Calcium Dynamics In Astrocytes Research Articles

Aβ-mediated synaptic glutamate dynamics and calcium dynamics in astrocytes associated with Alzheimer’s disease

Aβ-mediated synaptic glutamate dynamics and calcium dynamics in astrocytes associated with Alzheimer’s disease

Basolateral Amygdala Astrocytes Are Engaged by the Acquisition and Expression of a Contextual Fear Memory.

Astrocytes are key cellular regulators within the brain. The basolateral amygdala (BLA) is implicated in fear memory processing, yet most research has entirely focused on neuronal mechanisms, despite a significant body of work implicating astrocytes in learning and memory. In the present study, we used in vivo fiber photometry in C57BL/6J male mice to record from amygdalar astrocytes across fear learning, recall, and three separate periods of extinction. We found that BLA astrocytes robustly responded to foot shock during acquisition, their activity remained remarkably elevated across days in comparison to unshocked control animals, and their increased activity persisted throughout extinction. Further, we found that astrocytes responded to the initiation and termination of freezing bouts during contextual fear conditioning and recall, and this behavior-locked pattern of activity did not persist throughout the extinction sessions. Importantly, astrocytes do not display these changes while exploring a novel context, suggesting that these observations are specific to the original fear-associated environment. Chemogenetic inhibition of fear ensembles in the BLA did not affect freezing behavior or astrocytic calcium dynamics. Overall, our work presents a real-time role for amygdalar astrocytes in fear processing and provides new insight into the emerging role of these cells in cognition and behavior.SIGNIFICANCE STATEMENT We show that basolateral amygdala astrocytes are robustly responsive to negative experiences, like shock, and display changed calcium activity patterns through fear learning and memory. Additionally, astrocytic calcium responses become time locked to the initiation and termination of freezing behavior during fear learning and recall. We find that astrocytes display calcium dynamics unique to a fear-conditioned context, and chemogenetic inhibition of BLA fear ensembles does not have an impact on freezing behavior or calcium dynamics. These findings show that astrocytes play a key real-time role in fear learning and memory.

Modeling the relationship between neuronal activity and the BOLD signal: contributions from astrocyte calcium dynamics

Functional magnetic resonance imaging relies on the coupling between neuronal and vascular activity, but the mechanisms behind this coupling are still under discussion. Recent experimental evidence suggests that calcium signaling may play a significant role in neurovascular coupling. However, it is still controversial where this calcium signal is located (in neurons or elsewhere), how it operates and how relevant is its role. In this paper we introduce a biologically plausible model of the neurovascular coupling and we show that calcium signaling in astrocytes can explain main aspects of the dynamics of the coupling. We find that calcium signaling can explain so-far unrelated features such as the linear and non-linear regimes, the negative vascular response (undershoot) and the emergence of a (calcium-driven) Hemodynamic Response Function. These features are reproduced here for the first time by a single model of the detailed neuronal-astrocyte-vascular pathway. Furthermore, we analyze how information is coded and transmitted from the neuronal to the vascular system and we predict that frequency modulation of astrocytic calcium dynamics plays a key role in this process. Finally, our work provides a framework to link neuronal activity to the BOLD signal, and vice-versa, where neuronal activity can be inferred from the BOLD signal. This opens new ways to link known alterations of astrocytic calcium signaling in neurodegenerative diseases (e.g. Alzheimer’s and Parkinson’s diseases) with detectable changes in the neurovascular coupling.

Analysis of Network Models with Neuron-Astrocyte Interactions

Neural networks, composed of many neurons and governed by complex interactions between them, are a widely accepted formalism for modeling and exploring global dynamics and emergent properties in brain systems. In the past decades, experimental evidence of computationally relevant neuron-astrocyte interactions, as well as the astrocytic modulation of global neural dynamics, have accumulated. These findings motivated advances in computational glioscience and inspired several models integrating mechanisms of neuron-astrocyte interactions into the standard neural network formalism. These models were developed to study, for example, synchronization, information transfer, synaptic plasticity, and hyperexcitability, as well as classification tasks and hardware implementations. We here focus on network models of at least two neurons interacting bidirectionally with at least two astrocytes that include explicitly modeled astrocytic calcium dynamics. In this study, we analyze the evolution of these models and the biophysical, biochemical, cellular, and network mechanisms used to construct them. Based on our analysis, we propose how to systematically describe and categorize interaction schemes between cells in neuron-astrocyte networks. We additionally study the models in view of the existing experimental data and present future perspectives. Our analysis is an important first step towards understanding astrocytic contribution to brain functions. However, more advances are needed to collect comprehensive data about astrocyte morphology and physiology in vivo and to better integrate them in data-driven computational models. Broadening the discussion about theoretical approaches and expanding the computational tools is necessary to better understand astrocytes’ roles in brain functions.

Rescue of astrocyte activity by the calcium sensor STIM1 restores long-term synaptic plasticity in female mice modelling Alzheimer’s disease

Calcium dynamics in astrocytes represent a fundamental signal that through gliotransmitter release regulates synaptic plasticity and behaviour. Here we present a longitudinal study in the PS2APP mouse model of Alzheimer’s disease (AD) linking astrocyte Ca2+ hypoactivity to memory loss. At the onset of plaque deposition, somatosensory cortical astrocytes of AD female mice exhibit a drastic reduction of Ca2+ signaling, closely associated with decreased endoplasmic reticulum Ca2+ concentration and reduced expression of the Ca2+ sensor STIM1. In parallel, astrocyte-dependent long-term synaptic plasticity declines in the somatosensory circuitry, anticipating specific tactile memory loss. Notably, we show that both astrocyte Ca2+ signaling and long-term synaptic plasticity are fully recovered by selective STIM1 overexpression in astrocytes. Our data unveil astrocyte Ca2+ hypoactivity in neocortical astrocytes as a functional hallmark of early AD stages and indicate astrocytic STIM1 as a target to rescue memory deficits.

Hippocampal astrocytes encode reward location.

Astrocytic calcium dynamics has been implicated in the encoding of sensory information1-5, and modulation of calcium in astrocytes has been shown to affect behaviour6-10. However, longitudinal investigation of the real-time calcium activity of astrocytes in the hippocampus of awake mice is lacking. Here we used two-photon microscopy to chronically image CA1 astrocytes as mice ran in familiar or new virtual environments to obtain water rewards. We found that astrocytes exhibit persistent ramping activity towards the reward location in a familiar environment, but not in a new one. Shifting the reward location within a familiar environment also resulted in diminished ramping. After additional training, as the mice became familiar with the new context or new reward location, the ramping was re-established. Using linear decoders, we could predict the location of the mouse in a familiar environment from astrocyte activity alone. We could not do the same in a new environment, suggesting that the spatial modulation of astrocytic activity is experience dependent. Our results indicate that astrocytes can encode the expected reward location in spatial contexts, thereby extending their known computational abilities and their role in cognitive functions.

Amyloid pathology disrupts gliotransmitter release in astrocytes.

Accumulation of amyloid-beta (Aβ) is associated with synaptic dysfunction and destabilization of astrocytic calcium homeostasis. A growing body of evidence support astrocytes as active modulators of synaptic transmission via calcium-mediated gliotransmission. However, the details of mechanisms linking Aβ signaling, astrocytic calcium dynamics, and gliotransmission are not known. We developed a biophysical model that describes calcium signaling and the ensuing gliotransmitter release from a single astrocytic process when stimulated by glutamate release from hippocampal neurons. The model accurately captures the temporal dynamics of microdomain calcium signaling and glutamate release via both kiss-and-run and full-fusion exocytosis. We investigate the roles of two crucial calcium regulating machineries affected by Aβ: plasma-membrane calcium pumps (PMCA) and metabotropic glutamate receptors (mGluRs). When we implemented these Aβ-affected molecular changes in our astrocyte model, it led to an increase in the rate and synchrony of calcium events. Our model also reproduces several previous findings of Aβ associated aberrant calcium activity, such as increased intracellular calcium level and increased spontaneous calcium activity, and synchronous calcium events. The study establishes a causal link between previous observations of hyperactive astrocytes in Alzheimer's disease (AD) and Aβ-induced modifications in mGluR and PMCA functions. Analogous to neurotransmitter release, gliotransmitter exocytosis closely tracks calcium changes in astrocyte processes, thereby guaranteeing tight control of synaptic signaling by astrocytes. However, the downstream effects of AD-related calcium changes in astrocytes on gliotransmitter release are not known. Our results show that enhanced rate of exocytosis resulting from modified calcium signaling in astrocytes leads to a rapid depletion of docked vesicles that disrupts the crucial temporal correspondence between a calcium event and vesicular release. We propose that the loss of temporal correspondence between calcium events and gliotransmission in astrocytes pathologically alters astrocytic modulation of synaptic transmission in the presence of Aβ accumulation.

Toolbox for studying neurovascular coupling in vivo, with a focus on vascular activity and calcium dynamics in astrocytes.

.Significance: Insights into the cellular activity of each member of the neurovascular unit (NVU) is critical for understanding their contributions to neurovascular coupling (NVC)—one of the key control mechanisms in cerebral blood flow regulation. Advances in imaging and genetic tools have enhanced our ability to observe, manipulate and understand the cellular activity of NVU components, namely neurons, astrocytes, microglia, endothelial cells, vascular smooth muscle cells, and pericytes. However, there are still many unresolved questions. Since astrocytes are considered electrically unexcitable, signaling is the main parameter used to monitor their activity. It is therefore imperative to study astrocytic dynamics simultaneously with vascular activity using tools appropriate for the question of interest.Aim: To highlight currently available genetic and imaging tools for studying the NVU—and thus NVC—with a focus on astrocyte dynamics and vascular activity, and discuss the utility, technical advantages, and limitations of these tools for elucidating NVC mechanisms.Approach: We draw attention to some outstanding questions regarding the mechanistic basis of NVC and emphasize the role of astrocytic elevations in functional hyperemia. We further discuss commonly used genetic, and optical imaging tools, as well as some newly developed imaging modalities for studying NVC at the cellular level, highlighting their advantages and limitations.Results: We provide an overview of the current state of NVC research, focusing on the role of astrocytic elevations in functional hyperemia; summarize recent advances in genetically engineered indicators, fluorescence microscopy techniques for studying NVC; and discuss the unmet challenges for future imaging development.Conclusions: Advances in imaging techniques together with improvements in genetic tools have significantly contributed to our understanding of NVC. Many pieces of the puzzle have been revealed, but many more remain to be discovered. Ultimately, optimizing NVC research will require a concerted effort to improve imaging techniques, available genetic tools, and analytical software.

Bayesian inference of molecular kinetic parameters from astrocyte calcium imaging data

Model-based Bayesian inference from high-content data obtained on live specimens is a burgeoning field with demonstrated applications to neuroscience. In parallel, computer vision methods for extracting the calcium signaling information from imaging data have advanced in application to neuronal physiology. Here, we are describing in detail a method we have recently developed to study calcium dynamics in astrocytes, which combines computer vision with model-based Bayesian learning to deduce the most likely molecular kinetic parameters underlying the observed calcium activity. As reported in the companion experimental study, this method allowed us to identify the key molecular changes downstream of a multi-gene deletion modeling the human 22q11.2 deletion syndrome, the most common human microdeletion and the genetic factor with the highest penetrance for schizophrenia.•Methodological details are laid out, from our imaging approach to our adaptation of the VBA-CaBBI algorithm previously developed primarily for brain functional imaging data.•The analytical pipeline is suited for further applications to glial cells and adaptable to other cell types exhibiting complexcalcium dynamics.

Regional heterogeneity of astrocyte morphogenesis dictated by the formin protein, Daam2, modifies circuit function.

Astrocytes display extraordinary morphological complexity that is essential to support brain circuit development and function. Formin proteins are key regulators of the cytoskeleton; however, their role in astrocyte morphogenesis across diverse brain regions and neural circuits is unknown. Here, we show that loss of the formin protein Daam2 in astrocytes increases morphological complexity in the cortex and olfactory bulb, but elicits opposing effects on astrocytic calcium dynamics. These differential physiological effects result in increased excitatory synaptic activity in the cortex and increased inhibitory synaptic activity in the olfactory bulb, leading to altered olfactory behaviors. Proteomic profiling and immunoprecipitation experiments identify Slc4a4 as a binding partner of Daam2 in the cortex, and combined deletion of Daam2 and Slc4a4 restores the morphological alterations seen in Daam2 mutants. Our results reveal new mechanisms regulating astrocyte morphology and show that congruent changes in astrocyte morphology can differentially influence circuit function.

Modifying the Models of Calcium Dynamics in Astrocytes by Ryanodine Release

The influence of ryanodine channels on the cytosole Ca2+ dynamics was studied. We added the equations for ryanodine receptors and voltage-gated calcium channels into the original De Pitta et al. model of Ca2+. The derived model was shown to have significantly wider range of predictions: we derived the frequency of cytosole calcium spontaneous oscillations (which are absent in the original De Pitta et al. model) for various existing models of Ca2+ signalling in astrocytes. Particularly, the initial De Pitta et al. results can be converted to either Lavrentovich and Hemkin model or in the Dupont et al model predictions. The absence of the Ca2+ oscillations in astrocytes with the active ryanodine channels only was recently reported. This behaviour can be achieved in our model predictions for the certain values of parameters, which are supposedly responsible for the bifurcation landscape between the oscillatory and non-oscillatory dynamics of cytosol Ca2+ in astrocytes. We also investigated the interplay between the spontaneous and glutamate-triggered oscillations.

Astrocytic Calcium Dynamics Along the Pain Pathway.

Astrocytes, once thought to be passive cells merely filling the space between neurons in the nervous system, are receiving attention as active modulators of the brain and spinal cord physiology by providing nutrients, maintaining homeostasis, and modulating synaptic transmission. Accumulating evidence indicates that astrocytes are critically involved in chronic pain regulation. Injury induces astrocytes to become reactive, and recent studies suggest that reactive astrocytes can have either neuroprotective or neurodegenerative effects. While the exact mechanisms underlying the transition from resting astrocytes to reactive astrocytes remain unknown, astrocytic calcium increase, coordinated by inflammatory molecules, has been suggested to trigger this transition. In this mini review article, we will discuss the roles of astrocytic calcium, channels contributing to calcium dynamics in astrocytes, astrocyte activations along the pain pathway, and possible relationships between astrocytic calcium dynamics and chronic pain.

Calcium Dynamics in Astrocytes During Cell Injury.

The changes in intracellular calcium concentration ([Ca2+]) following laser-induced cell injury in nearby cells were studied in primary mouse astrocytes selectively expressing the Ca2+ sensitive GFAP-Cre Salsa6f fluorescent tandem protein, in an Ast1 astrocyte cell line, and in primary mouse astrocytes loaded with Fluo4. Astrocytes in these three systems exhibit distinct changes in [Ca2+] following induced death of nearby cells. Changes in [Ca2+] appear to result from release of Ca2+ from intracellular organelles, as opposed to influx from the external medium. Salsa6f expressing astrocytes displayed dynamic Ca2+ changes throughout the phagocytic response, including lamellae protrusion, cytosolic signaling during vesicle formation, vesicle maturation, and vesicle tract formation. Our results demonstrate local changes in [Ca2+] are involved in the process of phagocytosis in astrocytes responding to cell corpses and/or debris.

Stimulation of water and calcium dynamics in astrocytes with pulsed infrared light.

Astrocytes are non-neuronal cells that govern the homeostatic regulation of the brain through ions and water transport, and Ca2+ -mediated signaling. As they are tightly integrated into neural networks, label-free tools that can modulate cell function are needed to evaluate the role of astrocytes in brain physiology and dysfunction. Using live-cell fluorescence imaging, pharmacology, electrophysiology, and genetic manipulation, we show that pulsed infrared light can modulate astrocyte function through changes in intracellular Ca2+ and water dynamics, providing unique mechanistic insight into the effect of pulsed infrared laser light on astroglial cells. Water transport is activated and, IP3 R, TRPA1, TRPV4, and Aquaporin-4 are all involved in shaping the dynamics of infrared pulse-evoked intracellular calcium signal. These results demonstrate that astrocyte function can be modulated with infrared light. We expect that targeted control over calcium dynamics and water transport will help to study the crucial role of astrocytes in edema, ischemia, glioma progression, stroke, and epilepsy.

Circuit modelling of 2-AG indirect pathway via astrocyte as a catalyst for synaptic self repair

Recent neurophysiological findings illustrate that the brain detects the faulty synapses and is able to repair them to some extent. It was shown that retrograde signaling via astrocytes leads to modulation of synaptic activity. Specifically, indirect signaling pathways of endocannabinoids modulate the synaptic transmission probability. Given these observations, this paper presents a novel analog circuit realization of the bio-inspired neuron-Astrocyte using analog elements which designed and simulated in LTSPICE simulator (Linear Technology SPICE simulator). This is based on the principle of retrograde signaling in the astrocyte-neuron network using astrocytic calcium dynamics. The simulation results of the analog circuit, demonstrate that 2-AG indirect signaling via astrocytes increases the probability of neurotransmitter release at the damaged synapses. This result indicates a promising approach toward a new class of adaptive, distributed computing systems in which self-repairing mechanism actively contributes to fault diagnosis and modification which also can be extended to neuronal networks.

Characterization of Correlated Calcium Dynamics in Astrocytes in PCL Scaffold: Application of Wavelet Transform Coherence

While 2D culture models have been used extensively to elucidate the cell-to-cell communication, they do not recapitulate fully the 3D characteristics of microenvironment in vivo, e.g., polarized cell attachment and generally confer a considerably stiffer substrate than the endogenous extracellular matrix. Development of fibrous scaffolds that can better mimic the native microenvironment and improve the spatial arrangement of seeded cells should foster experimental strategies to monitor and determine the 3D cell-to-cell communication. In this study, poly(e- caprolactone) (PCL) fibers were fabricated in different sizes using a microfluidic platform and spatially arranged to create a suitable 3D microenvironment in order to investigate the cell viability and calcium signaling in mouse astrocytes. A powerful algorithm, referred to as wavelet transform coherence (WTC), was applied to establish the correlation between astrocytes that were seeded on the PCL fiber. As expected, two astrocytes that appeared to be in physical contact showed high correlation, whereas two astrocytes seeded within a few cell lengths but not in physical contact showed negligible correlation. The WTC correlation analysis of a cluster of six astrocytes seeded on a single PCL fiber led to surprising results that the cells can communicate over many cell lengths without being in physical contact. More systematic studies using spatially controlled 3D microenvironment will likely help unravel the intricate cell communication mechanisms.

Papers of note in Science 356 (6339)

This week’s articles identify a repulsive guidance system that controls the recruitment of T cells to germinal centers; a mechanism for controlling the transfer of nutrients from the placenta to early mouse embryos; and the subcellular location of calcium dynamics in astrocytes.

Mathematical investigation of IP3-dependent calcium dynamics in astrocytes.

We study evoked calcium dynamics in astrocytes, a major cell type in the mammalian brain. Experimental evidence has shown that such dynamics are highly variable between different trials, cells, and cell subcompartments. Here we present a qualitative analysis of a recent mathematical model of astrocyte calcium responses. We show how the major response types are generated in the model as a result of the underlying bifurcation structure. By varying key channel parameters, mimicking blockers used by experimentalists, we manipulate this underlying bifurcation structure and predict how the distributions of responses can change. We find that store-operated calcium channels, plasma membrane bound channels with little activity during calcium transients, have a surprisingly strong effect, underscoring the importance of considering these channels in both experiments and mathematical settings. Variation in the maximum flow in different calcium channels is also shown to determine the range of stable oscillations, as well as set the range of frequencies of the oscillations. Further, by conducting a randomized search through the parameter space and recording the resulting calcium responses, we create a database that can be used by experimentalists to help estimate the underlying channel distribution of their cells.

Functional imaging of neuron-astrocyte interactions in a compartmentalized microfluidic device.

Traditional approaches in cultivating neural cells in a dish without orienting their interactions have had only limited success in revealing neural network properties. To enhance the experimental capabilities of studying neural circuitry in vitro, we designed an experimental system combining concepts of micropatterned surfaces, microfluidic devices and genetically encoded biosensors. Micropatterning was used to position neurons and astrocytes in defined locations and guide interactions between the two cell types. Microfluidic chambers were placed atop micropatterned surfaces to allow delivery of different pharmacological agents or viral vectors to the desired cell types. In this device, astrocytes and neurons communicated through grooves molded into the floor of the microfluidic device. By combining microfluidics with genetically encoded calcium indicators as functional readouts, we further demonstrated the utility of this device for analyzing neuron–neuron and neuron–astrocyte interactions in vitro under both healthy and pathophysiological conditions. We found that both spontaneous and evoked calcium dynamics in astrocytes can be modulated by interactions with neurons. In the future, we foresee employing the microdevices described here for studying mechanisms of neurological disorders.

Subtle modulation of ongoing calcium dynamics in astrocytic microdomains by sensory inputs

Astrocytes communicate with neurons through their processes. In vitro experiments have demonstrated that astrocytic processes exhibit calcium activity both spontaneously and in response to external stimuli; however, it has not been fully determined whether and how astrocytic subcellular domains respond to sensory input in vivo. We visualized the calcium signals in astrocytes in the primary visual cortex of awake, head-fixed mice. Bias-free analyses of two-photon imaging data revealed that calcium activity prevailed in astrocytic subcellular domains, was coordinated with variable spot-like patterns, and was dominantly spontaneous. Indeed, visual stimuli did not affect the frequency of calcium domain activity, but it increased the domain size, whereas tetrodotoxin reduced the sizes of spontaneous calcium domains and abolished their visual responses. The “evoked” domain activity exhibited no apparent orientation tuning and was distributed unevenly within the cell, constituting multiple active hotspots that were often also recruited in spontaneous activity. The hotspots existed dominantly in the somata and endfeet of astrocytes. Thus, the patterns of astrocytic calcium dynamics are intrinsically constrained and are subject to minor but significant modulation by sensory input.