Biology subjects

Tavosanis, G.

Publications and source records attributed to Tavosanis, G..

10 recordsLinked to original sources

bioRxiv2025

Modeling AP2M1 Developmental and Epileptic Encephalopathy in Drosophila

Genetic defects in AP2M1, which encodes the -subunit of the adaptor protein complex 2 (AP-2) essential for clathrin-mediated endocytosis (CME), cause a rare form of developmental and epileptic encephalopathy (DEE). In this study, we modeled AP2M1-DEE in Drosophila melanogaster to gain deeper insights into the underlying disease mechanisms. Pan-neuronal knock-down of the Drosophila AP2M1 ortholog, AP-2{micro}, resulted in a consistent heat-sensitive paralysis phenotype and altered morphology in class IV dendritic arborization (c4da) neurons. Unexpectedly, affected flies were resistant to antiseizure medications and exhibited increased resistance to electrically induced seizures. A CRISPR-engineered fly line carrying the recurrent human disease variant p.Arg170Trp displayed a milder seizure resistance phenotype. While these findings contrast with the human phenotype, they align with previous studies on other CME-related genes in Drosophila. Our results suggest that hyperexcitability and seizures in AP2M1-DEE may stem from broader defects in neuronal development rather than direct synaptic dysfunction.

neuroscience↗

bioRxiv2025

Odour representations supporting ethology-relevant categorisation and discrimination in the Drosophila mushroom body

Neural representations of sensory stimuli serve multiple distinct purposes, from the rapid recognition of familiar environments, to the precise identification of individual salient cues. In the insect mushroom body (MB), odours are encoded by the activity of Kenyon cells (KCs). The random wiring of olfactory projection neurons (PNs) and KCs in the MB calyx is thought to enhance odour discrimination. Here, we examined the impact of deviations from random wiring and demonstrated their significant roles in shaping odour representations. We confirm that different KC types have distinct PN input biases correlated with the contextual relevance of the odour information delivered by the PNs. By recording the functional responses of different KC types to ethologically defined odour categories, we found that the {beta} and {beta} KCs produce segregated representations of relevant odour groups, potentially enhancing the categorisation of odours based on ethological relevance. Simultaneously, these same KC types displayed distinct representations for food-related odours, supporting precise discrimination. In contrast, {gamma} KCs lacked significant segregation of odour representations by ethological category. Computational simulations refined with our functional data indicated that the specific PN input connection pattern of individual KC types largely accounts for the observed representations. Taken together, we propose that individual KC types process odour information with distinct objectives, supporting both ethological categorisation and discrimination.

neuroscience↗

bioRxiv2024

Lipid Disbalance Affects Neuronal Dendrite Growth and Maintenance in a Human Ceramide Synthase Disease Model

The brain is susceptible to disturbances in lipid metabolism. Among the rare, genetically-linked epilepsies Progressive Myoclonic Epilepsy Type 8 (PME8), associated with the loss of Ceramide Synthase (CerS) activity, causes epileptic symptoms accompanied by neurodegenerative traits. We show that expression of a disease-causing cerS allele in Drosophila sensory neurons yielded developmental and degenerative dendrite loss. In cerS mutants, C18-C24 ceramides and membrane-forming complex sphingolipids, into which ceramides are converted, were reduced. At the same time bioactive signaling lipids including (dh)Sphingosine-1-P, deriving from the CerS substrate, were increased. To clarifying the etiology of PME8, we thus performed in vivo experiments to cell-autonomously rescue the individual metabolic alterations. We report that restoring specific long-chain ceramides while in parallel decreasing (dh)Sphingosine-1-P fully rescues the cerS mutant phenotype. Thus, despite the complex metabolic alterations, our data provide essential information about the metabolic origin of PME8 and delineate a potential therapy.

neuroscience↗

bioRxiv2024

Three-photon in vivo imaging of neurons and glia in the medial prefrontal cortex with sub-cellular resolution

The medial prefrontal cortex (mPFC) is important for higher cognitive functions, including working memory, decision making, and emotional control. In vivo recordings of neuronal activity in the mPFC have been achieved via invasive electrical and optical approaches. Here we apply low invasive three-photon in vivo imaging in the mPFC of the mouse at unprecedented depth. Specifically, we measure neuronal and astrocytic Ca2+-transient parameters in awake head-fixed mice up to a depth of 1700 {micro}m. Furthermore, we longitudinally record dendritic spine density (0.41 {+/-}0.07 {micro}m-1) deeper than 1 mm for a week. Using 1650 nm wavelength to excite red fluorescent microglia, we quantify their processes motility (58.9 {+/-}2% turnover rate) at previously unreachable depths (1100 {micro}m). We establish three-photon imaging of the mPFC enabling neuronal and glial recordings with subcellular resolution that will pave the way for novel discoveries in this brain region.

neuroscience↗

bioRxiv2021

MeDUsA: A novel system for automated axon quantification to evaluate neuroaxonal degeneration

BackgroundDrosophila is an excellent model organism for studying human neurodegenerative diseases (NDs), and the rough eye phenotype (REP) assay is a convenient experimental system for analysing the toxicity of ectopically expressed human disease genes. However, the association between REP and axonal degeneration, an early sign of ND, remains unclear. To address this question, we developed a method to evaluate axonal degeneration by quantifying the number of retinal R7 axons in Drosophila; however, it requires expertise and is time-consuming. Therefore, there is a need for an easy-to-use software that can automatically quantify the axonal degeneration. ResultWe created MeDUsA (a method for the quantification of degeneration using fly axons), which is a standalone executable computer program based on Python that combines a pre-trained deep-learning masking tool with an axon terminal counting tool. This software automatically quantifies the number of axons from a confocal z-stack image series. Using this software, we have demonstrated for the first time directly that axons degenerate when the causative factors of NDs (Syn, Tau, TDP-43, HTT) were expressed in the Drosophila eye. Furthermore, we compared axonal toxicity of the representative causative genes of NDs and their pathological alleles with REP and found no significant correlation between them. ConclusionsMeDUsA rapidly and accurately quantifies axons in Drosophila eye. By simplifying and automating time-consuming manual efforts requiring significant expertise, it enables large-scale, complex research efforts on axonal degeneration, such as screening to identify genes or drugs that mediate axonal toxicity caused by ND disease proteins.

pathology↗

bioRxiv2021

A quantitative model of sporadic axonal degeneration in the Drosophila visual system

In human neurodegenerative diseases, neurons undergo axonal degeneration months to years before they die. Here, we developed a system modelling early degenerative events in Drosophila adult photoreceptor cells. Thanks to the stereotypy of their axonal projections, this system delivers quantitative data on sporadic and progressive axonal degeneration of photoreceptor cells. Using this method, we show that exposure of adult flies to a constant light stimulation for several days overcomes the intrinsic resilience of R7 photoreceptors and leads to progressive axonal degeneration. This was not associated with apoptosis. We furthermore provide evidence that loss of synaptic integrity between R7 and a postsynaptic partner preceded axonal degeneration, thus recapitulating features of human neurodegenerative diseases. Finally, our experiments uncovered that neurotransmission to postsynaptic partners of R7 and their response are required to initiate degeneration, suggesting that postsynaptic cells signal back to the photoreceptor to maintain axonal structure. This model can be used to dissect cellular circuit mechanisms involved in the early events of axonal degeneration, allowing for a better understanding of how neurons cope with stress and lose their resilience capacities.

neuroscience↗

bioRxiv2021

The anterior paired lateral neuron normalizes odour-evoked activity at the mushroom body calyx

To identify and memorize discrete but similar environmental inputs, the brain needs to distinguish between subtle differences of activity patterns in defined neuronal populations. The Kenyon cells of the Drosophila adult mushroom body (MB) respond sparsely to complex olfactory input, a property that is thought to support stimuli discrimination in the MB. To understand how this property emerges, we investigated the role of the inhibitory anterior paired lateral neuron (APL) in the input circuit of the MB, the calyx. Within the calyx, presynaptic boutons of projection neurons (PNs) form large synaptic microglomeruli (MGs) with dendrites of postsynaptic Kenyon cells (KCs). Combining EM data analysis and in vivo calcium imaging, we show that APL, via inhibitory and reciprocal synapses targeting both PN boutons and KC dendrites, normalizes odour-evoked representations in MGs of the calyx. APL response scales with the PN input strength and is regionalized around PN input distribution. Our data indicate that the formation of a sparse code by the Kenyon cells requires APL-driven normalization of their MG postsynaptic responses. This work provides experimental insights on how inhibition shapes sensory information representation in a higher brain centre, thereby supporting stimuli discrimination and allowing for efficient associative memory formation.

animal behavior and cognition↗

bioRxiv2020

The branching code: a model of actin-driven dendrite arborisation

Dendrites display a striking variety of neuronal type-specific morphologies, but the mechanisms and principles underlying such diversity remain elusive. A major player in defining the morphology of dendrites is the neuronal cytoskeleton, including evolutionarily conserved actin-modulatory proteins (AMPs). Still, we lack a clear understanding of how AMPs might support developmental phenomena such as neuron-type specific dendrite dynamics. To address precisely this level of in vivo specificity, we concentrated on a defined neuronal type, the class III dendritic arborisation (c3da) neuron of Drosophila larvae, displaying actin-enriched short terminal branchlets (STBs). Computational modelling reveals that the main branches of c3da neurons follow a general growth model based on optimal wiring, but the STBs do not. Instead, model STBs are defined by a short reach and a high affinity to grow towards the main branches. We thus concentrated on c3da STBs and developed new methods to quantitatively describe dendrite morphology and dynamics based on in vivo time-lapse imaging of mutants lacking individual AMPs. In this way, we extrapolated the role of these AMPs in defining STB properties. We propose that dendrite diversity is supported by the combination of a common step, refined by a neuron type-specific second level. For c3da neurons, we present a molecular model of how the combined action of multiple AMPs in vivo define the properties of these second level specialisations, the STBs. In briefA quantitative morphological dissection of the concerted actin-modulatory protein actions provides a model of dendrite branchlet outgrowth. HighlightsO_LIActin organisation in small terminal branchlets of Drosophila class III dendritic arborisation neurons C_LIO_LISix actin-modulatory proteins individually control the characteristic morphology and dynamics of branchlets C_LIO_LIQuantitative tools for dendrite morphology and branch dynamics enable a comparative analysis C_LIO_LIA two-step computational growth model reproduces c3da dendrite morphology C_LI

neuroscience↗

bioRxiv2020

Circuit reorganization in the Drosophila mushroom body calyx accompanies memory consolidation

The capacity of utilizing past experience to guide future action is a fundamental and conserved function of the nervous system. Associative memory formation initiated by the coincident detection of a conditioned stimulus (CS, e.g. odour) and an unconditioned stimulus (US, e.g. sugar reward) can lead to a short-lived memory trace (STM) within distinct circuits [1-5]. Memories can be consolidated into long-term memories (LTM) through processes that are not fully understood, but depend on de-novo protein synthesis [6, 7], require structural modifications within the involved neuronal circuits and might lead to the recruitment of additional ones [8-17]. Compared to modulation of existing connections, the reorganization of circuits affords the unique possibility of sampling for potential new partners [18-20]. Nonetheless, only few examples of rewiring associated with learning have been established thus far [14, 21-24]. Here, we report that memory consolidation is associated with the structural and functional reorganization of an identified circuit in the adult fly brain. The formation and retrieval of olfactory associative memories in Drosophila requires the mushroom body (MB) [25]. We identified the individual synapses of olfactory projection neurons (PNs) that deliver a conditioned odour to the MB and reconstructed the complexity of the microcircuit they form. Combining behavioural experiments with high-resolution microscopy and functional imaging, we demonstrated that the consolidation of appetitive olfactory memories closely correlates with an increase in the number of synaptic complexes formed by the PNs that deliver the conditioned stimulus and their postsynaptic partners. These structural changes result in additional functional synaptic connections.

neuroscience↗

bioRxiv2020

A developmental stretch-and-fill process that optimises dendritic wiring

Circuit connectivity and computation depend on how dendrites branch and occupy space within neural tissue. While optimal wiring principles have long been known to constrain dendritic morphology and their scaling behaviour, the growth dynamics that produce such optimised structures remain unclear. Leveraging structural imaging across development, we identify two complementary growth strategies - inside-out versus outside-in - that together generate mature dendritic arbours. We formalise these dynamics in a mathematical model that captures the two growth modes and show that their interplay yields wiring-efficient, space-filling morphologies and class-specific developmental trajectories across species. This framework provides an algorithmic account of how local branching dynamics give rise to globally optimised architectures. By linking dendritic growth rules to functional design constraints, our theory offers a unifying description of dendritic differentiation and a basis for understanding how coverage and connectivity emerge during neural circuit formation. In briefWe derive a detailed mathematical model that describes long-term time-lapse data of growing dendrites; it optimises total wiring and space-filling. HighlightsO_LIFly neurons stretch and fill a given target area with precise scaling relations. C_LIO_LIWe observe a sequence of two growth strategies. C_LIO_LIEach growth type implements optimal wiring which leads to optimal space filling. C_LIO_LIA model combining these programs captures the development of dendritic structures. C_LI O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=200 SRC="FIGDIR/small/191064v2_ufig1.gif" ALT="Figure 1"> View larger version (50K): org.highwire.dtl.DTLVardef@1f2aed5org.highwire.dtl.DTLVardef@1b449edorg.highwire.dtl.DTLVardef@161a400org.highwire.dtl.DTLVardef@1562be5_HPS_FORMAT_FIGEXP M_FIG C_FIG

neuroscience↗