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Stefan, M. I.

Publications and source records attributed to Stefan, M. I..

7 recordsLinked to original sources

Resource competition controls the length-dependence of synaptic protein synthesis

Synaptic plasticity is dependent on the availability of specific proteins at the synapse with distinct requirements for potentiation and depression. Specifically, experimental evidence suggests an upregulation of short proteins and a downregulation for long proteins during long-term depression (LTD). The mechanism of this length-dependent protein synthesis regulation is yet unknown. In this work, we propose an computational model of ribosomal regulation of protein synthesis than can reproduce the length-dependent shift in protein expression seen during ribosomal elevations. By analysing the model dynamics, we discover that this effect is due to a shift from initiation-restricted to elongation-restricted protein synthesis. This works offers a novel explanation for the loss of ribosomal control of protein synthesis due to saturation in pathological ribosome enriched conditions. This new theoretical model of ribosomal regulation of protein synthesis is capable of explaining length-dependent shifts in protein synthesis.

neuroscience↗

One model to rule them all: unification of voltage-gated potassium channel models via deep non-linear mixed effects modelling

Ion channels are essential for signal processing and propagation in neural cells. Voltage-gated ion channels permeable to potassium (Kv) form one of the most prominent channel families. Techniques used to model the voltage-dependent gating of Kv channels date back to Hodgkin and Huxley (1952). Different Kv types can display radically different kinetic properties, requiring different mathematical models. However, the construction of Hodgkin-Huxley-like (HH-like) models is generally complex and time consuming due to the number of parameters, their tuning and having to choose functional forms to model gating. In addition to the between-Kv type heterogeneity, there can be significant within-Kv type kinetic heterogeneity between different cells with genetically identical channels. Since HH-like models do not account for such variability, extensions to it are necessary. We use scientific machine learning (SciML), the integration of machine learning methodologies with existing scientific models, and non-linear mixed effects (NLME) modelling to bypass the limitations of HH-like modelling. NLME is a modelling methodology that takes into account both within- and between-subject variability. These tools allowed us to complement the HH-like modelling and construct a unified SciML HH-like model that fits the recordings from 20 different Kv types. The unified SciML HH-like model produced closer fits to the data compared to a set of seven previous HH-like models and was able to represent the highly heterogeneous data from different cells. Our model may be the first step in producing a SciML foundation model for ion channels that would be capable of modelling the gating kinetics of any ion channel type. Author summaryIon channels are complex molecules embedded in the membranes of neurons - the cells responsible for signal propagation and processing in the brain. Ion channels can open and close in response to various types of stimuli, in particular the voltage difference across the cell membrane. Computational modelling, usage of mathematical techniques to represent a system and algorithmically solve for its dynamics, has been previously used to understand the dynamics of voltage-gated ion channels. However, computational modelling of voltage-gated ion channels requires costly and complex optimization routines to optimize their structure and parameters. We utilize two tools new to the modelling of voltage-gated ion channels - scientific machine learning and non-linear mixed effects modelling - to bypass some limitations associated with the existing methods. By using scientific machine learning and non-linear mixed effects modelling we were able to create a unified model capable of modelling the gating dynamics of 20 different ion channels. This is in stark contrast to the existing modelling approaches, where each channel requires its own model. Moreover, our unified model performed better than seven existing ion channel gating models. Therefore, the tools we used and the model we created is a significant step forward in facilitating the modelling of ion channel gating. Future work could include even more ion channels types within the scope of our unified model.

neuroscience↗

Fitting and comparison of calcium-calmodulin kinetic schemes to a common data set using non-linear mixed effects modelling

Calmodulin is a calcium binding protein that is essential in calcium signalling in the brain. There are many computational models of calcium-calmodulin binding that capture various calmodulin features. However, existing models have generally been fit to different data sets, with some publications not reporting their training and validation performance. Moreover, there is no model comparison using a common benchmark data set as is common practice in other modeling domains. Finally, some calmodulin models have been fit as a part of a larger kinetic scheme, which may have resulted in parameters being underdetermined. We address these three limitations of previous models by fitting the published calcium-calmodulin schemes to a common calcium-calmodulin data set comprising of equlibrium data from Shifman et al. and dynamical data from Faas et al. Due to technical limitations, the amount of uncaged calcium in Faas et al. data could not be predicted with certainty. To find good parameter fits, despite this uncertainty, we used non-linear mixed effects modelling as implemented in the Pumas.jl package. The Akaike information criterion values for our reaction rates were significantly lower than for the published parameters, indicating that the published parameters are suboptimal. Moreover, there were significant differences in calmodulin activation, both between the schemes and between our reaction rate and those previously published. A kinetic scheme with independent lobes and unique, rather than identical, binding sites fit the data best. Our results support two hypotheses: (1) partially bound calmodulin is important in cellular signalling; (2) calcium binding sites within a calmodulin lobe are kinetically distinct rather than identical. We conclude that more attention should be given to validation and comparison of models of individual molecules. Author summaryLearning and memory depend on changes in synapses, the connections between nerve cells. To understand how learning and memory work, it is important to understand how the proteins involved in these changes are regulated. Computational modelling of biochemical reactions has been used to understand the regulation processes involved in learning and memory. However, computational models often rely on simplifications of biochemical reaction schemes, and it can be difficult to tell which simplifications are "the best", i.e. capture important aspects of a proteins behaviour. It is also difficult to estimate the best model parameters, such as binding reaction rates or binding strengths. Modellers often base their estimates on known experimental results, but often not in a structured way. We examine and compare a variety of computational models of calmodulin, a protein necessary for changes in synaptic strength. We apply a new approach to infer the model parameters that best fit two published datasets, and compare our parameters to previously published ones. We find that some simplified reaction schemes are more successful than others in capturing the activation pattern of calmodulin. We also find that the parameter sets found through our approach outperform previously published parameter sets in fitting the experimental data.

systems biology↗

In Silico identification and modelling of FDA-approved drugs targeting T-type calcium channels

Studies have shown that inhibition of the Cav3.1 T-type calcium channel can prevent or suppress neurological diseases, such as epileptic seizures and diabetic neuropathy. In this study we aimed to use in silico simulations to identify a U.S. Food and Drug Administration (FDA)-approved drug that can bind to the Cav3.1 T-type calcium channel. We used the automated docking suite GOLD v5.5 with the genetic algorithm to simulate molecular docking and predict the protein-ligand binding modes, and the ChemPLP empirical scoring function to estimate the binding affinities of 2,115 FDA-approved drugs to the human Cav3.1 channel. Drugs with high binding affinity and appropriate pharmacodynamic and pharmacokinetic properties were selected for molecular mechanics Poisson-Boltzmann surface area (MMPBSA) and molecular mechanics generalised Born surface area (MMGBSA) binding free energy calculations, GROMACS molecular dynamics (MD) simulations and Monte Carlo Cell (MCell) simulations. The docking results indicated that the FDA-approved drug montelukast has a high binding affinity to Cav3.1, and data from the literature suggested that montelukast has the appropriate drug-like properties to cross the human blood-brain barrier and reach synapses in the central nervous system. MMPBSA, MMGBSA and MD simulations showed the high stability of the montelukast-Cav3.1 complex. MCell simulations indicated that the blockage of Cav3.1 by montelukast reduced the number of synaptic vesicles being released from the pre-synaptic region to the synaptic cleft, which may reduce the probability and amplitude of postsynaptic potentials. Author SummaryNew drugs for current illnesses or disorders are always necessary to enhance therapeutic outcomes or can be utilised for patients who have experienced side effects from current medicines. The Cav3.1 T-type calcium channel has been found to be linked to neurological diseases, such as epilepsy and diabetes. Consequently, identifying a drug that could bind to these receptors may yield therapeutic effects on these neurological conditions. This study utilises various computational methods to sift through 2,115 FDA-approved drugs that have been used for other diseases, not neurological ones, to determine whether some of these drugs could bind to the Cav3.1 T-type calcium channel and potentially produce therapeutic effects on neurological diseases. Our research indicates that the current asthma drug montelukast has the potential to bind to the calcium channel and is worthy of further investigation.

bioinformatics↗

The trade-off between temporal precision and effect amplitude of inhibitory plasticity regulation determines separability of learned representations

Synaptic plasticity, the process by which synapses change in an activity-dependent manner, is assumed to be the basis of learning. Experiments demonstrated that synaptic plasticity not only depends on excitatory activity but also on the rate and timing of inhibitory events. Hypothesising that the regulatory effect of inhibition is mediated by membrane potential hyperpolarisation, we identify fast fluctuation sensitivity as a contributing factor to the inhibitory regulation of plasticity. Fast fluctuation sensitivity characterises the influence of fast changes in the membrane potential on the plasticity model predictions, including (short) hyperpolarising events. Furthermore, by introducing a novel plasticity model, the Voltage-Dependent Pathway model, we show that fast fluctuation sensitivity enables a precise temporal regulation of plasticity via inhibition tightly locked to stimulus presentation. In recurrent networks, receptive fields develop independent of the degree of fluctuation sensitivity, yet the temporal precision of inhibitory regulation is critical for the heterogeneity and quality of the resulting representation. Author SummarySynaptic plasticity plays a central role in learning and enables us to interact within an ever-changing environment flexibly. The term governs all processes which leads to a change in connection strength between two neurons. Previously, studies of synaptic plasticity strongly focused on the role of excitatory activity. However, experiments excluding GABA blockers implicate inhibitory neurons to regulate synaptic plasticity. We further investigate the role of inhibitory neurons as regulators of synaptic plasticity in a model study. Specifically, we focus on the decrease in membrane potential due to inhibitory activity as a key actor in plasticity regulation. This comparative modelling study considers two well-established models and a novel model which differ in the order in which filtering and thresholding is applied to the membrane potential. We find that while all models develop connectivity structures resembling receptive fields characteristics of the visual cortex, the quality and heterogeneity of the developed feature detectors is model-specific. Heterogeneity and representation specificity is promoted by a high degree of fast fluctuation sensitivity of the model enabling temporally precise inhibitory regulation of plasticity determined by the models filter and threshold.

neuroscience↗

A multi-state model of the CaMKII dodecamer suggests a role for calmodulin in maintenance of autophosphorylation

Ca2+/calmodulin-dependent protein kinase II (CaMKII) accounts for up to 2 percent of all brain protein and is essential to memory function. CaMKII activity is known to regulate dynamic shifts in the size and signaling strength of neuronal connections, a process known as synaptic plasticity. Increasingly, computational models are used to explore synaptic plasticity and the mechanisms regulating CaMKII activity. Conventional modeling approaches may exclude biophysical detail due to the impractical number of state combinations that arise when explicitly monitoring the conformational changes, ligand binding, and phosphorylation events that occur on each of the CaMKII holoenzymes twelve subunits. To manage the combinatorial explosion without necessitating bias or loss in biological accuracy, we use a specialized syntax in the software MCell to create a rule-based model of the twelve-subunit CaMKII holoenzyme. Here we validate the rule-based model against previous measures of CaMKII activity and investigate molecular mechanisms of CaMKII regulation. Specifically, we explore how Ca2+/CaM-binding may both stabilize CaMKII subunit activation and regulate maintenance of CaMKII autophosphorylation. Noting that Ca2+/CaM and protein phosphatases bind CaMKII at nearby or overlapping sites, we compare model scenarios in which Ca2+/CaM and protein phosphatase do or do not structurally exclude each others binding to CaMKII. Our results suggest a functional mechanism for the so-called \"CaM trapping\" phenomenon, such that Ca2+/CaM structurally excludes phosphatase binding and thereby prolongs CaMKII autophosphorylation. We conclude that structural protection of autophosphorylated CaMKII by Ca2+/CaM may be an important mechanism for regulation of synaptic plasticity.\n\nAuthor summaryIn the hippocampus, the dynamic fluctuation in size and strength of neuronal connections is thought to underlie learning and memory processes. These fluctuations, called synaptic plasticity, are in-part regulated by the protein calcium/calmodulin-dependent kinase II (CaMKII). During synaptic plasticity, CaMKII becomes activated in the presence of calcium ions (Ca2+) and calmodulin (CaM), allowing it to interact enzymatically with downstream binding partners. Interestingly, activated CaMKII can phosphorylate itself, resulting in state changes that allow CaMKII to be functionally active independent of Ca2+/CaM. Phosphorylation of CaMKII at Thr-286/287 has been shown to be a critical component of learning and memory. To explore the molecular mechanisms that regulate the activity of CaMKII holoenzymes, we use a rule-based approach that reduces computational complexity normally associated with representing the wide variety of functional states that a CaMKII holoenzyme can adopt. Using this approach we observe regulatory mechanisms that might be obscured by reductive approaches. Our results newly suggest that CaMKII phosphorylation at Thr-286/287 is stabilized by a mechanism in which CaM structurally excludes phosphatase binding at that site.

neuroscience↗

A multi-scale model reveals cellular and physiological mechanisms underlying hyperpolarisation-gated synaptic plasticity

Neurons in the medial vestibular nucleus (MVN) display hyperpolarisation-gated synaptic plasticity, where inhibition believed to come from cerebellar cortical Purkinje cells can induce long-term potentiation (LTP) or long-term depression (LTD) of vestibular nerve afferent synapses. This phenomenon is thought to underlie the plasticity of the vestibulo-ocular reflex (VOR). The molecular and cellular mechanisms involved are largely unknown. Here we present a novel multi-scale computational model, which captures both electrophysiological and biochemical signalling at vestibular nerve synapses on proximal dendrites of the MVN neuron. We show that AMPA receptor phosphorylation at the vestibular synapse depends in complex ways on dendritic calcium influx, which is in turn shaped by patterns of post-synaptic hyperpolarisation and vestibular nerve stimulation. Hyperpolarisation-gated synaptic plasticity critically depends on the activation of LVA calcium channels and on the interplay between CaMKII and PP2B in dendrites of the post-synaptic MVN cell. The extent and direction of synaptic plasticity depend on the strength and duration of hyperpolarisation, and on the relative timing of hyperpolarisation and vestibular nerve stimulation. The multi-scale model thus enables us to explore in detail the interactions between electrophysiological activation and post-synaptic biochemical reaction systems. More generally, this model has the potential to address a wide range of questions about neural signal integration, post-synaptic biochemical reaction systems and plasticity.

neuroscience↗