title: Cellular migration associated with macular hole: a new method for comprehensive bird's-eye analysis of the internal limiting membrane.
abstract: OBJECTIVE
  To elucidate the pathogenesis of macular hole formation, focusing in particular on the possible role of cellular migration on the cortical vitreous and internal limiting membrane (ILM) around the macular hole.
  
  
  METHODS
  To gain a comprehensive overview of the ILM excised in macular hole surgery (n = 36), the ILMs were carefully unfolded and spread out onto glass slides as continuous flat sheets that each contained a macular hole. The specimens were observed by light microscopy and transmission electron microscopy (n = 9), and the cellular distribution was analyzed by scanning electron microscopy in a quantitative manner (n = 27). Immunohistochemistry for glial fibrillary acidic protein and cytokeratin 18 was carried out for cellular characterization. Cellular proliferation was assessed by immunohistochemistry for proliferating cell nuclear antigen and Ki-67.
  
  
  RESULTS
  Cellular migration was not apparent around the macular hole in the early stage of development of the macular hole (stage 2, 0 microm). As the macular hole passed through the later stages of development, cellular migration developed around the macular hole (stage 3, 84 microm) and the area of cellular migration gradually enlarged (stage 4, 420 microm). The immunophenotypic analysis showed that these cells were mainly glial fibrillary acidic protein-positive glial cells and cytokeratin 18-positive retinal pigment epithelial cells. The proliferating cell nuclear antigen and Ki-67 immunohistochemistry showed that some of these cells were proliferating on the ILM.
  
  
  CONCLUSIONS
  Cellular migration on the ILM is not necessary for the initial formation of a macular break. Cellular migration developed after the macular break occurred, and the migration and proliferation increased gradually from the macular hole.
  
  
  CONCLUSIONS
  This study provides a new method for understanding the ultrastructural analysis of the pathogenesis of the macular hole.
authors.affiliation: Department of Ophthalmology and Morphology Core, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan. hisatomi@med.kyushu-u.ac.jp
authors.forename: Toshio
authors.initials: T
authors.lastname: Hisatomi
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Enaida
authors.forename: Taiji
authors.initials: T
authors.lastname: Sakamoto
authors.forename: Takaaki
authors.initials: T
authors.lastname: Kanemaru
authors.forename: Tadahisa
authors.initials: T
authors.lastname: Kagimoto
authors.forename: Ichiro
authors.initials: I
authors.lastname: Yamanaka
authors.forename: Akifumi
authors.initials: A
authors.lastname: Ueno
authors.forename: Takao
authors.initials: T
authors.lastname: Nakamura
authors.forename: Yasuaki
authors.initials: Y
authors.lastname: Hata
authors.forename: Tatsuro
authors.initials: T
authors.lastname: Ishibashi
mesh_terms: D001485:Basement Membrane / Q000378:metabolism / Q000648:ultrastructure; D002465:Cell Movement; D049109:Cell Proliferation; D019773:Epiretinal Membrane / Q000378:metabolism / Q000473:pathology*; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D006801:Humans; D007633:Keratins / Q000378:metabolism; D019394:Ki-67 Antigen / Q000378:metabolism; D008855:Microscopy, Electron, Scanning; D016253:Microscopy, Immunoelectron; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D011446:Prospective Studies; D012167:Retinal Perforations / Q000378:metabolism / Q000473:pathology*
chemical_list: D005904:Glial Fibrillary Acidic Protein; D019394:Ki-67 Antigen; D018809:Proliferating Cell Nuclear Antigen; D007633:Keratins