Use of the MIB-1 antibody for detecting proliferating cells in the retina

Invest Ophthalmol Vis Sci. 1995 Mar;36(3):737-44.

Abstract

Purpose: To study intraretinal proliferation as a response to experimental retinal detachment using an antibody that recognizes the nuclear specific antigen Ki-67 in proliferating cells.

Methods: Experimental retinal detachments were produced in cats (1, 3, 7, and 28 days) and rabbits (1, 3, and 7 days). The animals were killed and the eyes were fixed and embedded in paraffin. Histologic sections were processed for immunohistochemistry using the MIB-1 antibody to detect the Ki-67 protein. Labeled cells were identified, and the proliferative response was quantified.

Results: In normal cat retina, approximately 0.05 cells per millimeter of retina are labeled. In cat retina detached for 1, 3, 7, or 28 days, the number of cells labeled by MIB-1 is 0.06, 5.03, 1.38, and 0.23 cells per millimeter of retina, respectively. MIB-1 labeling yields an approximate fivefold increase over the number of proliferating cells detected in retinal sections using 3H-thymidine autoradiography. Detachment of the rabbit retina elicits a similar response as measured by MIB-1 immunohistochemistry.

Conclusions: In contrast to 3H-thymidine, which labels cells in S-phase only, the MIB-1 antibody labels proliferating cells regardless of their location within the cell cycle. MIB-1 labeling, therefore, is a more accurate means of evaluating cellular proliferation in the retina and elsewhere in the central nervous system, and it is a relatively simple way of evaluating the effects of agents that may affect this response.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Antibodies, Monoclonal*
  • Autoradiography
  • Cats
  • Cell Count
  • Cell Division
  • DNA / biosynthesis
  • DNA Replication
  • Immunoenzyme Techniques
  • Ki-67 Antigen
  • Neoplasm Proteins*
  • Nuclear Proteins*
  • Rabbits
  • Retina / pathology*
  • Retinal Detachment / pathology*
  • Thymidine / metabolism

Substances

  • Antibodies, Monoclonal
  • Ki-67 Antigen
  • Neoplasm Proteins
  • Nuclear Proteins
  • DNA
  • Thymidine