Larval Nervous System Research Articles

Whole organism and tissue-specific analysis of pexophagy in Drosophila.

Peroxisomes are essential organelles involved in critical metabolic processes in animals such as fatty acid oxidation, ether phospholipid production and reactive oxygen species detoxification. We have generated transgenic Drosophila melanogaster models expressing fluorescent reporters for the selective autophagy of peroxisomes, a process known as pexophagy. We show that these reporters are colocalized with a peroxisomal marker and that they can reflect pexophagy induction by iron chelation and inhibition by depletion of the core autophagy protein Atg5. Using light sheet microscopy, we have been able to obtain a global overview of pexophagy levels across the entire organism at different stages of development. Tissue-specific control of pexophagy is exemplified by areas of peroxisome abundance but minimal pexophagy, observed in clusters of oenocytes surrounded by epithelial cells where pexophagy is much more evident. Enhancement of pexophagy was achieved by feeding flies with the iron chelator deferiprone, in line with past results using mammalian cells. Specific drivers were used to visualize pexophagy in neurons, and to demonstrate that specific depletion in the larval central nervous system of Hsc70-5, the Drosophila homologue of the chaperone HSPA9/mortalin, led to a substantial elevation in pexophagy.

LC-Orbitrap HRMS-Based Proteomics Reveals Novel Mitochondrial Dynamics Regulatory Proteins Associated with RasV12-Induced Glioblastoma (GBM) of Drosophila.

Glioblastoma multiforme (GBM) is the most prevalent and aggressive brain tumor found in adult humans with a poor prognosis and average survival of 14-15 months. In order to have a comprehensive understanding of proteome and identify novel therapeutic targets, this study focused mainly on the differentially abundant proteins (DAPs) of RasV12-induced GBM. RasV12 is a constitutively active Ras mutant form essential for tumor progression by continuously activating signaling pathways leading to uncontrolled tumor growth. This study used a transgenic Drosophila model with RasV12 overexpression using the repo-GAL4 driver line, specifically in glial cells, to study GBM. The high-resolution mass spectrometry (HRMS)-based proteomic analysis of the GBM larval central nervous system identified three novel DAPs specific to mitochondria. These DAPs, probable maleylacetoacetate isomerase 2 (Q9VHD2), bifunctional methylene tetrahydrofolate dehydrogenase (Q04448), and glutamine synthetase1 (P20477), identified through HRMS were further validated by qRT-PCR. The protein-protein interaction analysis revealed interactions between RasV12 and DAPs, with functional links to mitochondrial dynamics regulators such as Drp1, Marf, Parkin, and HtrA2. Notably, altered expressions of Q9VHD2, P20477, and Q04448 were observed during GBM progression, which offers new insights into the involvement of mitochondrial dynamic regulators in RasV12-induced GBM pathophysiology.

Comparison of neurogenesis in bivalves with different types of development

The presence of different types of larvae within the same class suggests a broad ecological diversification. A clear comparison of bivalve larval nervous systems would give a broader view on evolutionary and ecological picture of the clade in question. The present study focused on the neurodevelopment in two bivalve species with different larval types: pericalymma of Acila insignis (Bivalvia: Protobranchia) and veliger of Spisula sybillae (Bivalvia: Autobranchia). It was shown that the pioneer dorsal and ventral neurons in S. sybillae appear at the trochophore stage. Subsequently, future three paired ganglia are developed on the nerve cords in pediveliger. In the pericalymma of A. insignis, serotonin- and FMRFamide-positive cells are found in the apical organ (AO), as well as two pairs of FMRFamide positive neurons are detected on dorsal and posterior part of pericalymma. A comparative analysis showed significant differences in the larval neuromorphology between veliger and pericalymma. In contrast to the S. sybillae veliger, the nervous system of the A. insignis pericalymma is simple, likely due to its different lifestyle. The larval nervous system in the species under study has features characteristic of Lophotrochozoa and Spiralia.

Repellency, toxicity, and physiological actions of low molecular weight basic amines in mosquitoes.

This study investigated the behavioral responses and toxicity of three basic amines: 1-methylpiperazine, 1-methylpyrrolidine, and triethylamine (TEA), compounds suggested previously to be anosmic in vapor exposures to caged mosquitoes. These compounds showed repellency of Aedes aegypti mosquitoes, followed by flightlessness, knockdown, and paralysis, all increasing with exposure time and dosage. Electrophysiological experiments showed a blocking effect on nerve discharge of the Drosophila melanogaster larval central nervous system (CNS) with little evidence of hyperexcitation. Blockage of voltage-gated (Kv2) potassium channel currents under patch clamp occurred at similar concentrations. Involvement of K+ channels in the action of basic amines was supported by behavior and CNS recordings of a Shaker Kv1 mutant exposed to TEA, where instead of blockage, a hyperexcitation of nerve firing was observed. Experiments on cockroach leg mechanoreceptors demonstrated neuronal excitation and on mosquito antennae strong electroantennogram (EAG) signals with an augmentation of blank air responses after a single puff of basic amine. The neurophysiological effects of basic amines are consistent with K+ channel block, whereas the antennal EAG response was not obviously associated with anosmia. The low-dose effects of basic amines appear to be repellency and bradykinesia. Overall, the findings provide key insights into the mechanisms underlying the biological activity of basic amines. © 2024 Society of Chemical Industry.

Divergence in developmental patterns of larval and adult nervous systems in direct-developing holothuria Eupentacta fraudatrix suggests their independent evolution

Divergence in developmental patterns of larval and adult nervous systems in direct-developing holothuria Eupentacta fraudatrix suggests their independent evolution

Drosophila TET acts with PRC1 to activate gene expression independently of its catalytic activity.

Enzymes of the ten-eleven translocation (TET) family play a key role in the regulation of gene expression by oxidizing 5-methylcytosine (5mC), a prominent epigenetic mark in many species. Yet, TET proteins also have less characterized noncanonical modes of action, notably in Drosophila, whose genome is devoid of 5mC. Here, we show that Drosophila TET activates the expression of genes required for larval central nervous system (CNS) development mainly in a catalytic-independent manner. Genome-wide profiling shows that TET is recruited to enhancer and promoter regions bound by Polycomb group complex (PcG) proteins. We found that TET interacts and colocalizes on chromatin preferentially with Polycomb repressor complex 1 (PRC1) rather than PRC2. Furthermore, PRC1 but not PRC2 is required for the activation of TET target genes. Last, our results suggest that TET and PRC1 binding to activated genes is interdependent. These data highlight the importance of TET noncatalytic function and the role of PRC1 for gene activation in the Drosophila larval CNS.

Study of Axonal Injury and Degeneration in Drosophila.

A fundamental feature of nervous systems is a highly specified synaptic connectivity between cells and the ability to adaptively change this connectivity through plasticity mechanisms. Plasticity mechanisms are highly relevant for responding to nervous system damage, and studies using nervous system injury paradigms in Drosophila (as well as other model organisms) have revealed conserved molecular pathways that are triggered by axon damage. Simple assays that introduce injuries to axons in either adult flies or larvae have proven to be particularly powerful for uncovering mechanisms of axonal degeneration and clearance. They have also been used to reveal requirements for regrowth of axons and dendrites, as well as signaling pathways that regulate cellular responses to nerve injury. Here we review commonly used and simple to carry out techniques that enable experimenters to study responses to axonal damage in either adult flies (following antennal transection) or larvae (following nerve crush to segmental nerves). Because axons and dendrites in the larval peripheral nervous system can be readily visualized through the translucent cuticle, another versatile method to probe injury responses is to focus high-energy laser light to a small and specific location in the animal. We therefore discuss a method for immobilizing intact larvae for imaging through the cuticle to carry out injury by pulse dye laser, which can be used to generate many different kinds of injuries and directed ablations within intact larvae. These techniques, combined with powerful genetic tools in Drosophila, make the fruit fly an excellent model system for studying the effects of injury and the mechanisms of axon degeneration, synapse plasticity, and immune response.

The development of the adult nervous system in the annelid Owenia fusiformis

BackgroundThe evolutionary origins of animal nervous systems remain contentious because we still have a limited understanding of neural development in most major animal clades. Annelids — a species-rich group with centralised nervous systems — have played central roles in hypotheses about the origins of animal nervous systems. However, most studies have focused on adults of deeply nested species in the annelid tree. Recently, Owenia fusiformis has emerged as an informative species to reconstruct ancestral traits in Annelida, given its phylogenetic position within the sister clade to all remaining annelids.MethodsCombining immunohistochemistry of the conserved neuropeptides FVamide-lir, RYamide-lir, RGWamide-lir and MIP-lir with gene expression, we comprehensively characterise neural development from larva to adulthood in Owenia fusiformis.ResultsThe early larval nervous system comprises a neuropeptide-rich apical organ connected through peripheral nerves to a prototroch ring and the chaetal sac. There are seven sensory neurons in the prototroch. A bilobed brain forms below the apical organ and connects to the ventral nerve cord of the developing juvenile. During metamorphosis, the brain compresses, becoming ring-shaped, and the trunk nervous system develops several longitudinal cords and segmented lateral nerves.ConclusionsOur findings reveal the formation and reorganisation of the nervous system during the life cycle of O. fusiformis, an early-branching annelid. Despite its apparent neuroanatomical simplicity, this species has a diverse peptidergic nervous system, exhibiting morphological similarities with other annelids, particularly at the larval stages. Our work supports the importance of neuropeptides in animal nervous systems and highlights how neuropeptides are differentially used throughout development.

Development of Serotonergic and Dopaminergic Neuronal Networks of the Central Nervous System in King Crab, Paralithodes camtschaticus.

This article presents recent findings as regards distribution of cells producing serotonin and dopamine in the larval central nervous system at different developmental stages, including four pelagic larval stages (zoea I-IV), a semibenthic postlarval stage glaucothoe (megalopa), benthic juveniles, and adult red king crabs, Paralithodes camtschaticus, made by using immunocytochemistry and confocal laser scanning microscopy. We have shown that the serotonergic and dopaminergic neurons are present long before the onset of metamorphosis. In the red king crab b larval nervous system, the changes become particularly pronounced during the first metamorphosis from zoea IV to glaucothoe, which may be related to the development of the segmental appendages and maturation of motor behaviors in decapods. This work presents the distribution and dynamics of the development of serotonergic and dopaminergic neuronal networks in king crab show, the potential roles of serotonin and dopamine in the modulation of olfactory and visual processing in the early stages of larval development, and also the mechanosensory and chemosensory processing in the glaucothoe stage during settlement and in their transition from a pelagic to benthic lifestyle.

Palps across the tree - the neuronal innervation and development of sensory head appendages in Annelida.

Polychaetes inhabit a wide variety of habitats and show a great morphological diversity. In this context, a key morphological structure for adapting to their individual lifestyles and ecological niches are the prominent head appendages. In the last years more and more studies focused on the mainly sensory annelid head appendages - namely the antennae, palps, buccal lips and cirri - to unravel the evolutionary origin and phylogeny of Annelida. Unfortunately, comparable data for most of the polychaete families are lacking so far, especially when it comes to features of the larval anterior nervous system and the related innervation and potential homology of these head appendages. In this study, we therefore use an integrative morphological approach including immunohistochemistry and confocal laser scanning microscopy in combination with histological serial sections and 3D-visualizations. With special focus on the palp-like appendages, our data provides a closer look into the development of the larval anterior nervous system and the related sensory structures of three polychaete families representing major groups of the annelid tree of life. Hence, we investigate members of the palaeoannelid Magelonidae as well as basally-branching Amphinomidae, and the pleistoannelid Spionidae forming a taxon deeply nested within Sedentaria. Our comparative data of larval and adult neuronal features support the homology of feeding-palps across the annelid tree. Furthermore, our observations show that larval palps gradually transform into the adult ones while keeping a very similar neuronal innervation pattern. Solely for Amphinomidae a loss of larval palps during ontogenesis has to be assumed. Therefore, our investigations uncover important and so far unknown details in terms of structural homology across Annelida and provide important results necessary for our understanding of annelid evolution.

MoS2 2D materials induce spinal cord neuroinflammation and neurotoxicity affecting locomotor performance in zebrafish.

MoS2 nanosheets belong to an emerging family of nanomaterials named bidimensional transition metal dichalcogenides (2D TMDCs). The use of such promising materials, featuring outstanding chemical and physical properties, is expected to increase in several fields of science and technology, with an enhanced risk of environmental dispersion and associated wildlife and human exposures. In this framework, the assessment of MoS2 nanosheets toxicity is instrumental to safe industrial developments. Currently, the impact of the nanomaterial on the nervous tissue is unexplored. In this work, we use as in vivo experimental model the early-stage zebrafish, to investigate whether mechano-chemically exfoliated MoS2 nanosheets reach and affect, when added in the behavioral ambient, the nervous system. By high throughput screening of zebrafish larvae locomotor behavioral changes upon exposure to MoS2 nanosheets and whole organism live imaging of spinal neuronal and glial cell calcium activity, we report that sub-acute and prolonged ambient exposures to MoS2 nanosheets elicit locomotor abnormalities, dependent on dose and observation time. While 25 μg mL-1 concentration treatments exerted transient effects, 50 μg mL-1 ones induced long-lasting changes, correlated to neuroinflammation-driven alterations in the spinal cord, such as astrogliosis, glial intracellular calcium dysregulation, neuronal hyperactivity and motor axons retraction. By combining integrated technological approaches to zebrafish, we described that MoS2 2D nanomaterials can reach, upon water (i.e. ambient) exposure, the nervous system of larvae, resulting in a direct neurological damage.

Visualizing and Measuring Dendrite Arborization in Drosophila Somatosensory Neurons.

Dendrites of neurons receive synaptic or sensory inputs and are important sites of neuronal computation. The morphological features of dendrites not only are hallmarks of the neuronal type but also largely determine a neuron's function. Thus, dendrite morphogenesis has been a subject of intensive study in neuroscience. Quantification of dendritic morphology, which is required for accurate assessment of phenotypes, can often be a challenging task, especially for complex neurons. Because manual tracing of dendritic branches is labor-intensive and time-consuming, automated or semiautomated methods are required for efficient analysis of a large number of samples. A popular in vivo model system for studying the mechanisms of dendrite morphogenesis is dendritic arborization (da) neurons in the Drosophila larval peripheral nervous system. In this chapter, we introduce methods for visualizing and measuring the dendritic arbors of these neurons. We begin with an introduction of da neurons and an overview of the methods that have been used for measuring da neuron dendrites. We then discuss the techniques and detailed steps of neuron visualization and image acquisition. Finally, we provide example steps for dendrite tracing and measurement.

A description of the bat star nervous system throughout larval ontogeny.

Larvae represent a distinct life history stage in which animal morphology and behavior contrast strongly to adult organisms. This life history stage is a ubiquitous aspect of animal life cycles, particularly in the marine environment. In many species, the structure and function of the nervous system differ significantly between metamorphosed juveniles and larvae. However, the distribution and diversity of neural cell types in larval nervous systems remains incompletely known. Here, the expression of neurotransmitter and neuropeptide synthesis and transport genes in the bat star Patiria miniata is examined throughout larval development. This characterization of nervous system structure reveals three main neural regions with distinct but overlapping territories. These regions include a densely innervated anterior region, an enteric neural plexus, and neurons associated with the ciliary band. In the ciliary band, cholinergic cells are pervasive while dopaminergic, noradrenergic, and GABAergic cells show regional differences in their localization patterns. Furthermore, the distribution of some neural subtypes changes throughout larval development, suggesting that changes in nervous system structure align with shifting ecological priorities during different larval stages, before the development of the adult nervous system. While past work has described aspects of P. miniata larval nervous system structure, largely focusing on early developmental timepoints, this work provides a comprehensive description of neural cell type localization throughout the extensive larval period.

Repellency and Toxicity of Vapor-Active Benzaldehydes against Aedes aegypti.

Chemical screening efforts recently found that 3-phenoxybenzaldehyde, a breakdown product of alpha-cyano pyrethroids, was a potent spatial repellent against Aedes aegypti mosquitoes in a glass tube repellency assay. In order to characterize this molecule further and identify structure-activity relationships, a set of 12 benzaldehyde analogues were screened for their repellency and toxicity in vapor phase exposures at 100 μg/cm2. Dose-response analyses were performed for the most active compounds in order to better characterize their repellent potency and toxicity compared to those of other commercially available toxicants. The three most toxic compounds (LC50 values) were 3-chlorobenzaldehyde (CBA) (37 μg/cm2), biphenyl-3-carboxaldehyde (BCA) (48 μg/cm2), and 3-vinylbenzaldehyde (66 μg/cm2), which makes them less toxic than bioallethrin (6.1 μg/cm2) but more toxic than sandalwood oil (77 μg/cm2), a repellent/toxic plant essential oil. The most repellent analogues with EC50 values below 30 μg/cm2 were 3-phenoxybenzaldehyde (6.3 μg/cm2), isophthalaldehyde (23 μg/cm2), BCA (17 μg/cm2), and CBA (22 μg/cm2), which makes them about as active as N,N-diethyl-3-methylbenzamide (25.4 μg/cm2). We further investigated the activity of a select group of these benzaldehydes to block the firing of the central nervous system of A. aegypti larvae. Compounds most capable of repelling and killing mosquitoes in the vapor phase were also those most capable of blocking nerve firing in the larval mosquito nervous system. The results demonstrate that benzaldehyde analogues are viable candidate repellent and insecticidal molecules and may lead to the development of future repellent and vapor toxic vector control tools.

Preliminary study on toxicological mechanism of golden cuttlefish (Sepia esculenta) larvae exposed to cd

BackgroundCadmium (Cd) flows into the ocean with industrial and agricultural pollution and significantly affects the growth and development of economic cephalopods such as Sepia esculenta, Amphioctopus fangsiao, and Loligo japonica. As of now, the reasons why Cd affects the growth and development of S. esculenta are not yet clear.ResultsIn this study, transcriptome and four oxidation and toxicity indicators are used to analyze the toxicological mechanism of Cd-exposed S. esculenta larvae. Indicator results indicate that Cd induces oxidative stress and metal toxicity. Functional enrichment analysis results suggest that larval ion transport, cell adhesion, and some digestion and absorption processes are inhibited, and the cell function is damaged. Comprehensive analysis of protein-protein interaction network and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis was used to explore S. esculenta larval toxicological mechanisms, and we find that among the 20 identified key genes, 14 genes are associated with neurotoxicity. Most of them are down-regulated and enriched to the neuroactive ligand-receptor interaction signaling pathway, suggesting that larval nervous system might be destroyed, and the growth, development, and movement process are significantly affected after Cd exposure.ConclusionsS. esculenta larvae suffered severe oxidative damage after Cd exposure, which may inhibit digestion and absorption functions, and disrupt the stability of the nervous system. Our results lay a function for understanding larval toxicological mechanisms exposed to heavy metals, promoting the development of invertebrate environmental toxicology, and providing theoretical support for S. esculenta artificial culture.

Ectopic expression of matrix metalloproteinases and filopodia extension via JNK activation are involved in the invasion of blood tumor cells in Drosophila mxc mutant.

Drosophila mxcmbn1 mutant exhibits severe hyperplasia in larval hematopoietic tissue called the lymph glands (LGs). However, the malignant nature of these cells remains unknown. We aimed to identify if mxcmbn1 LG cells behave as malignant tumor cells and uncover the mechanism(s) underlying the malignancy of the mutant hemocytes. When mutant LG cells were allografted into normal adult abdomens, they continued to proliferate; however, normal LG cells did not proliferate. Mutant circulating hemocytes also attached to the larval central nervous system (CNS), where the basement membrane was disrupted. The mutant hemocytes displayed higher expression of matrix metalloproteinase (MMP) 1 and MMP2 and higher activation of the c-Jun N-terminal kinase (JNK) pathway than normal hemocytes. Depletion of MMPs or JNK mRNAs in LGs resulted in reduced numbers of hemocytes attached to the CNS, suggesting that the invasive phenotype involved elevated expression of MMPs via hyperactivation of the JNK pathway. Moreover, hemocytes with elongated filopodia and extra lamellipodia were frequently observed in the mutant hemolymph, which also depended on JNK signaling. Thus, the MMP upregulation and overextension of actin-based cell protrusions were also involved in hemocyte invasion in mxcmbn1 larvae. These findings contribute to the understanding of molecular mechanisms underlying mammalian leukemic invasion.

Unbiased characterization of the larval zebrafish enteric nervous system at a single cell transcriptomic level

The enteric nervous system (ENS) regulates many gastrointestinal functions including peristalsis, immune regulation and uptake of nutrients. Defects in theENS can lead to severe enteric neuropathies such as Hirschsprung disease (HSCR). Zebrafish have proven to be fruitful in the identification of genes involved in ENS development and HSCR pathogenesis. However, composition and specification of enteric neurons and glial subtypes at larval stages, remains mainly unexplored. Here, we performed single cell RNA sequencing of zebrafish ENS at 5days post-fertilization. We identified vagal neural crest progenitors, Schwann cell precursors, and four clusters of differentiated neurons. In addition, a previously unrecognized elavl3+/phox2bb-population of neurons and cx43+/phox2bb-enteric glia was found. Pseudotime analysis supported binary neurogenic branching of ENS differentiation, driven by a notch-responsive state. Taken together, we provide new insights on ENS development and specification, proving that the zebrafish is a valuable model for the study of congenital enteric neuropathies.

Using the Raspberry Pi Virtual Reality (PiVR) System to Study Drosophila Larval Chemotaxis with Real and Virtual Odor Gradients.

Here, we present a detailed protocol for the study of the orientation behavior of larvae of the fruit fly Drosophila melanogaster in response to both real and virtual odors (chemotaxis). An element common to the study of navigation directed by all sensory modalities is the need to correlate changes in behavioral states (e.g., crawling and turning) with temporal changes in the stimulus preceding these events. It has been shown recently that virtual odor landscapes, with any arbitrary geometry, can be created by combining a platform known as "Raspberry Pi virtual reality" (PiVR) with optogenetics. This methodology offers a technical foundation with which to characterize how the larval nervous system responds to stimulation by real and virtual odors. Furthermore, the experimental steps presented and discussed herein highlight important considerations that are needed to ensure experimental reproducibility. Finally, we believe that this framework can be easily adapted and generalized to allow investigators to study other sensory modalities in the Drosophila larva and in other animals.

Issue Information

Current ProtocolsVolume 3, Issue 2 e467 ISSUE INFORMATIONFree Access Issue Information First published: 25 February 2023 https://doi.org/10.1002/cpz1.467AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Graphical Abstract Cover: Synthetic assembly DNA cloning for Drosophila melanogaster is described in Venken et al. Different DNA parts, obtained from flies directly, other species, or through synthetic means, are put together using synthetic assembly DNA cloning to generate continuously growing plasmids of increasing complexity. The resulting end products can be two transgenes, a first transgene providing G418 sulfate selection (provided by the NeoR marker) and encoding the binary LexA transactivator driven by the R70B04 enhancer (i.e., the G418 sulfate-selectable LexA transactivator plasmid), and a second transgene providing Blasticidin S selection (provided by the BlastR marker) and encoding the binary LexA-Op responder, reporting green fluorescent protein (sfGFP) reporter expression driven by the LexA binary transactivator, whose expression is regulated by the R70B04 enhancer (i.e., the Blastidicin S-selectable LexA-Op responder plasmid). A double transgenic fly can then be obtained directly by injecting both transgenes (i.e., dual transgenesis) followed by co-selection using both G418 sulfate and Blasticidin S. The double transgenic fly can then be analyzed for expression patterns. Expression analysis of dual transgenic larval animals shows green fluorescent protein (sfGFP) reporter expression against the neuropil marker Discs Large (Dlg) in the larval central nervous system. View all latest articles RelatedInformation

NuRD‐independent Mi‐2 activity represses ectopic gene expression during neuronal maturation

During neuronal development, extensive changes to chromatin states occur to regulate lineage‐specific gene expression. The molecular factors underlying the repression of non‐neuronal genes in differentiated neurons are poorly characterised. The Mi2/NuRD complex is a multiprotein complex with nucleosome remodelling and histone deacetylase activity. Whilst NuRD has previously been implicated in the development of nervous system tissues, the precise nature of the gene expression programmes that it coordinates is ill‐defined. Furthermore, evidence from several species suggests that Mi‐2 may be incorporated into multiple complexes that may not possess histone deacetylase activity. We show that Mi‐2 activity is required for suppressing ectopic expression of germline genes in neurons independently of HDAC1/NuRD, whilst components of NuRD, including Mi‐2, regulate neural gene expression to ensure proper development of the larval nervous system. We find that Mi‐2 binding in the genome is dynamic during neuronal maturation, and Mi‐2‐mediated repression of ectopic gene expression is restricted to the early stages of neuronal development, indicating that Mi‐2/NuRD is required for establishing stable neuronal transcriptomes during the early stages of neuronal differentiation.