Relationship of Telomeres and p53 in Aging Bovine Corneal Endothelial Cell Cultures

• Published in: Investigative ophthalmology & visual science Vol. 41; no. 5; pp. 1070 - 1075
• Main Authors: Whikehart, David R, Register, Shilpa J, Chang, Qing, Montgomery, Brodrick
• Format: Journal Article
• Language: English
• Published: Rockville, MD ARVO 01.04.2000; Association for Research in Vision and Ophtalmology
• Subjects: Animals; beta-Galactosidase - metabolism; Biological and medical sciences; Cattle; Cells, Cultured; Cellular Senescence - physiology; DNA - analysis; Endothelium, Corneal - cytology; Endothelium, Corneal - physiology; Enzyme-Linked Immunosorbent Assay; Eye and associated structures. Visual pathways and centers. Vision; Fundamental and applied biological sciences. Psychology; Telomere - physiology; Tumor Suppressor Protein p53 - metabolism; Vertebrates: nervous system and sense organs; Eye; Cell culture; Endothelial cell; Cornea; Length; Evolution; Telomere; Age
• ISSN: 0146-0404; 1552-5783

To demonstrate a relationship between telomere lengths and levels of p53 in cultured bovine corneal endothelial cells (CECs) during aging. Bovine CECs were grown and aged as long-term cultures. Telomere lengths were determined directly on gels with 32P probes after treatment of isolated DNA with RsaI and HinfI. Protein p53 was determined using an enzyme-linked immunosorbent sandwich assay. Cellular aging and the development of replicative senescence were monitored by the appearance of senescent morphology and the beta-galactosidase assay. Bovine CEC telomeres lost 4 kb (from 12.8 to 8.8 kb) over 1 year (89 population doublings [PDs]). The p53 levels in bovine CECs were initially small (approximately 60 pg/million cells), but rose 3.5-fold by culture age of 260 days (64 PDs). On initiation, cultured bovine CECs did not stain for the senescent marker beta-galactosidase. However, these cells stained at 89 PDs and senescent morphology was observed in the cultures at 64 PDs. The data indicate an inverse relationship between telomere lengths (decreasing) and levels of p53 (increasing) in bovine CECs during aging. These properties may influence the ability of these cells to divide as they enter into replicative senescence.