Summary
Fluorochrome conjugated lectins were used to observe cell surface changes in the corneal endothelium during wound repair in the adult rat and during normal fetal development. Fluorescence microscopy of non-injured adult corneal endothelia incubated in wheat-germ agglutinin (WGA), Concanavalin A (Con A), and Ricinus communis agglutinin I (RCA), revealed that these lectins bound to cell surfaces. Conversely, binding was not observed for either Griffonia simplicifolia I (GS-I), soybean agglutinin (SBA) or Ulex europaeus agglutinin (UEA). Twenty-four hours after a circular freeze injury, endothelial cells surrounding the wound demonstrated decreased binding for WGA and Con A, whereas, RCA binding appeared reduced but centrally clustered on the apical cell surface. Furthermore, SBA now bound to endothelial cells adjacent to the wound area, but not to cells near the tissue periphery. Neither GS-I nor UEA exhibited any binding to injured tissue. By 48 h post-injury, the wound area repopulates and endothelial cells begin reestablishing the monolayer. These cells now exhibit increased binding for WGA, especially along regions of cell-to-cell contact, whereas, Con A, RCA and SBA binding patterns remain unchanged. Seventy-two hours after injury, the monolayer is well organized with WGA, Con A and RCA binding patterns becoming similar to those observed for non-injured tissue. However, at this time, SBA binding decreases dramatically. By 1 week post-injury, binding patterns for WGA, ConA and RCA closely resemble their non-injured counterparts while SBA continues to demonstrate low levels of binding. In early stages of its development, the endothelium actively proliferates and morphologically resembles adult tissue during wound repair. The 16-day fetal tissue is mitotically active, does not exhibit a well defined monolayer, and demonstrates weak fluorescence binding for WGA, Con A and RCA. Conversely, SBA binding is readily detected on many cell surfaces. By 19 days in utero, the endothelial monolayers becomes organized and cell proliferation greatly diminishes. WGA, Con A and RCA now exhibit binding similar to that seen in the adult tissue. SBA binding is not detected at this time. Thus, changes in lectin binding during wound repair of the adult rat corneal endothelium mimic changes in lectin binding seen during the development of the tissue.
Similar content being viewed by others
References
Borek C, Grob M, Burger MM (1973) Surface alterations in transformed epithelial and fibroblastic cells in culture: a disturbance of membrane degradation versus biosynthesis? Exp Cell Res 77:207–215
Calderó J, Campo E, Torra M (1988) Distribution and changes of glycoconjugates in rat colonic mucosa during development. A histochemical study using lectins. Histochemistry 90:261–270
Chi HH, Teng CC, Katzin HM (1960) Healing process in the mechanical denudation of the corneal endothelium. Arch Ophthalmol 49:693–703
Clark RAF (1988) Potential roles of fibronectin in cutaneous wound repair. Arch Dermatol 124:201–206
Cloyd MW, Bigner DD (1977) Surface morphology of normal and neoplastic rat cells. Am J Pathol 88:29–52
Doughman DJ, VanHorn DL, Rodman WP, Byrnes P, Lindstrom RL (1976) Human corneal endothelial layer repair during organ culture. Arch Ophthalmol 94:1791–1796
Dufour S, Duband JL, Thiery JP (1986) Role of major cell-substratum adhesion system in cell behavior and morphogenesis. Biol Cell 58:1–14
Edelman GM (1983) Cell adhesion molecules. Science 219:450–457
ffrench-Constant C, VanDeWater L, Dvorak HF, Hynes RO (1989) Reappearance of an embryonic pattern of fibronectin splicing during wound repair. J Cell Biol 109:903–914
Fisher C, Holbrook KA (1987) Cell surface and cytoskeletal changes associated with epidermal stratification and differentiation in organ cultures of embryonic human skin. Dev Biol 119:231–241
Fox TO, Sheppard JR, Burger MM (1971) Cyclic membrane changes in animal cells: Transformed cells permanently display a surface architecture detected only during mitosis. Proc Natl Acad Sci USA 68:244–247
Gabbiani G, Gabbiani F, Lombardi D, Schwartz SM (1983) Organization of actin cytoskeleton in normal and regenerating arterial endothelial cells. Proc Natl Acad Sci USA 80:2361–2364
Gipson IK, Kiorpes TC (1982) Epithelial sheet movement: protein and glycoprotein synthesis. Dev Biol 92:259–262
Gipson IK, Riddle CV, Kiorpes TC, Spurr SJ (1983) Lectin binding to cell surfaces: comparisons between normal and migrating corneal epithelium. Dev Biol 96:337–345
Gipson IK, Kiorpes TC, Brennan SJ (1984) Epithelial sheet movement: effects of tunicamycin on migration and glycoprotein synthesis. Dev Biol 101:212–220
Gordon SR (1988) Changes in distribution of extracellular matrix proteins during wound repair in corneal endothelium. J Histochem Cytochem 36:409–416
Gordon SR (1990) Changes in extracellular matrix proteins and actin during corneal endothelial growth. Invest Ophthalmol Vis Sci 31:94–101
Gordon SR, Rothstein H (1978) Studies on corneal endothelial growth and repair. I. Microfluorometric and autoradiographic analyses of DNA synthesis, mitosis and amitosis following freeze injury. Metab Ophthalmol 2:57–63
Gordon SR, Rothstein H (1982) Studies on corneal endothelial growth and repair. III. Effects of DNA and RNA synthesis inhibitors upon restoration of transparency following injury. Ophthalmic Res 14:195–209
Gordon SR, Staley CA (1990) Role of the cytoskeleton during injury-induced cell migration in corneal endothelium. Cell Motil Cytoskel 16:47–57
Gordon SR, Essner E, Rothstein H (1982) In situ demonstration of actin in normal and injured ocular tissues using 7-nitrobenz-2-oxa-1,3-diazole phallacidin. Cell Motil 2:343–354
Gordon SR, Rothstein H, Harding CV (1983) Studies on corneal endothelial growth and repair. IV. Changes in the surface during cell division as revealed by scanning electron microscopy. Eur J Cell Biol 31:26–33
Hebel R, Stromberg MW (1986) Anatomy and embryology of the laboratory rat. BioMed Verlag, Worthsee, pp 231–257
Hergott GJ, Sandig M, Kalnins VI (1989) Cytoskeletal organization of migrating retinal pigment epithelial cells during wound healing in organ culture. Cell Motil Cytoskel 13:83–93
Johnson GD, Araujo GMDN (1982) A simple method of reducing the fading of immunofluorescence during microscopy. J Immunol Methods 43:349–350
Kimber SJ (1989) Changes in cell-surface glycoconjugates during embryonic development demonstrated using lectins and other probes. Biochem Soc Trans 17:23–27
Lis H, Sharon N (1986) Lectins as molecules and as tools. Annu Rev Biochem 55:35–67
Lundgren E, Roos G (1976) Cell surface changes as an indication of cell cycle events. Cancer Res 36:4044–4051
Matsuda M, Sawa M, Edelhauser HF, Bartels SP, Neufeld AH, Kenyon KR (1985) Cellular migration and morphology in corneal endothelial repair. Invest Ophthalmol Vis Sci 26:443–449
Matsumura T, Sugimoto T, Sawada T, Amagai T, Negoro S, Kemshead JT (1987) Cell surface membrane antigen present on neuroblastoma cells but not fetal neuroblasts recognized by a monoclonal antibody (KP-NAC8). Cancer Res 47:2924–2930
Muramatsu T (1988) Alterations of cell-surface carbohydrates during differentiation and development. Biochimie 70:1587–1596
Nutall RP (1976) DNA synthesis during the development of the chick cornea. J Exp Zool 198:193–208
Oppenheimer SB (1978) Cell surface carbohydrates in adhesion and migration. Am Zool 18:13–23
Panjwani N, Baum J (1985) Rabbit corneal endothelial cell surface glycoproteins. Invest Ophthalmol Vis Sci 26:450–456
Podmaniczky E, Szende B, Lapis K, Ferenc G (1976) Cell surface changes observed in MC-29 virus infected chicken embryo fibroblast (CEC) cultures. Int J Cancer 18:536–539
Porter KR, Todaro GJ, Fonte V (1973) A scanning electron microscope study of surface features of viral and spontaneously transformants of mouse BALB/3T3 cells. J Cell Biol 59:633–642
Rauvala H (1983) Cell surface carbohydrates and cell ahdesion. Trends Biochem Sci 8:323–325
Rieber M, Rieber MS (1981) Metastatic potential correlates with cell-surface protein alterations in B-16 melanoma variants. Nature 293:74–76
Sabet MD, Gordon SR (1989) Ultrastructural immunocytochemical localization of fibronectin deposition during corneal endothelial wound repair. Evidence for cytoskeletal involvement. Biol Cell 65:171–179
Sallman L von, Caravaggio LL, Grimes P (1961) Studies on the corneal endothelium of the rabbit. I. Cell division and growth. Am J Ophthalmol 51:955–966
Sanger JM, Reingold AM, Sanger JW (1984) Cell surface changes during mitosis and cytokinesis of epithelial cells. Cell Tissue Res 237:409–417
Takashima A, Billingham RE, Grinnell F (1986) Activation of rabbit keratinocyte fibronectin receptor function in vivo during wound healing. J Invest Dermatol 86:585–590
Turner RS, Burger MM (1974) A cell surface change in mitotic fibroblasts monitored with lectins. Ann NY Acad Sci 234:332–347
Wieser RJ, Janik-Schmitt B, Renauer D, Schafer A, Heck R, Oesch F (1988) Contact-dependent inhibition of growth of normal diploid human fibroblasts by plasma membrane glycoproteins. Biochimie 70:1661–1671
Author information
Authors and Affiliations
Additional information
Supported by grant EY-06435 from The National Institutes of Health
Rights and permissions
About this article
Cite this article
Gordon, S.R., Marchand, J. Lectin binding to injured corneal endothelium mimics patterns observed during development. Histochemistry 94, 455–462 (1990). https://doi.org/10.1007/BF00272607
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00272607