Summary
The capacities of retinal and pigmented cells to regenerate histotypic in-vitro-retinae (IVR) in rotary culture were investigated by dividing the eye cups of 6-day-old chicken embryos into a central and a peripheral part; they were cut along the ora serrata, and the retinal and the pigmented constituents of both parts were isolated. The 4 dissociated cell populations were cultured separately and in all double combinations. Two different types of IVR's were generated; one developed from central or peripheral retinal cells, the other required the addition of pigmented cells from the ciliary margin of the eye. The shape of these IVR's was examined using scanning electron microscopy, and they were also characterized histologically. The acetylcholinesterase pattern marked the inner half of the retina; F11-antibody and a peanut agglutinin marker revealed both plexiform layers and a radial fiber system. In both types, organized histotypical areas consisted of complete sets of retinal layers. In the type containing pigmented cells from the eye periphery, the sequence of layers was identical with that of an in-situ-retina (“laminar IVR”). In IVR's derived from retinal cells only, the sequence of layers was reversed (“rosetted IVR”).
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References
Adler R (1973) Cell interactions and histogenesis in embryonic neural aggregates. Exp Cell Res 77:367–375
Adler R, Jerdan J, Hewitt AT (1985) Responses of cultured neural retinal cells to substratum-bound laminin and other extracellular matrix molecules. Dev Biol 112:100–114
Adler R, Teitelman G (1974) Aggregates formed by mixtures of embryonic neural cells: Activity of enzymes of the cholinergic system. Dev Biol 39:317–321
Akagawa K, Hicks D, Barnstable CJ (1987) Histiotypic organization and cell differentiation in rat retinal reaggregate cultures. Brain Res 437:298–308
Armstrong PB (1971) Light and electron microscopic studies of cell sorting in combinations of chick embryo neural retina and retinal pigment epithelium. Wilhelm Roux Archiv 168:125–141
Beebe DC, Reichman E (1988) The development of the ciliary epithelium and its role in eye growth. [Abstr] 8th Int Congr Eye Res, p 97
Bryan JA, Campochiaro PA (1986) A retinal pigment epithelial cell-derived growth factor(s). Arch Ophthalmol 104:422–425
Burck HC (1981) In: Histologische Technik. Thieme, Stuttgart-New York
Burke JM, Foster SJ (1985) Induction of DNA synthesis by coculture of retinal glia and pigment epithelium. Invest Ophthalmol Vis Sci 26:636–642
Coulombre JL, Coulombre AJ (1965) Regeneration of neural retina from pigmented epithelium in the chick embryo. Dev Biol 12:79–92
Coulombre JL, Coulombre AJ (1970) Influence of mouse neural retina on regeneration of chick neural retina from chick embryonic pigmented epithelium. Nature 228:559–560
Detwiler SR, VanDyke RH (1953) The induction of neural retina from the pigment epithelial layer of the eye. J Exp Zool 122:367–384
Dorris F (1938) Differentiation of the chick eye in vitro. J Exp Zool 78:386–407
Dragomirov N (1936) Über Induktion sekundärer Retina im transplantierten Augenbecher bei Triton und Pelobates. Wilhelm Roux Archiv 134:716–737
Dütting D, Gierer A, Hansmann G (1983) Self-renewal of stem cells and differentiation of nerve cells in the developing chick retina. Dev Brain Res 10:21–32
Fujisawa H (1971) A complete reconstruction of the neural retina of chick embryos grafted onto the chorio-allantoic membrane. Dev Growth Differentiation 13:25–36
Fujisawa H (1973) The process of reconstruction of histological architecture from dissociated retinal cells. Wilhelm Roux Archiv 171:312–330
Hewitt AT, Newsome DA (1988) Altered proteoglycans in cultured human retinitis pigmentosa retinal-pigment epithelium. Invest Ophthalmol Vis Sci 29:720–726
Itoh Y, Eguchi G (1986) In vitro analysis of cellular metaplasia from pigmented epithelial cells to lens phenotypes: a unique model system for studying cellular and molecular mechanisms of “transdifferentiation”. Dev Biol 115:353–362
Keefe JR (1973) An analysis of urodelian regeneration: I. Studies of the cellular source of retinal regeneration in Notophthalmus viridescens utilizing 3H-thymidine. J Exp Zool 184:185–206
Klein G, Langegger M, Timpl R, Ekblom P (1988) Role of laminin A chain in the development of epithelial cell polarity. Cell 55:331–341
Kugler O (1987) Improvement of the methods of Karnovsky and Roots for the histochemical demonstration of acetylcholinesterase. Histochemistry 86:531–532
Layer PG (1983) Comparative localization of acetylcholinesterase and pseudocholinesterase during morphogenesis of the chick brain. Proc Natl Acad Sci USA 80:6413–6417
Layer PG, Sporns O (1987) Spatiotemporal relationship of embryonic cholinesterases with cell proliferation in chicken brain and eye. Proc Natl Acad Sci USA 84:284–288
Linser P, Moscona AA (1979) Induction of glutamine synthetase in embryonic neural retina: Localization of Müller fibers and dependence on cell interactions. Proc Natl Acad Sci USA 76:6476–6480
Liu L, Layer PG, Gierer A (1983) Binding of FITC-coupled peanut-agglutinin (FITC-PNA) to embryonic chicken retinae reveals developmental spatio-temporal patterns. Dev Brain Res 8:223–229
Liu L, Cheng SH, Jiang LZ, Hansmann G, Layer PG (1988) The pigmented epithelium sustains cell growth and tissue differentiation of chicken retinal explants in vitro. Exp Eye Res 46:801–812
Martin GR, Timpl R (1987) Laminin and other basement membrane components. Annu Rev Cell Biol 3:57–85
Mayerson PL, Moscona AA (1979) Malformation of embryonic neural retina elicited by BrdU. Differentiation 13:173–184
Moscona AA (1952) Cell suspensions from organ rudiments of chick embryos. Exp Cell Res 3:535
Needham LK, Adler R, Hewitt AT (1988) Proteoglycan synthesis in flat cell-free cultures of chick embryo retinal neurons and photoreceptors. Dev Biol 126:304–314
Newsome DA, Hewitt AT, Huh W, Robey PG, Hassell JR (1987a) Detection of specific extracellular matrix molecules in drusen, Bruch's membrane, and ciliary body. Am J Ophthalmol 104:373–381
Newsome DA, Huh W, Green WR (1987b) Bruch's membrane age-related changes vary by region. Curr Eye Res 6:1211–1221
Newsome DA, Pfeffer BA, Hewitt AT, Robey PG, Hassell JR (1988) Detection of extracellular matrix molecules synthesized in vitro by monkey and human retinal pigment epithelium: influence of donor age and multiple passages. Exp Eye Res 46:305–321
Okada TS (1983) Recent progress in studies of the transdifferentiation of eye tissue in vitro. Cell Differ 13:177–183
Okada TS, Nomura K, Yasuda K (1983) Commitment to transdifferentiation into lens occurs in neural retina cells after brief spreading culture of the dissociated cells. Cell Differ 121:85–92
Orts-Llorca F, Genis-Galvez JM (1960) Experimental production of retinal septa in the chick embryo. Differentiation of pigment epithelium into neural retina. Acta Anat (Basel) 42:31–70
Rathjen FG, Wolff JM, Frank R, Bonhoeffer F, Rutishauser U (1987) Membrane glycoproteins involved in neurite fasciculation. J Cell Biol 104:343–353
Reh TA, Radke K (1988) A role for the extracellular matrix in retinal neurogenesis in vitro. Dev Biol 129:283–293
Reh TA, Nagy T, Gretton H (1987) Laminin promotes transdifferentiation of retinal pigment epithelial cells to neurons. Nature 330:68–71
Sheffield JB, Moscona AA (1969) Early stages in the reaggregation of embryonic chick neural retina cells. Exp Cell Res 57:462–466
Stroeva OG (1960) Experimental analysis of the eye morphogenesis in mammals. J Embryol Exp Morphol 8:349–368
Tawara A, Varner HH, Hollyfield JG (1988) Proteoglycans in the mouse interphotoreceptor matrix. I. Histochemical studies using cuprolinic blue. Exp Eye Res 46:689–704
Trinkaus JP (1963) Behavior of dissociated retinal pigmentepithelial cells in heterotypic cell aggregates. Ann NY Acad Sci 100:413–434
Tsunematsu Y, Coulombre AJ (1981) Demonstration of transdifferentiation of neural retina from pigmented retina in culture. Dev Growth Differ 23:297–311
Varner HH, Rayborn ME, Osterfeld AM, Hollyfield JG (1987) Localization of proteoglycan within the extracellular matrix sheath of cone photoreceptors. Exp Eye Res 44:633–642
Vollmer G, Layer PG (1986a) Reaggregation of chick retinal and mixtures of retinal and pigment epithelial cells: The degree of laminar organization is dependent on age. Neurosci Lett 63:91–95
Vollmer G, Layer PG (1986b) An in vitro model of proliferation and differentiation of the chick retina: Coaggregates of retinal and pigment epithelial cells. J Neurosci 6:1885–1896
Vollmer G, Layer PG (1987) Cholinesterases and cell proliferation in “nonstratified” and “stratified” cell aggregates from chicken retina and tectum. Cell Tissue Res 250:481–487
Vollmer G, Layer PG, Gierer A (1984) Reaggregation of embryonic chick retina cells: Pigment epithelial cells induce a high order of stratification. Neurosci Lett 48:191–196
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Layer, P.G., Willbold, E. Embryonic chicken retinal cells can regenerate all cell layers in vitro, but ciliary pigmented cells induce their correct polarity. Cell Tissue Res. 258, 233–242 (1989). https://doi.org/10.1007/BF00239443
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DOI: https://doi.org/10.1007/BF00239443