{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T02:21:20Z","timestamp":1760235680392,"version":"build-2065373602"},"reference-count":50,"publisher":"MDPI AG","issue":"19","license":[{"start":{"date-parts":[[2021,9,22]],"date-time":"2021-09-22T00:00:00Z","timestamp":1632268800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The retina is a complex neurological tissue and is extremely sensitive to an insufficient supply of oxygen. Hypoxia plays a major role in several retinal diseases, and often results in the loss of cells that are essential for vision. Cyclosporine A (CsA) is a widely used immunosuppressive drug. Furthermore, treatment with CsA has neuroprotective effects in several neurologic disorders. No data are currently available on the tolerated concentration of CsA when applied to the retina. To reveal the most effective dose, retinal explants from rat eyes were exposed to different CsA concentrations (1\u20139 \u00b5g\/mL). Immunohistochemistry with brain-specific homeobox\/POU domain protein 3a (Brn3a) and TUNEL staining was performed to determine the percentage of total and apoptotic retinal ganglion cells (RGCs), as well as the responses of micro- and macroglial cells. Furthermore, optical coherence tomography (OCT) scans were performed to measure the changes in retinal thickness, and recordings with multielectrode array (MEA) were performed to evaluate spontaneous RGC spiking. To examine the neuroprotective effects, retinas were subjected to a hypoxic insult by placing them in a nitrogen-streamed hypoxic chamber prior to CsA treatment. In the biocompatibility tests, the different CsA concentrations had no negative effect on RGCs and microglia. Neuroprotective effects after a hypoxic insult on RGCs was demonstrated at a concentration of 9 \u00b5g\/mL CsA. CsA counteracted the hypoxia-induced loss of RGCs, reduced the percentage of TUNEL+ RGCs, and prevented a decrease in retinal thickness. Taken together, the results of this study suggest that CsA can effectively protect RGCs from hypoxia, and the administered concentrations were well tolerated. Further in vivo studies are needed to determine whether local CsA treatment may be a suitable option for hypoxic retinal diseases.<\/jats:p>","DOI":"10.3390\/ijms221910196","type":"journal-article","created":{"date-parts":[[2021,9,24]],"date-time":"2021-09-24T20:15:32Z","timestamp":1632514532000},"page":"10196","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Cyclosporine A Protects Retinal Explants against Hypoxia"],"prefix":"10.3390","volume":"22","author":[{"given":"Sven","family":"Schnichels","sequence":"first","affiliation":[{"name":"Centre for Ophthalmology T\u00fcbingen, University Eye Hospital T\u00fcbingen, 72076 T\u00fcbingen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maximilian","family":"Schultheiss","sequence":"additional","affiliation":[{"name":"Clinic for Ophthalmology, University Medical Center Hamburg-Eppendorf (UKE), 20246 Hamburg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Patricia","family":"Klemm","sequence":"additional","affiliation":[{"name":"Centre for Ophthalmology T\u00fcbingen, University Eye Hospital T\u00fcbingen, 72076 T\u00fcbingen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Matthias","family":"Blak","sequence":"additional","affiliation":[{"name":"Centre for Ophthalmology T\u00fcbingen, University Eye Hospital T\u00fcbingen, 72076 T\u00fcbingen, Germany"},{"name":"Department of Ophthalmology, Klinikum Stuttgart, 70174 Stuttgart, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Thoralf","family":"Herrmann","sequence":"additional","affiliation":[{"name":"NMI Natural and Medical Sciences Institute, University of T\u00fcbingen, 72770 Reutlingen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marion","family":"Melchinger","sequence":"additional","affiliation":[{"name":"Centre for Ophthalmology T\u00fcbingen, University Eye Hospital T\u00fcbingen, 72076 T\u00fcbingen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Karl-Ulrich","family":"Bartz-Schmidt","sequence":"additional","affiliation":[{"name":"Centre for Ophthalmology T\u00fcbingen, University Eye Hospital T\u00fcbingen, 72076 T\u00fcbingen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marina","family":"L\u00f6scher","sequence":"additional","affiliation":[{"name":"Centre for Ophthalmology T\u00fcbingen, University Eye Hospital T\u00fcbingen, 72076 T\u00fcbingen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"G\u00fcnther","family":"Zeck","sequence":"additional","affiliation":[{"name":"Institute of Electrodynamics, Microwave and Circuit Engineering, TU Wien, 1040 Vienna, Austria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4588-1767","authenticated-orcid":false,"given":"Martin Stehphan","family":"Spitzer","sequence":"additional","affiliation":[{"name":"Centre for Ophthalmology T\u00fcbingen, University Eye Hospital T\u00fcbingen, 72076 T\u00fcbingen, Germany"},{"name":"Clinic for Ophthalmology, University Medical Center Hamburg-Eppendorf (UKE), 20246 Hamburg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8697-2569","authenticated-orcid":false,"given":"Jos\u00e9","family":"Hurst","sequence":"additional","affiliation":[{"name":"Centre for Ophthalmology T\u00fcbingen, University Eye Hospital T\u00fcbingen, 72076 T\u00fcbingen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,9,22]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1016\/j.preteyeres.2003.12.001","article-title":"Retinal ischemia: Mechanisms of damage and potential therapeutic strategies","volume":"23","author":"Osborne","year":"2004","journal-title":"Prog. Retin. Eye Res."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"S11","DOI":"10.1038\/sj.eye.6702880","article-title":"The neurobiology of cell death in glaucoma","volume":"21","author":"Vrabec","year":"2007","journal-title":"Eye"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"971","DOI":"10.1007\/s00125-009-1655-6","article-title":"Neurodegenerative influence of oxidative stress in the retina of a murine model of diabetes","volume":"53","author":"Sasaki","year":"2010","journal-title":"Diabetologia"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"124","DOI":"10.1016\/S0140-6736(09)62124-3","article-title":"Diabetic retinopathy","volume":"376","author":"Cheung","year":"2010","journal-title":"Lancet"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1016\/j.maturitas.2015.10.005","article-title":"Age-related eye disease and gender","volume":"83","author":"Zetterberg","year":"2016","journal-title":"Maturitas"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1684","DOI":"10.4103\/ijo.IJO_1446_18","article-title":"Central retinal artery occlusion","volume":"66","author":"Hayreh","year":"2018","journal-title":"Indian J. Ophthalmol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1901","DOI":"10.1001\/jama.2014.3192","article-title":"The pathophysiology and treatment of glaucoma: A review","volume":"311","author":"Weinreb","year":"2014","journal-title":"JAMA"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"692609","DOI":"10.1155\/2012\/692609","article-title":"Animal Models of Glaucoma","volume":"2012","author":"Bouhenni","year":"2012","journal-title":"J. Biomed. Biotechnol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"281214","DOI":"10.1155\/2015\/281214","article-title":"Experimentally Induced Mammalian Models of Glaucoma","volume":"2015","author":"Ishikawa","year":"2015","journal-title":"BioMed Res. Int."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1016\/j.optom.2016.02.003","article-title":"Glaucoma history and risk factors","volume":"10","author":"McMonnies","year":"2017","journal-title":"J. Optom."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1177\/026119291704500105","article-title":"A novel porcine ex vivo retina culture model for oxidative stress induced by H(2)O(2)","volume":"45","author":"Hurst","year":"2017","journal-title":"Altern. Lab Anim."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"557","DOI":"10.1177\/026119291604400608","article-title":"The novel induction of retinal ganglion cell apoptosis in porcine organ culture by NMDA\u2014An opportunity for the replacement of animals in experiments","volume":"44","author":"Kuehn","year":"2016","journal-title":"Altern. Lab. Anim."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1016\/j.exer.2017.01.003","article-title":"Degenerative effects of cobalt-chloride treatment on neurons and microglia in a porcine retina organ culture model","volume":"155","author":"Kuehn","year":"2017","journal-title":"Expe. Eye Res."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"32","DOI":"10.1016\/j.brainres.2004.05.056","article-title":"Combined cyclosporine-A and methylprednisolone treatment exerts partial and transient neuroprotection against ischemic stroke","volume":"1018","author":"Yu","year":"2004","journal-title":"Brain Res."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"58","DOI":"10.1016\/j.expneurol.2010.06.009","article-title":"Evaluation of cyclosporine A in a stroke model in the immature rat brain","volume":"230","author":"Leger","year":"2011","journal-title":"Exp. Neurol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1254\/jphs.FP0050799","article-title":"Effect of cyclosporin a on immediate early gene in rat global ischemia and its neuroprotection","volume":"100","author":"Yamaguchi","year":"2006","journal-title":"J. Pharmacol. Sci."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"226","DOI":"10.1006\/exnr.1999.7197","article-title":"Cyclosporin A attenuates acute mitochondrial dysfunction following traumatic brain injury","volume":"160","author":"Sullivan","year":"1999","journal-title":"Exp. Neurol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"411","DOI":"10.1517\/14740330903066742","article-title":"Cyclosporine A for neuroprotection: Establishing dosing guidelines for safe and effective use","volume":"8","author":"Cook","year":"2009","journal-title":"Expert Opin. Drug Saf."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"783","DOI":"10.1089\/neu.1999.16.783","article-title":"Cyclosporin A significantly ameliorates cortical damage following experimental traumatic brain injury in rodents","volume":"16","author":"Scheff","year":"1999","journal-title":"J. Neurotrauma"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"3686","DOI":"10.1073\/pnas.89.9.3686","article-title":"Calcineurin phosphatase activity in T lymphocytes is inhibited by FK 506 and cyclosporin A","volume":"89","author":"Fruman","year":"1992","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"359","DOI":"10.1016\/j.npep.2011.04.002","article-title":"Cyclosporine-A as a neuroprotective agent against stroke: Its translation from laboratory research to clinical application","volume":"45","author":"Osman","year":"2011","journal-title":"Neuropeptides"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"6125","DOI":"10.1523\/JNEUROSCI.16-19-06125.1996","article-title":"Mitochondrial dysfunction is a primary event in glutamate neurotoxicity","volume":"16","author":"Schinder","year":"1996","journal-title":"J. Neurosci."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"677","DOI":"10.1046\/j.1471-4159.1998.70020677.x","article-title":"N-methyl-D-aspartate receptor desensitisation is neuroprotective by inhibiting glutamate-induced apoptotic-like death","volume":"70","author":"Wood","year":"1998","journal-title":"J. Neurochem."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1016\/S0014-2999(00)00584-7","article-title":"Cyclosporin A targets involved in protection against glutamate excitotoxicity","volume":"404","author":"Ruiz","year":"2000","journal-title":"Eur. J. Pharmacol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"e1105","DOI":"10.1038\/cddis.2014.80","article-title":"Inhibition of cyclophilin D by cyclosporin A promotes retinal ganglion cell survival by preventing mitochondrial alteration in ischemic injury","volume":"5","author":"Kim","year":"2014","journal-title":"Cell Death Dis."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"219","DOI":"10.1097\/IJG.0000000000000040","article-title":"Cyclosporine a protects RGC-5 cells from excitotoxic cell death","volume":"23","author":"Schultheiss","year":"2014","journal-title":"J. Glaucoma"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1007\/978-1-4614-3209-8_19","article-title":"What is the nature of the RGC-5 cell line?","volume":"801","author":"Sippl","year":"2014","journal-title":"Adv. Exp. Med. Biol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"5712","DOI":"10.1167\/iovs.13-12085","article-title":"A forensic path to RGC-5 cell line identification: Lessons learned","volume":"54","author":"Krishnamoorthy","year":"2013","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"100880","DOI":"10.1016\/j.preteyeres.2020.100880","article-title":"Retina in a dish: Cell cultures, retinal explants and animal models for common diseases of the retina","volume":"81","author":"Schnichels","year":"2020","journal-title":"Prog. Retin. Eye Res."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1056","DOI":"10.1242\/bio.025429","article-title":"Establishment of a retinal hypoxia organ culture model","volume":"6","author":"Schnichels","year":"2017","journal-title":"Biol. Open"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1043","DOI":"10.1111\/ceo.13565","article-title":"Hypothermia protects retinal ganglion cells against hypoxia-induced cell death in a retina organ culture model","volume":"47","author":"Klemm","year":"2019","journal-title":"Clin. Exp. Ophthalmol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1016\/j.drudis.2007.11.002","article-title":"Current and future ophthalmic drug delivery systems: A shift to the posterior segment","volume":"13","author":"Urtti","year":"2008","journal-title":"Drug Discov. Today"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"5151","DOI":"10.1523\/JNEUROSCI.18-14-05151.1998","article-title":"Cyclosporin A, but not FK 506, protects mitochondria and neurons against hypoglycemic damage and implicates the mitochondrial permeability transition in cell death","volume":"18","author":"Friberg","year":"1998","journal-title":"J. Neurosci."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1016\/0006-8993(92)91465-Q","article-title":"Cyclosporin A protects against ischemia-reperfusion injury in the brain","volume":"595","author":"Shiga","year":"1992","journal-title":"Brain Res."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"997","DOI":"10.1038\/sj.bjp.0704177","article-title":"Treatment of hippocampal neurons with cyclosporin A results in calcium overload and apoptosis which are independent on NMDA receptor activation","volume":"133","author":"Kaminska","year":"2001","journal-title":"Br. J. Pharmacol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1016\/j.exer.2016.05.023","article-title":"N-methyl-N-nitrosourea-induced neuronal cell death in a large animal model of retinal degeneration in vitro","volume":"148","author":"Taylor","year":"2016","journal-title":"Exp. Eye Res."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"522","DOI":"10.1016\/j.brainres.2016.06.039","article-title":"Human neural progenitor cells decrease photoreceptor degeneration, normalize opsin distribution and support synapse structure in cultured porcine retina","volume":"1646","author":"Mollick","year":"2016","journal-title":"Brain Res."},{"key":"ref_38","first-page":"1","article-title":"Cyclosporine A Reduces Glial Scarring and Facilitates Functional Recovery Following Transient Focal Ischemia","volume":"2015","author":"Abeysinghe","year":"2015","journal-title":"J. Neurol. Neurophysiol."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Schultheiss, M., Schnichels, S., Hermann, T., Hurst, J., Feldkaemper, M., Arango-Gonzalez, B., Ueffing, M., Bartz-Schmidt, K.U., Zeck, G., and Spitzer, M.S. (2016). Hypothermia Protects and Prolongs the Tolerance Time of Retinal Ganglion Cells against Ischemia. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0148616"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"861","DOI":"10.1038\/nrn2735","article-title":"Plasticity during stroke recovery: From synapse to behaviour","volume":"10","author":"Murphy","year":"2009","journal-title":"Nat. Rev. Neurosci."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1161\/01.STR.8.1.51","article-title":"Cortical evoked potential and extracellular K+ and H+ at critical levels of brain ischemia","volume":"8","author":"Astrup","year":"1977","journal-title":"Stroke"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"64","DOI":"10.3171\/spi.2006.4.1.64","article-title":"A comparison of adenosine A2A agonism and methylprednisolone in attenuating neuronal damage and improving functional outcome after experimental traumatic spinal cord injury in rabbits","volume":"4","author":"Okonkwo","year":"2006","journal-title":"J. Neurosurg. Spine"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"4929","DOI":"10.1074\/jbc.M309767200","article-title":"Critical role of calpain-mediated cleavage of calcineurin in excitotoxic neurodegeneration","volume":"279","author":"Wu","year":"2004","journal-title":"J. Biol. Chem."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"461","DOI":"10.1038\/nature02229","article-title":"The ADP\/ATP translocator is not essential for the mitochondrial permeability transition pore","volume":"427","author":"Kokoszka","year":"2004","journal-title":"Nature"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"607","DOI":"10.1084\/jem.193.5.607","article-title":"Selective inhibition of vascular endothelial growth factor-mediated angiogenesis by cyclosporin A: Roles of the nuclear factor of activated T cells and cyclooxygenase 2","volume":"193","author":"Hernandez","year":"2001","journal-title":"J. Exp. Med."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"15809","DOI":"10.1073\/pnas.95.26.15809","article-title":"Systemic hypoxia changes the organ-specific distribution of vascular endothelial growth factor and its receptors","volume":"95","author":"Marti","year":"1998","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"707","DOI":"10.1172\/JCI17423","article-title":"Glomerular-specific alterations of VEGF-A expression lead to distinct congenital and acquired renal diseases","volume":"111","author":"Eremina","year":"2003","journal-title":"J. Clin. Investig."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1016\/j.exer.2016.04.011","article-title":"Ex-vivo-examination of ultrastructural changes in organotypic retina culture using near-infrared imaging and optical coherence tomography","volume":"147","author":"Schnichels","year":"2016","journal-title":"Exp. Eye Res."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"2777","DOI":"10.1167\/iovs.13-13372","article-title":"Loss of inner retinal neurons after retinal ischemia in rats","volume":"55","author":"Schmid","year":"2014","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"e45","DOI":"10.1093\/nar\/29.9.e45","article-title":"A new mathematical model for relative quantification in real-time RT-PCR","volume":"29","author":"Pfaffl","year":"2001","journal-title":"Nucleic Acids Res."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/19\/10196\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:03:16Z","timestamp":1760166196000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/19\/10196"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,9,22]]},"references-count":50,"journal-issue":{"issue":"19","published-online":{"date-parts":[[2021,10]]}},"alternative-id":["ijms221910196"],"URL":"https:\/\/doi.org\/10.3390\/ijms221910196","relation":{},"ISSN":["1422-0067"],"issn-type":[{"type":"electronic","value":"1422-0067"}],"subject":[],"published":{"date-parts":[[2021,9,22]]}}}