Biology subjects

Waddell, S.

Publications and source records attributed to Waddell, S..

At least 19 recordsLinked to original sources

bioRxiv2026

Dietary magnesium supplement enhances memory through Hippo signaling

Dietary magnesium (Mg2+) supplement enhances memory in mammals and Drosophila, but whether and how Mg2+ changes molecular states of neurons, remains unknown. Here we used single-cell RNA sequencing to map cell-type specific transcriptional responses to memory-enhancing dietary Mg2+. These analyses revealed changes in Hippo-pathway gene expression in memory-relevant {beta} Kenyon Cells (KCs) of the mushroom bodies. Targeted knock-down of Merlin, hippo (hpo) and warts demonstrated requirement in {beta} KCs for Mg2+-enhanced, but not baseline memory. Further study of Hippo signaling identified Yorkie (yki) transcriptional coactivator and DNA-binding partner Scalloped (sd) to be required for baseline and Mg2+-enhanced memory. Moreover, releasing Yki::Sd of inhibition from Tondu-domain containing growth inhibitor (Tgi) specifically in {beta} KCs enhanced memory, independent of diet. Analyses of Yki::Sd target genes identified several required in {beta} KCs for baseline memory, including crb and Glut1 which are also Mg2+-induced in a Hippo-dependent manner in {beta} KCs. These genes suggest that aiding the metabolic demands of long-term memory and facilitating structural neuronal plasticity are key mediators of Mg2+-induced memory enhancement. We propose pharmacological enhancement of Sd-dependent transcription, and activity of particular Sd-target genes may have therapeutic potential for memory disorders.

neuroscience↗

bioRxiv2026

Lateralized dopaminergic neuron activity encodes goal-directed turning

Animals navigate through dynamic environments in search of resources. Dopaminergic systems are known to control movement and assign motivational value to cues, actions and outcomes, but how they guide ongoing locomotion towards a goal remains unclear. Here, we demonstrate that Drosophila mushroom body innervating dopaminergic neurons encode steering signals in world-centred (allocentric) and body-centred (egocentric) coordinates during goal-directed navigation. Thirsty flies track water vapour through upwind directed turning and locomotion, that persists for several seconds after the vapour disappears. We identified a population of rewarding dopaminergic neurons to be specifically required for maintaining directed tracking following vapour offset. Recordings of these dopaminergic neurons during behaviour revealed delayed bilateral activity that followed upwind-directed turning in allocentric coordinates. Moreover, lateralized ipsilateral activity in a subset of these dopaminergic neurons encode whether the fly turns left or right in egocentric coordinates. Finally, these dopaminergic neurons also exhibited increased ipsilateral activity when turning in the direction where water vapour was previously encountered. Therefore, activity of mushroom body dopaminergic neurons represents steering and direction-specific actions that together maintain goal-directed tracking in turbulent environments.

neuroscience↗

bioRxiv2026

Hunger reconfigures a reward learning circuit into a memory competent mode

Internal states such as hunger dynamically reshape activity across circuits to support resource seeking. Neuromodulation provides a means of controlling such physiological properties of neurons, but how this flexibility regulates memory networks remains unclear. Here, we describe how hunger reconfigures a dopaminergic food-reward circuit between two physiological modes that support memory formation, in Drosophila. Starvation suppresses baseline dopaminergic activity through peptidergic signalling, enabling reward-evoked, large-amplitude dopamine neuron spikes to reinforce learning. This spiking mode can be engaged by sugar consumption and persists beyond feeding, reflecting the fly's satiety state. Persistent dopaminergic large-amplitude spiking reinforces learning and transitions the memory network into a mode that prioritizes consolidation of recently acquired memories. The transition from hunger to satiation is also reflected in the activity of postsynaptic output neurons that shift from a tonic, decorrelated mode that is responsive to dopamine into a bursting, correlated mode in which further dopaminergic modulation is occluded. Therefore, nutrient deprivation reconfigures the memory circuit into a learning competent dopamine-receptive mode, which persistent reinforcing dopamine then switches into a satiated mode driving memory consolidation. Together these processes mechanistically intertwine state-dependent reward-signalling with subsequent memory stabilization through transitions in physiological mode.

neuroscience↗

bioRxiv2026

Genetic sex of enteric neurons enables ovarian relaxin togate maternal gut plasticity

Animals must align intestinal plasticity and feeding with reproductive state, yet the checkpoint that gates these adaptations is unknown. Here we show that an ovary-to-enteric-neuron axis gates the onset of maternal gut plasticity in Drosophila. Genetic sex establishes endocrine competence in a subset of enteric neurons via the sex determination pathway, enabling female-specific expression of the relaxin-family receptor Lgr3. After mating, steroid signalling increases Lgr3 receptor expression, priming these neurons for reproductive adaptation. Once oocytes mature fully, follicle cells secrete the relaxin-like hormone dILP8, which activates Lgr3 to trigger gut enlargement and increased feeding. Disrupting the sex determination pathway in enteric neurons, Lgr3, or ovarian dILP8 prevents gut enlargement and reduces feeding. Thus, genetic sex establishes competence, steroid signalling primes it, and ovarian relaxin triggers it, defining a maternal intestinal plasticity checkpoint that ensures gut adaptations initiate only once reproduction is underway and energy demands peak. Our findings delineate an ovary-to-enteric-neuron axis that couples reproductive state to maternal gut plasticity.

physiology↗

bioRxiv2025

Visceral signaling of post-ingestive malaise directs memory updating in Drosophila

Consolidation is a time when labile memories transition to a stable form. Malaise learning in Drosophila reveals consolidation to also permit memory updating. Flies taught to associate one of two odors with toxin-tainted sugar initially express conditioned odor approach, that following consolidation switches to avoidance. Behavioral reversal emerges from dopaminergic update of parallel memories for the two trained odors. Differential serotoninergic modulation of specific aversive and rewarding dopaminergic neuron subtypes permits post-ingestive intoxication to suppress consolidation of initial odor-sugar memory and simultaneously invert reward memory plasticity into "safety" memory for the odor experienced without food. Fat body release of the Toll-ligand activating protease modSP, and resilience factor Turandot A, instruct malaise updates by triggering autocrine Toll signaling in the same brain dopaminergic neurons that form and consolidate initial sugar memory. This neural mechanism overcomes the credit assignment problem of delayed post-ingestive reinforcement by updating earlier memories of the trained odors.

neuroscience↗

bioRxiv2025

Connectomics Reveals a Feed-Forward Swallowing Circuit Driving Protein Appetite

Nutrient state shapes not only what animals eat, but how they eat it. We lack circuit-level mechanisms explaining how gustatory and internal state information regulates feeding at the motor control level. In Drosophila, protein deprivation prolongs protein-specific feeding bursts, yet the detailed motor mechanisms underlying this change remain unknown. Using EM connectomics, we identified a feed-forward pathway from proteinsensitive gustatory receptor neurons to swallowing motor neurons. At its core is the Sustain neuron, which coordinates multiple swallowing motor neurons to move food efficiently through the cibarium and pharynx. This nutrient-dependent facilitation of swallowing sustains long feeding bursts, directly linking internal state to the temporal structure of the feeding motor program. Our findings reveal how a dedicated sensorimotor circuit translates physiological need into precise motor control to drive nutrient-specific feeding appetite and highlights the power of combining EM connectomics with experimental circuit dissection.

neuroscience↗

bioRxiv2024

A modulatory attentional gate promotes recent memory expression in Drosophila

Using short-term memory (STM) requires that animals can distinguish memories of recent experience from those learned previously. In Drosophila the neuromodulator octopamine (OA) specifically affects STM through adrenergic-like receptors within neurons in the Mushroom Body (MB) network. However, why OA preferentially impacts STM remains unclear. A fly brain connectome reveals that two OA-VPM3 neurons provide most OA input to the MB network. Artificial OA-VPM3 activation during olfactory learning enhances STM and can divert odor salience. -adrenergic-like receptor knock-down in MB {beta} and {gamma} lobe MB Output Neurons (MBONs) generally impaired STM, while {beta}-adrenergic-like receptor loss from lobe MBONs enhanced memory. The identity and connectivity of the relevant MBONs suggests OA modulates an interaction between sites of STM and long-term memory (LTM) storage. Indeed, synthetic, odor, and learning-driven, OA-VPM3 activation temporarily blocks LTM expression. Therefore, OA reconfigures the MBON network so the fly prioritizes expression of recent over remote memories.

neuroscience↗

bioRxiv2023

Sexually dimorphic mechanisms of VGLUT-mediated protection from dopaminergic neurodegeneration

Parkinsons disease (PD) targets some dopamine (DA) neurons more than others. Sex differences offer insights, with females more protected from DA neurodegeneration. The mammalian vesicular glutamate transporter VGLUT2 and Drosophila ortholog dVGLUT have been implicated as modulators of DA neuron resilience. However, the mechanisms by which VGLUT2/dVGLUT protects DA neurons remain unknown. We discovered DA neuron dVGLUT knockdown increased mitochondrial reactive oxygen species in a sexually dimorphic manner in response to depolarization or paraquat-induced stress, males being especially affected. DA neuron dVGLUT also reduced ATP biosynthetic burden during depolarization. RNA sequencing of VGLUT+ DA neurons in mice and flies identified candidate genes that we functionally screened to further dissect VGLUT-mediated DA neuron resilience across PD models. We discovered transcription factors modulating dVGLUT-dependent DA neuroprotection and identified dj-1{beta} as a regulator of sex-specific DA neuron dVGLUT expression. Overall, VGLUT protects DA neurons from PD-associated degeneration by maintaining mitochondrial health.

neuroscience↗

bioRxiv2023

EfpA is required for re-growth of Mycobacterium tuberculosis following isoniazid exposure

Efflux of antibiotics is a survival strategy in bacteria. Mycobacterium tuberculosis has approximately sixty efflux pumps, but little is known about the role of each pump, or which moieties that they efflux. The putative efflux pump, EfpA, is a member of the major facilitator superfamily that has been shown to be essential by saturation transposon mutagenesis studies. It has been implicated in the efflux of the frontline drug isoniazid (INH) in M. tuberculosis. This is supported by evidence from transcriptional profiling that efpA is induced in response to INH exposure. However, its role in physiology and adaptation of M. tuberculosis to antibiotics, have yet to be determined. Here, we describe the repression of efpA using CRISPR interference and the direct effect of this on the ability of M. tuberculosis to survive exposure to INH over a 45-day time-course. We determined that wild-type levels of efpA were required for the recovery of M. tuberculosis cultures following INH exposure and that, after 45-days of INH exposure, no viable colonies were recoverable from efpA-repressed M. tuberculosis cultures. We postulate that EfpA is required for the recovery of M. tuberculosis following INH-exposure and that EfpA may have a role in the development of resistance, during treatment and contributes to relapse in patients.

microbiology↗

bioRxiv2022

Multisensory learning binds modality-specific neurons into a cross-modal memory engram

Withdrawal StatementThe authors have withdrawn this manuscript. After conducting replication experiments and reanalysing the original data, the authors discovered that the voltage imaging results are not reproducible. Additional voltage imaging work by the authors and their collaborators has not produced robust signals. Furthermore, errors in the pipeline originally used for data analysis could have led to the contamination of data. A complete reanalysis by the authors of the voltage imaging data independently from the original pipeline did not replicate the original analysis. Although the authors have replicated the behavioural and connectomic data and consider that the general conclusions of the article are substantiated, the authors have no confidence in the voltage imaging data. The authors would like to apologize for any inconvenience caused to their scientific colleagues and the readers of the journal. The authors would like to thank Waddell lab members, A. Cook, A. Miriyala and M. Klappenbach, who were involved in the replication experiments but are not authors of the original paper and M. Hoffmann, S. Thornquist and G. Maimon of Rockefeller University for identifying possible errors in the voltage data processing. All authors agree with the withdrawal, with the exception of P. F. Jacob who did not respond to correspondence. Therefore, the authors do not wish this work to be cited as reference for the project. If you have any questions, please contact the corresponding author.

neuroscience↗

bioRxiv2022

Gliotransmission of D-serine promotes thirst-directed behaviors in Drosophila

Thirst emerges from a range of cellular changes that ultimately motivate an animal to consume water. Although thirst-responsive neuronal signals have been reported, the full complement of brain responses is unclear. Here we identify molecular and cellular adaptations in the brain using single-cell sequencing of water deprived Drosophila. Water deficiency primarily altered the glial transcriptome. Screening the regulated genes revealed astrocytic expression of the astray-encoded phosphoserine phosphatase to bi-directionally regulate water consumption. Astray synthesizes the gliotransmitter D-serine and vesicular release from astrocytes is required for drinking. Moreover, dietary D-serine rescues aay-dependent drinking deficits while facilitating water consumption and expression of water-seeking memory. D-serine action requires binding to neuronal NMDA-type glutamate receptors. Fly astrocytes contribute processes to tripartite synapses and the proportion of astrocytes that are themselves activated by glutamate increases with water deprivation. We propose that thirst elevates astrocytic D-serine release, which awakens quiescent glutamatergic circuits to enhance water procurement.

neuroscience↗

bioRxiv2022

Different dopaminergic neurons signal absolute and relative aversive value in the Drosophila mushroom body.

Animals use prior experience to assign absolute (good or bad) and also relative (better or worse) value to new experience. These learned values guide appropriate later decision-making. While our understanding of how the dopaminergic system computes absolute value is relatively advanced, the mechanistic underpinnings of relative valuation are unclear. Here we reveal mechanisms of absolute and relative aversive valuation in Drosophila. Three types of punishment-sensitive dopaminergic neurons (DANs) drive intensity-scaled plasticity at their respective mushroom body output neuron (MBON) connections to code absolute aversive value. In contrast, by comparing current and previous aversive experiences the MBON-DAN network can code relative aversive value by recruiting a specific subtype of reward-coding dopaminergic neurons which assigns a better than value to the lesser of two aversive experiences. This study therefore provides an important functional consequence of having opposing populations of DANs and illustrates how these can operate together as a system within the MB network to code and compare sequential aversive experience to learn relative aversive value.

neuroscience↗

bioRxiv2021

In vivo modelling of patient genetic heterogeneity identifies concurrent Wnt and PI3K activity as a potent driver of invasive cholangiocarcinoma growth.

Intrahepatic cholangiocarcinoma (ICC) is an aggressive and lethal malignancy of the bile ducts within the liver characterised by high levels of genetic heterogeneity. In the context of such genetic variability, determining which oncogenic mutations drive ICC growth has been difficult and developing modes of patient stratification and targeted therapies remains challenging. As a result, survival rates following a diagnosis with ICC have remained static since the late 1970s, whilst incidence of ICC has increased. Here, we performed the first functional in vivo study into the role that genetic heterogeneity plays in drivinga ICC via modelling of interactions between rare mutations with more common driver genes. By leveraging human ICC sequencing data to stratify and then model genetic heterogeneity in the mouse, we uncovered numerous novel tumour suppressors which, when lost, cooperate with the RAS oncoprotein to drive ICC growth. In this study, we specifically focus on a set of driver mutations that interact with KRAS to initiate aggressive, sarcomatoid-type ICC. We show that tumour growth of this cancer relies on both Wnt and PI3K signalling to drive proliferation and suppress apoptosis. Finally, we demonstrate that pharmacological co-inhibition of Wnt and PI3K in vivo substantially impedes the growth of ICC, regardless of mutational profile. As such, Wnt and PI3K activity should be considered as a signature by which patients can be stratified for treatment and inhibitors of these pathways should be levied as a treatment for patients diagnosed with ICC.

cancer biology↗

bioRxiv2021

CMTr cap-adjacent 2`-O-ribose mRNA methyltransferases are required for reward learning and mRNA localization to synapses

Cap-adjacent nucleotides of animal, protist and viral mRNAs can be dynamically O-methylated at the 2 position of the ribose (cOMe). The functions of cOMe in animals, however, remain unknown. Here we show that the two cap methyltransferases (CMTr1 and CMTr2) of Drosophila can methylate the ribose of the first nucleotide in mRNA. Double-mutant flies lack cOMe but are viable. Consistent with prominent neuronal expression, they have a reward learning defect that can be rescued by conditional expression in mushroom body neurons before training. Among CMTr targets are cell adhesion and signaling molecules relevant for learning and cOMe is required for local translation of mRNAs at synapses. Hence, our study reveals a mechanism to co-transcriptionally prime mRNAs by cOMe for localized protein synthesis at synapses.

molecular biology↗

bioRxiv2021

An opposing self-reinforced odor pre-exposure memory produces latent inhibition in Drosophila

Prior experience of a stimulus can inhibit subsequent acquisition or expression of a learned association of that stimulus. However, the neuronal manifestations of this learning effect, named latent inhibition (LI), are poorly understood. Here we show that odor pre-exposure produces LI of appetitive olfactory memory performance in Drosophila. Behavioral expression of LI requires that the context during memory testing resembles that during the odor pre-exposures. Odor pre-exposure forms an aversive memory that requires dopaminergic neurons that innervate the {gamma}2'1 and 3 mushroom body compartments - those to 3 exhibit increasing odor-driven activity with successive pre-exposures. In contrast, odor-specific responses of the corresponding mushroom body output neurons are suppressed. Odor pre-exposure therefore recruits specific dopaminergic neurons that provide teaching signals that attach negative valence to the odor itself. LI of Drosophila appetitive memory consequently results from a temporary and context-dependent retrieval deficit imposed by competition with this short-lived aversive memory.

neuroscience↗

bioRxiv2020

Dendritic localization of mRNA in Drosophila Mushroom Body Output Neurons

Memory-relevant neuronal plasticity is believed to require local translation of new proteins at synapses. Understanding this process requires the visualization of the relevant mRNAs within these neuronal compartments. Here we used single-molecule fluorescence in situ hybridization (smFISH) to localize mRNAs at subcellular resolution in the adult Drosophila brain. mRNAs for subunits of nicotinic acetylcholine receptors and kinases could be detected within the dendrites of co-labelled Mushroom Body Output Neurons (MBONs) and their relative abundance showed cell-specificity. Moreover, aversive olfactory learning produced a transient increase in the level of CaMKII mRNA within the dendritic compartments of the {gamma}5{beta}'2a MBONs. Localization of specific mRNAs in MBONs before and after learning represents a critical step towards deciphering the role of dendritic translation in the neuronal plasticity underlying behavioural change in Drosophila.

neuroscience↗

bioRxiv2020

The connectome of the adult Drosophila mushroom body: implications for function

Making inferences about the computations performed by neuronal circuits from synapse-level connectivity maps is an emerging opportunity in neuroscience. The mushroom body (MB) is well positioned for developing and testing such an approach due to its conserved neuronal architecture, recently completed dense connectome, and extensive prior experimental studies of its roles in learning, memory and activity regulation. Here we identify new components of the MB circuit in Drosophila, including extensive visual input and MB output neurons (MBONs) with direct connections to descending neurons. We find unexpected structure in sensory inputs, in the transfer of information about different sensory modalities to MBONs, and in the modulation of that transfer by dopaminergic neurons (DANs). We provide insights into the circuitry used to integrate MB outputs, connectivity between the MB and the central complex and inputs to DANs, including feedback from MBONs. Our results provide a foundation for further theoretical and experimental work.

neuroscience↗

bioRxiv2020

Input connectivity reveals additional heterogeneity of dopaminergic reinforcement in Drosophila

Different types of Drosophila dopaminergic neurons (DANs) reinforce memories of unique valence and provide state-dependent motivational control [1]. Prior studies suggest that the compartment architecture of the mushroom body (MB) is the relevant resolution for distinct DAN functions [2, 3]. Here we used a recent electron microscope volume of the fly brain [4] to reconstruct the fine anatomy of individual DANs within three MB compartments. We find the 20 DANs of the {gamma}5 compartment, at least some of which provide reward teaching signals, can be clustered into 5 anatomical subtypes that innervate different regions within {gamma}5. Reconstructing 821 upstream neurons reveals input selectivity, supporting the functional relevance of DAN sub-classification. Only one PAM-{gamma}5 DAN subtype {gamma}5(fb) receives direct recurrent input from {gamma}5{beta}2a mushroom body output neurons (MBONs) and behavioral experiments distinguish a role for these DANs in memory revaluation from those reinforcing sugar memory. Other DAN subtypes receive major, and potentially reinforcing, inputs from putative gustatory interneurons or lateral horn neurons, which can also relay indirect feedback from MBONs. We similarly reconstructed the single aversively reinforcing PPL1-{gamma}1pedc DAN. The {gamma}1pedc DAN inputs mostly differ from those of {gamma}5 DANs and they cluster onto distinct dendritic branches, presumably separating its established roles in aversive reinforcement and appetitive motivation [5, 6]. Tracing also identified neurons that provide broad input to {gamma}5, {beta}2a and {gamma}1pedc DANs suggesting that distributed DAN populations can be coordinately regulated. These connectomic and behavioral analyses therefore reveal further complexity of dopaminergic reinforcement circuits between and within MB compartments. HighlightsO_LINanoscale anatomy reveals additional subtypes of rewarding dopaminergic neurons. C_LIO_LIConnectomics reveals extensive input specificity to subtypes of dopaminergic neurons. C_LIO_LIAxon morphology implies dopaminergic neurons provide subcompartment-level function. C_LIO_LIUnique dopaminergic subtypes serve aversive memory extinction and sugar learning. C_LI

neuroscience↗