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Kuhn, B.

Publications and source records attributed to Kuhn, B..

14 recordsLinked to original sources

How Bias Shapes the Leaderboard: Scoring Function Performance Under Scrutiny

Structure-based scoring functions leveraging machine learning have recently demonstrated superior performance over classical scoring functions, particularly on virtual screening benchmarks. However, due to the fundamental differences between their underlying model principles and architectures, it remains unclear to what extent performance stems from an understanding of molecular binding or from exploitation of systemic biases. Thus, disentangling the factors underlying benchmark performance is essential for determining whether a scoring function will generalize to novel chemical space and succeed in prospective drug discovery. To address this need, we present a case study investigating the nature and impact of systemic biases on benchmark comparisons between different scoring function paradigms. By systematically analyzing the evaluation workflows of prominent models, we reveal pocket bias, a form of spatial coordinate frame leakage arising from static binding pocket extraction, which artificially inflates benchmark performance. To progressively eliminate these sources of bias, we benchmarked two selected graph neural network scoring functions against two minmalist machine learning models and a classical scoring function under four increasingly stringent evaluation levels, successively removing pocket bias, reducing structural data leakage, and finally evaluating on out-of-distribution (OOD) protein targets. Upon removal of pocket bias and structural data leakage, the performance of all machine learning models dropped substantially. When evaluated on out-of-distribution protein families, the classical baseline AutoDock Vina outperformed the machine learning models in five of seven virtual screening tasks and dominated the docking power evaluation. Our findings indicate that benchmark performance can be heavily shaped by evaluation design and dataset artifacts, potentially overshadowing algorithmic improvements. While the tested machine learning models remain heavily dependent on encountering familiar data distributions to achieve competitive results, AutoDock Vina demonstrated superior generalization capacity on OOD targets. This work underscores the critical need for rigorous, artifact-free benchmarking protocols to guide the development of truly prospective machine learning models for virtual screening.

bioinformatics↗

Precision in Motion: Reactive and Anticipatory Control of Mouse Tongue Movement for Interception

Object interception requires integrating prediction and sensory feedback, making it a powerful model for studying sensorimotor transformations. However, traditional head-fixed mouse paradigms reduce behavior to stereotyped stimulus-response associations and lack key features of natural interception. To address this limitation, we developed a behavioral task in which head-fixed mice use their tongue to intercept a food pellet moving at one of seven constant speeds randomly selected on each trial. We characterized the 3D kinematics of discrete tongue-reaching movements, their distinct motor phases, and learning-dependent changes in movement timing and kinematics. Mice adapted both tongue kinematics and movement onset to pellet speed, initially relying on reactive control and later adopting an anticipatory strategy. Anticipatory control depended largely on vision and intact lateral cerebellar circuits. This paradigm provides a powerful platform for investigating the neural mechanisms underlying predictive sensorimotor control and object interception in head-fixed mice. Significance StatementIn natural environments, interception requires seamless sensory-motor transformation, yet traditional paradigms like classical conditioning and virtual reality fall short of capturing this complex behavior. We introduce a behavioral paradigm in which trained mice intercept food pellets moving at constant speeds that vary randomly across trials, with discrete tongue licks. Remarkably, mice exhibit a shift from a reactive to a more anticipatory motor strategy, adaptively tuning lick initiation and projection speed in a learning-dependent manner. This work establishes a simplified interception-like framework for studying sensorimotor dynamics in head-fixed mice.

neuroscience↗

Persistent cerebellar molecular layer interneuron activity facilitates anticipatory control of tongue movements

Precise, anticipatory movements depend on the brains ability to generate motor commands in advance of immediate sensory input. Although the cerebellar cortex receives abundant sensory input via the mossy-fiber pathway, the mechanisms by which continuous sensory signals are transformed or gated remain poorly understood. Here, we show that molecular layer interneurons acquire persistent sensory-related Ca2+ activity in mice that develop anticipatory behavior. Employing a targeted lick-interception task, we found that interneurons in lobule Crus I, but much less in Crus II, exhibit enhanced Ca2+ activity up to five seconds before movement onset, locked to the spatial position of a continuously moving target, independent of body movement. In more anticipatory mice, movement onset becomes linked to the dynamics of this persistent activity, whereas less anticipatory mice adjust their movements based on immediate sensory-related input. Our results indicate that anticipation in sensorimotor tasks is supported by dynamic sensory representations encoded by interneurons.

neuroscience↗

SARM1 base-exchange inhibitors induce SARM1 activation and neurodegeneration at low doses

SARM1 has emerged as a promising therapeutic target in neurology due to its central role in axonal degeneration and its amenability to different modes of small molecule inhibition. One chemical approach to modulate SARM1 involves orthosteric inhibition via a SARM1-mediated base-exchange reaction between a small molecule and nicotinamide adenine dinucleotide (NAD+), the substrate of SARM1, to generate the active inhibitor. Here, we report that subinhibitory concentrations of SARM1 base-exchange inhibitors (BEIs) paradoxically increase SARM1 activity and worsen SARM1-induced cell death and neuronal damage in vitro. Low dose administration of RO-7529, a SARM1 BEI, exacerbated experimental autoimmune encephalomyelitis (EAE)-induced neurodegeneration in vivo. Our data highlight a unique pharmacological feature of SARM1 BEIs that may limit their therapeutic application in disorders associated with SARM1 activation and axonal degeneration.

neuroscience↗

Long-access heroin self-administration induces region specific reduction of grey matter volume and microglia reactivity in the rat.

In opioid use disorder (OUD) patients, a decrease in brain grey matter volume (GMV) has been reported. It is unclear whether this is the consequence of prolonged exposure to opioids or is a predisposing causal factor in OUD development. To investigate this, we conducted a structural MRI longitudinal study in NIH Heterogeneous Stock rats exposed to heroin self-administration and age-matched naive controls housed in the same controlled environment. Structural MRI scans were acquired before (MRI1) and after (MRI2) a prolonged period of long access heroin self-administration resulting in escalation of drug intake. Heroin intake resulted in reduced GMV in various cortical and sub-cortical brain regions. In drug-naive controls no difference was found between MRI1 and MRI2. Notably, the degree of GMV reduction in the medial prefrontal cortex (mPFC) and the insula positively correlated with the amount of heroin consumed and the escalation of heroin use. In a preliminary gene expression analysis, we identified a number of transcripts linked to immune response and neuroinflammation. This prompted us to hypothesize a link between changes in microglia homeostasis and loss of GMV. For this reason, we analyzed the number and morphology of microglial cells in the mPFC and insula. The number of neurons and their morphology was also evaluated. The primary motor cortex, where no GMV change was observed, was used as negative control. We found no differences in the number of neurons and microglia cells following heroin. However, in the same regions where reduced GMV was detected, we observed a shift towards a rounder shape and size reduction in microglia, suggestive of their homeostatic change towards a reactive state. Altogether these findings suggest that escalation of heroin intake correlates with loss of GMV in specific brain regions and that this phenomenon is linked to changes in microglial morphology.

neuroscience↗

A screen of alkaline and oxidative formulations for their inactivation efficacy of metal surface adsorbed prions using a steel-bead seed amplification assay

Iatrogenic transmission of prions, the infectious agents of fatal Creutzfeldt-Jakob disease, through inefficiently decontaminated medical instruments remains a critical issue. Harsh chemical treatments are effective, but not suited for routine reprocessing of reusable surgical instruments in medical cleaning and disinfection processes due to material incompatibilities. The identification of mild detergents with activity against prions is therefore of high interest but laborious due to the low throughput of traditional assays measuring prion infectivity. Here, we report the development of TESSA (sTainlESs steel-bead Seed Amplification assay), a prion seed amplification assay that explores the propagation activity of prions with stainless steel beads. TESSA was applied for the screening of about 70 different commercially available and novel formulations and conditions for their prion inactivation efficacy. One hypochlorite-based formulation, two commercially available alkaline formulations and a manual alkaline pre-cleaner were found to be highly effective in inactivating prions under conditions simulating automated washer-disinfector cleaning processes. The efficacy of these formulations was confirmed in vivo in a murine prion infectivity bioassay, yielding a reduction of the prion titer for the bead surface adsorbed prions below detectability. Our data suggest that TESSA represents an effective method for a rapid screening of prion-inactivating detergents, and that alkaline and oxidative formulations are promising in reducing the risk of potential iatrogenic prion transmission through insufficiently decontaminated instrument surfaces.

neuroscience↗

TOB is an effector of the hippocampus-mediated acute stress response

Stress affects behavior and involves critical dynamic changes at multiple levels ranging from molecular pathways to neural circuits and behavior. Abnormalities at any of these levels lead to decreased stress resilience and pathological behavior. However, temporal modulation of molecular pathways underlying stress response remains poorly understood. Transducer of ErbB2.1, known as TOB, (TOB1) is involved in different physiological functions, including cellular stress and immediate response to stimulation. In this study, we investigated the role of TOB in the brains stress machinery at molecular, neural circuit, and behavioral levels. Interestingly, TOB protein levels increased after mice were exposed to acute stress. At the neural circuit level, functional magnetic resonance imaging (fMRI) suggested that intra-hippocampal and hippocampal-prefrontal connectivity were dysregulated in Tob knockout (Tob-KO) mice. Electrophysiological recordings in hippocampal slices showed increased postsynaptic AMPAR-mediated neurotransmission, accompanied by decreased GABA neurotransmission and subsequently altered Excitatory/Inhibitory balance after Tob deletion. At the behavioral level, Tob-KO mice show abnormal, hippocampus-dependent, contextual fear conditioning and extinction, and depression-like behaviors. On the other hand, increased anxiety observed in Tob-KO mice is hippocampus-independent. At the molecular level, we observed decreased stress-induced LCN2 expression and ERK phosphorylation, as well as increased MKP-1 expression. This study suggests that TOB serves as an important modulator in hippocampal stress signaling machinery. In summary, we show a molecular pathway and neural circuit mechanism by which TOB deletion contributes to expression of pathological stress-related behavior.

neuroscience↗

A potent and selective inhibitor for the modulation of MAGL activity in the neurovasculature

Chronic inflammation and blood-brain barrier dysfunction are key pathological hallmarks of neurological disorders such as multiple sclerosis, Alzheimers disease and Parkinsons disease. Major drivers of these pathologies include pro-inflammatory stimuli such as prostaglandins, which are produced in the central nervous system by the oxidation of arachidonic acid in a reaction catalyzed by the cyclooxygenases COX1 and COX2. Monoacylglycerol lipase hydrolyzes the endocannabinoid signaling lipid 2-arachidonyl glycerol, enhancing local pools of arachidonic acid in the brain and leading to cyclooxygenase-mediated prostaglandin production and neuroinflammation. Monoacylglycerol lipase inhibitors were recently shown to act as effective anti-inflammatory modulators, increasing 2-arachidonyl glycerol levels while reducing levels of arachidonic acid and prostaglandins, including PGE2 and PGD2. In this study, we characterized a novel, highly selective, potent and reversible monoacylglycerol lipase inhibitor (MAGLi 432) in a mouse model of lipopolysaccharide-induced blood-brain barrier permeability and in both human and mouse cells of the neurovascular unit: brain microvascular endothelial cells, pericytes and astrocytes. We confirmed the expression of monoacylglycerol lipase in specific neurovascular unit cells in vitro, with pericytes showing the highest expression level and activity. However, MAGLi 432 did not ameliorate lipopolysaccharide-induced blood-brain barrier permeability in vivo or reduce the production of pro-inflammatory cytokines in the brain. Our data confirm monoacylglycerol lipase expression in mouse and human cells of the neurovascular unit and provide the basis for further cell-specific analysis of MAGLi 432 in the context of blood-brain barrier dysfunction caused by inflammatory insults.

neuroscience↗

Software for Non-Parametric Image Registration of 2-Photon Imaging Data

Functional 2-photon microscopy is a key technology for imaging neuronal activity. The recorded image sequences, however, can contain non-rigid movement artifacts which requires high-accuracy movement correction. Variational optical flow (OF) estimation is a group of methods for motion analysis with established performance in many computer vision areas. However, it has yet to be adapted to the statistics of 2-photon neuroimaging data. In this work, we present the motion compensation method Flow-Registration that outperforms previous alignment tools and allows to align and reconstruct even low signal-to-noise ratio 2-photon imaging data and is able to compensate high-divergence displacements during local drug injections. The method is based on statistics of such data and integrates previous advances in variational OF estimation. Our method is available as an easy-to-use ImageJ / FIJI plugin as well as a MATLAB toolbox with modular, object oriented file IO, native multi-channel support and compatibility with existing 2-photon imaging suites.

neuroscience↗

Fast Variational Alignment of non-flat 1D Displacements for Applications in Neuroimaging

BackgroundIn the context of signal analysis and pattern matching, alignment of 1D signals for the comparison of signal morphologies is an important problem. For image processing and computer vision, 2D optical flow (OF) methods find wide application for motion analysis and image registration and variational OF methods have been continuously improved over the past decades. New MethodWe propose a variational method for the alignment and displacement estimation of 1D signals. We pose the estimation of non-flat displacements as an optimization problem with a similarity and smoothness term similar to variational OF estimation. To this end, we can make use of efficient optimization strategies that allow real-time applications on consumer grade hardware. ResultsWe apply our method to two applications from functional neuroimaging: The alignment of 2-photon imaging line scan recordings and the denoising of evoked and event-related potentials in single trial matrices. We can report state of the art results in terms of alignment quality and computing speeds. Existing MethodsExisting methods for 1D alignment target mostly constant displacements, do not allow native subsample precision or precise control over regularization or are slower than the proposed method. ConclusionsOur method is implemented as a MATLAB toolbox and is online available. It is suitable for 1D alignment problems, where high accuracy and high speed is needed and non-constant displacements occur.

neuroscience↗

Dendritic coincidence detection in Purkinje neurons of awake mice

Dendritic coincidence detection is thought fundamental to neuronal processing, yet the underlying dendritic voltage-calcium relationship remains unexplored in awake animals. Here, using simultaneous voltage and calcium two-photon imaging of Purkinje neuron spiny dendrites, we show how coincident sub- and suprathreshold synaptic inputs modulate dendritic calcium signaling during sensory stimulation in awake mice. Sensory stimulation evokes subthreshold excitatory and inhibitory post-synaptic potentials, that coincide with suprathreshold dendritic spikes triggered by climbing fiber and parallel fiber synaptic input. Purkinje neuron dendrites integrate these inputs in a time-dependent and non-linear fashion to enhance the sensory evoked dendritic calcium signal. Intrinsic supra-linear dendritic mechanisms, including voltage gated calcium channels and metabotropic glutamate receptors, are recruited cooperatively to expand the dynamic range of sensory evoked dendritic calcium signals. This establishes how dendrites use multiple interplaying mechanisms to perform coincidence detection, as a fundamental and ongoing feature of dendritic integration during behavior.

neuroscience↗

Control of cytokinesis by β-adrenergic receptors indicates an approach for regulating cardiomyocyte endowment

ABSTRACT/SUMMARYOne million patients with congenital heart disease (CHD) live in the US. They have a lifelong risk of developing heart failure. Current concepts do not sufficiently address mechanisms of heart failure development specifically for these patients. We show that cardiomyocyte cytokinesis failure is increased in tetralogy of Fallot with pulmonary stenosis (ToF/PS), a common form of CHD. Labeling of a ToF/PS baby with isotope-tagged thymidine showed cytokinesis failure after birth. We used single-cell transcriptional profiling to discover that the underlying mechanism is repression of the cytokinesis gene ECT2, and show that this is downstream of {beta}-adrenergic receptors ({beta}-AR). Inactivation of the {beta}-AR genes and administration of the {beta}-blocker propranolol increased cardiomyocyte division in neonatal mice, which increased the endowment and conferred benefit after myocardial infarction in adults. Propranolol enabled the division of ToF/PS cardiomyocytes. These results suggest that {beta}-blockers should be evaluated for increasing cardiomyocyte division in patients with ToF/PS and other types of CHD.

cell biology↗

The Paraventricular Thalamus is a Critical Mediator of Top-down Control of Cue-motivated Behavior

Cues in the environment can elicit complex emotional states, and thereby maladaptive behavior, as a function of their ascribed value. Here we capture individual variation in the propensity to attribute motivational value to reward-cues using the sign-tracker/goal-tracker animal model. Goal-trackers attribute predictive value to reward-cues, and sign-trackers attribute both predictive and incentive value. Using chemogenetics and microdialysis, we show that, in sign-trackers, stimulation of the neuronal pathway from the prelimbic cortex (PrL) to the paraventricular nucleus of the thalamus (PVT) decreases the incentive value of a reward-cue. In contrast, in goal-trackers, inhibition of the PrL-PVT pathway increases both the incentive value and dopamine levels in the nucleus accumbens shell. The PrL-PVT pathway, therefore, exerts top-down control over the dopamine-dependent process of incentive salience attribution. These results highlight PrL-PVT pathway as a potential target for treating psychopathologies associated with the attribution of excessive incentive value to reward-cues, including addiction.

neuroscience↗

Affective bonding explains post-traumatic behavioral development in adult mice

In human post-traumatic stress disorder (PTSD), a major psychiatry challenge is how diverse stress reactions emerge after a protracted symptom-free period. Here, we study the behavioral development in mice isolated after observing an aggressive encounter inflicted upon their pair-housed partners and compared the results with those in multiple control paradigms. Compared with mice plainly isolated, mice isolated following the acute witnessing social stress gradually developed a wide range of long-term differences of their physiological conditions, spontaneous behaviors, and social interactions, including paradoxical results if interpreted in traditional ways. To address this developmental diversity, we applied fine-scale behavioral analysis to standard behavioral tests and showed that the seemingly sudden emergent behavioral differences developed gradually. Mice showed different developmental patterns in different zones of a behavior testing apparatus. However, the results of the fine-scale analysis together with state-space behavioral characterization allow a consistent interpretation of the seemingly conflicting observations among multiple tests. Interestingly, these behavioral differences were not observed if the aggressive encounter happened to a stranger mouse. Additionally, traumatized mice showed rebound responses to their partners after the long separation. In contrast, mice pair-housed with their attacked partners after the aggressive encounters still showed a difference in social interactions, while a difference in spontaneous behaviors did not occur. Accordingly, we propose that social relationship is the single common factor underlying the otherwise independent development of behavioral differences in this mouse paradigm and that the gained insights could have parallels in human PTSD development.

neuroscience↗