@article{DillingerMayerSchneideretal., author = {Dillinger, Andrea Elisabeth and Mayer, Matthias and Schneider, Magdalena and Weber, Gregor R. and Goeppner, Corinna and Tamm, Ernst R. and Shamonin (Chamonine), Mikhail and Monkman, Gareth J. and Fuchshofer, Rudolf}, title = {Reactive changes in optic nerve astrocytes are mediated by CTGF, TGF beta 2 and increasing substratum stiffness}, series = {Investigative ophthalmology \& visual science}, volume = {56}, journal = {Investigative ophthalmology \& visual science}, number = {7}, publisher = {ASSOC RESEARCH VISION OPHTHALMOLOGY INC}, abstract = {Purpose: Patients with primary open-angle glaucoma (POAG) show a stiffer peripapillary sclera, reactive astrocytes and a remodeled lamina cribrosa (LC). The changes are thought to be mediated by TGFβ2 and its downstream mediator CTGF. Recently we developed a murine glaucoma model by overexpressing CTGF in the anterior eye (βb1CTGF). In this study we investigated the glial lamina region of βb1CTGF mice, and the changes of astrocytes in response to CTGF and TGFβ2 as well as increasing substratum stiffness. Methods: Tangential sections of the glial LC of 2-month-old βb1CTGF mice and their wild-type littermates (WT) were stained with phalloidin and antibodies against GFAP, CTGF and fibronectin (FN). Murine optic nerve (ON) astrocytes from CD1 mice were isolated, cultured and characterized by GFAP staining. The astrocytes were treated with TGFβ2 (1ng/ml) and CTGF (50ng/ml and 100ng/ml). In addition, the cells were seeded on PDMS substrata with different stiffness (10, 30 and 60 kPa). Treated cells were analyzed by Western blotting, real-time RT-PCR and immunohistochemistry. Wound healing assays were performed to analyze migration rate following growth factor treatment. Results: βb1CTGF mice showed a massive increase in CTGF and GFAP in the glial LC when compared with WT mice. Moreover, an increase in FN staining and phalloidin-labeled actin was observed in the peripapillary sclera. Murine ON astrocytes reacted on increased substrate stiffness by increasing the synthesis of GFAP, vimentin and CTGF. Treatment of the cells with TGFβ2 and CTGF led to an enhanced migration rate. In addition, treatment resulted in an increased expression and synthesis of ECM proteins, including FN, tropoelastin, collagen type I and III. The in vitro findings correlated with those seen in the glaucoma mouse model. Conclusions: We conclude that changes in the ECM of LC and peripapillary sclera alter their biomechanical properties and thereby induce reactive changes in resident astrocytes. The reactive changes induced by higher stiffness of their surrounding ECM give rise to a self-amplifying process that includes increased TGFβ2/CTGF signaling and leads to synthesis of ECM and cytoskeletal proteins, a process that in turn augments the stiffness at the optic nerve head (ONH). Such a scenario may finally result in a vicious circle as the causative mechanism for ONH deformation in POAG.}, language = {en} } @article{DillingerWeberMayeretal., author = {Dillinger, Andrea Elisabeth and Weber, Gregor R. and Mayer, Matthias and Schneider, Magdalena and G{\"o}ppner, Corinna and Ohlmann, Andreas and Shamonin (Chamonine), Mikhail and Monkman, Gareth J. and Fuchshofer, Rudolf and Keller, Kate and Lozano, Diana C. and Clark, Abbot}, title = {CCN2/CTGF-A Modulator of the Optic Nerve Head Astrocyte}, series = {Frontiers in cell and developmental biology (Front Cell Dev Biol.)}, volume = {10}, journal = {Frontiers in cell and developmental biology (Front Cell Dev Biol.)}, publisher = {frontiers}, doi = {10.3389/fcell.2022.864433}, pages = {864433}, abstract = {In primary open-angle glaucoma (POAG), a neurodegenerative disease of the optic nerve (ON) and leading cause of blindness, the optic nerve head (ONH) undergoes marked structural extracellular matrix (ECM) changes, which contribute to its permanent deformation and to degeneration of ON axons. The remodeling process of the ECM causes changes in the biomechanical properties of the ONH and the peripapillary sclera, which is accompanied by an increased reactivity of the resident astrocytes. The molecular factors involved in the remodeling process belong to the Transforming growth factor (TGF)-β superfamily, especially TGF-β2. In previous publications we showed that TGF-β2 induced ECM alterations are mediated by Cellular Communication Network Factor (CCN)2/Connective Tissue Growth Factor (CTGF) and recently we showed that CCN2/CTGF is expressed by astrocytes of the ON under normal conditions. In this study we wanted to get a better understanding of the function of CCN2/CTGF under normal and pathologic conditions. To this end, we analyzed the glial lamina and peripapillary sclera of CCN2/CTGF overexpressing mice and studied the effect of CCN2/CTGF and increasing substratum stiffness on murine ON astrocytes in vitro. We observed enhanced astrocyte reactivity in the ONH, increased ECM protein synthesis in the peripapillary sclera and increased Ccn2/Ctgf expression in the ONH during the pathologic development in situ. CCN2/CTGF treatment of primary murine ON astrocytes induced a higher migration rate, and increase of ECM proteins including fibronectin, elastin and collagen type III. Furthermore, the astrocytes responded to stiffer substratum with increased glial fibrillary acidic protein, vimentin, actin and CCN2/CTGF synthesis. Finally, we observed the reinforced appearance of CCN2/CTGF in the lamina cribrosa of glaucomatous patients. We conclude that reactive changes in ONH astrocytes, induced by the altered biomechanical characteristics of the region, give rise to a self-amplifying process that includes increased TGF-β2/CCN2/CTGF signaling and leads to the synthesis of ECM molecules and cytoskeleton proteins, a process that in turn augments the stiffness at the ONH. Such a scenario may finally result in a vicious circle in the pathogenesis of POAG. The transgenic CTGF-overexpressing mouse model might be an optimal model to study the chronic pathological POAG changes in the ONH.}, language = {en} } @misc{ScharfenbergMottokArtmannetal., author = {Scharfenberg, Georg and Mottok, J{\"u}rgen and Artmann, Christina and Hobelsberger, Martin and Paric, Ivan and Großmann, Benjamin and Pohlt, Clemens and Wackerbarth, Alena and Pausch, Uli and Heidrich, Christiane and Fadanelli, Martin and Elsner, Michael and P{\"o}cher, Daniel and Pittroff, Lenz and Beer, Stefan and Br{\"u}ckl, Oliver and Haslbeck, Matthias and Sterner, Michael and Thema, Martin and Muggenthaler, Nicole and Lenck, Thorsten and G{\"o}tz, Philipp and Eckert, Fabian and Deubzer, Michael and Stingl, Armin and Simsek, Erol and Kr{\"a}mer, Stefan and Großmann, Benjamin and Schlegl, Thomas and Niedersteiner, Sascha and Berlehner, Thomas and Joblin, Mitchell and Mauerer, Wolfgang and Apel, Sven and Siegmund, Janet and Riehle, Dirk and Weber, Joachim and Palm, Christoph and Zobel, Martin and Al-Falouji, Ghassan and Prestel, Dietmar and Scharfenberg, Georg and Mandl, Roland and Deinzer, Arnulf and Halang, W. and Margraf-Stiksrud, Jutta and Sick, Bernhard and Deinzer, Renate and Scherzinger, Stefanie and Klettke, Meike and St{\"o}rl, Uta and Wiech, Katharina and Kubata, Christoph and Sindersberger, Dirk and Monkman, Gareth J. and Dollinger, Markus and Dembianny, Sven and K{\"o}lbl, Andreas and Welker, Franz and Meier, Matthias and Thumann, Philipp and Swidergal, Krzysztof and Wagner, Marcus and Haug, Sonja and Vernim, Matthias and Seidenst{\"u}cker, Barbara and Weber, Karsten and Arsan, Christian and Schone, Reinhold and M{\"u}nder, Johannes and Schroll-Decker, Irmgard and Dillinger, Andrea Elisabeth and Fuchshofer, Rudolf and Monkman, Gareth J. and Shamonin (Chamonine), Mikhail and Geith, Markus A. and Koch, Fabian and {\"U}hlin, Christian and Schratzenstaller, Thomas and Saßmannshausen, Sean Patrick and Auchter, Eberhard and Kriz, Willy and Springer, Othmar and Thumann, Maria and Kusterle, Wolfgang and Obermeier, Andreas and Udalzow, Anton and Schmailzl, Anton and Hierl, Stefan and Langer, Christoph and Schreiner, Rupert}, title = {Forschungsbericht / Ostbayerische Technische Hochschule Regensburg}, editor = {Baier, Wolfgang}, address = {Regensburg}, organization = {Ostbayerische Technische Hochschule Regensburg}, isbn = {978-3-00-048589-3}, doi = {10.35096/othr/pub-1386}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:898-opus4-13867}, language = {de} }