Vesicular HMGB1 release from neurons stressed with spreading depolarization enables confined inflammatory signaling to astrocytes
Abstract The role of high mobility group box 1 (HMGB1) in inflammation is well characterized in the immune system and in response to tissue injury. More recently, HMGB1 was also shown to initiate an “inflammatory signaling cascade” in the brain parenchyma after a mild and brief disturbance, such as...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2023-12-01
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| Series: | Journal of Neuroinflammation |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12974-023-02977-6 |
| _version_ | 1797388161179975680 |
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| author | Zeynep Kaya Nevin Belder Melike Sever-Bahcekapili Buket Donmez-Demir Şefik Evren Erdener Naz Bozbeyoglu Canan Bagci Emine Eren-Kocak Muge Yemisci Hulya Karatas Esra Erdemli Ihsan Gursel Turgay Dalkara |
| author_facet | Zeynep Kaya Nevin Belder Melike Sever-Bahcekapili Buket Donmez-Demir Şefik Evren Erdener Naz Bozbeyoglu Canan Bagci Emine Eren-Kocak Muge Yemisci Hulya Karatas Esra Erdemli Ihsan Gursel Turgay Dalkara |
| author_sort | Zeynep Kaya |
| collection | DOAJ |
| description | Abstract The role of high mobility group box 1 (HMGB1) in inflammation is well characterized in the immune system and in response to tissue injury. More recently, HMGB1 was also shown to initiate an “inflammatory signaling cascade” in the brain parenchyma after a mild and brief disturbance, such as cortical spreading depolarization (CSD), leading to headache. Despite substantial evidence implying a role for inflammatory signaling in prevalent neuropsychiatric disorders such as migraine and depression, how HMGB1 is released from healthy neurons and how inflammatory signaling is initiated in the absence of apparent cell injury are not well characterized. We triggered a single cortical spreading depolarization by optogenetic stimulation or pinprick in naïve Swiss albino or transgenic Thy1-ChR2-YFP and hGFAP-GFP adult mice. We evaluated HMGB1 release in brain tissue sections prepared from these mice by immunofluorescent labeling and immunoelectron microscopy. EzColocalization and Costes thresholding algorithms were used to assess the colocalization of small extracellular vesicles (sEVs) carrying HMGB1 with astrocyte or microglia processes. sEVs were also isolated from the brain after CSD, and neuron-derived sEVs were captured by CD171 (L1CAM). sEVs were characterized with flow cytometry, scanning electron microscopy, nanoparticle tracking analysis, and Western blotting. We found that HMGB1 is released mainly within sEVs from the soma of stressed neurons, which are taken up by surrounding astrocyte processes. This creates conditions for selective communication between neurons and astrocytes bypassing microglia, as evidenced by activation of the proinflammatory transcription factor NF-ĸB p65 in astrocytes but not in microglia. Transmission immunoelectron microscopy data illustrated that HMGB1 was incorporated into sEVs through endosomal mechanisms. In conclusion, proinflammatory mediators released within sEVs can induce cell-specific inflammatory signaling in the brain without activating transmembrane receptors on other cells and causing overt inflammation. |
| first_indexed | 2024-03-08T22:36:50Z |
| format | Article |
| id | doaj.art-d89093d484494452821fd6d02092bc64 |
| institution | Directory Open Access Journal |
| issn | 1742-2094 |
| language | English |
| last_indexed | 2024-03-08T22:36:50Z |
| publishDate | 2023-12-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Neuroinflammation |
| spelling | doaj.art-d89093d484494452821fd6d02092bc642023-12-17T12:25:20ZengBMCJournal of Neuroinflammation1742-20942023-12-0120111610.1186/s12974-023-02977-6Vesicular HMGB1 release from neurons stressed with spreading depolarization enables confined inflammatory signaling to astrocytesZeynep Kaya0Nevin Belder1Melike Sever-Bahcekapili2Buket Donmez-Demir3Şefik Evren Erdener4Naz Bozbeyoglu5Canan Bagci6Emine Eren-Kocak7Muge Yemisci8Hulya Karatas9Esra Erdemli10Ihsan Gursel11Turgay Dalkara12Institute of Neurological Sciences and Psychiatry, Hacettepe UniversityInstitute of Neurological Sciences and Psychiatry, Hacettepe UniversityInstitute of Neurological Sciences and Psychiatry, Hacettepe UniversityInstitute of Neurological Sciences and Psychiatry, Hacettepe UniversityInstitute of Neurological Sciences and Psychiatry, Hacettepe UniversityDepartment of Molecular Biology and Genetics, Science Faculty, Bilkent UniversityInstitute of Neurological Sciences and Psychiatry, Hacettepe UniversityInstitute of Neurological Sciences and Psychiatry, Hacettepe UniversityInstitute of Neurological Sciences and Psychiatry, Hacettepe UniversityInstitute of Neurological Sciences and Psychiatry, Hacettepe UniversityDepartment of Histology and Embryology, Faculty of Medicine, Ankara UniversityDepartment of Molecular Biology and Genetics, Science Faculty, Bilkent UniversityInstitute of Neurological Sciences and Psychiatry, Hacettepe UniversityAbstract The role of high mobility group box 1 (HMGB1) in inflammation is well characterized in the immune system and in response to tissue injury. More recently, HMGB1 was also shown to initiate an “inflammatory signaling cascade” in the brain parenchyma after a mild and brief disturbance, such as cortical spreading depolarization (CSD), leading to headache. Despite substantial evidence implying a role for inflammatory signaling in prevalent neuropsychiatric disorders such as migraine and depression, how HMGB1 is released from healthy neurons and how inflammatory signaling is initiated in the absence of apparent cell injury are not well characterized. We triggered a single cortical spreading depolarization by optogenetic stimulation or pinprick in naïve Swiss albino or transgenic Thy1-ChR2-YFP and hGFAP-GFP adult mice. We evaluated HMGB1 release in brain tissue sections prepared from these mice by immunofluorescent labeling and immunoelectron microscopy. EzColocalization and Costes thresholding algorithms were used to assess the colocalization of small extracellular vesicles (sEVs) carrying HMGB1 with astrocyte or microglia processes. sEVs were also isolated from the brain after CSD, and neuron-derived sEVs were captured by CD171 (L1CAM). sEVs were characterized with flow cytometry, scanning electron microscopy, nanoparticle tracking analysis, and Western blotting. We found that HMGB1 is released mainly within sEVs from the soma of stressed neurons, which are taken up by surrounding astrocyte processes. This creates conditions for selective communication between neurons and astrocytes bypassing microglia, as evidenced by activation of the proinflammatory transcription factor NF-ĸB p65 in astrocytes but not in microglia. Transmission immunoelectron microscopy data illustrated that HMGB1 was incorporated into sEVs through endosomal mechanisms. In conclusion, proinflammatory mediators released within sEVs can induce cell-specific inflammatory signaling in the brain without activating transmembrane receptors on other cells and causing overt inflammation.https://doi.org/10.1186/s12974-023-02977-6Spreading depolarizationNeuronal extracellular vesiclesHMGB1Inflammation |
| spellingShingle | Zeynep Kaya Nevin Belder Melike Sever-Bahcekapili Buket Donmez-Demir Şefik Evren Erdener Naz Bozbeyoglu Canan Bagci Emine Eren-Kocak Muge Yemisci Hulya Karatas Esra Erdemli Ihsan Gursel Turgay Dalkara Vesicular HMGB1 release from neurons stressed with spreading depolarization enables confined inflammatory signaling to astrocytes Journal of Neuroinflammation Spreading depolarization Neuronal extracellular vesicles HMGB1 Inflammation |
| title | Vesicular HMGB1 release from neurons stressed with spreading depolarization enables confined inflammatory signaling to astrocytes |
| title_full | Vesicular HMGB1 release from neurons stressed with spreading depolarization enables confined inflammatory signaling to astrocytes |
| title_fullStr | Vesicular HMGB1 release from neurons stressed with spreading depolarization enables confined inflammatory signaling to astrocytes |
| title_full_unstemmed | Vesicular HMGB1 release from neurons stressed with spreading depolarization enables confined inflammatory signaling to astrocytes |
| title_short | Vesicular HMGB1 release from neurons stressed with spreading depolarization enables confined inflammatory signaling to astrocytes |
| title_sort | vesicular hmgb1 release from neurons stressed with spreading depolarization enables confined inflammatory signaling to astrocytes |
| topic | Spreading depolarization Neuronal extracellular vesicles HMGB1 Inflammation |
| url | https://doi.org/10.1186/s12974-023-02977-6 |
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