cellular diversity and structural organisation in the adult brain, such as the establishment of the correct neuroblast number, the spatial and temporal control of neuroblast proliferation, cell fate determination, and the generation of the precise pattern of neuronal connectivity, are largely unknown. In a screen for viable mutations affecting adult central brain structures, we isolated the mushroom bodies tiny (mbt) gene of Drosophila, which encodes a protein related to p21-activated kinase (PAK). We show that mutations in mbt primarily interfere with the generation or survival of the intrinsic cells (Kenyon cells) of the mushroom body, a paired neuropil structure in the adult brain involved in learning and memory [4–6].

Passive avoidance performance of mice fed marginally or severely zinc deficient diets during post-embryonic brain development

Swiss-Webster outbred mice were fed marginally or severely zinc deficient diets (9 or 2 ppm zinc) from day 16 gestation to day 15 postnatal. Control mice were fed a 100 ppm diet either ad lib or in amounts equal to the diet intake of deprived mice (pair fed controls). Male and female offspring were tested at 70 days of age in a one-trial passive avoidance task with a 30 min train-test interval. Both marginally and severely zinc deprived offspring (but not pair fed controls) had shorter avoidance latencies than offspring of ad lib fed zinc replete controls. Zinc deprived offspring did not differ from control mice when either baseline or “stressed” (exposure to novel environment) plasma corticosterone levels were quantitated. Further, zinc staining patterns of the hippocampus (Timm-sulfide stain) were not altered in the nutritionally deprived offspring. Thus marginal dietary zinc deficiency during development can lead to impaired passive avoidance performance in adult mice. This behavioral effect is not associated with altered pituitary-adrenocortical activity or with a permanent reduction in hippocampal zinc staining. This result has significant implications for the influence of zinc deprivation in utero and in the neonatal animal on adult behavior characteristics.

Maze exploration in juvenile rats treated with corticosteroids during development

The possibility of permanent damage to brain function after developmental corticosteroid treatment has been raised in connection with therapeutic use of potent synthetic corticosteroids during the period of brain development in the human fetus and infant. In the present study, brain function was evaluated in rats treated during brain development with triamcinolone acetonide (TAC). A subtoxic dose (0.9 mg/kg), which did not retard growth, was used. TAC treatment at two periods of post-embryonic brain development (day 0 and day 10 postnatal) led to deviations from normal exploration strategies in a radial maze. Treatment on day 16 gestation was effective only in males. Neonatal TAC-treatment also affected spontaneous alternation in a T-maze. Measures which reflected arousal-activity levels during behavioral tests, such as time to initiate exploration and time to complete exploration, were not affected by treatment. These results suggest that the development of complex brain function in rats can be altered by subtoxic corticosteroid treatment in the perinatal period.

Postembryonic brain development in the monarch butterfly,Danaus plexippus plexippus, L. : I. Cellular events during brain morphogenesis.

1. Cellular morphogenesis during postembryonic brain development inDanaus plexippus plexippus L. was examined using histological techniques including radioautography. 2. The production of new neurones is continuous throughout larval and pupal stages and shows no fluctuations corresponding to ecdysis. Glial cell production, on the other hand, occurs at the time of molting. 3. New ganglion cells are formed by the division of neuroblasts found in aggregates or isolated among larval ganglion cells. Asymmetrical neuroblast divisions yield one neuroblast and one ganglion-mother cell which then divides at least once to form the new ganglion cells. Such divisions begin earlier inDanaus than in other investigated Lepidoptera. Symmetrical divisions yielding two neuroblasts also occur, but only among aggregated neuroblasts. 4. Radioautographs of brains fixed at progressive intervals after Tritiated Thymidine (H3TdR) injection have permitted description of the basic pattern by which cells of the adult brain cortex are laid out and progressive changes in the relationship of new ganglion cells derived from a single neuroblast. Ganglion-mother cells are deposited between the neuroblast and the neuropile, thus forming a row of cells which move the neuroblast progressively farther from the neuropile. New ganglion cells produced by ganglion-mother cell mitoses, which usually are oriented at 45° angles to the neuropile, expand the cell cluster. Differentiating fibers of these cells are apparent within a few days of their production and seem to enter the neuropile in one bundle. Later with increased neuropile volume and further cell differentiation the cells are no longer clumped and thus are not recognizable as offspring of a single neuroblast. 5. Neuroblasts found scattered among the larval ganglion cells arise from cells near the neuropile. These cells, at first indistinguishable from their neighbors, gradually assume the size and ready stainability of neuroblasts and subsequently divide according to the pattern described above. 6. Scattered neuroblasts degenerate beginning shortly after pupation and have completely disappeared by the end of the fourth day. 7. Except in the developing optic lobe, glial cell numbers increase through the proliferation of already existing glial cells. All glial cells show H3TdR uptake during a 12 hour period surrounding each larval-larval molt and for a somewhat longer period after pupation. However, in the larval stages mitotic figures were seen only among glial I, II, and IV. Glial I cells divide through the entire last larval stage and for two days following pupation. Large irregular mitoses seen among glial III cells at pupation indicate that these cells are probably polyploid. 8. In the newly forming adult optic lobe glial II, III, and IV cells appear to develop from preganglion cells or cells indistinguishable from them. These cells gradually stain more and more darkly, segregate into the normal glial positions, and subsequently divide in accord with other glial cells. 9. At the end of the fifth instar the perineurium (glial I cells), which begins to thicken during the third larval instar, is multilayered and contains many vacuolar cells. Just prior to pupation the neurilemma begins to disintegrate and during the next five days all but the cells closest to the brain disappear. Hemocytes are seen to engulf portions of the disintegrating neurilemma and already degenerating perineurial cells, but do not seem to engulf live cells. The glial I cells remaining adjacent to the brain secrete a new neurilemma. 10. There is no evidence for mass destruction of larval ganglion cells by either autolysis or phagocytosis, and only in the antennal center is there evidence of degeneration of larval cells (NORDLANDER andEDWARDS, in press).