Protocol for Co-culture of Microglia with Axons

Fujita, Yuki; Yamashita, Toshio

doi:10.1016/j.xpro.2020.100111

Cited by 3 publications

(1 citation statement)

References 11 publications

(12 reference statements)

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“…The technology consisted in a disposable, pre-assembled platform, suitable for human stem-cell-derived neurons providing better cell attachment and long-term growth. The research group further published a protocol describing the procedure for modeling the accumulation of microglia toward neuronal axons, using the abovementioned axon isolation culture device [ 210 ].…”

Section: Microfluidics Technologies For Microglia–neuron Interactimentioning

confidence: 99%

Epigenetics and Communication Mechanisms in Microglia Activation with a View on Technological Approaches

et al. 2021

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Microglial cells, the immune cells of the central nervous system (CNS), play a crucial role for the proper brain development and function and in CNS homeostasis. While in physiological conditions, microglia continuously check the state of brain parenchyma, in pathological conditions, microglia can show different activated phenotypes: In the early phases, microglia acquire the M2 phenotype, increasing phagocytosis and releasing neurotrophic and neuroprotective factors. In advanced phases, they acquire the M1 phenotype, becoming neurotoxic and contributing to neurodegeneration. Underlying this phenotypic change, there is a switch in the expression of specific microglial genes, in turn modulated by epigenetic changes, such as DNA methylation, histones post-translational modifications and activity of miRNAs. New roles are attributed to microglial cells, including specific communication with neurons, both through direct cell–cell contact and by release of many different molecules, either directly or indirectly, through extracellular vesicles. In this review, recent findings on the bidirectional interaction between neurons and microglia, in both physiological and pathological conditions, are highlighted, with a focus on the complex field of microglia immunomodulation through epigenetic mechanisms and/or released factors. In addition, advanced technologies used to study these mechanisms, such as microfluidic, 3D culture and in vivo imaging, are presented.

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Section: Microfluidics Technologies For Microglia–neuron Interactimentioning

confidence: 99%

Epigenetics and Communication Mechanisms in Microglia Activation with a View on Technological Approaches

et al. 2021

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Analgecine regulates microglia polarization in ischemic stroke by inhibiting NF-κB through the TLR4 MyD88 pathway

Gong

Chen

Wang

et al. 2021

International Immunopharmacology

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Spatial transcriptomics and neurofilament light chain reveal changes in lesion patterns in murine autoimmune neuroinflammation

Brummer,

Schillner,

Steffen

et al. 2023

J Neuroinflammation

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Objective Ongoing neuroaxonal damage is a major contributor to disease progression and long-term disability in multiple sclerosis. However, spatio-temporal distribution and pathophysiological mechanisms of neuroaxonal damage during acute relapses and later chronic disease stages remain poorly understood. Methods Here, we applied immunohistochemistry, single-molecule array, spatial transcriptomics, and microglia/axon co-cultures to gain insight into spatio-temporal neuroaxonal damage in experimental autoimmune encephalomyelitis (EAE). Results Association of spinal cord white matter lesions and blood-based neurofilament light (sNfL) levels revealed a distinct, stage-dependent anatomical pattern of neuroaxonal damage: in chronic EAE, sNfL levels were predominately associated with anterolateral lumbar lesions, whereas in early EAE sNfL showed no correlation with lesions in any anatomical location. Furthermore, neuroaxonal damage in late EAE was largely confined to white matter lesions but showed a widespread distribution in early EAE. Following this pattern of neuroaxonal damage, spatial transcriptomics revealed a widespread cyto- and chemokine response at early disease stages, whereas late EAE was characterized by a prominent glial cell accumulation in white matter lesions. These findings were corroborated by immunohistochemistry and microglia/axon co-cultures, which further revealed a strong association between CNS myeloid cell activation and neuroaxonal damage both in vivo and in vitro. Interpretation Our findings indicate that CNS myeloid cells may play a crucial role in driving neuroaxonal damage in EAE. Moreover, neuroaxonal damage can progress in a stage-dependent centripetal manner, transitioning from normal-appearing white matter to focal white matter lesions. These insights may contribute to a better understanding of neurodegeneration and elevated sNfL levels observed in multiple sclerosis patients at different disease stages. Graphical Abstract

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Protocol for Co-culture of Microglia with Axons

Cited by 3 publications

References 11 publications

Epigenetics and Communication Mechanisms in Microglia Activation with a View on Technological Approaches

Epigenetics and Communication Mechanisms in Microglia Activation with a View on Technological Approaches

Analgecine regulates microglia polarization in ischemic stroke by inhibiting NF-κB through the TLR4 MyD88 pathway

Spatial transcriptomics and neurofilament light chain reveal changes in lesion patterns in murine autoimmune neuroinflammation

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