title: Complement expression in the retina is not influenced by short-term pressure elevation.
abstract: OBJECTIVE
  To determine whether short-term pressure elevation affects complement gene expression in the retina in vitro and in vivo.
  
  
  METHODS
  Muller cell (TR-MUL5) cultures and organotypic retinal cultures from adult mice and monkeys were subjected to either 24-h or 72-h of pressure at 0, 15, 30, and 45 mmHg above ambient. C57BL/6 mice were subjected to microbead-induced intraocular pressure (IOP) elevation for 7 days. RNA and protein were extracted and used for analysis of expression levels of complement component genes and complement component 1, q subcomponent (C1q) and complement factor H (CFH) immunoblotting.
  
  
  RESULTS
  mRNA levels of complement genes and C1q protein levels in Muller cell cultures remained the same for all pressure levels after exposure for either 24 or 72 h. In primate and murine organotypic cultures, pressure elevation did not produce changes in complement gene expression or C1q and CFH protein levels at either the 24-h or 72-h time points. Pressure-related glial fibrillary acidic protein (GFAP) mRNA expression changes were detected in primate retinal organotypic cultures (analysis of variance [ANOVA]; p<0.05). mRNA expression of several other genes changed as a result of time in culture. Eyes subjected to microbead-induced IOP elevation had no differences in mRNA expression of complement genes and C1q protein levels (ANOVA; p>0.05 for both) with contralateral control and naïve control eyes.
  
  
  CONCLUSIONS
  Short-term elevation of pressure in vitro as well as short-term (1 week) IOP elevation in vivo does not seem to dramatically alter complement system gene expression in the retina. Prolonged expression to elevated pressure may be necessary to affect the complement system expression.
authors: Department of Cell Biology, SUNY Downstate Medical Center, Brooklyn, NY.
authors: Konstantin
authors: K
authors: Astafurov
authors: Department of Ophthalmology, SUNY Downstate Medical Center, Brooklyn, NY.
authors: Cecilia Q
authors: CQ
authors: Dong
authors: Department of Cell Biology, SUNY Downstate Medical Center, Brooklyn, NY.
authors: Lampros
authors: L
authors: Panagis
authors: Department of Ophthalmology, Mount Sinai School of Medicine, New York, NY.
authors: Gautam
authors: G
authors: Kamthan
authors: Department of Cell Biology, SUNY Downstate Medical Center, Brooklyn, NY.
authors: Lizhen
authors: L
authors: Ren
authors: Department of Biological Sciences, CUNY Kingsborough Community College, Brooklyn, NY.
authors: Anna
authors: A
authors: Rozenboym
authors: Department of Psychiatry, Columbia University Medical Center and New York State Psychiatric Institute, New York, NY.
authors: Tarique D
authors: TD
authors: Perera
authors: Department of Psychiatry, SUNY Downstate Medical Center, Brooklyn, NY.
authors: Jeremy D
authors: JD
authors: Coplan
authors: Department of Cell Biology, SUNY Downstate Medical Center, Brooklyn, NY ; Department of Ophthalmology, SUNY Downstate Medical Center, Brooklyn, NY.
authors: John
authors: J
authors: Danias
mesh_terms: D000818:Animals; D002469:Cell Separation; D002478:Cells, Cultured; D003165:Complement System Proteins / Q000378:metabolism*; D005110:Extracellular Space / Q000378:metabolism; D007267:Injections; D007429:Intraocular Pressure / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008863:Microspheres; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D012160:Retina / Q000378:metabolism*; D013696:Temperature; D013997:Time Factors
chemical_list: D012333:RNA, Messenger; D003165:Complement System Proteins