YB-1 expression analysis in the developing mouse eye by immunohistochemistry

YB-1 expression analysis in the developing mouse eye by immunohistochemistry | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Short Report YB-1 expression analysis in the developing mouse eye by immunohistochemistry Alexander Nass, Hella Wolf, Saadettin Sel, Thomas Kalinski, Norbert Nass This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4164659/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Objective: Cold shock proteins such as YB-1 (ybx1) function in the regulation of transcription, mRNA stability, and translation. Consequently, YB-1 contributes to differentiation, stress responses and oncogenesis. Eye development is a complex process involving the differentiation of a signifiant number of cell-types with distinct functions. Additionally, the adult eye is exposed to UV-radiation causing significant oxidative stress. We therefore hypothesized that YB-1 plays a role in eye development as well as stress defence. As a first step to understand YB-1 function in this context, we analyzed its expression in the developing and adult mouse eye by immunohistochemistry. Results: Expression of the YB-1 protein in the developing mouse eye at stages (E12, E15 and E18) and in adult eyes (P14) was detected in all retinal cells and in cells of the cornea and the lens epithelium at all stages investigated. These findings support a significant function of YB-1 in the eye, may be related to development and differentiation. eye development YB-1 immunohistochemistry cornea retina Figures Figure 1 Figure 2 Introduction The mammalian eye is a highly dedicated organ, involving several specialized tissues. It develops from cells of the mesenchyme, ectoderm and the neuronal crest ( 1 ). In particular, the retina, iris, ciliary body and nerves derive from cells of the neural epithelium ( 2 ), whereas cornea and lens originate from the surface ectoderm ( 3 ). Vitreous and sclera are formed from mesenchymal cells ( 4 , 5 ). The whole process is considered mainly as a result of sequential induction events. One major and well understood event in this process is the expression of the master control gene pax6 ( 6 , 7 ). The vertebrate neuroretina is a multi-layered tissue and responsible for light perception and initial signal processing ( 2 ). Six retinal cell types contribute to its structure, the photoreceptors, (classified as rod and cone cells), amacrine cells, horizontal cells, bipolar cells and ganglion cells as well as three cell types of glia cells namely astrocytes, Müller cells and resident microglia. These cells form a highly ordered structure with three nuclear layers separated by two plexiform sheets. The inner nuclear layer (INL) contains the nuclei of the amacrine-, Müller glia, horizontal and bipolar cells ( 8 , 9 ). The outer nuclear layer (ONL) comprises the nuclei of the photoreceptor cells. The ganglion cell layer (GCL) is made up of displaced amacrine- and ganglion cells. These three layers are separated by the inner (IPL) and outer plexiform (OPL) layers, which contain axons and dendrites connecting the neuronal cells of the nuclear layers. The whole structure develops from a pool of pluripotent stem cells in a chronological order ( 10 , 11 ). First, retinal ganglion cells are formed, followed by overlapping stages where horizontal cells, cone photoreceptors, amacrine cells, rod photoreceptors, bipolar and Müller glia cells differentiate ( 12 ). The cornea derives from cells of the surface ectoderm ( 13 ). This transparent tissue consists of the corneal epithelium (CEpi), the endothelium (CEndo) and the stroma. The corneal stroma (CS) is further embedded between two acellular structures called Bowman´s and Descemet´s membranes ( 14 ). The lens is also a derivative of the surface epithelium. During mice development at around day ten after fertilization (E10), the lens vesicle invaginates ( 15 ). The overlying surface epithelium of this vesicle forms the corneal stroma and the endothelium ( 16 ). The cornea, the lens and the vitreous are transparent structures allowing light to reach the retinal tissue. As a result, the biomolecules in these tissues are exposed to constant stress especially due to reactive oxygen species that result from absorbed photons ( 17 ). This results in oxidative modifications as well as glycation especially of the persisting proteins. As defence, vitreous and lens contains significant amounts of ascorbic acid and other antioxidants ( 18 ). Although ascorbate can act as an effective antioxidant, it is also capable of forming reactive intermediates that contribute to the formation of glycated proteins ( 19 ). Glycation results in inactivation of enzymes, cross links in matrix proteins and inflammation mediated by the receptor for AGEs (RAGE) ( 20 ). The YB-1 protein is the product of the ybx1 gene. It is a member of the large cold shock protein family that is characterized by the highly conserved cold shock domain ( 21 ). This domain functions in DNA and RNA binding. The protein is capable of regulating transcription as well as translation. It influences DNA repair, drug resistance and stress responses. The protein is mostly localized in the cytosol but can migrate into the nucleus, often depending on modifications such as phosphorylation and acetylation ( 22 , 23 ). YB-1 can also be found extracellular and contributes to the formation of exosomes ( 24 ). The expression of YB-1 mRNA in the eye has already been reported ( 25 ), but no further, detailed expression analysis in the developing eye has been published yet. We were therefore eager to investigate the expression pattern in the developing mouse eye. As a first step, we here investigated the expression of YB-1 during the mouse embryonic eye development and in adult eyes by immunohistochemistry. Methods All tissues were collected as described earlier ( 26 – 28 ). Mice (C57Bl/6N) were bread in the animal facility of the Medical Faculty of the Martin-Luther University Halle-Wittenberg. The study was approved by the institutional animal care and use committee in accordance to the ARVO (Association for Research in Vision and Ophthalmology) statement for the use of animals in ophthalmic and vision research. For studies on developing eyes, the day of vaginal plug formation was determined and noon was set to embryo stage E0.5. Day of birth was considered as P0. Tissues from stages E12, E15, E18, P7 and P14 were investigated. Tissues were fixed overnight in 4% formaldehyde in phosphate buffered saline (PBS). The fixed tissues were either stored in 70% ethanol or directly embedded in paraffin. For immunohistochemistry, blocks were sectioned in 2 µm thick slices and transferred to frosted glass slides. After deparaffinization by xylole and rehydration, epitope demasking was achieved by autoclaving in citrate buffer (pH = 6.0). 3% hydrogenperoxide was applied for 10 min to inactivate endogenous peroxidase activity. Primary rabbit monoclonal antibody (abcam 76149) was applied in antibody dilution reagent (Ventana, Mannheim, Germany) 1:400 over-night at 4°C. After three washes in TBS containing Tween 20 (0.05%) staining was performed using a secondary, HRP conjugated antibody (Dianova, Hamburg, Germany) and a DAB detection system (Vector Laboratories, via Biozol, Eching, Germany). Nuclei were counterstained using Mayer's hemalaun solution (Roth, Germany). Then, slides were mounted and scanned using a Hamamatsu Nanozoomer scanner (Hamamatsu-Photonics, Japan) using a 20 x lens. Negative controls were performed by omitting the primary antibody. Mouse liver and kidney tissue were used as positive controls. Results YB-1 is expressed in the adult retina and during mouse retinal development We performed an immunohistochemical analysis to examine the temporal and spatial expression pattern of YB-1 in the adult and developing neuroepithelium. Developmental stages from E12 to P14 were investigated. YB-1 expression could already be detected in stage E12 and was continuously seen in the adult retina. Positive staining was localized mainly to the cytoplasm (Fig. 1). YB-1 is expressed in the developing and adult cornea and lens epithelium We also detected YB-1 protein in the presumptive epithelium of the lens and corneal epithelium cells, corneal fibroblasts as well as corneal endothelial cells in all stages analyzed. This expression is basically maintained in the mature tissues of stage P14 (Fig. 2). Discussion YB-1 function is related to developmental processes such as proliferation and differentiation. On the molecular level, YB-1 is associated with RNA translation and stability. Especially under severe stress, this protein is located in the so-called stress granules ( 29 , 30 ), which represent a storage compartment for RNA ( 31 , 32 ). YB-1 also associates with proteins having functions in protein biosynthesis, e.g. eukaryotic initiation factors ( 33 , 34 ). Concerning developmental processes, YB-1 regulates neuronal cell renewal and differentiation e.g. by interaction with polycomb repressive complex 2 in the developing mouse brain. ( 35 ). Thus, we have proposed its expression in the developing eye, likely showing a distinct spatial and temporal pattern especially in cells of the developing neuroretina. However, we found the YB-1 protein expressed throughout the development of the eye already at E12 and its expression is maintained in adult eyes (P14) in the cytoplasm of essentially all cells observed. Our protein expression data are consistent with in-situ hybridization data ( 36 ). Thus, the expression of the YB-1 protein was neither restricted to neuronal nor to differentiating or proliferating cells. Based on these data, a distinct function for the cell differentiation might be debated. However, a function for stress responses still seems likely. The adult eye is exposed to radiation, causing continuous oxidative stress. One important defence mechanism is the high concentration of ascorbic acid, especially in the vitreous. Nevertheless, the lens as well as the vitreous are subject to the accumulation of modified proteins including advanced glycation end products ( 37 ). Such modifications can be caused by reducing sugars and also by ascorbic acid ( 19 ). All these protein modifications are postulated to contribute to several eye diseases ( 38 , 39 ). AGEs activate their receptor RAGE ( 40 ), which contributes to inflammation, a process that is also influenced by YB-1 ( 41 – 43 ). Therefore, there might be a functional correlation of these two processes. YB-1 expression is similar to NNAT-expression, which we have investigated earlier ( 26 ). NNAT and YB-1 are both genes involved in neuronal development and oncogenesis. Both proteins might therefore be essential for eye development and physiology. Limitations A major limitation of our study is that we investigated the presence of the protein only. Therefore, further studies on activation state mediated by phosphorylation or acetylation might provide further insight into the function of this protein in the developing eye. A histological analysis of the retina architecture of YB-1 knock-out mice ( 44 ) may well provide further inside into a function of YB-1 in development of the murine eye. In conclusion, the ubiquitous expression of the YB-1 protein in the developing and adult eye supports the idea that this protein is necessary for eye development and stress responses. Declarations Ethics approval and consent to participate Not applicable. Consent for publication Not applicable. Availability of data and materials All data generated or analysed during this study are included in this published article. Competing interests The authors declare that they have no competing interests Funding This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors. Authors´ contributions AFN, HW: Investigation; NN: Conceptualisation, Writing original draft, Methodology; SS: Methodology, Writing Review and Editing; TK: Conceptualisation, Writing original draft, Supervision Acknowledgements The authors thank the immunohistochemistry laboratory of the Institute of Pathology, Magdeburg for preparing the slides for staining. References Graw J. Eye development. Curr Top Dev Biol. 2010;90:343–86. Hoon M, Okawa H, Della Santina L, Wong ROL. Functional architecture of the retina: development and disease. Prog Retin Eye Res. September 2014;42:44–84. Chang W, Zhao Y, Rayêe D, Xie Q, Suzuki M, Zheng D, u. a. Dynamic changes in whole genome DNA methylation, chromatin and gene expression during mouse lens differentiation. Epigenetics Chromatin. 25. Januar 2023;16(1):4. Balazs EA, Toth LZ, Ozanics V. Cytological studies on the developing vitreous as related to the hyaloid vessel system. Albrecht Von Graefes Arch Klin Exp Ophthalmol Albrecht Von Graefes Arch Clin Exp Ophthalmol. 1980;213(2):71–85. Ito M, Nakashima M, Tsuchida N, Imaki J, Yoshioka M. Histogenesis of the intravitreal membrane and secondary vitreous in the mouse. Invest Ophthalmol Vis Sci. Mai 2007;48(5):1923–30. Favor J, Gloeckner CJ, Neuhäuser-Klaus A, Pretsch W, Sandulache R, Saule S, u. a. Relationship of Pax6 activity levels to the extent of eye development in the mouse, Mus musculus. Genetics. Juli 2008;179(3):1345–55. van Heyningen V, Williamson KA. PAX6 in sensory development. Hum Mol Genet. 15. Mai 2002;11(10):1161–7. Byerly MS, Blackshaw S. Vertebrate retina and hypothalamus development. Wiley Interdiscip Rev Syst Biol Med. 2009;1(3):380–9. Nakhai H, Sel S, Favor J, Mendoza-Torres L, Paulsen F, Duncker GIW, u. a. Ptf1a is essential for the differentiation of GABAergic and glycinergic amacrine cells and horizontal cells in the mouse retina. Dev Camb Engl. März 2007;134(6):1151–60. Cayouette M, Poggi L, Harris WA. Lineage in the vertebrate retina. Trends Neurosci. Oktober 2006;29(10):563–70. Cepko C. Intrinsically different retinal progenitor cells produce specific types of progeny. Nat Rev Neurosci. September 2014;15(9):615–27. Marquardt T, Gruss P. Generating neuronal diversity in the retina: one for nearly all. Trends Neurosci. Januar 2002;25(1):32–8. Swamynathan SK. Ocular surface development and gene expression. J Ophthalmol. 2013;2013:103947. Pajoohesh-Ganji A, Stepp MA. In search of markers for the stem cells of the corneal epithelium. Biol Cell. April 2005;97(4):265–76. Graw J. Chapter Ten - Eye Development. In: Koopman P, Herausgeber. Current Topics in Developmental Biology [Internet]. Academic Press; 2010 [zitiert 17. Januar 2024]. S. 343–86. (Organogenesis in Development; Bd. 90). Verfügbar unter: https://www.sciencedirect.com/science/article/pii/S0070215310900100 Kondoh H. 21 - Development of the Eye. In: Rossant J, Tam PPL, Herausgeber. Mouse Development [Internet]. San Diego: Academic Press; 2002 [zitiert 17. Januar 2024]. S. 519–38. Verfügbar unter: https://www.sciencedirect.com/science/article/pii/B9780125979511500251 Böhm EW, Buonfiglio F, Voigt AM, Bachmann P, Safi T, Pfeiffer N, u. a. Oxidative stress in the eye and its role in the pathophysiology of ocular diseases. Redox Biol. Dezember 2023;68:102967. Ankamah E, Sebag J, Ng E, Nolan JM. Vitreous Antioxidants, Degeneration, and Vitreo-Retinopathy: Exploring the Links. Antioxidants. Januar 2020;9(1):7. Sadowska-Bartosz I, Bartosz G. Ascorbic acid and protein glycation in vitro. Chem Biol Interact. 5. Oktober 2015;240:154–62. Taguchi K, Fukami K. RAGE signaling regulates the progression of diabetic complications. Front Pharmacol. 2023;14:1128872. Dn L, Ia E, Lp O. YB-1 protein: functions and regulation. Wiley Interdiscip Rev RNA [Internet]. Februar 2014 [zitiert 17. Januar 2024];5(1). Verfügbar unter: https://pubmed.ncbi.nlm.nih.gov/24217978/ Lindquist JA, Mertens PR. Cold shock proteins: from cellular mechanisms to pathophysiology and disease. Cell Commun Signal CCS. 26. September 2018;16(1):63. Shah A, Lindquist JA, Rosendahl L, Schmitz I, Mertens PR. Novel Insights into YB-1 Signaling and Cell Death Decisions. Cancers. 1. Juli 2021;13(13):3306. Xue X, Huang J, Yu K, Chen X, He Y, Qi D, u. a. YB-1 transferred by gastric cancer exosomes promotes angiogenesis via enhancing the expression of angiogenic factors in vascular endothelial cells. BMC Cancer. 14. Oktober 2020;20(1):996. Shaughnessy M, Wistow G. Absence of MHC gene expression in lens and cloning of dbpB/YB-1, a DNA-binding protein expressed in mouse lens. Curr Eye Res. Februar 1992;11(2):175–81. Sel S, Patzel E, Poggi L, Kaiser D, Kalinski T, Schicht M, u. a. Temporal and spatial expression pattern of Nnat during mouse eye development. Gene Expr Patterns. Januar 2017;23–24:7–12. Sel S, Kaiser M, Nass N, Trau S, Roepke A, Storsberg J, u. a. Oligophrenin-1 (Ophn1) is expressed in mouse retinal vessels. Gene Expr Patterns GEP. Februar 2012;12(1–2):63–7. Sel S, Kalinski T, Enssen I, Kaiser M, Nass N, Trau S, u. a. Expression analysis of ADAM17 during mouse eye development. Ann Anat Anat Anz Off Organ Anat Ges. Juli 2012;194(4):334–8. Guarino AM, Mauro GD, Ruggiero G, Geyer N, Delicato A, Foulkes NS, u. a. YB-1 recruitment to stress granules in zebrafish cells reveals a differential adaptive response to stress. Sci Rep. 21. Juni 2019;9(1):9059. Lyons SM, Achorn C, Kedersha NL, Anderson PJ, Ivanov P. YB-1 regulates tiRNA-induced Stress Granule formation but not translational repression. Nucleic Acids Res. 19. August 2016;44(14):6949–60. Somasekharan SP, El-Naggar A, Leprivier G, Cheng H, Hajee S, Grunewald TGP, u. a. YB-1 regulates stress granule formation and tumor progression by translationally activating G3BP1. J Cell Biol. 30. März 2015;208(7):913–29. Tanaka T, Ohashi S, Kobayashi S. Roles of YB-1 under arsenite-induced stress: translational activation of HSP70 mRNA and control of the number of stress granules. Biochim Biophys Acta. März 2014;1840(3):985–92. Ivanova IG, Park CV, Yemm AI, Kenneth NS. PERK/eIF2α signaling inhibits HIF-induced gene expression during the unfolded protein response via YB1-dependent regulation of HIF1α translation. Nucleic Acids Res. 4. Mai 2018;46(8):3878–90. Vo DK, Engler A, Stoimenovski D, Hartig R, Kaehne T, Kalinski T, u. a. Interactome Mapping of eIF3A in a Colon Cancer and an Immortalized Embryonic Cell Line Using Proximity-Dependent Biotin Identification. Cancers. 14. März 2021;13(6):1293. Evans MK, Matsui Y, Xu B, Willis C, Loome J, Milburn L, u. a. Ybx1 fine-tunes PRC2 activities to control embryonic brain development. Nat Commun. 13. August 2020;11(1):4060. Blackshaw S, Harpavat S, Trimarchi J, Cai L, Huang H, Kuo WP, u. a. Genomic Analysis of Mouse Retinal Development. PLOS Biol. 29. Juni 2004;2(9):e247. Kandarakis SA, Piperi C, Topouzis F, Papavassiliou AG. Emerging role of advanced glycation-end products (AGEs) in the pathobiology of eye diseases. Prog Retin Eye Res. September 2014;42:85–102. Oshitari T. Advanced Glycation End-Products and Diabetic Neuropathy of the Retina. Int J Mol Sci. 2. Februar 2023;24(3):2927. Zhou Q, Yang L, Wang Q, Li Y, Wei C, Xie L. Mechanistic investigations of diabetic ocular surface diseases. Front Endocrinol. 2022;13:1079541. Zong H, Ward M, Stitt AW. AGEs, RAGE, and diabetic retinopathy. Curr Diab Rep. August 2011;11(4):244–52. Hanssen L, Alidousty C, Djudjaj S, Frye BC, Rauen T, Boor P, u. a. YB-1 is an early and central mediator of bacterial and sterile inflammation in vivo. J Immunol Baltim Md 1950. 1. September 2013;191(5):2604–13. Rybalkina EY, Moiseeva NI. Role of YB-1 Protein in Inflammation. Biochem Biokhimiia. Januar 2022;87(Suppl 1):S94-202. Wang J, Djudjaj S, Gibbert L, Lennartz V, Breitkopf DM, Rauen T, u. a. YB-1 orchestrates onset and resolution of renal inflammation via IL10 gene regulation. J Cell Mol Med. Dezember 2017;21(12):3494–505. Bernhardt A, Häberer S, Xu J, Damerau H, Steffen J, Reichardt C, u. a. High salt diet-induced proximal tubular phenotypic changes and sodium-glucose cotransporter-2 expression are coordinated by cold shock Y-box binding protein-1. FASEB J Off Publ Fed Am Soc Exp Biol. Oktober 2021;35(10):e21912. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4164659","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Short Report","associatedPublications":[],"authors":[{"id":284599007,"identity":"44b68ea7-3805-4d1e-b433-db3741a5e6c8","order_by":0,"name":"Alexander Nass","email":"","orcid":"","institution":"University Hospital Brandenburg/Havel","correspondingAuthor":false,"prefix":"","firstName":"Alexander","middleName":"","lastName":"Nass","suffix":""},{"id":284599011,"identity":"d96a2ef4-9fd5-4884-a66c-e76345853daf","order_by":1,"name":"Hella Wolf","email":"","orcid":"","institution":"University Hospital Brandenburg/Havel","correspondingAuthor":false,"prefix":"","firstName":"Hella","middleName":"","lastName":"Wolf","suffix":""},{"id":284599016,"identity":"8f4b9359-fd66-42af-96ca-ecb835a33b19","order_by":2,"name":"Saadettin Sel","email":"","orcid":"","institution":"University of Heidelberg","correspondingAuthor":false,"prefix":"","firstName":"Saadettin","middleName":"","lastName":"Sel","suffix":""},{"id":284599020,"identity":"f52cd652-b629-4f9e-a28f-ff9d85cccaba","order_by":3,"name":"Thomas Kalinski","email":"","orcid":"","institution":"University Hospital Brandenburg/Havel","correspondingAuthor":false,"prefix":"","firstName":"Thomas","middleName":"","lastName":"Kalinski","suffix":""},{"id":284599022,"identity":"5b8d81b7-89b5-44ec-a878-2d0a2930d787","order_by":4,"name":"Norbert Nass","email":"data:image/png;base64,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","orcid":"","institution":"University Hospital Brandenburg/Havel","correspondingAuthor":true,"prefix":"","firstName":"Norbert","middleName":"","lastName":"Nass","suffix":""}],"badges":[],"createdAt":"2024-03-25 16:24:50","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4164659/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4164659/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":53840170,"identity":"4a27c842-a9cd-4f84-92f0-3a2528c93f14","added_by":"auto","created_at":"2024-04-01 07:08:11","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":6850758,"visible":true,"origin":"","legend":"\u003cp\u003eExpression of YB-1 in embryonal and postnatal neuroretina at stage E12, E18 and P14 as shown by immunohistochemistry. A, C, E hematoxylin/eosin staining. B, D, F immunostaining for YB-1. Retinal pigment epithelium: RPE, outer nuclear layer: ONL, outer plexiform layer: OPN, inner nuclear layer: INL, inner plexiform layer: IPL, ganglion cell layer: GCL, neuroblastic layer: NPL. Scale bar indicates 100 µm.\u003c/p\u003e","description":"","filename":"Fig1RetinagesamtB.png","url":"https://assets-eu.researchsquare.com/files/rs-4164659/v1/4721333db6c1b79e17cdd3f8.png"},{"id":53840171,"identity":"88180a9e-eb5b-4155-b799-e504e7a21e7e","added_by":"auto","created_at":"2024-04-01 07:08:11","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":2609281,"visible":true,"origin":"","legend":"\u003cp\u003eExpression of YB-1 protein in cornea and lens epithelium at stage E12 and P14 as shown by immunohistochemistry. A, C hematoxylin/eosin staining. B, D immunostaining for YB-1. Scale bar indicates 100 µm.\u003c/p\u003e","description":"","filename":"Fig2CorneaA.png","url":"https://assets-eu.researchsquare.com/files/rs-4164659/v1/44fdad2ee6e34b0cc67c6fcf.png"},{"id":78189021,"identity":"372f03a4-e213-4c5e-92c4-ef1725d1f870","added_by":"auto","created_at":"2025-03-10 19:37:18","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":14406319,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4164659/v1/59f9ee20-f09f-4184-ae32-8465d32ad5cd.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"YB-1 expression analysis in the developing mouse eye by immunohistochemistry","fulltext":[{"header":"Introduction","content":"\u003cp\u003eThe mammalian eye is a highly dedicated organ, involving several specialized tissues. It develops from cells of the mesenchyme, ectoderm and the neuronal crest (\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e). In particular, the retina, iris, ciliary body and nerves derive from cells of the neural epithelium (\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e), whereas cornea and lens originate from the surface ectoderm (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e). Vitreous and sclera are formed from mesenchymal cells (\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e, \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e). The whole process is considered mainly as a result of sequential induction events. One major and well understood event in this process is the expression of the master control gene pax6 (\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e). The vertebrate neuroretina is a multi-layered tissue and responsible for light perception and initial signal processing (\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e). Six retinal cell types contribute to its structure, the photoreceptors, (classified as rod and cone cells), amacrine cells, horizontal cells, bipolar cells and ganglion cells as well as three cell types of glia cells namely astrocytes, M\u0026uuml;ller cells and resident microglia. These cells form a highly ordered structure with three nuclear layers separated by two plexiform sheets. The inner nuclear layer (INL) contains the nuclei of the amacrine-, M\u0026uuml;ller glia, horizontal and bipolar cells (\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e, \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e). The outer nuclear layer (ONL) comprises the nuclei of the photoreceptor cells. The ganglion cell layer (GCL) is made up of displaced amacrine- and ganglion cells. These three layers are separated by the inner (IPL) and outer plexiform (OPL) layers, which contain axons and dendrites connecting the neuronal cells of the nuclear layers. The whole structure develops from a pool of pluripotent stem cells in a chronological order (\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e). First, retinal ganglion cells are formed, followed by overlapping stages where horizontal cells, cone photoreceptors, amacrine cells, rod photoreceptors, bipolar and M\u0026uuml;ller glia cells differentiate (\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e). The cornea derives from cells of the surface ectoderm (\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e). This transparent tissue consists of the corneal epithelium (CEpi), the endothelium (CEndo) and the stroma. The corneal stroma (CS) is further embedded between two acellular structures called Bowman\u0026acute;s and Descemet\u0026acute;s membranes (\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e). The lens is also a derivative of the surface epithelium. During mice development at around day ten after fertilization (E10), the lens vesicle invaginates (\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e). The overlying surface epithelium of this vesicle forms the corneal stroma and the endothelium (\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e). The cornea, the lens and the vitreous are transparent structures allowing light to reach the retinal tissue. As a result, the biomolecules in these tissues are exposed to constant stress especially due to reactive oxygen species that result from absorbed photons (\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e). This results in oxidative modifications as well as glycation especially of the persisting proteins. As defence, vitreous and lens contains significant amounts of ascorbic acid and other antioxidants (\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e). Although ascorbate can act as an effective antioxidant, it is also capable of forming reactive intermediates that contribute to the formation of glycated proteins (\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e). Glycation results in inactivation of enzymes, cross links in matrix proteins and inflammation mediated by the receptor for AGEs (RAGE) (\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e). The YB-1 protein is the product of the ybx1 gene. It is a member of the large cold shock protein family that is characterized by the highly conserved cold shock domain (\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e). This domain functions in DNA and RNA binding. The protein is capable of regulating transcription as well as translation. It influences DNA repair, drug resistance and stress responses. The protein is mostly localized in the cytosol but can migrate into the nucleus, often depending on modifications such as phosphorylation and acetylation (\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e, \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e). YB-1 can also be found extracellular and contributes to the formation of exosomes (\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e). The expression of YB-1 mRNA in the eye has already been reported (\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e), but no further, detailed expression analysis in the developing eye has been published yet.\u003c/p\u003e \u003cp\u003eWe were therefore eager to investigate the expression pattern in the developing mouse eye. As a first step, we here investigated the expression of YB-1 during the mouse embryonic eye development and in adult eyes by immunohistochemistry.\u003c/p\u003e"},{"header":"Methods","content":"\u003cp\u003eAll tissues were collected as described earlier (\u003cspan additionalcitationids=\"CR27\" citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e). Mice (C57Bl/6N) were bread in the animal facility of the Medical Faculty of the Martin-Luther University Halle-Wittenberg. The study was approved by the institutional animal care and use committee in accordance to the ARVO (Association for Research in Vision and Ophthalmology) statement for the use of animals in ophthalmic and vision research. For studies on developing eyes, the day of vaginal plug formation was determined and noon was set to embryo stage E0.5. Day of birth was considered as P0. Tissues from stages E12, E15, E18, P7 and P14 were investigated. Tissues were fixed overnight in 4% formaldehyde in phosphate buffered saline (PBS). The fixed tissues were either stored in 70% ethanol or directly embedded in paraffin. For immunohistochemistry, blocks were sectioned in 2 \u0026micro;m thick slices and transferred to frosted glass slides. After deparaffinization by xylole and rehydration, epitope demasking was achieved by autoclaving in citrate buffer (pH\u0026thinsp;=\u0026thinsp;6.0). 3% hydrogenperoxide was applied for 10 min to inactivate endogenous peroxidase activity. Primary rabbit monoclonal antibody (abcam 76149) was applied in antibody dilution reagent (Ventana, Mannheim, Germany) 1:400 over-night at 4\u0026deg;C. After three washes in TBS containing Tween 20 (0.05%) staining was performed using a secondary, HRP conjugated antibody (Dianova, Hamburg, Germany) and a DAB detection system (Vector Laboratories, via Biozol, Eching, Germany). Nuclei were counterstained using Mayer's hemalaun solution (Roth, Germany). Then, slides were mounted and scanned using a Hamamatsu Nanozoomer scanner (Hamamatsu-Photonics, Japan) using a 20 x lens. Negative controls were performed by omitting the primary antibody. Mouse liver and kidney tissue were used as positive controls.\u003c/p\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003eYB-1 is expressed in the adult retina and during mouse retinal development\u003c/h2\u003e \u003cp\u003eWe performed an immunohistochemical analysis to examine the temporal and spatial expression pattern of YB-1 in the adult and developing neuroepithelium. Developmental stages from E12 to P14 were investigated. YB-1 expression could already be detected in stage E12 and was continuously seen in the adult retina. Positive staining was localized mainly to the cytoplasm (Fig.\u0026nbsp;1).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eYB-1 is expressed in the developing and adult cornea and lens epithelium\u003c/h2\u003e \u003cp\u003eWe also detected YB-1 protein in the presumptive epithelium of the lens and corneal epithelium cells, corneal fibroblasts as well as corneal endothelial cells in all stages analyzed. This expression is basically maintained in the mature tissues of stage P14 (Fig.\u0026nbsp;2).\u003c/p\u003e \u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eYB-1 function is related to developmental processes such as proliferation and differentiation. On the molecular level, YB-1 is associated with RNA translation and stability. Especially under severe stress, this protein is located in the so-called stress granules (\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e, \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e), which represent a storage compartment for RNA (\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e, \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e). YB-1 also associates with proteins having functions in protein biosynthesis, e.g. eukaryotic initiation factors (\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e, \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e). Concerning developmental processes, YB-1 regulates neuronal cell renewal and differentiation e.g. by interaction with polycomb repressive complex 2 in the developing mouse brain. (\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e). Thus, we have proposed its expression in the developing eye, likely showing a distinct spatial and temporal pattern especially in cells of the developing neuroretina. However, we found the YB-1 protein expressed throughout the development of the eye already at E12 and its expression is maintained in adult eyes (P14) in the cytoplasm of essentially all cells observed. Our protein expression data are consistent with \u003cem\u003ein-situ\u003c/em\u003e hybridization data (\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e). Thus, the expression of the YB-1 protein was neither restricted to neuronal nor to differentiating or proliferating cells. Based on these data, a distinct function for the cell differentiation might be debated. However, a function for stress responses still seems likely. The adult eye is exposed to radiation, causing continuous oxidative stress. One important defence mechanism is the high concentration of ascorbic acid, especially in the vitreous. Nevertheless, the lens as well as the vitreous are subject to the accumulation of modified proteins including advanced glycation end products (\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e). Such modifications can be caused by reducing sugars and also by ascorbic acid (\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e). All these protein modifications are postulated to contribute to several eye diseases (\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e, \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e). AGEs activate their receptor RAGE (\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e), which contributes to inflammation, a process that is also influenced by YB-1 (\u003cspan additionalcitationids=\"CR42\" citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e). Therefore, there might be a functional correlation of these two processes. YB-1 expression is similar to NNAT-expression, which we have investigated earlier (\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e). NNAT and YB-1 are both genes involved in neuronal development and oncogenesis. Both proteins might therefore be essential for eye development and physiology.\u003c/p\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eLimitations\u003c/h2\u003e \u003cp\u003eA major limitation of our study is that we investigated the presence of the protein only. Therefore, further studies on activation state mediated by phosphorylation or acetylation might provide further insight into the function of this protein in the developing eye. A histological analysis of the retina architecture of YB-1 knock-out mice (\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e) may well provide further inside into a function of YB-1 in development of the murine eye.\u003c/p\u003e \u003cp\u003eIn conclusion, the ubiquitous expression of the YB-1 protein in the developing and adult eye supports the idea that this protein is necessary for eye development and stress responses.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e "},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll data generated or analysed during this study are included in this published article.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no competing interests\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors\u0026acute; contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAFN, HW: Investigation; NN: Conceptualisation, Writing original draft, Methodology; SS: Methodology, Writing Review and Editing; TK: Conceptualisation, Writing original draft, Supervision\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors thank the immunohistochemistry laboratory of the Institute of Pathology, Magdeburg for preparing the slides for staining.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eGraw J. Eye development. Curr Top Dev Biol. 2010;90:343\u0026ndash;86. \u003c/li\u003e\n\u003cli\u003eHoon M, Okawa H, Della Santina L, Wong ROL. Functional architecture of the retina: development and disease. Prog Retin Eye Res. September 2014;42:44\u0026ndash;84. \u003c/li\u003e\n\u003cli\u003eChang W, Zhao Y, Ray\u0026ecirc;e D, Xie Q, Suzuki M, Zheng D, u. a. Dynamic changes in whole genome DNA methylation, chromatin and gene expression during mouse lens differentiation. Epigenetics Chromatin. 25. Januar 2023;16(1):4. \u003c/li\u003e\n\u003cli\u003eBalazs EA, Toth LZ, Ozanics V. Cytological studies on the developing vitreous as related to the hyaloid vessel system. Albrecht Von Graefes Arch Klin Exp Ophthalmol Albrecht Von Graefes Arch Clin Exp Ophthalmol. 1980;213(2):71\u0026ndash;85. \u003c/li\u003e\n\u003cli\u003eIto M, Nakashima M, Tsuchida N, Imaki J, Yoshioka M. Histogenesis of the intravitreal membrane and secondary vitreous in the mouse. Invest Ophthalmol Vis Sci. Mai 2007;48(5):1923\u0026ndash;30. \u003c/li\u003e\n\u003cli\u003eFavor J, Gloeckner CJ, Neuh\u0026auml;user-Klaus A, Pretsch W, Sandulache R, Saule S, u. a. Relationship of Pax6 activity levels to the extent of eye development in the mouse, Mus musculus. Genetics. Juli 2008;179(3):1345\u0026ndash;55. \u003c/li\u003e\n\u003cli\u003evan Heyningen V, Williamson KA. PAX6 in sensory development. Hum Mol Genet. 15. Mai 2002;11(10):1161\u0026ndash;7. \u003c/li\u003e\n\u003cli\u003eByerly MS, Blackshaw S. Vertebrate retina and hypothalamus development. Wiley Interdiscip Rev Syst Biol Med. 2009;1(3):380\u0026ndash;9. \u003c/li\u003e\n\u003cli\u003eNakhai H, Sel S, Favor J, Mendoza-Torres L, Paulsen F, Duncker GIW, u. a. Ptf1a is essential for the differentiation of GABAergic and glycinergic amacrine cells and horizontal cells in the mouse retina. Dev Camb Engl. M\u0026auml;rz 2007;134(6):1151\u0026ndash;60. \u003c/li\u003e\n\u003cli\u003eCayouette M, Poggi L, Harris WA. Lineage in the vertebrate retina. Trends Neurosci. Oktober 2006;29(10):563\u0026ndash;70. \u003c/li\u003e\n\u003cli\u003eCepko C. Intrinsically different retinal progenitor cells produce specific types of progeny. Nat Rev Neurosci. September 2014;15(9):615\u0026ndash;27. \u003c/li\u003e\n\u003cli\u003eMarquardt T, Gruss P. Generating neuronal diversity in the retina: one for nearly all. Trends Neurosci. Januar 2002;25(1):32\u0026ndash;8. \u003c/li\u003e\n\u003cli\u003eSwamynathan SK. Ocular surface development and gene expression. J Ophthalmol. 2013;2013:103947. \u003c/li\u003e\n\u003cli\u003ePajoohesh-Ganji A, Stepp MA. In search of markers for the stem cells of the corneal epithelium. Biol Cell. April 2005;97(4):265\u0026ndash;76. \u003c/li\u003e\n\u003cli\u003eGraw J. Chapter Ten - Eye Development. In: Koopman P, Herausgeber. Current Topics in Developmental Biology [Internet]. Academic Press; 2010 [zitiert 17. Januar 2024]. S. 343\u0026ndash;86. (Organogenesis in Development; Bd. 90). Verf\u0026uuml;gbar unter: https://www.sciencedirect.com/science/article/pii/S0070215310900100\u003c/li\u003e\n\u003cli\u003eKondoh H. 21 - Development of the Eye. In: Rossant J, Tam PPL, Herausgeber. Mouse Development [Internet]. San Diego: Academic Press; 2002 [zitiert 17. Januar 2024]. S. 519\u0026ndash;38. Verf\u0026uuml;gbar unter: https://www.sciencedirect.com/science/article/pii/B9780125979511500251\u003c/li\u003e\n\u003cli\u003eB\u0026ouml;hm EW, Buonfiglio F, Voigt AM, Bachmann P, Safi T, Pfeiffer N, u. a. Oxidative stress in the eye and its role in the pathophysiology of ocular diseases. Redox Biol. Dezember 2023;68:102967. \u003c/li\u003e\n\u003cli\u003eAnkamah E, Sebag J, Ng E, Nolan JM. Vitreous Antioxidants, Degeneration, and Vitreo-Retinopathy: Exploring the Links. Antioxidants. Januar 2020;9(1):7. \u003c/li\u003e\n\u003cli\u003eSadowska-Bartosz I, Bartosz G. Ascorbic acid and protein glycation in vitro. Chem Biol Interact. 5. Oktober 2015;240:154\u0026ndash;62. \u003c/li\u003e\n\u003cli\u003eTaguchi K, Fukami K. RAGE signaling regulates the progression of diabetic complications. Front Pharmacol. 2023;14:1128872. \u003c/li\u003e\n\u003cli\u003eDn L, Ia E, Lp O. YB-1 protein: functions and regulation. Wiley Interdiscip Rev RNA [Internet]. Februar 2014 [zitiert 17. Januar 2024];5(1). Verf\u0026uuml;gbar unter: https://pubmed.ncbi.nlm.nih.gov/24217978/\u003c/li\u003e\n\u003cli\u003eLindquist JA, Mertens PR. Cold shock proteins: from cellular mechanisms to pathophysiology and disease. Cell Commun Signal CCS. 26. September 2018;16(1):63. \u003c/li\u003e\n\u003cli\u003eShah A, Lindquist JA, Rosendahl L, Schmitz I, Mertens PR. Novel Insights into YB-1 Signaling and Cell Death Decisions. Cancers. 1. Juli 2021;13(13):3306. \u003c/li\u003e\n\u003cli\u003eXue X, Huang J, Yu K, Chen X, He Y, Qi D, u. a. YB-1 transferred by gastric cancer exosomes promotes angiogenesis via enhancing the expression of angiogenic factors in vascular endothelial cells. BMC Cancer. 14. Oktober 2020;20(1):996. \u003c/li\u003e\n\u003cli\u003eShaughnessy M, Wistow G. Absence of MHC gene expression in lens and cloning of dbpB/YB-1, a DNA-binding protein expressed in mouse lens. Curr Eye Res. Februar 1992;11(2):175\u0026ndash;81. \u003c/li\u003e\n\u003cli\u003eSel S, Patzel E, Poggi L, Kaiser D, Kalinski T, Schicht M, u. a. Temporal and spatial expression pattern of Nnat during mouse eye development. Gene Expr Patterns. Januar 2017;23\u0026ndash;24:7\u0026ndash;12. \u003c/li\u003e\n\u003cli\u003eSel S, Kaiser M, Nass N, Trau S, Roepke A, Storsberg J, u. a. Oligophrenin-1 (Ophn1) is expressed in mouse retinal vessels. Gene Expr Patterns GEP. Februar 2012;12(1\u0026ndash;2):63\u0026ndash;7. \u003c/li\u003e\n\u003cli\u003eSel S, Kalinski T, Enssen I, Kaiser M, Nass N, Trau S, u. a. Expression analysis of ADAM17 during mouse eye development. Ann Anat Anat Anz Off Organ Anat Ges. Juli 2012;194(4):334\u0026ndash;8. \u003c/li\u003e\n\u003cli\u003eGuarino AM, Mauro GD, Ruggiero G, Geyer N, Delicato A, Foulkes NS, u. a. YB-1 recruitment to stress granules in zebrafish cells reveals a differential adaptive response to stress. Sci Rep. 21. Juni 2019;9(1):9059. \u003c/li\u003e\n\u003cli\u003eLyons SM, Achorn C, Kedersha NL, Anderson PJ, Ivanov P. YB-1 regulates tiRNA-induced Stress Granule formation but not translational repression. Nucleic Acids Res. 19. August 2016;44(14):6949\u0026ndash;60. \u003c/li\u003e\n\u003cli\u003eSomasekharan SP, El-Naggar A, Leprivier G, Cheng H, Hajee S, Grunewald TGP, u. a. YB-1 regulates stress granule formation and tumor progression by translationally activating G3BP1. J Cell Biol. 30. M\u0026auml;rz 2015;208(7):913\u0026ndash;29. \u003c/li\u003e\n\u003cli\u003eTanaka T, Ohashi S, Kobayashi S. Roles of YB-1 under arsenite-induced stress: translational activation of HSP70 mRNA and control of the number of stress granules. Biochim Biophys Acta. M\u0026auml;rz 2014;1840(3):985\u0026ndash;92. \u003c/li\u003e\n\u003cli\u003eIvanova IG, Park CV, Yemm AI, Kenneth NS. PERK/eIF2\u0026alpha; signaling inhibits HIF-induced gene expression during the unfolded protein response via YB1-dependent regulation of HIF1\u0026alpha; translation. Nucleic Acids Res. 4. Mai 2018;46(8):3878\u0026ndash;90. \u003c/li\u003e\n\u003cli\u003eVo DK, Engler A, Stoimenovski D, Hartig R, Kaehne T, Kalinski T, u. a. Interactome Mapping of eIF3A in a Colon Cancer and an Immortalized Embryonic Cell Line Using Proximity-Dependent Biotin Identification. Cancers. 14. M\u0026auml;rz 2021;13(6):1293. \u003c/li\u003e\n\u003cli\u003eEvans MK, Matsui Y, Xu B, Willis C, Loome J, Milburn L, u. a. Ybx1 fine-tunes PRC2 activities to control embryonic brain development. Nat Commun. 13. August 2020;11(1):4060. \u003c/li\u003e\n\u003cli\u003eBlackshaw S, Harpavat S, Trimarchi J, Cai L, Huang H, Kuo WP, u. a. Genomic Analysis of Mouse Retinal Development. PLOS Biol. 29. Juni 2004;2(9):e247. \u003c/li\u003e\n\u003cli\u003eKandarakis SA, Piperi C, Topouzis F, Papavassiliou AG. Emerging role of advanced glycation-end products (AGEs) in the pathobiology of eye diseases. Prog Retin Eye Res. September 2014;42:85\u0026ndash;102. \u003c/li\u003e\n\u003cli\u003eOshitari T. Advanced Glycation End-Products and Diabetic Neuropathy of the Retina. Int J Mol Sci. 2. Februar 2023;24(3):2927. \u003c/li\u003e\n\u003cli\u003eZhou Q, Yang L, Wang Q, Li Y, Wei C, Xie L. Mechanistic investigations of diabetic ocular surface diseases. Front Endocrinol. 2022;13:1079541. \u003c/li\u003e\n\u003cli\u003eZong H, Ward M, Stitt AW. AGEs, RAGE, and diabetic retinopathy. Curr Diab Rep. August 2011;11(4):244\u0026ndash;52. \u003c/li\u003e\n\u003cli\u003eHanssen L, Alidousty C, Djudjaj S, Frye BC, Rauen T, Boor P, u. a. YB-1 is an early and central mediator of bacterial and sterile inflammation in vivo. J Immunol Baltim Md 1950. 1. September 2013;191(5):2604\u0026ndash;13. \u003c/li\u003e\n\u003cli\u003eRybalkina EY, Moiseeva NI. Role of YB-1 Protein in Inflammation. Biochem Biokhimiia. Januar 2022;87(Suppl 1):S94-202. \u003c/li\u003e\n\u003cli\u003eWang J, Djudjaj S, Gibbert L, Lennartz V, Breitkopf DM, Rauen T, u. a. YB-1 orchestrates onset and resolution of renal inflammation via IL10 gene regulation. J Cell Mol Med. Dezember 2017;21(12):3494\u0026ndash;505. \u003c/li\u003e\n\u003cli\u003eBernhardt A, H\u0026auml;berer S, Xu J, Damerau H, Steffen J, Reichardt C, u. a. High salt diet-induced proximal tubular phenotypic changes and sodium-glucose cotransporter-2 expression are coordinated by cold shock Y-box binding protein-1. FASEB J Off Publ Fed Am Soc Exp Biol. Oktober 2021;35(10):e21912. \u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"eye, development, YB-1, immunohistochemistry, cornea, retina","lastPublishedDoi":"10.21203/rs.3.rs-4164659/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4164659/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eObjective:\u003c/h2\u003e \u003cp\u003eCold shock proteins such as YB-1 (ybx1) function in the regulation of transcription, mRNA stability, and translation. Consequently, YB-1 contributes to differentiation, stress responses and oncogenesis. Eye development is a complex process involving the differentiation of a signifiant number of cell-types with distinct functions. Additionally, the adult eye is exposed to UV-radiation causing significant oxidative stress. We therefore hypothesized that YB-1 plays a role in eye development as well as stress defence. As a first step to understand YB-1 function in this context, we analyzed its expression in the developing and adult mouse eye by immunohistochemistry.\u003c/p\u003e\u003ch2\u003eResults:\u003c/h2\u003e \u003cp\u003eExpression of the YB-1 protein in the developing mouse eye at stages (E12, E15 and E18) and in adult eyes (P14) was detected in all retinal cells and in cells of the cornea and the lens epithelium at all stages investigated. These findings support a significant function of YB-1 in the eye, may be related to development and differentiation.\u003c/p\u003e","manuscriptTitle":"YB-1 expression analysis in the developing mouse eye by immunohistochemistry","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-04-01 07:08:06","doi":"10.21203/rs.3.rs-4164659/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"2658804d-3cb2-4944-8340-7c09ac2ea623","owner":[],"postedDate":"April 1st, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-03-10T18:51:02+00:00","versionOfRecord":[],"versionCreatedAt":"2024-04-01 07:08:06","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4164659","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4164659","identity":"rs-4164659","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}