AOHC : Vol. 64 (2001) , No. 4 p.379-391

Novel Use of Bovine Zeta-Crystallin as a Conformational DNA Probe to Characterize a Phase Transition Zone and Terminally Differentiating Fiber Cells in the Adult Canine Ocular Lens

Claude E. GAGNA¹⁾²⁾³⁾, Hon-Reen KUO¹⁾, Nick AGOSTINO³⁾, Douglas RIZZO³⁾, Irwin R. ISQUITH³⁾, Joslyn MATHEW²⁾, Jubina MOHAMMED²⁾, Sheldon HOO³⁾ and W. Clark LAMBERT¹⁾

1) Department of Pathology and Laboratory Medicine, University of Medicine and Dentistry of New Jersey-Medical School
2) School of Allied Health and Life Sciences, New York Institute of Technology
3) School of Natural Sciences, Fairleigh Dickinson University

(Received April 9, 2001)

Summary. Using a novel immunocytochemical staining method, we aimed to characterize the phase transition zone (PTZ) (∼100 μm) in adult ocular lenses and the process of terminal differentiation (denucleation) within normal fiber cells. The binding to DNA of zeta(ζ)-crystallin (Z-DNA-binding protein) and anti-double-stranded (ds-)-B-DNA antibody probes was found to decline gradually throughout denucleating fibers, with a precipitous decrease occurring at about 100 μm (PTZ). Nuclei of superficial fiber cells (in front of the PTZ) showed the highest DNA probe-binding values, followed by middle fibers (MF) and deep fibers (DF). With the use of ζ-crystallin, anti-ds-B-DNA antibody, and anti-single stranded (ss-) DNA antibody probes, it was possible to reveal a loss of reactivity of fiber cell ds-DNA. Ss-DNA antibody binding was seen initially in the MF and reached its highest intensity level in the DF. The pattern of ζ-crystallin probe-DNA reactivity correlates with the loss of anti-B-DNA antibody staining and decreased eosin-protein staining. These data suggest that a reorganization of DNA and intracellular protein supramolecular order in normal adult lenses occurs at a depth of about 100 μm (PTZ).

	To cite this article:
	Claude E. GAGNA, Hon-Reen KUO, Nick AGOSTINO, Douglas RIZZO, Irwin R. ISQUITH, Joslyn MATHEW, Jubina MOHAMMED, Sheldon HOO and W. Clark LAMBERT: “Novel Use of Bovine Zeta-Crystallin as a Conformational DNA Probe to Characterize a Phase Transition Zone and Terminally Differentiating Fiber Cells in the Adult Canine Ocular Lens”. Arch. Histol. Cytol. Vol. 64: 379-391 (2001) .

	doi:10.1679/aohc.64.379
	JOI JST.JSTAGE/aohc/64.379