Biology subjects

Szyszka, P.

Publications and source records attributed to Szyszka, P..

12 recordsLinked to original sources

bioRxiv2026

Evidence that Dogs Can Use Temporal Difference in Odorant Arrival to Discriminate Odorant Mixtures

In turbulent environments, odorants from different sources arrive at different times, potentially providing cues for odor source segregation. In several invertebrate species, short differences in odorant onset enable freely moving animals to discriminate odorant mixtures. In vertebrates, however, studies of sensitivity to odorant onset asynchrony have been conducted under highly constrained sampling conditions, such as with odor delivery tightly coupled to respiration. In this study, we investigated whether domestic dogs could detect odorant onset asynchrony in odorant mixtures under conditions that preserve key features of natural odor sampling. Dogs performed a discrimination task in which odor stimuli were presented as ongoing pulse trains that began independently of animal behavior, avoiding artificial synchronization of odor delivery with sniff cycles. Dogs were trained to discriminate between mixtures of two odorants with synchronous onsets and mixtures with asynchronous onsets. Of the dogs trained, one was able to discriminate odorant onset asynchronies as short as 633 ms. Dogs also displayed sensitivity to auditory stimulus onset asynchrony, discriminating auditory asynchronies as short as 30 ms. These results provide the first demonstration of temporal sensitivity in canine olfaction and the first evidence that vertebrates can use odorant onset asynchrony under conditions that permit free odor sampling.

animal behavior and cognition↗

bioRxiv2025

Seizures, increased interhemispheric synchrony, altered brain transcriptomics and a leaky blood-brain barrier result from loss of ap3b2 in a CRISPR tadpole model of DEE48

Loss-of-function variants in AP3B2, a neuronal adaptor protein required for synaptic vesicle formation, cause a severe early-onset neurodevelopmental epilepsy known as Developmental and Epileptic Encephalopathy 48 (DEE48). To investigate how AP3B2 loss alters brain development, leading to increased seizure susceptibility, we generated a Xenopus laevis model by targeting the orthologous gene using CRISPR/Cas9. Ap3b2.S-/- (mosaic) F0 tadpoles displayed increased locomotor activity with frequent seizure-like episodes when compared to sibling controls. Visualisation of forebrain and midbrain activity using the genetically encoded Ca2+ sensor GCaMP6s detected spontaneous, large amplitude, prolonged and widespread neural activity, alongside increased interhemispheric synchrony of both regions. Comparison of whole-brain transcriptomes from ap3b2 CRISPants and unedited sibling controls detected mainly downregulation of brain expressed genes, with significant over-representation of pathways involved in ion transport, axon formation and guidance, inhibitory (GABA) neurotransmission, and transport across the blood-brain barrier (BBB). In a simple assay for BBB integrity, CRISPant tadpoles were confirmed to have faster leakage of sodium fluoresceinAcute exposure to the angiotensin receptor blocker losartan significantly reduced locomotor hyperactivity, and CRISPant cohorts treated with losartan tended to have lower neural activity, indicating incomplete rescue of the ap3b2.S CRISPant phenotype. These findings demonstrate how AP3B2 loss of function alters brain development and the establishment of the BBB, with the resulting alterations in neurotransmitter pathways predisposing the brain to spontaneous seizures. Our results suggest that traditional anti-seizure medications designed to alter ion transport and GABA metabolism could be augmented with drugs targeting neuroinflammation, as adjunct seizure control options in infants with DEE48.

developmental biology↗

bioRxiv2023

Knockdown of NeuroD2 leads to seizure-like behaviour, brain neuronal hyperactivity and a leaky blood-brain barrier in a Xenopus laevis tadpole model of DEE75

Developmental and Epileptic Encephalopathies (DEE) are a genetically diverse group of severe, early onset seizure disorders. DEE are normally identified clinically in the first six months of life by the presence of frequent, difficult to control seizures and accompanying stalling or regression of development. DEE75 results from de novo mutations of the NEUROD2 gene that result in loss of activity of the encoded transcription factor, and the seizure phenotype was shown to be recapitulated in Xenopus tropicalis tadpoles. We used CRISPR/Cas9 to make a DEE75 model in Xenopus laevis, to further investigate the developmental aetiology. NeuroD2.S CRISPR/Cas9 edited tadpoles were more active, swam faster on average, and had more unprovoked escape responses (C-starts) than their sibling controls. Live imaging of Ca2+ signalling revealed prolongued, strong signals sweeping through the brain, indicative of neuronal hyperactivity. While the resulting tadpole brain appeared grossly normal, the blood-brain barrier was found to be leakier than that of controls. Additionally, the TGF{beta} antagonist Losartan was shown to have a short-term protective effect, reducing neuronal hyperactivity and reducing permeability of the blood- brain barrier. Severity of the behavioral phenotype correlated with increased with editing efficiency. Our results support a haploinsufficiency model of DEE75 resulting from reduced NeuroD2 activity during vertebrate brain development, and indicate that a leaky blood- brain barrier contributes to epileptogenesis.

neuroscience↗

bioRxiv2023

Ephaptic coupling between olfactory receptor neurons is sensitive to relative stimulus timing: Implications for odour source discrimination

Insect olfactory receptor neurons (ORNs) are often co-localized within sensilla and exhibit non-synaptic reciprocal inhibition through ephaptic coupling. It has been postulated that this inhibition aids odour source discrimination, as synchronous arrival of different odour molecules (odorants) from a single source should increase ephaptic inhibition, whereas asynchronous arrival of odorants from different sources should decrease ephaptic inhibition. However, it was as yet unknown whether temporal arrival patterns of different odorants indeed modulate ephaptic inhibition, since past studies have focused on ephaptic inhibition of sustained ORN responses to prolonged and constant odour stimuli. However, most natural odour stimuli are not constant but rather transient and fluctuate as a result of dispersion in turbulent plumes in the air. To investigate the role of ephaptic inhibition in olfaction within turbulent environments, we recorded co-localized ORNs in the fruit fly Drosophila melanogaster exposed to dynamic odorant mixtures. We found that ephaptic inhibition does modulate transient ORN responses, and the strength of ephaptic inhibition decreases as the synchrony between arriving odorants decreases. These results support the hypothesis that ephaptic inhibition aids odour source discrimination.

neuroscience↗

bioRxiv2022

Expression of mRNA encoding two gain-of-function cyfip2 variants associated with DEE65 results in spontaneous seizures in Xenopus laevis tadpoles

Developmental and epileptic encephalopathies (DEE) are a genetically diverse group of disorders with similar early clinical presentations. DEE65 is caused by de novo, non-synonymous, gain-of-function mutations in CYFIP2. It presents in early infancy as hypotonia, epileptic spasms and global developmental delay. While modelling loss-of-function mutations can be done using knockdown or knockout techniques to reduce the amount of functional protein, modelling gain-of-function mutations requires different approaches. Here, we show that transient ectopic expression of the Arg87Cys pathogenic variant of cyfip2 mRNA in Xenopus laevis tadpoles resulted in increased seizure-related behaviours such as rapid darting and swimming in circles. In contrast, expression of a second pathological variant, Tyr108Cys, did not alter tadpole behaviour. Expression of either pathogenic variant resulted in spontaneous epileptic activity in the brain. For both variants, neuronal hyperactivity was reduced by treating the tadpole with 5 mM of the anti-seizure drug valproate (VPA). mRNA overexpression of gain-of-function variants in X. laevis tadpoles may be useful both for understanding the aetiology of DEE and for pre-clinical drug testing.

genetics↗

bioRxiv2022

Characterisation and automated quantification of induced seizure-related behaviours in Xenopus laevis tadpoles

Epilepsy, a clinical diagnosis characterized by paroxysmal episodes known as seizures, affects 1% of people worldwide. Safe and patient-specific treatment is vital and can be achieved by the development of rapid pre-clinical models of for identified epilepsy genes. Epilepsy can result from either brain injury or gene mutations, and can also be induced chemically. Xenopus laevis tadpoles could be a useful model for confirmation of variants of unknown significance found in epilepsy patients, and for drug re-purposing screens that could eventually lead to benefits for patients. Here, we characterise and quantify seizure-related behaviours in X. laevis tadpoles arrayed in 24-well plates. To provoke acute seizure behaviours, tadpoles were chemically induced with either pentylenetetrazole (PTZ) or 4-aminopyridine (4-AP). To test the capacity to adapt this method for drug testing, we also exposed induced tadpoles to the anti-seizure drug valproate (VPA). Four induced seizure-like behaviours were described and manually quantified, and two of these (darting, circling) could be accurately detected automatically, using the video analysis software TopScan. Additionally, we recorded swimming trajectories and mean swimming velocity. Automatic detection showed that either PTZ or 4-AP induced darting behaviour and increased mean swimming velocity compared to untreated controls. Both parameters were significantly reduced in the presence of VPA. In particular, darting behaviour was a shown to be a sensitive measure of epileptic seizure activity. While we could not automatically detect the full range of seizure behaviours, this method shows promise for future studies, since X. laevis is a well-characterised and genetically tractable model organism.

neuroscience↗

bioRxiv2022

Sugar alcohols have the potential as bee-safe feeding stimulants for wasp control

Pest insects are often baited with poisoned feeding stimulants, the most common of which are sugars. However, sugars are attractive for most animal species, which makes it difficult to target only a specific pest insect species. Here, we assessed different sugar alcohols for their potential as more species-selective feeding stimulants for pest insects. We tested the attractiveness of the sugar alcohols sorbitol, xylitol and erythritol with a capillary feeder assay in wasps (as potential pest insects, because introduced wasps are a pest in many regions) and bees (as non-target insects). For the common wasp (Vespula vulgaris), sorbitol and xylitol acted as nutritive feeding stimulants, and erythritol acted as a non-nutritive feeding stimulant. For the buff-tailed bumble bee (Bombus terrestris), sorbitol acted as a feeding stimulant, while for the honey bee (Apis mellifera), none of the sugar alcohols acted as feeding stimulant. The species-specific preferences for sugar alcohols suggest their potential as species-selective insect baits. The wasp-specific preference for xylitol suggests its potential as bee-safe alternative to sugar-containing bait for wasp pest control. GRAPHICAL ABSTRACT O_FIG_DISPLAY_L [Figure 1] M_FIG_DISPLAY We tested the attractiveness of sugar alcohols with a capillary feeder assay in wasps and bees. Species-specific preferences suggests that sugar alcohols have the potential for being used as bee-safe insect baits. C_FIG_DISPLAY

ecology↗

bioRxiv2020

Adaptive evolution of olfactory degeneration in recently flightless insects

Fast-moving animals need fast-acting sensory systems. Flying insects have thus evolved exceptionally quick visual (1) and olfactory processing ability (2). For example, flighted insects can track the temporal structure of turbulent odor plumes at rates above 100 Hz (3). The evolutionary lability of such sensory systems, however, remains unknown. We test for rapid evolutionary shifts in olfactory processing speed associated with flight loss, through neurobiological comparisons of sympatric flighted versus flightless lineages within a wing-polymorphic stonefly species. Our analyses of sensory responses reveal that recently-evolved flightless lineages have substantially degraded olfactory acuity. By comparing flighted versus flightless ecotypes with similar genetic backgrounds (4), we eliminate other confounding factors that might have affected the evolution of their olfactory reception mechanisms. Our detection of different patterns of degraded olfactory sensitivity and speed in independently wing-reduced lineages highlights parallel evolution of sensory degeneration. These reductions in sensory ability also echo the rapid vestigialization of wings themselves (4, 5), and represent a neurobiological parallel to the convergent phenotypic shifts seen under sharp selective gradients in other systems (e.g. parallel loss of vision in diverse cave fauna (6)). Our study provides the first direct evidence for the hypothesis that flight poses a selective pressure on the speed of olfactory receptor neurons. Our findings also emphasize the energetic costs of rapid olfaction, and the key role of natural selection in shaping dramatic neurobiological shifts. Significance StatementFlying insects move fast and have therefore evolved exceptionally quick-acting sensory systems. The speed with which such neurobiological shifts can evolve, however, remains unclear. Under the use it or lose it hypothesis, loss of flight should lead to degradation of this fast sensory processing ability. We test for evolutionary reductions in olfactory acuity linked to flight loss, through neurobiological comparisons of flightless versus flighted lineages within a wing-polymorphic insect. Our analyses reveal that newly wing-reduced populations have substantially degraded olfactory acuity, with parallel reductions in this sensory ability detected in independently flightless lineages. These findings reveal that flight poses strong selective pressure for rapid olfaction, and highlight the potential of natural selection in rapidly shaping adaptive shifts in animal sensory systems.

neuroscience↗

bioRxiv2019

Social foraging extends associative odor-food memory expression in fruit flies (Drosophila melanogaster)

Animals socially interact during foraging and share information about the quality and location of food sources. The mechanisms of social information transfer during foraging have been mostly studied at the behavioral level, and its underlying neural mechanisms are largely unknown. The fruit fly Drosophila melanogaster has become a model for studying the neural bases of social information transfer, as fruit flies show a rich repertoire of social behaviors and provide a well-developed genetic toolbox to monitor and manipulate neuronal activity. Social information transfer has already been characterized for fruit flies egg laying, mate choice, foraging and aversive associative learning, however the role of social information transfer on associative odor-food learning during foraging are unknown. Here we present an automated learning and memory assay for walking flies that allows studying the effect of group size on social interactions and on the formation and expression of associative odor-food memories. We found that both inter-fly attraction and the duration of odor-food memory expression increase with group size. We discuss possible behavioral and neural mechanisms of this social effect on odor-food memory expression. This study opens up opportunities to investigate how social interactions are relayed in the neural circuitry of learning and memory expression.

neuroscience↗

bioRxiv2019

Alpha oscillations govern interhemispheric spike timing coordination in the honey bee brain

In 1929 Hans Berger discovered the alpha oscillations: a prominent, ongoing 10 Hz rhythm in the human electroencephalogram (EEG). These alpha oscillations are amongst the most widely studied cerebral signals, related to cognitive phenomena such as attention, memory, and consciousness. However, the mechanisms by which alpha oscillations affect human cognition await demonstration. Here we suggest the honey bee brain as an experimentally more accessible model system for investigating the functional roles of alpha oscillations. We found a prominent alpha wave-like spontaneous neural activity (~ 18 Hz) that is reduced in amplitude upon stimulus presentation. The phase of this alpha activity biased both timing of neuronal spikes and amplitude of high-frequency gamma activity (~ 30 Hz). These results suggest a common role of oscillatory neuronal activity across phyla and provide an unprecedented new venue for causal studies on the relationship between neuronal spikes, brain oscillations, and cognition.

neuroscience↗

bioRxiv2019

Odor-background segregation of unknown odorants based on stimulus onset asynchrony in honey bees

Animals use olfaction to search for distant objects. Unlike vision, where objects are spaced out, olfactory information mixes when it reaches olfactory organs. Therefore, efficient olfactory search requires segregating odors that are mixed with background odors. Animals can segregate known target odors by detecting short differences in the arrival of odorants from different sources (stimulus onset asynchrony). However, it is unclear whether animals can also use stimulus onset asynchrony to segregate previously unknown odorants that have no innate or learned relevance. Using behavioral experiments in honey bees, we here show that stimulus onset asynchrony also improves odor-background segregation of unknown odorants. The stimulus onset asynchrony necessary to behaviorally segregate unknown odorants is in the range of seconds, which is two orders of magnitude larger than the previously reported stimulus asynchrony sufficient for segregating known odorants. We propose that for unknown odorants, odor-background segregation requires sensory adaptation to the background stimulus.

neuroscience↗

bioRxiv2018

Olfactory object recognition based on fine-scale stimulus timing in Drosophila

Odorants of behaviorally relevant objects (e.g., food sources) intermingle with those from other sources. Therefore, to sniff out whether an odor source is good or bad - without actually visiting it - animals first need to segregate the odorants from different sources. To do so, animals could use temporal cues, since odorants from one source exhibit correlated fluctuations, while odorants from different sources are less correlated. However, it remains unclear whether animals can rely solely on temporal cues for odor source segregation. Here we show that 1) flies can use a few milliseconds differences in odorant arrival to segregate a target odorant from a binary mixture, 2) segregation does not improve when the target odorant arrives first, and 3) segregation works for odorants with innate, as well as learned valences. These properties of odor segregation parallel those of concurrent sound segregation and figure-ground segregation by onset asynchrony in humans.

animal behavior and cognition↗