Activity changes in neuron-astrocyte networks in culture under the effect of norepinephrine

El, Yasmin Bar; Kanner, Sivan; Barzilai, Ari; Hanein, Yael

doi:10.1371/journal.pone.0203761

Cited by 20 publications

(27 citation statements)

References 88 publications

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“…Here we report that NE suppresses ammonium‐induced [Ca 2+ ] i oscillations in neurons and evokes Ca 2+ response in astrocytes. The same findings were reported in the work (Bar El, Kanner, Barzilai, & Hanein, ). The authors showed that NE induced Ca 2+ response in astrocytes and suppressed the activity of neurons in cortical mixed neuroglial cell cultures.…”

Section: Discussionsupporting

confidence: 91%

Activation of alpha‐2 adrenergic receptors stimulates GABA release by astrocytes

Gaidin

Зинченко

Сергеев

et al. 2019

Glia

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Norepinephrine is one of the key neurotransmitters in the hippocampus, but its role in the functioning of the neuroglial networks remains unclear. Here we show that norepinephrine suppresses NH4Cl‐induced oscillations of the intracellular Ca2+ concentration ([Ca2+]i) in hippocampal neurons. We found that the inhibitory effect of norepinephrine against ammonium‐induced [Ca2+]i oscillations is mediated by activation of alpha‐2 adrenergic receptors. Furthermore, UK 14,304, an agonist of alpha‐2 adrenergic receptors, evokes a biphasic [Ca2+]i elevation in a minor population of astrocytes. This elevation consists of an initial fast, peak‐shaped [Ca2+]i rise, mediated by Giβγ subunit and subsequent PLC‐induced mobilization of Ca2+ from internal stores, and a plateau phase, mediated by a Ca2+ influx from the extracellular medium through store‐operated and TRPC3 channels. We show the correlation between the Ca2+ response in astrocytes and suppression of [Ca2+]i oscillations in neurons. The inhibitory effect of UK 14,304 is abolished in the presence of gallein, an inhibitor of Gβγ‐signaling. In turn, application of the agonist in the presence of the PLC inhibitor decreases the frequency and amplitude of [Ca2+]i oscillations in neurons but does not suppress them. The same effect is observed in the presence of bicuculline, a GABA(A) receptor antagonist. We demonstrate that UK 14,304 application increases the frequency and amplitude of slow outward chloride currents in neurons, indicating the release of GABA by astrocytes. Thus, our findings indicate that the activation of astrocytic alpha‐2 adrenergic receptors stimulates GABA release from astrocytes via Giβγ subunit‐associated signaling pathway, contributing to the suppression of neuronal activity.

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Section: Discussionsupporting

confidence: 91%

Activation of alpha‐2 adrenergic receptors stimulates GABA release by astrocytes

Gaidin

Зинченко

Сергеев

et al. 2019

Glia

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“…We also found that astrocyte maturation was time dependent and marker dependent, as shown by the profile of maturation of “immature” hiPSC‐A (i.e., hiPSC‐A cultured for 60DIV prior to use) in coculture, which was less marked than 90DIV hiPSC‐A, with, however, the exception of two markers, Cx43 and AQP4. These early changes in AQP4 and Cx43 profiles suggest a neuronal influence on their expression not related to astrocyte maturation, and parallels previous literature on rodent astrocytes showing elevations in calcium intercellular signaling and changes in astrocyte volume in response to neuronal activity.…”

Section: Discussionsupporting

confidence: 87%

Role of Human-Induced Pluripotent Stem Cell-Derived Spinal Cord Astrocytes in the Functional Maturation of Motor Neurons in a Multielectrode Array System

Taga

Dastgheyb

Habela

et al. 2019

Stem Cells Translational Medicine

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The ability to generate human-induced pluripotent stem cell (hiPSC)-derived neural cells displaying region-specific phenotypes is of particular interest for modeling central nervous system biology in vitro. We describe a unique method by which spinal cord hiPSC-derived astrocytes (hiPSC-A) are cultured with spinal cord hiPSC-derived motor neurons (hiPSC-MN) in a multielectrode array (MEA) system to record electrophysiological activity over time. We show that hiPSC-A enhance hiPSC-MN electrophysiological maturation in a time-dependent fashion. The sequence of plating, density, and age in which hiPSC-A are cocultured with MN, but not their respective hiPSC line origin, are factors that influence neuronal electrophysiology. When compared to coculture with mouse primary spinal cord astrocytes, we observe an earlier and more robust electrophysiological maturation in the fully human cultures, suggesting that the human origin is relevant to the recapitulation of astrocyte/motor neuron crosstalk. Finally, we test pharmacological compounds on our MEA platform and observe changes in electrophysiological activity, which confirm hiPSC-MN maturation. These findings are supported by immunocytochemistry and real-time PCR studies in parallel cultures demonstrating human astrocyte mediated changes in the structural maturation and protein expression profiles of the neurons. Interestingly, this relationship is reciprocal and coculture with neurons influences astrocyte maturation as well. Taken together, these data indicate that in a human in vitro spinal cord culture system, astrocytes support hiPSC-MN maturation in a time-dependent and species-specific manner and suggest a closer approximation of in vivo conditions. STEM CELLS TRANSLATIONAL MEDICINE 2019;8:1272-1285 SIGNIFICANCE STATEMENTThis study develops a method by which human-induced pluripotent stem cell-derived astrocytes (hiPSC-A) with distinct spinal cord identity are cocultured with spinal cord motor neurons (hiPSC-MN) for multielectrode array (MEA) recordings. It also demonstrates that hiPSC-A influence the morphological, molecular, and electrophysiological maturation of hiPSC-MN. Similarly, this study shows that hiPSC-A maturation is enhanced by the coculture with hiPSC-MN. This fully human, spinal cord-specific, coculture platform with MEA analyses provides a new tool for investigating astrocyte/MN interactions and has the potential to more accurately model human diseases with spinal cord pathology, including spinal muscular atrophy and amyotrophic lateral sclerosis.

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“…In this respect, and beyond our preliminary observations based on RNA levels, previous literature on rodent astrocytes had already shown that Cx43 and AQP4 function is modulated by neuron-astrocyte interactions, as suggested by elevations in calcium intercellular signaling 46,47 and changes in astrocyte volume 48,49 , respectively, in response to neuronal activity.…”

Section: Discussionsupporting

confidence: 50%

Role of human induced pluripotent stem cell-derived spinal cord astrocytes in the functional maturation of motor neurons in a multielectrode array system

Taga

Dastgheyb

Habela

et al. 2019

Preprint

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The ability to generate human induced pluripotent stem cell (hiPSC)-derived neural cells displaying region-specific phenotypes is of particular interest for modeling central nervous system (CNS) biology in vitro. We describe a unique method by which spinal cord hiPSC-derived astrocytes (hiPSC-A) are cultured with spinal cord hiPSC-derived motor neurons (hiPSC-MN) in a multielectrode array (MEA) system to record electrophysiological activity over time. We show that hiPSC-A enhance hiPSC-MN electrophysiological maturation in a time-dependent fashion.The sequence of plating, density, and age in which hiPSC-As are co-cultured with MN, but not their respective hiPSC line origin, are factors that influence neuronal electrophysiology. When compared to co-culture with mouse primary spinal cord astrocytes, we observe an earlier and more robust electrophysiological maturation in the fully human cultures, suggesting that the human origin is relevant to the recapitulation of astrocyte/motor neuron cross-talk. Finally, we test pharmacological compounds on our MEA platform and observe changes in electrophysiological activity which confirm hiPSC-MN maturation. These findings are supported by immunocytochemistry and real time PCR studies in parallel cultures demonstrating human astrocyte mediated changes in the structural maturation and protein expression profiles of the neurons. Interestingly, this relationship is reciprocal and co-culture with neurons influences astrocyte maturation as well. Taken together these data indicate that in a human in vitro spinal cord culture system, astrocytes alter hiPSC-MN maturation in a time-dependent and species specific manner and suggest a closer approximation of in vivo conditions. Main Points1. We developed a method for the co-culture of human iPSC-A/MN for multielectrode array recordings.2. The morphological, molecular, pharmacological, and electrophysiological characterization of the co-cultures suggests bidirectional maturation.

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Activity changes in neuron-astrocyte networks in culture under the effect of norepinephrine

Cited by 20 publications

References 88 publications

Activation of alpha‐2 adrenergic receptors stimulates GABA release by astrocytes

Activation of alpha‐2 adrenergic receptors stimulates GABA release by astrocytes

Role of Human-Induced Pluripotent Stem Cell-Derived Spinal Cord Astrocytes in the Functional Maturation of Motor Neurons in a Multielectrode Array System

Role of human induced pluripotent stem cell-derived spinal cord astrocytes in the functional maturation of motor neurons in a multielectrode array system

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