Brillouin Microscopic Assessment of Retinal Stiffness During Reactive Müller Gliosis
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ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE302395 | GEO | 2026/02/23
REPOSITORIES: GEO
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Project description:Brillouin Microscopic Assessment of Retinal Stiffness During Reactive Müller Gliosis
| PRJNA1290320 | ENA
Project description:Purpose: Müller glia play dual roles in glaucoma, contributing to both retinal homeostasis and neuroinflammation; their activation by elevated intraocular pressure through the mechanosensitive channel TRPV4 promotes a reactive state that drives retinal ganglion cell (RGC) loss. Lipoxin B4 (LXB4), an endogenous lipid mediator produced by retinal astrocytes, has been shown to suppress glial reactivity and directly protect RGCs. This study investigated whether LXB4 modulates TRPV4-driven Müller glial activation and inflammation and whether Müller glia themselves contribute to this retinal lipoxin pathway. Methods: Ocular hypertension (OHT) was induced in mice via a silicone oil model, and reactive Müller glia were isolated via magnetic sorting for transcriptomic analysis. In vitro, primary and immortalized Müller glia were treated with a TRPV4 agonist with or without LXB4. Glial reactivity was assessed by flow cytometry, immunostaining, qPCR, and western blotting. LC–MS/MS-based lipidomics was used to quantify lipoxin pathway metabolites, and single-cell RNA-seq was used to examine transcriptional responses to LXB4 treatment. GFAP and TRPV4 expression was evaluated via immunohistochemistry in retinal sections. Results: RNA bulk-sequencing analysis and qPCR revealed that Müller glia express both 5- and 15-lipoxygenase. Lipidomic analysis confirmed that the lipoxin pathway is functional and that Müller glia endogenously produce LXB4, establishing this essential cell type as a source of anti-inflammatory and neuroprotective LXB4 in the retina. TRPV4 activation induced a reactive glial phenotype characterized by increased GFAP and IL6 expression, increased STAT3 phosphorylation, and increased production of lipoxin pathway metabolites, suggesting that biomechanical stress simultaneously triggers both gliosis and lipid signaling. Exogenous LXB4 suppressed TRPV4-induced gliosis in vitro by downregulating IL6 and inhibiting STAT3 activation, and in vivo treatment during OHT reduced the expression of Stat3, Il6, and TNF-α while attenuating TRPV4 upregulation in Müller glia. Conclusion: These findings identify a self-regulating lipid circuit in Müller glia and support the targeting of the TRPV4–lipoxin pathway to inhibit gliosis and protect against neurodegeneration in glaucoma.
2026-02-11 | GSE306785 | GEO
Project description:Irreversible loss of retinal neurons is the leading cause of blindness. Harnessing the latent stem cell potential of mammalian Müller glia offers a promising strategy for endogenous retinal regeneration. Shifting the Müller glia response from gliosis to regeneration after retinal injury is a critical step in promoting the orderly entry of Müller glia into a regenerative program. However, the molecular switch governing this divergent fate decision remains elusive. Here, through RNA-seq and scRNA-seq, we confirm the upstream determinant of Müller glia fate after retinal injury.
2026-01-18 | GSE316279 | GEO
Project description:Irreversible loss of retinal neurons is the leading cause of blindness. Harnessing the latent stem cell potential of mammalian Müller glia offers a promising strategy for endogenous retinal regeneration. Shifting the Müller glia response from gliosis to regeneration after retinal injury is a critical step in promoting the orderly entry of Müller glia into a regenerative program. However, the molecular switch governing this divergent fate decision remains elusive. Here, through RNA-seq and scRNA-seq, we confirm the upstream determinant of Müller glia fate after retinal injury.
2026-01-18 | GSE316280 | GEO
Project description:To address the function of Rax in differentiated Müller glial cells, we generated Rax tamoxifen-induced conditional knock-out (Rax iCKO) mice, where Rax can be depleted in mTFP-labeled Müller glial cells upon tamoxifen treatment, by crossing Raxflox/flox mice with Rlbp1-CreERT2 mice and found histological characteristic of reactive gliosis and an enhanced gliosis of Müller glial cells in the Rax iCKO retina under normal and stress conditions, respectively. RNA sequencing using isolated Müller glial cells from the Rax iCKO retina by fluorescence-activated cell sorting demonstrated that reduced expression of suppressor of cytokine signaling-3 (Socs3), whose depletion leads to reactive gliosis in Müller glial cells. Reporter gene assays showed that Rax directly transactivates Socs3 and we observed decreased expression of Socs3 in Müller glial cells of the Rax iCKO retina by immunostaining. Taken together, these results suggest that the Rax homeoprotein suppresses reactive gliosis in Müller glial cells by transactivating Socs3 in vivo. Our results may shed light on the transcriptional regulatory mechanisms underlying Müller glial cell homeostasis.
2023-12-20 | GSE237650 | GEO
Project description:PURPOSE. During retinal degeneration, Müller glia cells respond to photoreceptor loss by undergoing reactive gliosis, with both detrimental and beneficial effects. Increasing our knowledge of the complex molecular response of Müller cells to retinal degeneration is thus essential for the development of new therapeutic strategies. The purpose of this work was to identify new factors involved in Müller cell response to photoreceptor cell death. METHODS. Whole transcriptome sequencing was performed from wild-type and degenerating rd10 mouse retinas at P30. The changes in mRNA abundance for several deregulated genes were assessed by RT-qPCR. Protein expression level and retinal cellular localization were determined by western-blot and immunohistochemistry, respectively. RESULTS. Pathway-level analysis from whole transcriptomic data revealed the Hippo/YAP pathway as one of the main signaling pathways altered in response to photoreceptor degeneration in rd10 retinas. We found that downstream effectors of this pathway, YAP and TEAD1, are specifically expressed in Müller cells and that their expression, at both the mRNA and protein levels, is increased in rd10 reactive Müller glia after the onset of photoreceptor degeneration. The expression of Ctgf and Cyr61, two target genes of the transcriptional YAP/TEAD complex, is also upregulated following photoreceptor loss. CONCLUSIONS. This work reveals for the first time that YAP and TEAD1, key downstream effectors of the Hippo pathway, are specifically expressed in Müller cells. We also uncovered a deregulation of the expression and activity of Hippo/YAP pathway components in reactive Müller cells under pathological conditions.
2017-05-29 | GSE94534 | GEO
Project description:Contrasting with fish or amphibian, retinal regeneration from Müller glial cells is largely limited in mammals. In our quest towards the identification of molecular cues that may boost their stemness potential, we investigated the involvement of the Hippo pathway effector YAP, which we previously found to be upregulated in Müller cells following retinal injury. We report that conditional Yap deletion in Müller cells prevents the upregulation of cell cycle genes that normally accompanies reactive gliosis upon photoreceptor cell death. This occurs as a consequence of defective EGFR signaling. Consistent with a function of YAP in triggering Müller glia cell cycle re-entry, we further show that in Xenopus, a species endowed with efficient regenerative capacity, YAP is required for their injury-dependent proliferative response. Finally, and noteworthy, we reveal that YAP overactivation in mouse Müller cells is sufficient to induce their reprogramming into highly proliferative cells. Overall, we unravel a pivotal role for YAP in tuning Müller cell response to injury and highlight a novel YAP-EGFR axis by which Müller cells exit their quiescence state, a critical step towards regeneration.
2019-06-01 | GSE121858 | GEO
Project description:Rat Retinal Müller cells from diabetic rats (diabetes duration 6 months) compared to Rat Retinal Müller cells from healthy rats. Diabetes was induced by streptozotozine. Diabetic rats were treated with small doses of insulin to prevent catabolism. Keywords: ordered
2004-11-17 | E-GEOD-1979 | biostudies-arrayexpress
Project description:Purpose. Immune privilege in the eye is a compilation of anti-inflammatory mechanisms that protect vision from the damaging sequelae of intense immune responses. Although the breakdown of privilege can lead to ocular disease, little is known about how these mechanisms are regulated. Since retinal Müller glial cells and autophagy also have anti-inflammatory properties, we tested the idea that Müller cells utilize autophagy to support immune privilege. Methods. The essential autophagy regulator Atg5 was deleted in retinal Muller cells using a tamoxifen inducible GlastCre ERT X Atg5f/f strain. Intraocular inflammation was induced by intravitreal injection of LPS and monitored by H&E staining, immunofluorescent confocal microscopy, and flow cytometry. scRNA-seq was performed on retinal Müller cells isolated from control and Atg5-deficient mice. Additionally, markers of Müller cell gliosis and function were assessed by western blotting and cytokines were detected by Luminex cytokine/chemokine arrays. In cultured Müller cells siRNA knockdown techniques were used to examine the role of autophagy in regulation of LPS-induced inflammatory pathways. Results. We observed increased and prolonged intraocular inflammation when Müller cells were autophagy (Atg5) deficient. Müller cell gliosis (as measured by Gfap expression) was significantly increased and inflammation was sustained over 5 days post-LPS injection compared to control. The retinae with Atg5-deficient Müller cells also contained increased inflammatory mediators and neuroprotective molecules. Gene expression analysis revealed a heterogeneous response to LPS in Müller glial cell population that revealed 2 states of activation. The normal retinae contain Müller cells in both a basal and an activated state, while autophagy deficient retinae (Atg5iΔMüller) contained only activated Müller cells. Analysis of gliosis markers Gfap and Lcn2 confirmed this heterogeneity showing that in control eyes basal and activated (gliotic) Müller glia were observed; however, with autophagy deficiency nearly all Müller cells were gliotic. Interestingly, activated cells were largely indistinguishable between autophagy sufficient and deficient Müller cells. In cultured Müller cells, siRNA knockdown of autophagy genes resulted in heightened mTOR activation, increased Gfap expression, and upregulated cytokine/chemokine production. Conclusions. Autophagy deficiency in Müller cells leads to enhanced intraocular inflammation though activation of the mTOR pathway. Our data suggests that autophagy defines a novel perspective on Müller glial heterogeneity based on their activation state. Thus, autophagy restrains cellular activation and inflammation, supporting immune privilege by preventing excessive and potentially destructive immune responses.
2025-09-10 | GSE293132 | GEO
Project description:The prevalence of myopia is rapidly increasing, significantly impacting the quality of life for affected individuals. Prior research in our group observed reactive gliosis in Müller cells within myopia-induced retinas, prompting further investigation into their role in myopia. In this study, we analyzed protein expression changes in CD29+ Müller cells, isolated using magnetic activated cell sorting (MACS), from form deprivation myopia-induced rabbit retina. These cells, the principal glial cells in the retina, showed significant alterations in metabolic pathways, particularly in glycolysis and angiogenesis, including the upregulation of glycolytic enzymes such as lactate dehydrogenase A and pyruvate kinase. These enzymes are implicated in adapting to increased metabolic demands under myopic stress. Additionally, a decrease in proteins associated with oxygen transport suggests an enhanced vulnerability to oxidative stress. Our findings highlight the proactive role of CD29+ Müller cells in modifying the retinal environment in response to myopic stress, suggesting potential therapeutic targets to mitigate myopia’s progression.
2025-05-07 | PXD053804 | Pride