bims-microg 2025-09-28 papers

bims-microg

Biomed News

on Microglia in health and disease

Issue of 2025–09–28
twenty-six papers selected by
Marcus Karlstetter, Universität zu Köln

Cholesterol fuels microglia in chronic stroke.
Cholesterol metabolic reprogramming mediates microglia-induced chronic neuroinflammation and hinders neurorestoration following stroke.
Copper overload worsens the inflammatory response of microglia to amyloid beta (Aβ) by impairing phagocytosis and promoting mitochondrial DNA-mediated NLRP3 inflammasome activation.
Microglia/macrophage-specific deletion of TLR-4 protects against neural effects of diet-induced obesity in a sexually dimorphic manner.
Macrophage expression of P2X7 controls autoimmune uveitis.
Macrophage/microglia-dependent mechanisms drive retinal pigment epithelium regeneration in zebrafish.
Hypoxia reverses the early anti-inflammatory microglial response to the β-amyloid peptide: mitochondrial involvement and beneficial roles of melatonin and naringenin.
Loss of DJ-1 alleviates microglia-mediated neuroinflammation in Parkinson's disease via autophagy-lysosomal degradation of NLRP3.
Inhibition of the HMGB1-RAGE Axis Attenuates Microglial Inflammation and Ameliorates Hypoxia-Induced Cognitive Impairment.
Characterizing Microglia Morphology in the Frontal Cortex of Pair-Bonded and Unpaired Prairie Voles (Microtus ochrogaster).
Astrocyte-microglia crosstalk in the hippocampus mediates cognitive impairments induced by chronic intermittent hypoxia.
P2RX7 modulates the function of human microglia-like cells and mediates the association of IL18 with Alzheimer's disease traits.
The alternative complement pathway drives neuroinflammation and neurodegeneration in mouse models of glaucoma and optic nerve injury.
Evaluating the Optic Nerve Crush Model to Understand the Function of Microglia in Glaucoma Neuroprotection.
The role of microglia and complement C5/C5a in the pathogenesis of rhegmatogenous retinal detachment with choroidal detachment.
Microglia reactivity is brain region and sex specific in the context of chronic stress.
Investigating the role of the ALPK1 signaling pathway in the pathogenesis of diabetic retinopathy.
GCN5L1 Aggravates Postherpetic Neuralgia Through Regulating Microglial Mitochondrial Fission-Fusion Homeostasis.
Enhanced M2 Polarization of Retinal Microglia in Streptozotocin-Induced Diabetic Mice upon Autoimmune Stimulation.
Axonal degeneration and retinal neurovascular dysfunction in the TgF344-AD rat model of Alzheimer's disease.
Growth hormone reduces retinal inflammation and preserves microglial morphology after optic nerve crush in male rats.
Microglia Activation and Migration in Retina during Acute High-altitude Exposure.
Diet gel-based oral drug delivery system for controlled dosing of small molecules for microglia depletion and inducible Cre recombination in mice.
Bibliometric Mapping of Research Trends and Hotspots of Microglia in Spinal Cord Injury (2000-2024).
Protocol for pooled FACS-based CRISPR knockout screening in human iPSC-derived microglia.
Petunidin attenuates lipopolysaccharide-induced retinal microglia inflammatory response in diabetic retinopathy by targeting OGT/NF-κB/LCN2 axis.

Nat Metab. 2025 Sep 23.

Cholesterol fuels microglia in chronic stroke.

Stefano Pluchino, Cory M Willis.

DOI: https://doi.org/10.1038/s42255-025-01362-2
Nat Metab. 2025 Sep 23.

Cholesterol metabolic reprogramming mediates microglia-induced chronic neuroinflammation and hinders neurorestoration following stroke.

Qiang Zhao, Jiajian Li, Jingjing Feng, Xin Wang, Yueting Liu, Fei Wang, Liang Liu, Bingxue Jin, Ming Lin, Ya-Chao Wang, Xiuhua Guo, Jieli Chen, Junwei Hao.

Chronic neuroinflammation is a major obstacle to post-stroke recovery, yet the underlying mechanisms, particularly the link between prolonged microglial activation and cholesterol metabolism, are not fully known. Here we show that ischaemic injury induces persistent microglial activation that perpetuates chronic inflammation, leading to microglial cholesterol accumulation and metabolic reprogramming. Using single-cell RNA sequencing, we identified distinct stroke-associated foamy microglia clusters characterized by extensive reprogramming of cholesterol metabolism. Furthermore, direct intracerebral free cholesterol or cholesterol crystal infusion recapitulated sustained microglial activation, directly linking aberrant cholesterol metabolism to prolonged neuroinflammatory responses. Therapeutically, we demonstrate that reducing microglial cholesterol overload through genetic or pharmacological activation of CYP46A1 in male mice promotes white matter repair and functional recovery. These findings highlight microglial cholesterol metabolism as a key driver of post-stroke inflammation, offering therapeutic strategies targeting cholesterol metabolism to mitigate long-term brain damage and promote neurorestoration, potentially improving stroke-related disability outcomes.

DOI: https://doi.org/10.1038/s42255-025-01379-7
Redox Biol. 2025 Sep 17. pii: S2213-2317(25)00385-4. [Epub ahead of print]87 103872

Copper overload worsens the inflammatory response of microglia to amyloid beta (Aβ) by impairing phagocytosis and promoting mitochondrial DNA-mediated NLRP3 inflammasome activation.

Marlene Zubillaga, Xenia Abadin, Elia Ivars, Margalida Puigròs, Ramon Trullas, M José Bellini, Nathalie Arnal, Anna Colell.

  Microglia play a significant role in the development and progression of Alzheimer's disease (AD). These brain-resident immune cells efficiently clear neurotoxic amyloid beta (Aβ) peptides; however, chronic activation may overwhelm their protective abilities, resulting in persistent neuroinflammation. The causes of aberrant microglial activation in AD remain elusive. Emerging evidence indicates that copper (Cu) accumulation, which can arise from prolonged exposure to various environmental sources, modifies the innate immune response in AD. Here, we sought to explore the mechanisms by which Cu overload regulates the microglial phenotype when exposed to Aβ. Our findings showed that exposure to sublethal doses of Cu led to the accumulation of this transition metal in the mitochondria. Elevated mitochondrial Cu (mtCu) levels were accompanied by reduced mitochondrial glutathione (mtGSH) and high oxidative stress, leading to Aβ-induced inflammasome activation through the release of oxidized mitochondrial DNA (ox-mtDNA). Moreover, increased intracellular Cu levels enhanced cholesterol biosynthesis and facilitated its transport to mitochondria. The combination of elevated cholesterol and mitochondrial oxidative stress hindered the ability of microglia to phagocytose Aβ effectively. As expected, conditioned medium from Cu-activated microglia reduced neuronal viability. The neurotoxicity caused by Cu-overloaded microglia was prevented by inhibiting inflammasome activation and restoring mtGSH levels. In conclusion, our study outlines a mechanistic pathway by which chronic exposure to environmental Cu may lead to neuroinflammation and Aβ accumulation in AD, underscoring the crucial role of mitochondrial oxidative stress.

Keywords:  ABCA7; Alzheimer's disease; Cholesterol; Mitochondrial oxidative stress; Neuroinflammation

DOI:  https://doi.org/10.1016/j.redox.2025.103872
J Neuroinflammation. 2025 Sep 26. 22(1): 214

Microglia/macrophage-specific deletion of TLR-4 protects against neural effects of diet-induced obesity in a sexually dimorphic manner.

Jiahui Liu, Ali Zaidi, Christian J Pike.

Obesity is associated with numerous adverse neural effects, including reduced neurogenesis, cognitive impairment, and increased risks for developing Alzheimer's disease (AD) and vascular dementia. Obesity is also characterized by chronic, low-grade inflammation that is implicated in mediating negative consequences body-wide. Toll-like receptor 4 (TLR4) signaling from peripheral macrophages is implicated as an essential regulator of the systemic inflammatory effects of obesity. In the brain, obesity drives chronic neuroinflammation that involves microglial activation, however the contributions of microglia-derived TLR4 signaling to the consequences of obesity are poorly understood. To investigate this issue, we first generated mice that carry an inducible, microglia/macrophage-specific deletion of TLR4 that yields long-term TLR4 knockout only in brain indicating microglial specificity. Next, we analyzed the effects of microglia/macrophage TLR4 deletion on systemic and neural effects of a 16-week of exposure to control versus obesogenic high-fat diets. In male mice, TLR4 deletion generally yielded limited effects on diet-induced systemic metabolic dysfunction but significantly reduced neuroinflammation and impairments in neurogenesis and cognitive performance. In female mice maintained on obesogenic diet, TLR4 deletion partially protected against weight gain, adiposity, and metabolic impairments. Compared to males, females showed milder diet-induced neural consequences, against which TLR4 deletion was protective. Collectively, these findings demonstrate a central role of microglia/macrophage TLR4 signaling in mediating the neural effects of obesogenic diet and highlight sexual dimorphic responses to both diet and TLR4.

DOI: https://doi.org/10.1186/s12974-025-03534-z
J Neuroinflammation. 2025 Sep 25. 22(1): 212

Macrophage expression of P2X7 controls autoimmune uveitis.

Paul-Alexandre Déchelle-Marquet, Yueshen Che, Camille Roux, Frédéric Blond, Kaitryn E Ronning, Sébastien Augustin, Pauline Lagouge-Roussey, Caroline Nous, Sara Touhami, Bahram Bodaghi, Jean Kanellopoulos, Sahil Adriouch, Xavier Guillonneau, Florian Sennlaub, Cécile Delarasse.

  The release of danger-associated molecular patterns (DAMPs) such as ATP from stressed or damaged cells is a key initiator of sterile inflammation. In autoimmune diseases, extracellular ATP acts as a potent pro-inflammatory signal by activating the purinergic receptor P2X7, which is expressed on both mononuclear phagocytes (MPs) and T cells-key drivers of pathological processes. While it is well established that P2X7 mediates ATP-dependent immune activation, its cell-specific contributions to innate versus adaptive immunity in autoimmune conditions remain unclear, especially in autoimmune uveitis. Here we used the experimental autoimmune uveoretinitis (EAU) mouse model to delineate the cellular mechanisms underlying P2X7's role in autoimmune responses. Using a combination of multiple cell-specific conditional models and transcriptomic approaches, we showed a pivotal role forP2X7 expressed by MPs in orchestrating T-cell mediated autoimmune responses. P2X7 deficiency in MPs decreased disease severity. Additionally, cell-specific transcriptomic analyses, including single-cell analyses, revealed that P2X7 exerted distinct modulatory effects across monocyte-derived macrophages (MdM) versus microglia. In MdM, lack of P2X7 was associated with reduced expression of genes related to the inflammasome, phagocytosis pathways, and components of the complement system, leading to a marked decrease in pathogenic Th17 cell frequency in the retina. In microglia, P2X7 deficiency instead particularly impacted an IFN-responsive microglial subset that is normally characteristic of EAU. By specifically deleting P2X7 in microglia, we demonstrate its role in driving pathogenic processes in this cell population. These findings suggest that inhibition of P2X7 could be a promising therapeutic strategy in autoimmune neuroinflammatory disorders.

Keywords:  Autoimmunity; Macrophage; Microglia; Neuroinflammation; Purinergic receptor; Uveitis

DOI:  https://doi.org/10.1186/s12974-025-03529-w
Cell Rep. 2025 Sep 19. pii: S2211-1247(25)01096-4. [Epub ahead of print]44(10): 116325

Macrophage/microglia-dependent mechanisms drive retinal pigment epithelium regeneration in zebrafish.

Lyndsay L Leach, Rebecca G Gonzalez, Sayuri U Jayawardena, Jeffrey M Gross.

  The retinal pigment epithelium (RPE) surrounds the posterior eye and maintains the health and function of the photoreceptors. Consequently, RPE dysfunction or damage has a devastating impact on vision. Due to complex etiologies, there are currently no cures for patients with RPE degenerative diseases, which remain some of the most prevalent causes of vision loss worldwide. Further, owing to a limited capacity for mammalian tissue repair, we know little about how the RPE regenerates. Here, we utilize zebrafish to uncover mechanisms driving intrinsic RPE regeneration. We show that microglia are indispensable for repair and present evidence that supports the role of interleukin-34 in recruiting macrophages/microglia to the RPE injury site. Further, we identify aberrant RPE injury site debris accumulation due to decreased macrophage/microglia localization, and we find that phagocytosis is an important mechanism driving debris clearance. Together, our results identify new regenerative functions of macrophages/microglia after RPE damage.

Keywords:  CP: Immunology; CSF1R; interleukin-34; macrophage; microglia; regeneration; retinal pigment epithelium; zebrafish

DOI:  https://doi.org/10.1016/j.celrep.2025.116325
J Transl Med. 2025 Sep 24. 23(1): 1012

Hypoxia reverses the early anti-inflammatory microglial response to the β-amyloid peptide: mitochondrial involvement and beneficial roles of melatonin and naringenin.

Cristiana Lucia Rita Lipari, Aurora Patti, Stefano Conti-Nibali, Angela Anna Messina, Andrea Magrì, Maria Angela Sortino, Sara Merlo.

   BACKGROUND: Early Alzheimer's disease (AD) is characterized by anti-inflammatory microglial responses to the beta amyloid peptide (Aβ), which later switch to pro-inflammatory. Such transition is relevant to disease progression and can be affected by concurrent insults, such as hypoxia (HY). This study explored whether a mild hypoxic stimulus could anticipate the microglial phenotypic switch, focusing in particular on involvement of SIRT1 and mitochondrial function.
METHODS: HMC3 human microglia were polarized to an anti-inflammatory phenotype by 3 h of exposure to 0.2 μM of Aβ42 to mimic early AD and transferred to a hypoxic chamber with 3% of O2 for 1 h. Effects on microglial activation were investigated by analysis of the SIRT1-BDNF axis activation and enzymatic and ELISA assays of inflammatory markers. Mitochondrial function and morphology were analyzed by high resolution respirometry and laser scanning confocal microscopy.
RESULTS: Hypoxia (HY) prevented the Aβ42-induced early induction of SIRT1 translocation and BDNF release and significantly increased caspase 1 and NF-kB activity. Moreover, mitochondrial oxygen flows evaluated by high resolution respirometry were significantly reduced, while mitochondrial area, perimeter and branching were increased by Aβ42 + HY, compared to Aβ alone. These changes were contrasted by both melatonin (1 μM) and naringenin (10 μM), natural substances able to induce SIRT1. However, use of the selective SIRT1 inhibitor EX-527 (5 μM) suggested only a partial involvement for SIRT1 in the observed effects, prevalent for naringenin.
CONCLUSIONS: Our results suggest that mild hypoxic insults during early asymptomatic stages of AD can pose as a risk factor for an accelerated progression of the disease and show the benefits of SIRT1 induction strategies, including use of natural substances like melatonin and naringenin.

Keywords:  Alzheimer’s disease; High resolution respirometry; Hypoxia; Microglial HMC3 cells; Mitochondrial dynamics; Natural substances; Neuroinflammation; SIRT1

DOI:  https://doi.org/10.1186/s12967-025-07044-7
Front Immunol. 2025 ;16 1656729

Loss of DJ-1 alleviates microglia-mediated neuroinflammation in Parkinson's disease via autophagy-lysosomal degradation of NLRP3.

Qianqian Miao, Tiantian Wang, Haoran Wang, Yanxia Yu, Xing Jin.

   Objective: This study aimed to investigate the role and underlying mechanism of DJ-1 in regulating NLRP3 inflammasome-mediated neuroinflammation during Parkinson's disease.
Methods: We used LPS to stimulate primary microglia in vitro and performed stereotactic injection of LPS into the substantia nigra of mice in vivo to investigate changes in DJ-1 expression following inflammatory stimulation. To evaluate the functional effects of DJ-1 on NLRP3 inflammasome activation, we used siRNA to knock down DJ-1 in primary microglia or BMDMs and analyzed downstream inflammatory responses as well as the specificity of this regulation. In vivo, we used microglia-specific AAV to selectively silence DJ-1 in the substantia nigra to further evaluate the anti-inflammatory effect of DJ-1 deficiency. To validate the direct interaction between DJ-1 and NLRP3, we performed co-immunoprecipitation and proximity ligation assay. We used the autophagy inhibitor 3-MA and activator rapamycin to investigate how NLRP3 is degraded upon DJ-1 deficiency in CRISPR-Cas9-engineered DJ-1 knockout HEK-293 cells.
Results: DJ-1 were significantly upregulated following LPS or LPS plus ATP stimulation in primary microglia. Similarly, after stereotactic LPS injection into the substantia nigra, we observed a significant upregulation of DJ-1 expression. Knockdown of microglial DJ-1 markedly suppressed NLRP3 inflammasome activation, as evidenced by reduced mature caspase-1 and decreased IL-1β secretion. We confirmed this phenomenon in BMDM and found that DJ-1 knockdown specifically inhibited NLRP3 inflammasome activation, with no effect on NLRC4 or AIM2 inflammasomes. In vivo, microglia-specific DJ-1 knockdown significantly attenuated microglial NLRP3 inflammasome activation in the substantia nigra and exerted neuroprotective effects after LPS treatment. Furthermore, DJ-1 was found to directly bind NLRP3 and stabilize its conformation, thereby preventing autophagic degradation.
Conclusion: This study demonstrates that DJ-1 deficiency in microglia inhibits NLRP3-driven inflammation by promoting NLRP3 degradation through the autophagy-lysosome pathway. Future studies should focus on identifying the specific binding sites and structure of DJ-1 with NLRP3, as well as investigating whether inhibiting DJ-1 in microglia could serve as a potential therapeutic target for suppressing neuroinflammation in Parkinson's disease.

Keywords:  DJ-1 (PARK7); NLRP3 inflammasome; Parkinson’s disease; microglia; neuroinflammation

DOI:  https://doi.org/10.3389/fimmu.2025.1656729
Int J Mol Sci. 2025 Sep 09. pii: 8782. [Epub ahead of print]26(18):

Inhibition of the HMGB1-RAGE Axis Attenuates Microglial Inflammation and Ameliorates Hypoxia-Induced Cognitive Impairment.

Chenlin Liu, Haowei Zhang, Ruili Guan, Yuankang Zou, Mengyu Chen, Mingrui Du, Wenjing Luo, Jianbin Zhang.

  The mechanisms underlying the abnormal activation of microglia affecting cognitive function under high-altitude hypobaric hypoxia (HAHH) have not been fully elucidated. This study aims to investigate the effects of HAHH on the expression of the receptor for advanced glycation end-products (RAGE) in hippocampal microglia of mice and to explore the role of RAGE inhibitors in alleviating HAHH-induced microglial inflammation and cognitive impairment. Mice were exposed to HAHH via a multi-environment simulation chamber, and RNA sequencing, qPCR, WB, flow cytometry and immunohistochemistry showed that HAHH exposome significantly increased RAGE expression in hippocampal microglia of mice (p < 0.001 vs. normoxia), which was closely related to microglial neuroinflammatory responses. RAGE inhibitor (FPS-ZM1) alleviated HAHH-induced microglial inflammation (TNF-α decreased by 64%, p < 0.001; CD86+ cells decreased by 42%, p < 0.001) and improved cognitive function in mice (Y-maze novel arm time: 28.08 ± 5.14 s vs. hypoxia 19.67 ± 4.68 s, p = 0.016; NORT recognition index: 0.52 ± 0.05 vs. hypoxia 0.33 ± 0.07, p < 0.001). Mechanistic studies revealed that RAGE inhibitors reduced microglial inflammation by inhibiting the MAPK pathway and decreasing nuclear translocation of NF-κB p65. Furthermore, high-mobility group box 1 (HMGB1) expression increased under hypoxic conditions (p < 0.001 vs. normoxia) and positively regulated RAGE expression. HMGB1 inhibitors reduced RAGE expression and attenuated HAHH-induced microglial inflammation. Overall, the HAHH exposome induces microglial inflammation via the HMGB1-RAGE-NF-κB pathway. RAGE and HMGB1 inhibitors may serve as novel therapeutic strategies to mitigate HAHH-induced cognitive impairment, providing a theoretical basis for the treatment of cognitive impairment.

Keywords:  HMGB1; RAGE; cognitive function; high-altitude hypobaric hypoxia (HAHH); inflammation; microglia

DOI:  https://doi.org/10.3390/ijms26188782
Int J Mol Sci. 2025 Sep 15. pii: 8966. [Epub ahead of print]26(18):

Characterizing Microglia Morphology in the Frontal Cortex of Pair-Bonded and Unpaired Prairie Voles (Microtus ochrogaster).

Tori Keefauver, Kyle L Gobrogge.

  Microtus ochrogaster, monogamous prairie voles, serve as translational animal models for studying monogamy and pair bonding. Microglia, the resident immune cells of the brain, are one of several cell types still poorly understood in non-classical animal models including prairie voles. Microglia are known to play mechanistic roles in mediating social behaviors using inflammatory signaling, but the relationship between microglia reactivity and pair bonding has not yet been investigated. The present study first developed a robust protocol for quantitative histological visualization of microglia in Microtus ochrogaster. Second, it investigated differences in microglia morphology, a reliable index of microglia reactivity and function, in pair-bonded vs. unpaired voles. Sections containing prefrontal cortex (PFC) and anterior cingulate cortex (ACC) were stained for ionized calcium-binding adaptor molecule I (Iba1) using immunohistochemistry (IHC). IHC results provided evidence for the successful use of murine histological protocols in prairie voles. Quantification results revealed a sexually dimorphic effect of pair bonding on microglia: somas were significantly larger in pair-bonded vs. unpaired females, and somas were significantly smaller in pair-bonded vs. unpaired males. Additionally, somas were significantly larger in unpaired males than females, with larger somas indicating higher microglia reactivity. While conclusions are limited due to the small sample size, results provide novel characterization of microglia morphology in the frontal cortex and elucidate how pair bonding may influence microglia function in a sexually dimorphic manner.

Keywords:  Microtus ochrogaster; glia; microglia; morphology; prairie vole; sex differences

DOI:  https://doi.org/10.3390/ijms26188966
Neurobiol Dis. 2025 Sep 22. pii: S0969-9961(25)00334-1. [Epub ahead of print] 107117

Astrocyte-microglia crosstalk in the hippocampus mediates cognitive impairments induced by chronic intermittent hypoxia.

Zhen-Huan Wu, Ming-Rui Zhai, Yu-Rong Wang, Long Ren, Jie Pan, Lei Xiao, Yue-Hua Liu.

  Obstructive sleep apnea (OSA), characterized by chronic intermittent hypoxia (CIH), is a common systemic disease with a high-risk factor for developing cognitive impairment. However, the possible mechanism(s) underlying the cognitive function impairment in CIH remain largely unknown. In this study, our results reveal that 8-week CIH reliably induces significant cognitive impairment, synaptic deficits, and pronounced microglial activation characterized by excessive synaptic phagocytosis. Pharmacological depletion of microglia using PLX5622 ameliorated these CIH-induced impairments. Furthermore, CIH enhanced the interaction between activated astrocytes and microglia, accompanied by upregulation of complement C3 in astrocytes and C3aR in microglia. Notably, blocking C3aR with SB290157 attenuated microglial overactivation, reduced aberrant synaptic engulfment, and improved cognitive performance in CIH-exposed mice. Collectively, these findings demonstrate that C3/C3aR-mediated astrocyte-microglia crosstalk contributes to CIH-induced cognitive dysfunction by activating microglia to excessive phagocytosis of synapses.

Keywords:  Chronic intermittent hypoxia; Cognitive impairment; Complement component; Microglia phagocytosis; Synaptic connection

DOI:  https://doi.org/10.1016/j.nbd.2025.107117
Neurobiol Dis. 2025 Sep 21. pii: S0969-9961(25)00323-7. [Epub ahead of print]216 107106

P2RX7 modulates the function of human microglia-like cells and mediates the association of IL18 with Alzheimer's disease traits.

Kelsey S Heavener, Kirstin A Tamucci, Annie J Lee, Khushbu Kabra, Zena K Chatila, Maedot Yidenk, David A Bennett, Badri N Vardarajan, Elizabeth M Bradshaw.

  The immune system plays a dynamic role in neurodegenerative diseases, and purinergic receptors allow immune cells to recognize neuronal signaling, cell injury, or stress. Purinergic Receptor 7 (P2RX7) can modulate inflammatory cascades, and its expression is upregulated in Alzheimer's disease (AD) brain tissue. P2RX7 expression is enriched in microglia, and elevated levels are found in microglia surrounding amyloid-beta (Aβ) plaques in the brain. Despite evidence linking P2RX7 to AD, the mechanisms by which it shapes microglial responses and contributes to AD remain poorly defined. Here, we utilize a human monocyte-derived microglia-like cell model (MDMi) to interrogate P2RX7 activation and downstream consequences on microglial function. Specifically, we measured IL1β and IL18 production and Aβ1-42 uptake following ATP-induced P2RX7 activation. Our results show that ATP-stimulation of MDMi triggers upregulation of IL1β and IL18 expression, which is blocked with the A740003 P2RX7 antagonist. Elevated extracellular ATP also impaired Aβ1-42 uptake, an effect reversed by P2RX7 inhibition with A740003. In addition, pretreatment of MDMi with IL-1Ra limited ATP-driven IL1β and IL18 gene expression upregulation, indicating that ATP immunomodulation of P2RX7 is IL-1R dependent. Critically, analysis of postmortem human brain revealed that P2RX7 expression significantly mediated the association between IL18, but not IL1β, and multiple AD traits, including amyloid load, tau tangle density, global AD pathology burden, cortical and neocortical Lewy bodies, cognitive decline, and clinical dementia. Consistent with these findings, in our MDMi model, P2RX7 gene expression correlated with IL18 but not IL1β. Together, these findings highlight P2RX7 as a critical integrator of extracellular danger signals and microglial immune function, underscoring its potential as a therapeutic target in neurodegeneration.

Keywords:  ATP; Alzheimer's disease; Amyloid-beta; IL-1Ra; Neurodegeneration; P2RX7

DOI:  https://doi.org/10.1016/j.nbd.2025.107106
Neurobiol Dis. 2025 Sep 24. pii: S0969-9961(25)00336-5. [Epub ahead of print] 107119

The alternative complement pathway drives neuroinflammation and neurodegeneration in mouse models of glaucoma and optic nerve injury.

Cindy Hoppe, Ryo Mukai, Nasrin Refaian, Margarete Karg, Shintaro Shirahama, Maleeka Shrestha, Yinjie Guo, Amarachi Nwogu, Drenushe Krasniqi-Vanmeter, Volha V Malechka, Bruce R Ksander, Kip Connor, Meredith Gregory-Ksander.

  Glaucoma is a leading cause of irreversible blindness worldwide, characterized by progressive retinal ganglion cell (RGC) loss and optic nerve degeneration. Although elevated intraocular pressure (IOP) is a major risk factor, disease progression can occur despite normal IOP, highlighting the need for neuroprotective strategies beyond IOP reduction. Neuroinflammation has been implicated in glaucomatous neurodegeneration through complement system activation via the classical and lectin pathways. However, the role of the alternative pathway (AP), which functions as an amplification loop for central complement component 3 (C3) activation, in glaucoma is unclear. In this study, we investigated the role of the AP in glaucoma using a microbead-induced mouse model of glaucoma in mice deficient in either complement factor B (Cfb-/-), to selectively block the AP, or in C3 (C3-/-) to block all three complement pathways. Our results indicate that the AP is critical for glaucoma development, and blocking this pathway resulted in significant neuroprotection, preventing loss of RGCs, axons, and visual acuity, which coincided with reduced glial activation and inflammatory signaling. Blocking the AP provided comparable neuroprotection to blocking all three complement pathways, indicating that the AP amplification loop is an essential component of destructive neuroinflammation in glaucoma. Furthermore, blocking the AP also conferred neuroprotection in the optic nerve crush model, suggesting a broader role for AP in optic neuropathies. These findings establish the AP as a key driver of complement-mediated neurodegeneration in glaucoma and highlight the therapeutic potential of targeting the AP in glaucoma and other neurodegenerative diseases.

Keywords:  Complement system; Glaucoma; Inflammation; Microglia; Neuroprotection

DOI:  https://doi.org/10.1016/j.nbd.2025.107119
Invest Ophthalmol Vis Sci. 2025 Sep 02. 66(12): 56

Evaluating the Optic Nerve Crush Model to Understand the Function of Microglia in Glaucoma Neuroprotection.

Xiaowu Gu, Tom Truong, Tiffany Heaster-Ford, Tae-Hoon Kim, Gyeong Jin Kang, Joanna Yung, Miriam Baca, Shawnta Y Chaney, Jeffrey W Hofmann, Jeffrey Eastham, Marion Jeanne.

Purpose: Microgliosis is a key neuroinflammatory feature in human glaucomatous retinas, believed to contribute to disease progression. This study aims to characterize changes in microglia and intra-retinal axons following optic nerve crush (ONC) and investigate microglial involvement in retinal ganglion cell (RGC) and axonal degeneration.
Methods: Using the CD11c.YFP.Venus.Tg mouse line, we tracked microglial activation and assessed the spatiotemporal changes in TUJ1+ intraretinal axons over a 2-week period post-ONC. Microglial function was examined by depleting microglia with the CSF1R inhibitor PLX5622 and using Trem2-deficient mice with dampened microglial activation.
Results: Activated microglia accumulated significantly in the retina from day 4 post-ONC, peaking at day 7. Retinal microglia became hypertrophic by day 1 and started proliferating. Axon beading occurred primarily in the peripheral retina by day 2 post-ONC, with more beaded axons appearing along long axonal bundles toward the optic nerve head (ONH) by day 7 and day 14. There was a significant reduction in overall TUJ1 expression and axonal bundle thickness during this period. Despite complete microglial depletion and significantly reduced activation, no differences were observed in the RGC count or the extent of optic nerve damage following ONC.
Conclusions: Microglial activation is secondary to axonal injury and plays a bystander role in the ONC model. Robust RGC and axonal degeneration appear unaffected by activated microglia. This finding challenges the utility of the ONC model for evaluating microglia-based glaucoma treatments. Additionally, the study reaffirms the value of combining fluorescent reporter mouse lines with noninvasive ocular imaging for streamlining future research.

DOI: https://doi.org/10.1167/iovs.66.12.56
Comput Struct Biotechnol J. 2025 ;27 3804-3813

The role of microglia and complement C5/C5a in the pathogenesis of rhegmatogenous retinal detachment with choroidal detachment.

Huiyan Xu, Qiuhong Wang, Xuan Chen, Qingyu Huang, Shasha Xu, Zhifeng Wu.

   Background: Rhegmatogenous retinal detachment with choroidal detachment (RRDCD) is an uncommon and sight-threatening disorder marked by fast development and significant inflammation. This study aimed to identify cellular and molecular signatures distinguishing RRDCD from typical rhegmatogenous retinal detachment (RRD) and to investigate the roles of microglia and the complement C5/C5a pathway in disease pathogenesis.
Methods: Single-cell RNA sequencing (scRNA-seq) was employed to analyze vitreous samples from patients with RRD and RRDCD to characterize the cellular composition and molecular pathways. In vitro co-culture experiments were performed to investigate the functional impact of complement C5 on primary retinal microglia, and their subsequent effects on RF/6 A endothelial cells and ARPE-19 epithelial cells.
Results: Our findings revealed a distinct cellular landscape in RRDCD, characterized by enhanced connectivity between microglia and dendritic cells, alongside a significant upregulation of the complement C5-C5AR1 interaction. In vitro experiments indicated that treatment with complement C5 enhanced microglial metabolic activity and activation, induced apoptosis in RF/6 A endothelial cells, and led to disruption of tight junction protein ZO-1 localization in ARPE-19 epithelial cells, suggesting a role in blood-retina barrier dysfunction.
Conclusion: The findings substantiate the inflammatory hypothesis regarding the pathogenesis of RRDCD, emphasizing the critical functions of microglia and the complement C5/C5a pathway in intensifying retinal inflammation and undermining vascular integrity.

Keywords:  Blood-retina barrier; Choroid detachment; Complement system; Inflammation; Microglia; Rhegmatogenous retinal detachment

DOI:  https://doi.org/10.1016/j.csbj.2025.08.019
Sci Rep. 2025 Sep 26. 15(1): 33285

Microglia reactivity is brain region and sex specific in the context of chronic stress.

Ariel Y Zhang, Elias Elias, Abigail G White, Laura G Jones, Taleen Hamad, Melissa T Manners.

  Chronic stress has been associated with an increased inflammatory profile in the brain, linking inflammation to the development of stress-related disorders. Although the detailed mechanism connecting chronic stress to inflammation remains unclear, as primary mediators of the immune response, it has been established that microglia play a role. We further investigated the effect of chronic stress on microglia reactivity by incorporating sex and multiple brain regions as variables in our analysis. We utilized the unpredictable chronic mild stress (UCMS) paradigm and then quantified the number of microglia, process arborization, process length, and the number of processes of microglia in the prefrontal cortex (PFC), nucleus accumbens (NAC), hypothalamus (HYPO), amygdala (AMY), and hippocampal CA1 and CA3. We did not observe a stress-induced change in the number of microglia in each region; however, chronic stress reduced microglia arborization, length, and number of processes in a brain region and sex-specific manner. Independent of stress, microglia exhibited a region-dependent reactive phenotype. Together, chronic stress affects microglia reactivity uniquely based on sex and brain region, while the different reactivity profile of microglia in males and females might underlie the sex-specific mechanism of the diseases.

Keywords:  Chronic stress; Microglia reactivity; Sex difference

DOI:  https://doi.org/10.1038/s41598-025-18000-2
Sci Rep. 2025 Sep 26. 15(1): 33103

Investigating the role of the ALPK1 signaling pathway in the pathogenesis of diabetic retinopathy.

Ting Wu, Yin-Kun Guo, Fang-Yi Long, Shang-Shang Duan, Xiao-Hong Li, Guo Liu, Jun-Rong Du, Nai-Hong Yan.

  Diabetic retinopathy (DR) is a key eye-related complication linked to diabetes. Chronic inflammation is recognized as the main pathological mechanism leading to retinal damage during the onset and progression of DR. Alpha kinase 1 (ALPK1), a member of a recently discovered serine/threonine protein kinase family, is associated with the onset of multiple inflammatory conditions. Dominant mutations in the ALPK1 gene can result in a rare genetic disorder, known as ROSAH syndrome, which is characterized by ocular inflammatory responses. The role of ALPK1 in DR remains largely unclear and necessitates additional research for a better understanding. Therefore, investigating the function and underlying mechanisms of ALPK1 in DR is highly necessary. In this research, ALPK1 expression and inflammatory cytokines levels were examined in db/db diabetic mice. Additionally, BV2 mouse microglial cells were utilized to establish a hypoxia model that mimicked DR conditions, and the influence of ALPK1 knockdown on these conditions was assessed. The effects of ALPK1 knockdown on retinal damage in db/db diabetic mice were also evaluated. In db/db mice, our findings indicated a decrease in retinal cell numbers and irregular retinal blood vessel formation, along with increased levels of ALPK1 and inflammatory markers TNF-α, IL-1β, IL-6, and IL-18. ALPK1 expression was found to be associated with the TIFA/TRAF6 signaling pathway. Knockdown of ALPK1 was shown to reduce the levels of these inflammatory cytokines, diminish GSDMD-mediated pyroptosis signaling, and mitigate the DR-induced retinal cell damage in BV2 cells. Furthermore, reducing ALPK1 expression in the retinas of db/db diabetic mice was observed to slow down retinal cell degeneration and alleviate microvascular injury within the retina. This research indicated the substantial involvement of ALPK1 in the diabetic retinopathy-linked inflammatory cascade and cell pyroptosis. Targeting the ALPK1 signaling pathway could open avenues for novel therapeutic strategies to manage DR.

Keywords:   Db/db diabetic mice; ALPK1; Pyroptosis; diabetic retinopathy

DOI:  https://doi.org/10.1038/s41598-025-17122-x
J Cell Mol Med. 2025 Sep;29(18): e70861

GCN5L1 Aggravates Postherpetic Neuralgia Through Regulating Microglial Mitochondrial Fission-Fusion Homeostasis.

Wang Li, Xin Cao, Shenghan Wang, Xuedong Jin, Hongqian Wang.

  Postherpetic neuralgia (PHN) is a debilitating chronic pain condition following varicella-zoster virus (VZV) reactivation, characterised by persistent neuroinflammation. However, the intracellular mechanisms that drive microglial activation and sustained pain sensitisation remain poorly understood. Due to mice having no VZV infection receptor, herpes simplex virus type 1 (HSV-1) infection is a well-established PHN mice model. Here, we identified GCN5L1, a mitochondrial acetylation modulator, as a critical regulator of microglial mitochondrial dynamics and a key contributor to PHN pathogenesis. We found that GCN5L1 was markedly upregulated in the spinal dorsal horn after PHN, particularly located in microglia. Microglial Gcn5l1 deficiency attenuated HSV-1-induced neuroinflammatory responses and alleviated mechanical allodynia, whereas Gcn5l1 overexpression exacerbated neuroinflammatory responses both in vivo and in vitro. Mechanistically, GCN5L1 promoted mitochondrial fission and impaired oxidative metabolism by enhancing DRP1 acetylation, without altering the expression of canonical fission-fusion regulators. Restoration of mitochondrial fission using MFI8 intrathecally reversed the anti-inflammatory and analgesic effects of Gcn5l1 deficiency, confirming that GCN5L1 mediated pain sensitisation through mitochondrial fission-fusion in PHN. Finally, inhibiting GCN5L1 by AAV-shGCN5L1 intrathecally suppressed neuroinflammation and mechanical allodynia in PHN mice. These findings uncovered that GCN5L1 aggravated neuroinflammation and PHN through regulating microglial mitochondrial fission-fusion homeostasis, offering new insights and potential feasibility in clinical translation for PHN management.

Keywords:  GCN5L1; microglia; mitochondrial fission–fusion; neuroinflammation; postherpetic neuralgia

DOI:  https://doi.org/10.1111/jcmm.70861
Biomedicines. 2025 Aug 22. pii: 2049. [Epub ahead of print]13(9):

Enhanced M2 Polarization of Retinal Microglia in Streptozotocin-Induced Diabetic Mice upon Autoimmune Stimulation.

Yoshiaki Nishio, Hideaki Someya, Kozo Harimoto, Tomohito Sato, Masataka Ito, Masaru Takeuchi.

  Background: This study aimed to investigate the impact of the diabetic environment on the development of experimental autoimmune uveoretinitis (EAU) and the activation status of microglia in the eye. Methods: EAU was induced in wild-type (WT) and streptozotocin (STZ)-induced diabetic mice (STZ-EAU mice). Disease severity was assessed using funduscopy, optical coherence tomography (OCT), and histopathological analysis. The proportions of Th1, Th17, and regulatory T cells in the spleen were analyzed by flow cytometry. Retinal microglia were quantified using immunohistochemistry. To further characterize retinal cell populations and gene expression profiles, single-cell RNA sequencing (scRNA-seq) was performed. Results: STZ-EAU mice exhibited significant reductions in both the incidence and severity of EAU compared with WT-EAU mice. These were accompanied by a decreased proportion of Th1 cells, which are crucial for EAU pathogenesis, in the spleens of STZ-EAU mice. Retinal microglial accumulation was markedly reduced in STZ-EAU mice compared with WT-EAU mice. scRNA-seq analysis revealed a significant change in the microglial phenotype in STZ-EAU mice, characterized by decreased expression of MHC class I/II and the suppression of antigen presentation signaling pathways. Activated microglia in STZ-EAU mice showed reduced gene expression of M1 markers (CD68, CD74, and IL1B) and increased gene expression of M2 markers (MSR1, CD163, and MRC1), suggesting a shift toward an anti-inflammatory M2 phenotype. Conclusions: EAU is suppressed in STZ-induced diabetic mice, likely due to alterations in microglial polarization toward an M2 phenotype. These results suggest a decrease in T cell responses to pathogens in a diabetic environment, which could be one of the underlying factors for the increased susceptibility to infection in diabetic patients. Inhibiting the M2 polarization of microglia may reduce the susceptibility to infection in patients with diabetes.

Keywords:  autoimmune disease; autoimmune uveitis; diabetes; experimental autoimmune uveoretinitis; microglia

DOI:  https://doi.org/10.3390/biomedicines13092049
J Alzheimers Dis. 2025 Sep 23. 13872877251379844

Axonal degeneration and retinal neurovascular dysfunction in the TgF344-AD rat model of Alzheimer's disease.

Tienan Qi, Yuan Feng, Natalia Kononenko, Christina Ising, Tim van Beers, Thibaut Sesia, Claus Cursiefen, Verena Prokosch, Hanhan Liu.

  BackgroundRecent research suggests that retinal changes occur at early stages of Alzheimer's disease (AD). However, gaps remain in understanding the detailed pathophysiological processes and their molecular underpinnings.ObjectiveThis study investigates alterations in the retinal neurovascular unit (NVU) and associated proteomic changes in the TgF344-AD rat model to determine how these changes relate to brain and known AD pathology.MethodsTransgenic rats and age-matched wild-type rats were studied. Proteomic analysis was conducted to identify changes in critical signaling pathways across the retina, the visual sensory thalamus (the lateral geniculate body, LGN) and hippocampus. Immunofluorescence staining was used to detect amyloid-β, glial, neuronal and vascular markers in the retina, and p-phenylenediamine staining examined axons of retinal ganglion cells.ResultsSignificant axonal degeneration in the optic nerve and optic tract of AD rats was detected, while axonal integrity in the optic chiasm, retinal ganglion cell numbers and retinal layer thickness remained unaffected. Proteomics showed a general downregulation of pathways essential for neural survival, glial function and vascular stability, with striking similarities between the retina and LGN. A significant reduction in amacrine cell numbers, increased microglial reactivity, decreased Müller cell immunoreactivity and reduced retinal pericyte density were also observed.ConclusionsThese findings suggest that retrograde and anterograde axonal degeneration, coupled with NVU dysfunction, are key features of retinal pathology in AD. The TgF344-AD rat model provides valuable insights into these changes, highlighting the retina as a potential site for early AD detection, monitoring and intervention.

Keywords:  Alzheimer's disease; amacrine cells; blood-retinal barrier; lateral geniculate body; neuroglia; proteomics; retinal ganglion cells

DOI:  https://doi.org/10.1177/13872877251379844
Front Cell Neurosci. 2025 ;19 1636399

Growth hormone reduces retinal inflammation and preserves microglial morphology after optic nerve crush in male rats.

Jerusa E Balderas-Márquez, David Epardo, Lourdes Siqueiros-Márquez, Martha Carranza, Maricela Luna, José Luis Quintanar, Carlos Arámburo, Carlos G Martínez-Moreno.

   Introduction: This study investigates the neuroprotective role of growth hormone (GH) in modulating retinal inflammation and microglial responses following optic nerve crush (ONC) in male rats.
Methods: Retinal inflammation and microglial activation were assessed at 24 h and 14 days post-ONC, with or without GH treatment (0.5 mg/kg, subcutaneously, every 12 h). Gene and protein expression of inflammatory markers (e.g., IL-6, TNFα, Iba1, CD86, CD206) were evaluated using qPCR, ELISA, and Western blotting. Microglial morphology was quantified using skeleton and fractal analysis of Iba1-stained retinal sections. Retinal structure and function were assessed via fundus imaging and optomotor reflex testing.
Results: ONC induced significant increases in proinflammatory cytokines (IL-6, TNFα, IL-18) and microglial activation, characterized by reduced branching complexity and increased cell density. GH treatment significantly decreased proinflammatory cytokine levels, modulated microglial phenotype (CD86/CD206 expression), and preserved microglial morphology in the retina. Using the SIM-A9 microglial cell line, we further demonstrated that GH reduces NFκB pathway activation and suppresses LPS-induced proinflammatory cytokine production. At 14 days post-injury, GH-treated retinas exhibited reduced optic nerve size and improved optomotor responses, indicating both structural neuroprotection and functional recovery.
Discussion: Overall, GH mitigates ONC-induced retinal inflammation by reducing proinflammatory signaling and preserving microglial architecture, thereby protecting retinal integrity and function. These findings highlight the potential of GH as a therapeutic agent for retinal neurodegenerative conditions.

Keywords:  growth hormone (GH); microglia; neuroinflammation; neuroprotection; optic nerve crush (ONC); proinflammatory cytokines; retina

DOI:  https://doi.org/10.3389/fncel.2025.1636399
Exp Eye Res. 2025 Sep 23. pii: S0014-4835(25)00425-7. [Epub ahead of print] 110653

Microglia Activation and Migration in Retina during Acute High-altitude Exposure.

Yuting Li, Cong Han, Jianping Zhang, Songjian Huang, Xin Zhao, Jiarui Zhu, Qian Niu, Yi Yang, Wenfang Zhang.

  High-altitude retinopathy (HAR), caused by hypobaric hypoxia, leads to retinal dysfunction. However, its pathogenic mechanism remains elusive. Microglia, the resident immune cells of the retina, play crucial roles in various retinal disorders. Here, we investigated the functional alterations and underlying mechanisms of microglial activation in mouse models exposed to high-altitude (5000m HAE). Hematoxylin and eosin (H&E) staining showed edema in the entire, inner, and outer retinal layers after high-altitude exposure (HAE). Electroretinogram (ERG) testing revealed impaired retinal function under hypobaric hypoxia. Immunofluorescence staining confirmed a time-dependent increase in microglial numbers within the retina following HAE, with activated microglia migrating during persistent hypoxic injury. Furthermore, these activated microglia predominantly differentiated into pro-inflammatory subtypes under acute high-altitude conditions. Notably, Connexin43 (Cx43) immunoreactivity increased, while interleukin-1β (IL-1β) levels were markedly elevated at different time points after HAE. In this study, we confirmed that microglia activation and migration are involved in retinal edema and functional injury induced by hypobaric hypoxia. Additionally, the dynamic changes in Cx43 and up-regulation of IL-1β might be related to the inflammatory activation of microglia under acute HAE.

Keywords:  Acute High-altitude Exposure; Inflammation; Microglia

DOI:  https://doi.org/10.1016/j.exer.2025.110653
Lab Anim (NY). 2025 Sep 26.

Diet gel-based oral drug delivery system for controlled dosing of small molecules for microglia depletion and inducible Cre recombination in mice.

Joel Jovanovic, Megan L Stone, Samantha R Dooyema, Yuankai K Tao, Sabine Fuhrmann, Edward M Levine.

Small molecules such as PLX5622 for microglia depletion and tamoxifen for inducible Cre recombination are commonly used in mouse research. Traditional application methods such as drug-infused chow, oral gavage or injections have limitations, including uncontrolled dosing (chow) or risk of injury and/or stress (gavage or injections). Here, to address these issues, we have developed an alternative oral drug delivery system using a gel-based rodent maintenance diet that allows for controlled consumption and adjustment of dosage and is suitable for water-insoluble small molecules. We tested DietGel 93M (93M) infused with PLX5622 (0.8 mg/g and 2.0 mg/g) in the Cx3cr1gfp/+ retinal microglia reporter mouse and tamoxifen-infused 93M (0.3125 mg/g) in the Rlbp1-CreERT2;Rosaai14 mouse with an inducible tdTomato reporter in retinal Müller glia. Mice were single caged and received daily batches of PLX5622-infused 93M over 14 days or tamoxifen-infused 93M for 1 or 3 days followed by a 14-day observation period. Longitudinal scanning laser ophthalmoscopy in vivo and fixed-tissue imaging were used to track GFP and tdTomato expression. Following evaluation of a suitable 93M consumption rate to sustain body weight, the PLX5622-93M diet at both concentrations tested showed a 94% microglia depletion rate at 3 days and >99% after 1 and 2 weeks. The tamoxifen-93M diet confirmed suitability for inducible Cre recombination, with significant treatment time-dependent efficacy and a positive correlation between total tamoxifen dose and tdTomato expression. This study demonstrates that a diet gel-based drug delivery system offers a minimally invasive alternative to current drug application methods for PLX5622 and tamoxifen. This approach could be useful for other drugs or tissues beyond the retina.

DOI: https://doi.org/10.1038/s41684-025-01617-1
Brain Behav. 2025 Sep;15(9): e70881

Bibliometric Mapping of Research Trends and Hotspots of Microglia in Spinal Cord Injury (2000-2024).

Ziming Cai, Gongpeng Xiong, Jintao Wu, Hanjun Zhang, Jian Huang, Qinghe Yu, Wenping Lin.

   INTRODUCTION: Spinal cord injury (SCI), acknowledged as the most severe complication arising from spinal trauma, pertains to the dysfunction of the spinal cord due to traumatic events or other pathological conditions. Extensive research has elucidated a substantial correlation between SCI and inflammatory processes, highlighting the critical involvement of microglia in orchestrating neuroinflammatory responses. Moreover, a growing body of evidence has demonstrated a strong connection between microglial activation and both the pathogenesis and progression of SCI.
OBJECTIVE: We chose bibliometric analysis to comprehensively summarize the research progress of microglia in SCI, aiming to provide researchers with current trends and future research directions.
METHODS: All articles and reviews addressing microglia in SCI were systematically retrieved from the Web of Science Core Collection database, spanning publications from 2000 to 2024. Subsequent bibliometric analysis was conducted utilizing four analytical tools: VOSviewer (version 1.6.20), R software (package bibliometrix), the Biblioshiny web interface, and CiteSpace (version 6.2.R4), ensuring comprehensive examination of publication patterns and research trends.
RESULTS: A total of 2428 publications were ultimately included in this bibliometric analysis. The annual publication count demonstrated a consistent upward trajectory. China is the country with the most published articles, and Ohio State University ranks first in institutional publications. Experimental Neurology is the journal with the most published articles, while Journal of Neuroscience is the journal with the most cited articles. Popovich Pg is the author with the highest productivity and co-citation. Cluster analysis yielded a total of 15 different co-citation clusters. Time analysis shows explosive citation outbreaks in 2006, 2009, and 2011. Keyword analysis revealed inflammation, expression, activation, and central nervous system as the most frequently occurring terms. Recent keyword trends feature emerging terms like exosomes, extracellular vesicles, and nanoparticles. Keyword bursts revealed promotes, extracellular vesicle, recovery, neuroinflammation, therapy, polarization, and pathway are the hotspots of research at the present stage and are likely to continue. These findings provide critical insights for developing microglia-targeted therapeutic strategies and prioritizing research directions in neuroinflammatory modulation to improve functional recovery after SCI.
CONCLUSION: Emerging research frontiers prominently feature exosomes, gut microbiota, and nanoparticles. The interplay between microglia-mediated neuroinflammation and SCI has emerged as a critical focal point in current scientific investigations and is anticipated to remain central to forthcoming scientific inquiries.

Keywords:  CiteSpace; VOSviewer; bibliometrics; microglia; spinal cord injury

DOI:  https://doi.org/10.1002/brb3.70881
STAR Protoc. 2025 Sep 20. pii: S2666-1667(25)00517-9. [Epub ahead of print]6(4): 104111

Protocol for pooled FACS-based CRISPR knockout screening in human iPSC-derived microglia.

Sam J Washer, Elena Navarro-Guerrero, Sally A Cowley, Daniel V Ebner, Andrew R Bassett.

  Here, we present a protocol for CRISPR knockout screening in human induced pluripotent stem cell (hiPSC)-derived microglia (iMGL) using lentiviral delivery of CRISPR-Cas9 and co-transduction of VPX virus-like particles (VPX-VLPs). We first describe large-scale production of iMGL from hiPSCs, production of the lentiviral and VPX-VLP libraries, and titration. Next, we describe how to perform a pooled CRISPR screen for phagocytosis including the computational analysis pipeline of CRISPR screening data. For complete details on the use and execution of this protocol, please refer to Perez-Alcantara et al.1.

Keywords:  CRISPR; cell culture; high-throughput screening; sequencing; stem cells

DOI:  https://doi.org/10.1016/j.xpro.2025.104111
Open Med (Wars). 2025 ;20(1): 20251274

Petunidin attenuates lipopolysaccharide-induced retinal microglia inflammatory response in diabetic retinopathy by targeting OGT/NF-κB/LCN2 axis.

Mengxi Yu.

   Background: Diabetic retinopathy (DR), as a frequent complication of diabetes mellitus, is a common cause of vision impairment and blindness. Microglial activation plays an important role in the pathological cascade of DR, and novel potential therapeutics is needed to interfere with this process. Petunidin (PET) is one of the most abundant natural anthocyanins with the biological activities of anti-inflammation, anti-cancer, and anti-microbial.
Objective: The purpose of this study is to investigate the effect of PET against lipopolysaccharide (LPS)-induced retinal microglia inflammatory response as well as the underpinning mechanism.
Methods: Cell viability was determined using MTT assay. Cytokines secretion was determined using ELISA assay. iTRAQ-based proteomic analysis was used for the identification of differentially expressed proteins. Western blot analysis, Co-IP and immunofluorescence analysis were applied in the study.
Results: The results showed that PET pre-treatment significantly reduced LPS-induced cytokines secretion in BV2 cells and primary retinal microglia, as well as lipocalin 2 (LCN2) upregulation in BV2 cells by suppressing activation of O-GlcNAc modification and activation of NF‑κB. Further study revealed that PET inactivated NF‑κB by down-regulating OGT in BV2 cells, indicating that the protective effect of PET against LPS-induced retinal microglia inflammatory response was achieved by regulating OGT/NF-κB/LCN2 axis.
Conclusion: Our findings may contribute to the potential clinical use of PET in treating DR and suggest OGT/NF-κB/LCN2 axis may be the potential therapeutic target of this disease.

Keywords:  diabetes mellitus; diabetic retinopathy; lipopolysaccharide

DOI:  https://doi.org/10.1515/med-2025-1274