Abstract
The distribution as well as the ultrastructural and biochemical characteristics of proliferating cells in the human eye were investigated in five conceptuses of 5–9 postovulatory weeks, using morphological techniques and Ki-67 immunostaining. The Ki-67 nuclear protein was used as a proliferation marker because of its expression in all phases of the cell cycle except the resting phase (G0). The labelling indices of Ki-67-positive cells were analysed by means of the Kruskal-Wallis ANOVA test and the Wilcoxon matched-pairs test. In the 5th week, mitotic cells were the most numerous between the two layers of the optic cup, the optic cup and stalk, and between the lens pit and the surface ectoderm. During the 6th week, cells were observed in the lens epithelium covering the whole cavity of the lens vesicle as well as in the neuroblast zone and the pigmented epithelium of the retina. At later stages (7th–9th weeks), Ki-67-positive cells were restricted to the anterior lens epithelium, the outer neuroblast zone, and the pigmented retina. Throughout all stages examined, mitotic figures were found lying exclusively adjacent to the intraretinal space. Early in the lens pit, they were confined to the free epithelial surface, and later were facing the cavity of the lens vesicle. The proliferative activity was the most intensive in the 6th week, whereas it decreased significantly in the later stages. Additionally, when proliferative activities were compared, the peripheral retina appeared to be less mature than the central before the 9th week. In the earliest analysed stage, cell proliferation might be associated with the sculpturing of the optic cup and stalk, the cornea, and the lens. In the 6th week, the most intensive proliferation seems to be involved not only in the further morphogenesis of the optic cup and the lens vesicle but also in the retinal neurogenesis. At later stages, the decreased proliferation might participate in the neurogenesis of the outer neuroblast zone and the secondary lens fibre formation.
Similar content being viewed by others
References
Ábrahám H, Tornóczky T, Kosztolányi G, Seress L (2001) Cell formation in the cortical layers of the developing human cerebellum. Int J Devel Neurosci 19:53–62
Blanks JC, Bok D (1977) An autoradiographic analysis of postnatal cell proliferation in the normal and degenerative mouse retina. J Comp Neur 174:317–328
Božanić D, Tafra R, Saraga-Babić M (2003) Role of apoptosis and mitosis during human eye development. Eur J Cell Biol 82:421–429
Braun N, Brendel P, Zimmermann H (1995) Distribution of 5’-nucleotidase in the developing mouse retina. Dev Brain Res 88:79–86
Buse E, Eichmann T, deGroot H, Leker A (1993) Differentiation of the mammalian retinal pigment epithelium in vitro: influence of presumptive retinal neuroepithelium and head mesenchyme. Anat Embryol 187:259–268
Caley DW, Johnson C, Liebelt RA (1972) The postnatal development of the retina in the normal and rodless CBA mouse: a light and electron microscopic study. Am J Anat 133:179–212
Cattoretti G, Becker MHG, Key G, Duchrow M, Schlüter C, Galle J, Gerdes J (1992) Monoclonal antibodies against recombinant parts of the Ki-67 antigen (MIB 1 and MIB 3) detect proliferating cells in microwave-processed formalin-fixed paraffin sections. J Pathol 168:357–363
Endl E, Gerdes J (2000) The Ki-67 protein: fascinating forms and an unknown function. Exp Cell Res 257:231–237
Fitzgerald MJT, Fitzgerald M (1994) Head and neck: the eye. In: WB Saunders (ed) Human embriology. Baillière Tindall, London, pp 186–191
Hinds JW, Hinds PL (1974) Early ganglion cell differentiation in the mouse retina: an electron microscopic analysis utilizing serial sections. Dev Biol 37:381–416
Isayama T, McLaughlin PJ, Zagon IS (1991) Endogenous opioids regulate cell proliferation in the retina of developing rat. Brain Res 544:79–85
Karlsson M, Boeryd B, Carstensen J, Frånlund B, Gustafsson B, Kågedal B, Sun X-F, Wingren S (1996) Correlations of Ki-67 and PCNA to DNA ploidy, S-phase fraction and survival in uveal melanoma. Eur J Cancer 32A:357–362
Kee N, Sivalingam S, Boonstra R, Wojtowicz JM (2002) The utility of Ki-67 and BrdU as proliferative markers of adult neurogenesis. J Neurosci Methods 115:97–105
Key G, Kubbutat MHG, Gerdes J (1994) Assessment of cell proliferation by means of an enzyme-linked immunosorbent assay based on the detection of the Ki-67 protein. J Immunol Methods 177:113–117
Klein CL, Wagner M, Kirkpatrick CJ, Van Kooten TG (2000) A new quantitative test method for cell proliferation based on detection of the Ki-67 protein. J Mater Sci-Mater M 11:125–132
Kreitz S, Fackelmayer FO, Gerdes J, Knippers R (2000) The proliferation-specific human Ki-67 protein is a constituent of compact chromatin. Exp Cell Res 261:284–292
Laemle LK, Puszkarczuk M, Feinberg RN (1999) Apoptosis in early ocular morphogenesis in the mouse. Dev Brain Res 112:129–133
Laprie C, Abadie J, Amardeilh MF, Raymond I, Delverdier M (1998) Detection of the Ki-67 proliferation associated nuclear epitope in normal canine tissues using the monoclonal antibody MIB-1. Anat Histol Embryol 27:251–256
Lopez F, Belloc F, Lacombe F, Dumain P, Reiffers J, Bernard P, Boisseau MR (1994) The labeling of proliferating cells by Ki67 and MIB-1 antibodies depends on the binding of a nuclear protein to the DNA. Exp Cell Res 210:145–153
MacCallum DE, Peter AH (1999) Biochemical characterization of pKi67 with the identification of a mitotic-specific form associated with hyperphosphorylation and altered DNA binding. Exp Cell Res 252:186–198
McAvoy JW (1978) Cell division, cell elongation and the co-ordination of crystallin gene expression during lens morphogenesis in the rat. J Embryol Exp Morph 45:271–281
Moore KL (1989) The eye and ear. In: Wonsiewicz M (ed) Before we are born: basic embriology and birth defects. WB Saunders, Philadelphia, pp 269–274
Moser B (1995) A silver stain for the detection of apoptosis at the light microscope. Microsc Anal: 21–23
Müller F, O’Rahilly R (1985) The first appearance of the neural tube and optic primordium in the human embryo at stage 10. Anat Embryol 172:157–169
Nguyen MM, Potter SJ, Griep AE (2002) Deregulated cell cycle control in lens epithelial cells by expression of inhibitors of tumor supressor function. Mech Develop 112:101–113
Ogilvie JM, Speck JD, Lett JM, Fleming TT (1999) A reliable method for organ culture of neonatal mouse retina with long-term survival. J Neurosci Methods 87:57–65
O’Rahilly R (1983) The timing and sequence of events in the development of the human eye and ear during the embryonic period proper. Anat Embryol 168:87–99
O’Rahilly R, Gardner R (1971) The timing and sequence of events in the development of the human nervous system during the embryonic period proper. Z Anat Entwickl Gesch 134:1-12
Pimentel B, Rodríguez-Borlado L, Hernández C, Carrera AC (2002) A role for phosphoinositide 3-kinase in the control of cell division and survival during retinal development. Dev Biol 247:295–306
Robinson G (1982) Electron microscopy 2: transmission (a) tissue preparation; (b) sectioning and staining. In: Bancroft JD, Stevens A (eds) Theory and practice of histological techniques. Churchill Livingstone, Edinburgh, pp 482–518
Sadler TW (1985) Eye. In: Tracy TM (ed) Langman’s medical embryology. Williams & Wilkins, Baltimore, pp 320–328
Schwarting R, Gerdes J, Niehus J, Jaeschke L, Stein H (1986) Determination of the growth fraction in cell suspensions by flow cytometry using the monoclonal antibody Ki-67. J Immunol Methods 90:65–70
Seress L, Ábrahám H, Tornóczky T, Kosztolányi GY (2001) Cell formation in the human hippocampal formation from mid-gestation to the late postnatal period. Neuroscience 105:831–843
Sharma RK, Ehinger B (1997) Cell proliferation in retinal transplants. Cell Transplant 6:141–148
Sivak B, Sivak J (2000) Vertebrate eye development and refractive function: an overview. In: Fini ME (ed) Vertebrate eye development. Springer, Berlin, Heidelberg, New York, pp 1–14
Spira AW, Hollenberg MJ (1973) Human retinal development: ultrastructure of the inner retinal layers. Dev Biol 31:1-21
Welsch U (2002) Zytologie. In: Welsch U (ed) Sobotta Atlas Histologie. Urban & Fischer, Munich, pp 44–47
West-Mays JA, Coyle BM, Piatigorsky J, Papagiotas S, Libby D (2002) Ectopic expression of AP-2α transcription factor in the lens disrupts fiber cell differentiation. Dev Biol 245:13–27
Wigle JT, Chowdhury K, Gruss P, Oliver G (1999) Prox1 function is crucial for mouse lens-fibre elongation. Nat Genet 21:318–322
Wride MA (1996) Cellular and molecular features of lens differentiation: a review of recent advances. Differentiation 61:77–93
Yamaguchi K, Tomita H, Sugano E, Nakazawa T, Tamai M (2002) Mitogen-activated protein kinase inhibitor, PD98059, inhibits rat retinal pigment epithelial cell replication by cell cycle arrest. Jpn J Ophthalmol 46:634–639
Yang P, Seiler MJ, Aramant RB, Whittemore SR (2002) In vitro isolation and expansion of human retinal progenitor cells. Exp Neurol 177:326–331
Young RW (1985a) Cell proliferation during postnatal development of the retina in the mouse. Dev Brain Res 21:229–239
Young RW (1985b) Cell differentiation in the retina of the mouse. Anat Rec 212:199–205
Zelenka PS, Gao C-Y, Rampalli A, Arora J, Chauthaiwale V, He H-Y (1997) Cell cycle regulation in the lens: proliferation, quiescence, apoptosis and differentiation. Prog Retin Eye Res 16:303–322
Zwaan J, Kirkland BM (1975) Malorientation of mitotic figures in the early lens rudiment of aphakia mouse embryos. Anat Rec 182:345–354
Acknowledgements
We are grateful to Mrs. Asja Miletić for her skillful technical assistance. This work is supported by the Ministry of Science and Technology of the Republic of Croatia (grant no. 021 6002).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Božanić, D., Saraga-Babić, M. Cell proliferation during the early stages of human eye development. Anat Embryol 208, 381–388 (2004). https://doi.org/10.1007/s00429-004-0410-5
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00429-004-0410-5