{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T04:34:45Z","timestamp":1787027685971,"version":"build-2736575974"},"reference-count":43,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2024,5,10]],"date-time":"2024-05-10T00:00:00Z","timestamp":1715299200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"NIH","award":["R01EY029913"],"award-info":[{"award-number":["R01EY029913"]}]},{"name":"Jane and Roland Blumberg Professorship in Biology (AEC)","award":["R01EY029913"],"award-info":[{"award-number":["R01EY029913"]}]},{"name":"South Texas Center for Emerging Infectious Diseases Ph.D. scholarship (CR)","award":["R01EY029913"],"award-info":[{"award-number":["R01EY029913"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>Diabetic retinopathy (DR) affects over 140 million people globally. The mechanisms that lead to blindness are still enigmatic but there is evidence that sustained inflammation and hypoxia contribute to vascular damage. Despite efforts to understand the role of inflammation and microglia in DR\u2019s pathology, the contribution of astrocytes to hypoxic responses is less clear. To investigate the role of astrocytes in hypoxia-induced retinopathy, we utilized a 7-day systemic hypoxia model using the GFAP-CreERT2:Rosa26iDTR transgenic mouse line. This allows for the induction of inflammatory reactive astrogliosis following tamoxifen and diphtheria toxin administration. We hypothesize that DTx-induced astrogliosis is neuroprotective during hypoxia-induced retinopathy. Glial, neuronal, and vascular responses were quantified using immunostaining, with antibodies against GFAP, vimentin, IBA-1, NeuN, fibrinogen, and CD31. Cytokine responses were measured in both the brain and serum. We report that while both DTx and hypoxia induced a phenotype of reduced microglia morphological activation, DTx, but not hypoxia, induced an increase in the M\u00fcller glia marker vimentin. We did not observe that the combination of DTx and hypoxic treatments exacerbated the signs of reactive glial cells, nor did we observe a significant change in the expression immunomodulatory mediators IL-1\u03b2, IL2, IL-4, IL-5, IL-6, IL-10, IL-18, CCL17, TGF-\u03b21, GM-CSF, TNF-\u03b1, and IFN-\u03b3. Overall, our results suggest that, in this hypoxia model, reactive astrogliosis does not alter the inflammatory responses or cause vascular damage in the retina.<\/jats:p>","DOI":"10.3390\/biom14050567","type":"journal-article","created":{"date-parts":[[2024,5,14]],"date-time":"2024-05-14T08:59:37Z","timestamp":1715677177000},"page":"567","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Astrogliosis in the GFAP-CreERT2:Rosa26iDTR Mouse Model Does Not Exacerbate Retinal Microglia Activation or M\u00fcller Cell Gliosis under Hypoxic Conditions"],"prefix":"10.3390","volume":"14","author":[{"given":"Colin","family":"Rorex","sequence":"first","affiliation":[{"name":"Molecular Microbiology and Immunology, College of Sciences, The University of Texas at San Antonio, San Antonio, TX 78249, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9128-6742","authenticated-orcid":false,"given":"Sandra M.","family":"Cardona","sequence":"additional","affiliation":[{"name":"Molecular Microbiology and Immunology, College of Sciences, The University of Texas at San Antonio, San Antonio, TX 78249, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3982-8476","authenticated-orcid":false,"given":"Kaira A.","family":"Church","sequence":"additional","affiliation":[{"name":"Molecular Microbiology and Immunology, College of Sciences, The University of Texas at San Antonio, San Antonio, TX 78249, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1354-9735","authenticated-orcid":false,"given":"Derek","family":"Rodriguez","sequence":"additional","affiliation":[{"name":"Molecular Microbiology and Immunology, College of Sciences, The University of Texas at San Antonio, San Antonio, TX 78249, USA"},{"name":"Integrative Biology, The University of Texas at San Antonio, San Antonio, TX 78249, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Difernando","family":"Vanegas","sequence":"additional","affiliation":[{"name":"Molecular Microbiology and Immunology, College of Sciences, The University of Texas at San Antonio, San Antonio, TX 78249, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Reina","family":"Saldivar","sequence":"additional","affiliation":[{"name":"Molecular Microbiology and Immunology, College of Sciences, The University of Texas at San Antonio, San Antonio, TX 78249, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brianna","family":"Faz","sequence":"additional","affiliation":[{"name":"Integrative Biology, The University of Texas at San Antonio, San Antonio, TX 78249, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5093-8078","authenticated-orcid":false,"given":"Astrid E.","family":"Cardona","sequence":"additional","affiliation":[{"name":"Molecular Microbiology and Immunology, College of Sciences, The University of Texas at San Antonio, San Antonio, TX 78249, USA"},{"name":"South Texas Center for Emerging Infectious Diseases, The University of Texas at San Antonio, San Antonio, TX 78249, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,5,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"104517","DOI":"10.1016\/j.mvr.2023.104517","article-title":"Hypoxia-induced vascular remodeling responses in the brain are much more robust than other organs","volume":"148","author":"Sapkota","year":"2023","journal-title":"Microvasc. Res."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"218","DOI":"10.1038\/s41392-022-01080-1","article-title":"Hypoxia signaling in human health and diseases: Implications and prospects for therapeutics","volume":"7","author":"Luo","year":"2022","journal-title":"Signal Transduct. Target. Ther."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Shinojima, A., Lee, D., Tsubota, K., Negishi, K., and Kurihara, T. (2021). Retinal Diseases Regulated by Hypoxia-Basic and Clinical Perspectives: A Comprehensive Review. J. Clin. Med., 10.","DOI":"10.3390\/jcm10235496"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"70","DOI":"10.1080\/13510002.2022.2050086","article-title":"The evolving role of long noncoding RNA HIF1A-AS2 in diabetic retinopathy: A cross-link axis between hypoxia, oxidative stress and angiogenesis via MAPK\/VEGF-dependent pathway","volume":"27","author":"Atef","year":"2022","journal-title":"Redox Rep."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1167\/iovs.65.1.35","article-title":"Evaluation of Hypoxia and Microcirculation Factors in the Progression of Diabetic Retinopathy","volume":"65","author":"Majidova","year":"2024","journal-title":"Investig. Opthalmology Vis. Sci."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1007\/s10633-006-9038-5","article-title":"Hypobaric hypoxia reduces the amplitude of oscillatory potentials in the human ERG","volume":"114","author":"Benedek","year":"2007","journal-title":"Doc. Ophthalmol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"564077","DOI":"10.3389\/fimmu.2020.564077","article-title":"Microglia and Inflammatory Responses in Diabetic Retinopathy","volume":"11","author":"Kinuthia","year":"2020","journal-title":"Front. Immunol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"e00312","DOI":"10.1016\/j.neurot.2023.e00312","article-title":"Recombinant soluble form of receptor for advanced glycation end products ameliorates microcirculation impairment and neuroinflammation after subarachnoid hemorrhage","volume":"21","author":"Yang","year":"2024","journal-title":"Neurotherapeutics"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1016\/j.abb.2019.05.011","article-title":"STAT1 drives M1 microglia activation and neuroinflammation under hypoxia","volume":"669","author":"Butturini","year":"2019","journal-title":"Arch. Biochem. Biophys."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1122","DOI":"10.1097\/WNR.0000000000000236","article-title":"Hypoxia-inducible factor-1\u03b1 upregulation in microglia following hypoxia protects against ischemia-induced cerebral infarction","volume":"25","author":"Huang","year":"2014","journal-title":"Neuroreport"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"50","DOI":"10.1186\/s12987-021-00284-x","article-title":"The impact of chronic mild hypoxia on cerebrovascular remodelling; uncoupling of angiogenesis and vascular breakdown","volume":"18","author":"Halder","year":"2021","journal-title":"Fluids Barriers CNS"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1173","DOI":"10.1038\/s41590-023-01522-0","article-title":"Defining blood-induced microglia functions in neurodegeneration through multiomic profiling","volume":"24","author":"Mendiola","year":"2023","journal-title":"Nat. Immunol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2137","DOI":"10.1002\/glia.23467","article-title":"Prokineticin-2 promotes chemotaxis and alternative A2 reactivity of astrocytes","volume":"66","author":"Neal","year":"2018","journal-title":"Glia"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"5475","DOI":"10.1073\/pnas.1113363109","article-title":"Prokineticin 2 is an endangering mediator of cerebral ischemic injury","volume":"109","author":"Cheng","year":"2012","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1807","DOI":"10.1007\/s12035-023-03662-8","article-title":"Hypoxia-Induced miR-101 Impairs Endothelial Barrier Integrity Through Altering VE-Cadherin and Claudin-5","volume":"61","author":"Shukla","year":"2024","journal-title":"Mol. Neurobiol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"175","DOI":"10.1186\/s40478-020-01051-z","article-title":"Mild hypoxia triggers transient blood-brain barrier disruption: A fundamental protective role for microglia","volume":"8","author":"Halder","year":"2020","journal-title":"Acta Neuropathol. Commun."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"583687","DOI":"10.3389\/fimmu.2020.583687","article-title":"The Role of Inflammation in Diabetic Retinopathy","volume":"11","author":"Forrester","year":"2020","journal-title":"Front. Immunol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"917","DOI":"10.1038\/nn1715","article-title":"Control of microglial neurotoxicity by the fractalkine receptor","volume":"9","author":"Cardona","year":"2006","journal-title":"Nat. Neurosci."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1759091415608204","DOI":"10.1177\/1759091415608204","article-title":"Disruption of Fractalkine Signaling Leads to Microglial Activation and Neuronal Damage in the Diabetic Retina","volume":"7","author":"Cardona","year":"2015","journal-title":"ASN Neuro"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1130735","DOI":"10.3389\/fimmu.2023.1130735","article-title":"Pharmacological depletion of microglia alleviates neuronal and vascular damage in the diabetic CX3CR1-WT retina but not in CX3CR1-KO or hCX3CR1(I249\/M280)-expressing retina","volume":"14","author":"Church","year":"2023","journal-title":"Front. Immunol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"300","DOI":"10.1186\/s12974-022-02659-9","article-title":"Models of microglia depletion and replenishment elicit protective effects to alleviate vascular and neuronal damage in the diabetic murine retina","volume":"19","author":"Church","year":"2022","journal-title":"J. Neuroinflamm."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Rodriguez, D., Church, K.A., Smith, C.T., Vanegas, D., Cardona, S.M., Muzzio, I.A., Nash, K.R., and Cardona, A.E. (2024). Therapeutic Delivery of Soluble Fractalkine Ameliorates Vascular Dysfunction in the Diabetic Retina. Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms25031727"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Pawelec, P., Ziemka-Nalecz, M., Sypecka, J., and Zalewska, T. (2020). The Impact of the CX3CL1\/CX3CR1 Axis in Neurological Disorders. Cells, 9.","DOI":"10.3390\/cells9102277"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"419","DOI":"10.1038\/nmeth762","article-title":"A Cre-inducible diphtheria toxin receptor mediates cell lineage ablation after toxin administration","volume":"2","author":"Buch","year":"2005","journal-title":"Nat. Methods"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1555","DOI":"10.1038\/s41593-020-00735-y","article-title":"Severe reactive astrocytes precipitate pathological hallmarks of Alzheimer's disease via H2O2\u2212 production","volume":"23","author":"Chun","year":"2020","journal-title":"Nat. Neurosci."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"676","DOI":"10.1038\/nmeth.2019","article-title":"Fiji: An open-source platform for biological-image analysis","volume":"9","author":"Schindelin","year":"2012","journal-title":"Nat. Methods"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"269","DOI":"10.1002\/(SICI)1098-1136(199612)18:4<269::AID-GLIA2>3.0.CO;2-T","article-title":"Effects of GM-CSF and ordinary supplements on the ramification of microglia in culture: A morphometrical study","volume":"18","author":"Fujita","year":"1996","journal-title":"Glia"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"329","DOI":"10.1167\/iovs.03-0518","article-title":"Glial remodeling and neural plasticity in human retinal detachment with proliferative vitreoretinopathy","volume":"46","author":"Sethi","year":"2005","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Kugler, E.C., Greenwood, J., and MacDonald, R.B. (2021). The \"Neuro-Glial-Vascular\" Unit: The Role of Glia in Neurovascular Unit Formation and Dysfunction. Front. Cell Dev. Biol., 9.","DOI":"10.3389\/fcell.2021.732820"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"154","DOI":"10.1111\/j.1365-2567.2009.03225.x","article-title":"Inflammation in neurodegenerative diseases","volume":"129","author":"Amor","year":"2010","journal-title":"Immunology"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"3182","DOI":"10.1111\/ene.15978","article-title":"Serum and cerebrospinal fluid brain damage markers neurofilament light and glial fibrillary acidic protein correlate with tick-borne encephalitis disease severity-a multicentre study on Lithuanian and Swedish patients","volume":"30","author":"Veje","year":"2023","journal-title":"Eur. J. Neurol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1748","DOI":"10.1038\/s12276-021-00693-w","article-title":"M\u00fcller glial responses compensate for degenerating photoreceptors in retinitis pigmentosa","volume":"53","author":"Tomita","year":"2021","journal-title":"Exp. Mol. Med."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"2215","DOI":"10.1523\/JNEUROSCI.21-07-02215.2001","article-title":"Propagation of intercellular calcium waves in retinal astrocytes and M\u00fcller cells","volume":"21","author":"Newman","year":"2001","journal-title":"J. Neurosci."},{"key":"ref_34","first-page":"101","article-title":"Oxygen-induced retinopathy in the mouse","volume":"35","author":"Smith","year":"1994","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_35","first-page":"3500","article-title":"Retinal vascular development is mediated by endothelial filopodia, a preexisting astrocytic template and specific R-cadherin adhesion","volume":"43","author":"Aguilar","year":"2002","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"982","DOI":"10.1038\/nmeth.3125","article-title":"Neuronal morphometry directly from bitmap images","volume":"11","author":"Ferreira","year":"2014","journal-title":"Nat. Methods"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"815347","DOI":"10.3389\/fnagi.2022.815347","article-title":"Microglia Polarization From M1 to M2 in Neurodegenerative Diseases","volume":"14","author":"Guo","year":"2022","journal-title":"Front. Aging Neurosci."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"2699","DOI":"10.1172\/JCI39506","article-title":"Hypoxia-inducible factor 2alpha regulates macrophage function in mouse models of acute and tumor inflammation","volume":"120","author":"Imtiyaz","year":"2010","journal-title":"J. Clin. Investig."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"491","DOI":"10.1101\/gad.1881410","article-title":"Differential activation and antagonistic function of HIF-\u03b1 isoforms in macrophages are essential for NO homeostasis","volume":"24","author":"Takeda","year":"2010","journal-title":"Genes Dev."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"993","DOI":"10.1111\/febs.13572","article-title":"HIF3\u03b1: The little we know","volume":"283","author":"Ravenna","year":"2016","journal-title":"FEBS J."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"4","DOI":"10.1136\/svn-2022-001659","article-title":"CCL17 exerts neuroprotection through activation of CCR4\/mTORC2 axis in microglia after subarachnoid haemorrhage in rats","volume":"8","author":"Zhang","year":"2022","journal-title":"Stroke Vasc. Neurol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"3946320231177189","DOI":"10.1177\/03946320231177189","article-title":"Expression profile of cytokines and chemokines in a mouse high-altitude cerebral edema model","volume":"37","author":"Shi","year":"2023","journal-title":"Int. J. Immunopathol. Pharmacol."},{"key":"ref_43","unstructured":"Shukla, U.V., and Tripathy, K. (2024). StatPearls, StatPearls Publishing."}],"container-title":["Biomolecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2218-273X\/14\/5\/567\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T14:43:33Z","timestamp":1760107413000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2218-273X\/14\/5\/567"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,5,10]]},"references-count":43,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2024,5]]}},"alternative-id":["biom14050567"],"URL":"https:\/\/doi.org\/10.3390\/biom14050567","relation":{},"ISSN":["2218-273X"],"issn-type":[{"value":"2218-273X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,5,10]]}}}