March 1995
Volume 36, Issue 3
Free
Articles  |   March 1995
Use of the MIB-1 antibody for detecting proliferating cells in the retina.
Author Affiliations
  • S F Geller
    Neuroscience Research Institute, University of California, Santa Barbara 93106-5060.
  • G P Lewis
    Neuroscience Research Institute, University of California, Santa Barbara 93106-5060.
  • D H Anderson
    Neuroscience Research Institute, University of California, Santa Barbara 93106-5060.
  • S K Fisher
    Neuroscience Research Institute, University of California, Santa Barbara 93106-5060.
Investigative Ophthalmology & Visual Science March 1995, Vol.36, 737-744. doi:
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      S F Geller, G P Lewis, D H Anderson, S K Fisher; Use of the MIB-1 antibody for detecting proliferating cells in the retina.. Invest. Ophthalmol. Vis. Sci. 1995;36(3):737-744.

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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.

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