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The mixture was then transferred in a Precellys-Tube for homogenization using the Precellys Tissue Homogenizer tubes filled with small balls and 2 big  ceramic beads (Soft tissue homogenizing CK14,  – 0,2 mL; Bertin, Montigny-le-Bretonneux, FR) for tissue disruption. The samples were homogenized by shaking twice for 20 seconds at 5500 cycles per minute in a Precellys 24 homogenizer (Bertin, Montigny-le-Bretonneux, FR) . Lysates were centrifuged for 10 minutes at 10 000g and supernatant transferred into a fresh tube. Protein content was determined by Bradford assay (Biorad, Hercules, CA, USA). Equal amounts of tissue lysates were subjected to tryptic digest applying a modified filter aided sample preparation (FASP) procedure as described in (32). Peptides were collected by centrifugation (10 min at 14 000 g), acidified with trifluoroacetic acid (TFA), and stored at -20°C.The MS data were acquired in DIA mode on a Q Exactive HF mass spectrometer (Thermo Fisher Scientific Inc., Waltham, MA, USA). Equal amounts of peptides were automatically loaded to the online coupled RSLC (Ultimate 3000, Thermo Fisher Scientific Inc.) HPLC system. A Nano-Trap column was used (300-µm inner diameter (ID) × 5 mm, packed with Acclaim PepMap100 C18, 5µm, 100 Å from LC Packings, Sunnyvale, CA, USA, before separation by reversed-phase chromatography (Acquity UPLC M-Class HSS T3 Column 75µm ID × 250 mm, 1.8µm from Waters, Eschborn, Germany) at 40°C. Peptides were eluted from the column at 250 nl/min using increasing ACN concentration in 0.1% formic acid from 3 to 40% over a 90-min gradient. The DIA method consisted of a survey scan from 300 to 1650 m/z at 120000 resolution and an automatic gain control (AGC) target of 3e6 or 100 ms maximum injection time. Fragmentation was performed via higher-energy collisional dissociation with a target value of 3e6 ions determined with predictive AGC. Precursor peptides were isolated with 37 variable windows spanning from 300 to 1650 m/z at 30000 resolution with an AGC target of 3e6 and automatic injection time. The normalized collision energy was 28, and the spectra were recorded in profile type."],"repository":["pride"],"quantification_method":[""],"modification":[""],"data_protocol":["DIA files were processed with Spectronaut (Version 15, Biognosys) as direct DIA analysis against a SwissProt mouse database (17081 sequences; release 2020_02 including spike proteins), using BSG factory settings for Pulsar with variable modifications set to acetylation of protein N-terminus, deamidation (NQ) and Oxidation (M). For DIA analysis, default settings were applied with the following changes: For quantification, precursor filtering was set on Qvalue and proteotypicity filter was set on only proteotypic, LFQ method was set to Quant 2.0 algorithm, quantity MS level was MS2, quantity type was area, cross run normalization was set to local normalization and major group quantity was calculated as mean TOP3 peptide quantity. Data were exported without cross run normalization for further analysis.  All subsequent analyses were conducted in R v4.2.1. After removal of non-unique identifiers, the raw protein LFQ intensities of 3809 proteins were normalized using a variance stabilizing normalization (vsn) (33). We used euclidian distance and principal component analysis to analyze sample relationships and defined one   outlier sample, which was removed from the following analysis. This left N = 6 for both light-damage groups and N = 5 for both dark-adapted groups. Next, we corrected for the sex of the animals using limma (34). Dysregulation was computed using DEqMS (35) with a contrast table and the number of peptides per protein as a coefficient during model fitting of the vsn-normalized, sex-corrected LFQ intensities. DEqMS analysis resulted in a total of 3729 proteins, as some proteins were removed during the analysis due to too many missing data points per group."],"omics_type":["Proteomics"],"labhead":["Barbara M. Braunger"],"instrument_platform":[""],"submission_type":["PARTIAL"],"labhead_affiliation":["Barbara M. Braunger, Institute of Neuroanatomy, University Clinic Hamburg-Eppendorf, Hamburg, Germany"],"species":["Mus Musculus (mouse)"],"publication":["42276988 Bielmeier CB, Schmitt SI, Lehr V, Grundl A, Dillinger AE, Jägle H, von Toerne C, Hauck SM, Ergün S, Tamm ER, Schlecht A, Neueder A, Braunger BM. VEGFR2 deletion increases susceptibility to photoreceptor degeneration through glial-neuronal interaction. Cell Death Dis. 2026 17(1):564 10.1038/s41419-026-08963-z"],"submitter_mail":["vontoerne@helmholtz-muenchen.de"],"submitter_affiliation":["Helmholtz Zentrum München"],"submitter_country":["Germany"],"pubmed_abstract":["Intraocular anti-VEGFA injections are frequently used to counteract neovascularization in diseases such as the neovascular form of age-related macular degeneration (AMD). However, in the clinical context, patients with atrophic (dry) and neovascular (wet) AMD in the same eye will receive anti-VEGFA injections, but there are reports that this severely promotes the development of atrophic areas in dry AMD. To study this, we used mice with a tamoxifen-dependent deletion of Vegfr2 in the eye (Vegfr2<sup>Δeye</sup>) and the light damage model to mimic certain aspects of dry AMD. We examined retinal morphology and function, the degree of photoreceptor degeneration, and alterations of the retinal proteome. While steady-state retinal morphology and function were not altered due to VEGF signaling deficiency, light-induced photoreceptor degeneration was drastically exacerbated in Vegfr2<sup>Δeye</sup> retinae, concomitant with attenuated activation of the AKT kinase pathway. Furthermore, using single nuclei RNA sequencing data, we showed that in humans, Vegfr2 is predominantly expressed in macroglial cells of healthy, dry- and wet AMD retinae. This suggests a VEGF-dependent neuroprotective crosstalk from macroglial cells to photoreceptors and poses promising therapeutic options to attenuate photoreceptor degeneration in humans. Yet, our data also indicate that anti-VEGF therapy should be carefully considered in the presence of neurodegenerative comorbidities."],"pubmed_title":["VEGFR2 deletion increases susceptibility to photoreceptor degeneration through glial-neuronal interaction."],"pubmed_authors":["Bielmeier Christina B CB, Schmitt Sabrina I SI, Lehr Verena V, Grundl Anita A, Dillinger Andrea E AE, Jägle Herbert H, von Toerne Christine C, Hauck Stefanie M SM, Ergün Süleyman S, Tamm Ernst R ER, Schlecht Anja A, Neueder Andreas A, Braunger Barbara M BM"],"additional_accession":[]},"is_claimable":false,"name":"LC-MSMS on mouse retina to investigate photoreceptor degeneration","description":"Intraocular anti-VEGFA injections are frequently used to counteract neovascularization in diseases such as the neovascular (wet) form of age-related macular degeneration (AMD) or proliferative diabetic retinopathy. However, in the clinical context, patients with atrophic (dry) and wet AMD in the same eye will receive anti-VEGFA injections, but there are reports that this severely promotes the development of atrophic areas in dry AMD. To study this, we used mice with a tamoxifen-dependent deletion of Vegfr2 in the eye (Vegfr2Δeye) and the light damage model to induce degeneration of photoreceptors and to mimic certain aspects of dry AMD. We examined retinal morphology and function, the degree of photoreceptor degeneration, as well as alterations of the retinal proteome. While, steady-state retinal morphology and function was not altered due to VEGF signaling deficiency, light-induced photoreceptor degeneration was drastically exacerbated in VEGF signaling deficient retinae concomitant with attenuated activation of the AKT kinase pathway. Furthermore, using single nuclei RNA sequencing data, we show that in humans Vegfr2 is predominantly expressed in macroglia cells of healthy retinae, dry and wet AMD patients, suggesting a VEGF-dependent neuroprotective crosstalk from macroglia cells to photoreceptors which might pose promising therapeutic options to attenuate photoreceptor degeneration in humans. Yet, our data also indicate that the use of anti-VEGF therapy should be carefully considered in the presence of neurodegenerative comorbidities, since deletion of VEGF signaling dramatically promoted neurodegeneration","dates":{"publication":"2026-06-15","submission":"2025-07-17"},"accession":"PXD066263","cross_references":{"TAXONOMY":["NEWT:108018","NEWT:377960","NEWT:1131","NEWT:190486","NEWT:2737160","NEWT:2042546","NEWT:148942","NEWT:295546","NEWT:258594","NEWT:1126","NEWT:1129","NEWT:89187","NEWT:309807","NEWT:89184","NEWT:309800","NEWT:156563","NEWT:1323429","NEWT:876138","NEWT:44271","NEWT:193516","NEWT:2315688","NEWT:44270","NEWT:592321","NEWT:1073882","NEWT:6819","NEWT:93217","NEWT:428386","NEWT:1117","NEWT:498257","NEWT:10036","NEWT:32264","NEWT:10034","NEWT:661410","NEWT:488003","NEWT:224326","NEWT:376619","NEWT:3765","NEWT:2433","NEWT:3767","NEWT:433727","NEWT:135622","NEWT:672127","NEWT:656061","NEWT:37965","NEWT:656064","NEWT:3760","NEWT:36630","NEWT:381198","NEWT:1176302","NEWT:418106","NEWT:10029","NEWT:913645","NEWT:2492962","NEWT:641809","NEWT:44250","NEWT:77151","NEWT:317447","NEWT:698761","NEWT:5530","NEWT:868565","NEWT:135674","NEWT:1562698","NEWT:1177","NEWT:11308","NEWT:12637","NEWT:2164133","NEWT:1180","NEWT:2767846","NEWT:1343074","NEWT:314344","NEWT:108061","NEWT:224308","NEWT:3134131","NEWT:49928","NEWT:160621","NEWT:110368","NEWT:1916073","NEWT:1303873","NEWT:1310161","NEWT:1230383","NEWT:77133","NEWT:160620","NCBITaxon:77643","NEWT:1310165","NEWT:114727","NEWT:5516","NEWT:498211","NEWT:347256","NEWT:5518","NEWT:398007","NEWT:1527468","NEWT:498217","NEWT:498216","NEWT:11320","NEWT:1410384","NEWT:1310173","NEWT:1072569","NEWT:1072568","NEWT:161934","NEWT:156587","NEWT:1148","NEWT:5508","NEWT:135666","NEWT:419481","NEWT:5507","NEWT:1140","NCBITaxon:2157","NEWT:2066563","NEWT:1143","NEWT:1336794","NEWT:1462472","NEWT:1336795","NEWT:644042","NEWT:709964","NEWT:3710","NEWT:3712","NEWT:3711","NEWT:3714","NEWT:65121","NEWT:3716","NEWT:3715","NEWT:2065263","NEWT:1549675","NEWT:177437","NEWT:9014","NEWT:10891","NEWT:118221","NEWT:34869","NEWT:226126","NEWT:1616117","NEWT:34865","NEWT:28174","NEWT:550537","NEWT:284812","NEWT:284811","NEWT:2902907","NEWT:3701","NEWT:3702","NEWT:1245466","NEWT:3704","NEWT:1246791","NEWT:34875","NEWT:10882","NEWT:1431297","NEWT:9005","NEWT:34871","NEWT:9006","NEWT:33548","NEWT:1883426","NEWT:657321","NEWT:1423687","NEWT:657322","NEWT:10886","NEWT:3707","NEWT:3708","NEWT:536231","NEWT:219813","NEWT:1301262","NEWT:319224","NEWT:2305479","NEWT:215451","NEWT:460519","NEWT:272843","NEWT:272844","NEWT:1348799","NEWT:55784","NEWT:485","NEWT:223074","NEWT:2499525","NEWT:467144","NEWT:142786","NEWT:28196","NEWT:2853422","NEWT:487","NEWT:1679718","NEWT:387344","NEWT:657318","NEWT:480","NEWT:67767","NEWT:46835","NEWT:216778","NEWT:376686","NEWT:237610","EFO:0001352","NEWT:219809","NEWT:219808","NEWT:119562","NEWT:1883446","NEWT:1233435","NEWT:118223","NEWT:141469","NEWT:272831","NEWT:987059","NEWT:491","NEWT:422203","NEWT:460523","NEWT:2423","NEWT:85694","NEWT:85692","NEWT:911045","NEWT:3750","NEWT:123606","NEWT:85698","NEWT:291146","NEWT:233271","NEWT:3745","NEWT:1715989","NCBITaxon:4751","NEWT:3747","NEWT:278161","NEWT:415978","NEWT:410289","NEWT:191701","NEWT:9054","NEWT:472","NEWT:29459","NEWT:34839","NEWT:476","NEWT:1380096","NEWT:28128","NEWT:470","NEWT:41122","NEWT:1328300","NEWT:748693","NEWT:106299","NCBITaxon:50557","NEWT:63363","NEWT:3735","NEWT:29491","NEWT:911022","NEWT:446","NEWT:153481","NEWT:714485","NEWT:445","NEWT:2003758","NEWT:29486","NEWT:63366","NEWT:64697","NEWT:63367","NEWT:29488","NEWT:215402","NEWT:3726","NEWT:1862567","NEWT:591090","NEWT:30455","NEWT:28141","NEWT:9030","NEWT:9031","NEWT:3728","NEWT:108931","NEWT:219799","NEWT:1325292","NEWT:219798","NEWT:548","NEWT:150475","NEWT:546","NEWT:1431340","NEWT:256737","NEWT:51350","NEWT:8694","NEWT:8692","NEWT:542","NEWT:394503","NEWT:13076","NEWT:219785","NEWT:451515","NEWT:8673","NEWT:550","NEWT:10804","NEWT:52694","NEWT:554","NEWT:451516","NEWT:552","NEWT:1325291","NEWT:42410","NEWT:40686","NEWT:1225786","NEWT:771870","NEWT:8665","NEWT:51338","NEWT:377142","NEWT:9995","NEWT:28112","NEWT:49012","NEWT:28110","NEWT:10832","NEWT:311339","NEWT:507601","NEWT:520","NEWT:64657","NEWT:186441","NEWT:2585209","NEWT:643680","NEWT:214092","NEWT:40697","NEWT:13095","NEWT:162425","NEWT:242124","NEWT:9986","NEWT:8654","NEWT:536","NEWT:8658","NEWT:1268063","NEWT:8657","NEWT:9983","NEWT:8655","NEWT:80833","NEWT:479096","NEWT:28108","NEWT:28104","NEWT:1114957","NEWT:568703","NEWT:568708","NEWT:51315","NEWT:52641","NEWT:353152","NEWT:116597","NEWT:353153","NEWT:116598","NEWT:227377","NEWT:1774284","NEWT:27689","NEWT:207769","NEWT:40674","NEWT:51329","NCBITaxon:1763","NEWT:443906","NEWT:6055","NEWT:13037","NEWT:518","NEWT:519","NEWT:941638","NEWT:511","NEWT:7394","NEWT:6063","NEWT:404330","NEWT:1328388","NEWT:13064","NEWT:529120","NEWT:7375","NEWT:300641","NEWT:39655","NEWT:38323","NEWT:206411","NEWT:80869","NEWT:214053","NEWT:80863","NEWT:665890","NEWT:52639","NEWT:52638","NEWT:8697","NEWT:861109","NEWT:8695","NEWT:1196866","NEWT:6039","NEWT:37003","NEWT:1045775","NEWT:510516","NEWT:1123869","NCBITaxon:1335626",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