title: Immediate upregulation of proteins belonging to different branches of the apoptotic cascade in the retina after optic nerve transection and optic nerve crush.
abstract: OBJECTIVE
  To further investigate the molecular signals underlying optic nerve (ON) injury, the authors analyzed in adult control, ON-transected, and ON-crushed retinas the expression pattern and time-course regulation of the following proteins, all of which are linked to apoptosis through different pathways: Stat 1, caspase 11 (inflammation and death), cathepsins C and B (lysosomal death pathway), calpain 1 (endoplasmic reticulum stress), calreticulin (apoptosis marker), Jun (early response), and aryl hydrocarbon receptor (cell cycle arrest).
  
  
  METHODS
  Adult female rats were subjected to intraorbital optic nerve transection (IONT) or intraorbital optic nerve crush (IONC). Protein from naive and ON-injured adult rat retinas was extracted at different times postlesion, and Western blotting experiments were performed. For immunohistofluorescence analyses, retinal ganglion cells (RGCs) were retrogradely identified with fluorogold applied to the superior colliculi 1 week before injury.
  
  
  RESULTS
  Western blotting analyses revealed upregulation of all the analyzed proteins as early as 12 hours postlesion (hpl), peaking at 48 hpl, in agreement with our previous RNA study findings. Furthermore, immunohistofluorescence to radial sections showed that all but Stat 1 were expressed by the primarily injured neurons, the RGCs, as seen by colocalization with fluorogold.
  
  
  CONCLUSIONS
  All analyzed proteins were upregulated in the retina after IONT or IONC as early as 12 hpl, indicating that ON injury regulates several branches of the apoptotic cascade and suggesting that commitment to death might be an earlier event than previously anticipated.
authors.affiliation: Departamento de Oftalmología, Facultad de Medicina, Universidad de Murcia, Murcia, Spain. martabar@um.es
authors.forename: Marta
authors.initials: M
authors.lastname: Agudo
authors.forename: Maria C
authors.initials: MC
authors.lastname: Pérez-Marín
authors.forename: Paloma
authors.initials: P
authors.lastname: Sobrado-Calvo
authors.forename: Ulrika
authors.initials: U
authors.lastname: Lönngren
authors.forename: Manuel
authors.initials: M
authors.lastname: Salinas-Navarro
authors.forename: Isabel
authors.initials: I
authors.lastname: Cánovas
authors.forename: Francisco M
authors.initials: FM
authors.lastname: Nadal-Nicolás
authors.forename: Jaime
authors.initials: J
authors.lastname: Miralles-Imperial
authors.forename: Finn
authors.initials: F
authors.lastname: Hallböök
authors.forename: Manuel
authors.initials: M
authors.lastname: Vidal-Sanz
mesh_terms: D000818:Animals; D017209:Apoptosis; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism*; D015153:Blotting, Western; D064032:Calbindin 2; D020169:Caspases / Q000378:metabolism; D002401:Cathepsin B / Q000378:metabolism; D020789:Cathepsin C / Q000378:metabolism; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D009409:Nerve Crush; D020221:Optic Nerve Injuries / Q000378:metabolism*; D016755:Proto-Oncogene Proteins c-jun / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D018336:Receptors, Aryl Hydrocarbon / Q000378:metabolism; D012160:Retina / Q000378:metabolism*; D012165:Retinal Ganglion Cells / Q000378:metabolism / Q000473:pathology; D064030:S100 Calcium Binding Protein G / Q000378:metabolism; D050794:STAT1 Transcription Factor / Q000378:metabolism; D013997:Time Factors; D015854:Up-Regulation
chemical_list: D051017:Apoptosis Regulatory Proteins; D064032:Calbindin 2; D016755:Proto-Oncogene Proteins c-jun; D018336:Receptors, Aryl Hydrocarbon; D064030:S100 Calcium Binding Protein G; D050794:STAT1 Transcription Factor; C494081:Stat1 protein, rat; D020789:Cathepsin C; C533920:Ctsc protein, rat; C491071:Casp4 protein, rat; D020169:Caspases; D002401:Cathepsin B; C533916:Ctsb protein, rat