title: Novel MYOC gene mutation, Phe369Leu, in Japanese patients with primary open-angle glaucoma detected by denaturing high-performance liquid chromatography.
abstract: OBJECTIVE
  To screen for mutations in the MYOC gene in Japanese patients with primary open-angle glaucoma (POAG) using denaturing high-performance liquid chromatography (DHPLC).
  
  
  METHODS
  Blood samples were collected from 171 patients with POAG and 100 controls from seven institutions in Japan. For high-throughput analysis, seven exonic regions were amplified by polymerase chain reaction using DNA pooled from three patients; each DNA pool was then analyzed chromatographically. For analysis of a small number of samples, 7 exonic regions were amplified separately but simultaneously with annealing at 58 degrees C in each patient and then chromatographed, using 7 wells of the same 96-well plate per sample. When chromatographic patterns were abnormal by either method, the PCR products of the individual samples were sequenced.
  
  
  RESULTS
  Four glaucoma-causing mutations were identified in five POAG patients (2.9%). One missense mutation, Phe369Leu, is new; and three others, Ile360Asn, Ala363Thr, and Thr448Pro, have been reported in Japanese patients. Phe369Leu was associated with adult onset POAG.
  
  
  CONCLUSIONS
  Mutations in the MYOC gene were demonstrated chromatographically in 2.9% of our Japanese POAG patients. The use of pooled DNAs with DHPLC analysis is a time- and labor-saving technique. All mutations detected appear to be specific to Japanese patients.
authors.affiliation: Department of Ophthalmology, Keio University School of Medicine, Tokyo, Japan.
authors.forename: Karin
authors.initials: K
authors.lastname: Ishikawa
authors.forename: Tomoyo
authors.initials: T
authors.lastname: Funayama
authors.forename: Yuichiro
authors.initials: Y
authors.lastname: Ohtake
authors.forename: Tomihiko
authors.initials: T
authors.lastname: Tanino
authors.forename: Daijiro
authors.initials: D
authors.lastname: Kurosaka
authors.forename: Kotaro
authors.initials: K
authors.lastname: Suzuki
authors.forename: Hidenao
authors.initials: H
authors.lastname: Ideta
authors.forename: Takuro
authors.initials: T
authors.lastname: Fujimaki
authors.forename: Hidenobu
authors.initials: H
authors.lastname: Tanihara
authors.forename: Ryo
authors.initials: R
authors.lastname: Asaoka
authors.forename: Nobuhisa
authors.initials: N
authors.lastname: Naoi
authors.forename: Noriko
authors.initials: N
authors.lastname: Yasuda
authors.forename: Takeshi
authors.initials: T
authors.lastname: Iwata
authors.forename: Yukihiko
authors.initials: Y
authors.lastname: Mashima
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000595:Amino Acid Sequence; D044466:Asian People / Q000235:genetics*; D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003598:Cytoskeletal Proteins; D004247:DNA; D005136:Eye Proteins / Q000235:genetics; D005820:Genetic Testing; D005902:Glaucoma, Open-Angle / Q000235:genetics* / Q000473:pathology / Q000503:physiopathology; D006023:Glycoproteins / Q000235:genetics; D006801:Humans; D007930:Leucine; D008875:Middle Aged; D020125:Mutation, Missense; D009898:Optic Disk / Q000473:pathology; D010649:Phenylalanine; D016133:Polymerase Chain Reaction / Q000379:methods; D014794:Visual Fields; D000099179:Myocilin
chemical_list: D003598:Cytoskeletal Proteins; D004247:DNA; D005136:Eye Proteins; D006023:Glycoproteins; D007930:Leucine; D010649:Phenylalanine; D000099179:Myocilin