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Bruzzone, M.

Publications and source records attributed to Bruzzone, M..

8 recordsLinked to original sources

Repellent olfactory cues influence the optomotor response modulation in Drosophila melanogaster

Animals need to precisely perceive and integrate the environmental cues to orient and select the appropriate motor responses required for navigating. This is the case, for instance, of the optokinetic reflex (OKR) and the optomotor response (OMR) in Drosophila melanogaster, where optic flow stimulation modulates the head or the body and legs motor activity respectively. Despite large bodies of literature on both the OKR and the OMR, there is still a limited understanding, in flies, of the impact on these responses of concomitant, and potentially conflicting, sensory inputs. To investigate this aspect, we used fruit flies walking on a sphere, presented with optic flow stimulation leading to the OMR together with the simultaneous exposure to olfactory stimulation, either using established repellent or masking compounds. We analysed the effect of different substances, and of their concentration, on the dynamics of the flies response to moving gratings, evaluating the fly walking path as well as average speed and duration. This analysis revealed several alterations between the compounds tested, in agreement with reported data on the simpler OKR. In conclusion, we show that concomitant exposure to repellents and maskers may consistently affect fundamental processes (the OKR and OMR) available to insects for informing themselves while navigating through the environment.

animal behavior and cognition↗

Motor history influences behavioral strategy more than response outcome in Drosophila melanogaster

In an ever-changing environment, animals continuously implement decision-making processes. This mechanism primarily relies on gathering sensory information through evidence accumulation, which enables the optimization of strategies in uncertain conditions. According to this scheme, the higher the evidence level, the greater the level of accuracy. However, it is not yet clear which, apart from direct sensory inputs, are the other factors that potentially impact strategy selection and their corresponding temporal operating windows in invertebrates. Moreover, it is not guaranteed that maximal accuracy could be the only parameter dictating the decision-making process. To address these questions, we employed a visual stimulation paradigm based on the random dot motion kinetogram (RDMk) with varying coherence levels, applied to tethered Drosophila melanogaster. In this scenario, flies tend to turn their heads according to the optic flows strength and direction. We observed stereotyped head saccadic-like movements across all the RDMk coherence levels, with greater coherence leading to a higher total saccade number and an increased proportion of correct turns. Importantly, logistic regression analysis revealed that, besides sensory inputs, the behavioral outcome and, consequently, the maximal accuracy, are influenced by other factors, in particular the past motor activity. In conclusion, our data support a scenario where accuracy and reaction time are balanced to reach an optimal response condition.

animal behavior and cognition↗

Optokinetic response in D. Melanogaster reveals the nature of common repellent odorants.

Animals ability to orient and navigate relies on selecting an appropriate motor response based on the perception and integration of the environmental information. This is the case, for instance, of the optokinetic response (OKR) in Drosophila melanogaster, where optic flow visual stimulation modulates the walking or flying patterns. Despite a large body of literature on the OKR, theres still a limited understanding of the impact on OKR of concomitant, potentially conflicting, inputs. To evaluate the impact of this multimodal integration, we combined in D. melanogaster, while flying in a tethered condition, the optic flow stimulation leading to OKR with the simultaneous presentation of olfactory cues, based on repellent or masking compounds typically used against noxious insect species. First, this approach al lowed us to directly quantify the effect of several substances and their concentration, on the dynamics of the flies OKR in response to moving gratings by evaluating the number of saccades and the velocity of its slow phase. Subsequently, this analysis was capable of easily revealing the actual effect, i.e. masking vs repellent, of the compound tested. In conclusion, we show that D. melanogaster, a cost-affordable species, represents a viable option for studying the effects of several compounds on the navigational abilities of insects.

animal behavior and cognition↗

Simultaneous detection of membrane contact dynamics and associated Ca2+ signals by reversible chemogenetic reporters

Membrane contact sites (MCSs) are hubs allowing various cell organelles to coordinate their activities. The dynamic nature of these sites and their small size hinder analysis by current imaging techniques. To overcome these limitations, we here designed a series of reversible chemogenetic reporters incorporating improved, low-affinity variants of splitFAST, and studied the dynamics of different MCSs at high spatiotemporal resolution, both in vitro and in vivo. We demonstrated that these versatile reporters suit different experimental setups well, allowing one to address challenging biological questions. Using these novel probes, we identified a hitherto unknown pathway, in which calcium (Ca2+) signalling dynamically regulates endoplasmic reticulum-mitochondria juxtaposition, characterizing the underlying mechanism. Finally, by integrating Ca2+-sensing capabilities into the splitFAST technology, we introduced PRINCESS (PRobe for INterorganelle Ca2+-Exchange Sites based on SplitFAST), an unprecedented class of reporters to simultaneously detect MCSs and measure the associated Ca2+ dynamics using a single biosensor.

cell biology↗

A network-based method for extracting the organization of brain-wide circuits from reconstructed connectome datasets

Functional brain activity is supported by specific circuit wiring diagrams. When the synaptic annotation is available, the analysis of high-resolution connectomes allows for unraveling the circuit architectures. Despite the continuous technological and computational improvements, obtaining a whole-brain bauplan based on synaptic annotation remains a demanding effort. As an alternative, we present here an approach to extract an approximated brain connectome starting from libraries of single cell anatomical reconstructions aligned on the same anatomical reference brain. Our approach relies on the identification of neurite terminal nodes based on Strahler numbering of the cell morphology, and the adoption of a proximity range criterion, so as to infer, in the absence of synapse information, approximated brain network architectures. As an initial benchmark we used information theory metrics to confront our approach against a synaptically-annotated EM dataset of Drosophila melanogaster hemibrain. The comparison with our approach revealed a general agreement in the organization of the extracted connectivity modules measured in terms of Normalized Mutual Information, Adjusted Rand Index and Pearson correlation. Moreover, we show that the modules identified, along with their organization, can capture known circuit motives. We then applied this approach to a light microscopy dataset of the zebrafish larval brain composed of about 3000 neuronal skeletonizations. We show that the approximated connectome and the resulting modular organization is capable of capturing specific and topographically organized connection patterns as well as known functional circuit architectures. In conclusion, we present a scalable, from-circuit-to-brain range, approach to reveal approximated neuronal architectures supporting brain mechanisms, potentially suitable for hypothesis generation and for guiding the exploration of and integration with EM connectomes. Author SummaryUnderstanding how the brain works requires detailed maps of its neural connections, known as connectomes. While advanced techniques like electron microscopy (EM) can map these connections at the level of individual synapses, they are time consuming and resource intensive, limiting their use. As an alternative, we developed a computational method that approximates brain connectivity using existing datasets of neuron shapes (morphologies) without requiring synaptic annotations. Our approach identifies potential connections between neurons based on the proximity of their terminal branches regions where synapses are likely to form within a shared 3D reference brain. We validated our method using a high-resolution EM connectome of the fruit fly brain, demonstrating that it captures broad organizational patterns, such as clusters of densely interconnected neurons, despite moderate agreement at the synaptic level. Applying the method to a light microscopy dataset of the zebrafish larva brain ([~]3,000 neurons), we successfully reconstructed large-scale networks that recapitulated known functional circuits. This approach offers a scalable way to extract brain-wide connectivity principles from existing datasets, bridging the gap between cellular anatomy and circuit function. It can guide targeted experiments and complement future EM studies, making neuroscience data more accessible for hypothesis generation.

neuroscience↗

Generation of Bessel beam lattices by a single metasurface for neuronal activity recording in zebrafish larva

Bessel Beams (BBs) and BB lattices are structured-light excitation profiles frequently applied in material processing, nonlinear spectroscopy and in many fluorescence microscopy methods such as Light Sheet Microscopy (LSM). In LSM, BBs and BB-lattices offer wider excitation profiles, higher acquisition rate, enhanced resolution, and improved signal-to-noise ratio, while reducing the overall phototoxicity. However, this performance improvement typically comes at the cost of layout complexity and spatial constraints, originating from the optical arrangement required for obtaining BB features and for multiplexing the BB in a lattice of beamlets. Here, we introduce a novel method for encoding in a single flat element all the optical operations required to generate a BB lattice, including those of the excitation objective. We assessed the effective capabilities of this approach, using Meta-Surface (MS) technology to fabricate the corresponding flat optical element and to characterize its optical figures. Finally, we demonstrated its actual application in LSM, recording neuronal activity at cellular resolution in the zebrafish larval brain using fluorescence based neuronal activity reporters. In perspective, this approach, applied here for LSM, prompts a step forward in the BB versatility and in the BB application scenarios.

bioengineering↗

Use of Therapeutic Drug Monitoring to Characterize Cefepime-Induced Neurotoxicity

BackgroundThe incidence of cefepime-induced neurotoxicity (CIN) in hospitalized patients is highly variable. Although greater cefepime exposures incite neurotoxicity, data evaluating trough thresholds associated with CIN remains limited. The objectives of this study were to evaluate the incidence of CIN, assess the relationship between cefepime trough concentrations and CIN, investigate clinical factors associated with CIN, and describe electroencephalogram (EEG) abnormalities in CIN. MethodsThis was a retrospective study of adult patients who had received [&ge;] 5 days of cefepime with [&ge;] 1 trough concentration > 25 mg/L. Potential CIN cases were identified utilizing neurological symptoms, neurologist assessments, EEG findings and improvement of neurotoxicity after cefepime discontinuation. ResultsOne-hundred and forty-two patients were included. The incidence of CIN was 13% (18/142). The mean cefepime trough concentration in CIN patients was significantly greater than the non-neurotoxicity group (74.2 mg/L {+/-} 41.1 vs. 46.6 mg/L {+/-} 23, p=0.015). Lower renal function (creatinine clearance < 30 ml/min), greater time to therapeutic drug monitoring (TDM) ([&ge;]72 hours), and each 1 mg/mL rise in cefepime trough were independently associated with increased risk of CIN. Moderate generalized slowing of the background rhythm was the most common EEG pattern associated with CIN. ConclusionCefepime should be used cautiously in hospitalized patients due to the risk of neurotoxicity. Patients with greater renal function and those who had early cefepime TDM ([&le;] 72 hours) had lower risk of CIN.

pharmacology and toxicology↗

EEG power spectra and subcortical pathology in chronic disorders of consciousness

ObjectiveTo determine (i) the association between long-term impairment of consciousness after severe brain injury, spontaneous brain oscillations, and underlying subcortical damage, and (ii) whether such data can be used to aid patient diagnosis, a process known to be susceptible to high error rates. MethodsCross-sectional observational sample of 116 patients with a disorder of consciousness secondary to brain injury, collected prospectively at a tertiary center between 2011 and 2013. Multimodal analyses relating clinical measures of impairment, electroencephalographic measures of spontaneous brain activity, and magnetic resonance imaging data of subcortical atrophy were conducted in 2018. ResultsIn the final analyzed sample of 61 patients, systematic associations were found between electroencephalographic power spectra and subcortical damage. Specifically, the ratio of beta-to-delta relative power was negatively associated with greater atrophy in regions of the bilateral thalamus and globus pallidus (both left > right) previously shown to be preferentially atrophied in chronic disorders of consciousness. Power spectrum total density was also negatively associated with widespread atrophy in regions of the left globus pallidus, right caudate, and in brainstem. Furthermore, we showed that the combination of demographics, encephalographic, and imaging data in an analytic framework can be employed to aid behavioral diagnosis. ConclusionsThese results ground, for the first time, electroencephalographic presentation detected with routine clinical techniques in the underlying brain pathology of disorders of consciousness and demonstrate how multimodal combination of clinical, electroencephalographic, and imaging data can be employed in potentially mitigating the high rates of misdiagnosis typical of this patient cohort. Search termsdisorders of consciousness, subcortical pathology, EEG, MRI.

neuroscience↗