title: Bone morphogenetic protein 7 regulates reactive gliosis in retinal astrocytes and Müller glia.
abstract: OBJECTIVE
  The focus of this study was to determine whether bone morphogenetic proteins (BMPs) trigger reactive gliosis in retinal astrocytes and/or Müller glial cells.
  
  
  METHODS
  Retinal astrocytes and the Müller glial cell line MIO-M1 were treated with vehicle, BMP7, or BMP4. Samples from the treated cells were analyzed for changes in gliosis markers using reverse transcriptase - quantitative PCR (RT-qPCR) and western blotting. To determine potential similarities and differences in gliosis states, control and BMP-treated cells were compared to cells treated with sodium peroxynitrite (a strong oxidizing agent that will bring about some aspects of gliosis). Last, mature mice were microinjected intravitreally with BMP7 and analyzed for changes in gliosis markers using RT-qPCR, western blotting, and immunohistochemistry.
  
  
  RESULTS
  Treatment of retinal astrocyte cells and Müller glial cells with BMP7 regulated various reactive gliosis markers. When compared to the response of cells treated with sodium peroxynitrite, the profiles of gliosis markers regulated due to exposure to BMP7 were similar. However, as expected, the profiles including the oxidative agent and growth factor were not identical. Treatment of cells with BMP4, however, showed an attenuated response in comparison to peroxynitrite and BMP7 treatment. Injection of BMP7 into the mouse retina also triggered a reactive gliosis response 7 days after injection.
  
  
  CONCLUSIONS
  BMP7 induced changes in levels of mRNA and protein markers typically associated with reactive gliosis in retinal astrocytes and Müller glial cells, including glial fibrillary acidic protein (GFAP), glutamine synthetase (GS), a subset of chondroitin sulfate proteoglycans (CSPGs), matrix metalloproteinases (MMPs), and other molecules.
authors.affiliation: Department of Biology, Indiana University-Purdue University Indianapolis,, Indianapolis, IN ; Center for Regenerative Biology and Medicine, Indiana University- Purdue University Indianapolis, Indianapolis, IN.
authors.forename: Subramanian
authors.initials: S
authors.lastname: Dharmarajan
authors.affiliation: Department of Ophthalmology and Visual Sciences, University of Wisconsin School of Medicine and Public Health, Madison, WI.
authors.forename: Zafer
authors.initials: Z
authors.lastname: Gurel
authors.affiliation: Department of Ophthalmology and Visual Sciences, University of Wisconsin School of Medicine and Public Health, Madison, WI.
authors.forename: Shoujian
authors.initials: S
authors.lastname: Wang
authors.affiliation: Department of Pediatrics, University of Wisconsin School of Medicine and Public Health, Madison, WI ; McPherson Eye Research Institute, University of Wisconsin School of Medicine and Public health, Madison, WI.
authors.forename: Christine M
authors.initials: CM
authors.lastname: Sorenson
authors.affiliation: Department of Ophthalmology and Visual Sciences, University of Wisconsin School of Medicine and Public Health, Madison, WI ; McPherson Eye Research Institute, University of Wisconsin School of Medicine and Public health, Madison, WI.
authors.forename: Nader
authors.initials: N
authors.lastname: Sheibani
authors.affiliation: Department of Biology, Indiana University-Purdue University Indianapolis,, Indianapolis, IN ; Center for Regenerative Biology and Medicine, Indiana University- Purdue University Indianapolis, Indianapolis, IN.
authors.forename: Teri L
authors.initials: TL
authors.lastname: Belecky-Adams
mesh_terms: D030201:Activin Receptors, Type I / Q000378:metabolism; D000818:Animals; D001253:Astrocytes / Q000187:drug effects / Q000473:pathology / Q000502:physiology*; D055415:Bone Morphogenetic Protein 4 / Q000494:pharmacology / Q000502:physiology; D055419:Bone Morphogenetic Protein 7 / Q000008:administration & dosage / Q000494:pharmacology / Q000502:physiology*; D052005:Bone Morphogenetic Protein Receptors, Type I / Q000378:metabolism; D011508:Chondroitin Sulfate Proteoglycans / Q000235:genetics / Q000378:metabolism; D063928:Ependymoglial Cells / Q000187:drug effects / Q000473:pathology / Q000502:physiology*; D005904:Glial Fibrillary Acidic Protein; D005911:Gliosis / Q000209:etiology* / Q000473:pathology / Q000503:physiopathology; D005974:Glutamate-Ammonia Ligase / Q000235:genetics / Q000378:metabolism; D058449:Intravitreal Injections; D020782:Matrix Metalloproteinases / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D055351:Retinal Neurons / Q000187:drug effects / Q000473:pathology / Q000502:physiology; D015398:Signal Transduction / Q000187:drug effects; D051785:Smad Proteins / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000378:metabolism
chemical_list: C525810:Bmp4 protein, mouse; D055415:Bone Morphogenetic Protein 4; D055419:Bone Morphogenetic Protein 7; D011508:Chondroitin Sulfate Proteoglycans; D005904:Glial Fibrillary Acidic Protein; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; D051785:Smad Proteins; D016212:Transforming Growth Factor beta; C525820:bmp7 protein, mouse; C528900:glial fibrillary astrocytic protein, mouse; D030201:Activin Receptors, Type I; C498943:Acvr1 protein, mouse; C498212:Bmpr1a protein, mouse; C498215:Bmpr1b protein, mouse; D052005:Bone Morphogenetic Protein Receptors, Type I; D020782:Matrix Metalloproteinases; D005974:Glutamate-Ammonia Ligase