Cell commitment and differentiation in explants of embryonic rat neural retina. Comparison with the developmental potential of dissociated retina

Janet R. Sparrow, David Hicks, Colin Barnstable

Research output: Contribution to journalArticle

106 Citations (Scopus)

Abstract

The differentiation of presumptive neural retina following its isolation from rat embryos and growth in explant and monolayer culture has been studied to obtain information regarding the extent to which factors extrinsic and intrinsic to the retina participate in determining molecular and cytological differentiation. Explanted retinal epithelium retained the capacity for mitosis, as shown by [3H]thymidine incorporation, and from the undifferentiated neuroepithelium, retinal cell-types emerged and acquired a laminar organization resembling that in vivo. Characterization of rod photoreceptor cells at both the light and electron microscopic level showed that these cells exhibit differentiated structural features including inner segments, connecting cilia and membranous expansions suggestive of forming outer segments. Immunofluorescent labeling with an antibody to a synaptic vesicle protein, and electron microscopic identification of synaptic elements showed formation of synapses by the photoreceptor cells within the explant. Neurites extending from the explants exhibited growth on laminin, fibronectin and collagen substrates. Since the neurites immunolabeled with antibodies to the 140 kDa subunit of neurofilament and with antibodies to Thy-1, they could be identified as axons of ganglion cells. Antibodies to a variety of cell-type specific antigens showed that the cells expressed molecules associated with the fully differentiated cell. Furthermore, since our approach has been to explant embryonic retina at an age when the antigens are not yet expressed in vivo, the appearance of the antigens in culture represented de novo expression. In contrast, neural retinal cells in dissociated cultures did not exhibit de novo expression of differentiated molecular properties. Collectively, these results indicate that the isolated neural retina is capable of cell birth, commitment and differentiation when the cells are maintained together in explant culture, whereas the environment of monolayer cultures is apparently unable to promote the generation of mature neuronal phenotypes.

Original languageEnglish (US)
Pages (from-to)69-84
Number of pages16
JournalDevelopmental Brain Research
Volume51
Issue number1
DOIs
StatePublished - Jan 1 1990

Fingerprint

Retina
Cell Differentiation
Antibodies
Neurites
Antigens
Electrons
Retinal Rod Photoreceptor Cells
Photoreceptor Cells
Intrinsic Factor
Intermediate Filaments
Cilia
Synaptic Vesicles
Laminin
Growth
Fibronectins
Mitosis
Ganglia
Synapses
Thymidine
Axons

All Science Journal Classification (ASJC) codes

  • Developmental Neuroscience
  • Developmental Biology

Cite this

@article{98889cd40f8149e3ad26a004483e1064,
title = "Cell commitment and differentiation in explants of embryonic rat neural retina. Comparison with the developmental potential of dissociated retina",
abstract = "The differentiation of presumptive neural retina following its isolation from rat embryos and growth in explant and monolayer culture has been studied to obtain information regarding the extent to which factors extrinsic and intrinsic to the retina participate in determining molecular and cytological differentiation. Explanted retinal epithelium retained the capacity for mitosis, as shown by [3H]thymidine incorporation, and from the undifferentiated neuroepithelium, retinal cell-types emerged and acquired a laminar organization resembling that in vivo. Characterization of rod photoreceptor cells at both the light and electron microscopic level showed that these cells exhibit differentiated structural features including inner segments, connecting cilia and membranous expansions suggestive of forming outer segments. Immunofluorescent labeling with an antibody to a synaptic vesicle protein, and electron microscopic identification of synaptic elements showed formation of synapses by the photoreceptor cells within the explant. Neurites extending from the explants exhibited growth on laminin, fibronectin and collagen substrates. Since the neurites immunolabeled with antibodies to the 140 kDa subunit of neurofilament and with antibodies to Thy-1, they could be identified as axons of ganglion cells. Antibodies to a variety of cell-type specific antigens showed that the cells expressed molecules associated with the fully differentiated cell. Furthermore, since our approach has been to explant embryonic retina at an age when the antigens are not yet expressed in vivo, the appearance of the antigens in culture represented de novo expression. In contrast, neural retinal cells in dissociated cultures did not exhibit de novo expression of differentiated molecular properties. Collectively, these results indicate that the isolated neural retina is capable of cell birth, commitment and differentiation when the cells are maintained together in explant culture, whereas the environment of monolayer cultures is apparently unable to promote the generation of mature neuronal phenotypes.",
author = "Sparrow, {Janet R.} and David Hicks and Colin Barnstable",
year = "1990",
month = "1",
day = "1",
doi = "10.1016/0165-3806(90)90259-2",
language = "English (US)",
volume = "51",
pages = "69--84",
journal = "Developmental Brain Research",
issn = "0165-3806",
publisher = "Elsevier BV",
number = "1",

}

Cell commitment and differentiation in explants of embryonic rat neural retina. Comparison with the developmental potential of dissociated retina. / Sparrow, Janet R.; Hicks, David; Barnstable, Colin.

In: Developmental Brain Research, Vol. 51, No. 1, 01.01.1990, p. 69-84.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Cell commitment and differentiation in explants of embryonic rat neural retina. Comparison with the developmental potential of dissociated retina

AU - Sparrow, Janet R.

AU - Hicks, David

AU - Barnstable, Colin

PY - 1990/1/1

Y1 - 1990/1/1

N2 - The differentiation of presumptive neural retina following its isolation from rat embryos and growth in explant and monolayer culture has been studied to obtain information regarding the extent to which factors extrinsic and intrinsic to the retina participate in determining molecular and cytological differentiation. Explanted retinal epithelium retained the capacity for mitosis, as shown by [3H]thymidine incorporation, and from the undifferentiated neuroepithelium, retinal cell-types emerged and acquired a laminar organization resembling that in vivo. Characterization of rod photoreceptor cells at both the light and electron microscopic level showed that these cells exhibit differentiated structural features including inner segments, connecting cilia and membranous expansions suggestive of forming outer segments. Immunofluorescent labeling with an antibody to a synaptic vesicle protein, and electron microscopic identification of synaptic elements showed formation of synapses by the photoreceptor cells within the explant. Neurites extending from the explants exhibited growth on laminin, fibronectin and collagen substrates. Since the neurites immunolabeled with antibodies to the 140 kDa subunit of neurofilament and with antibodies to Thy-1, they could be identified as axons of ganglion cells. Antibodies to a variety of cell-type specific antigens showed that the cells expressed molecules associated with the fully differentiated cell. Furthermore, since our approach has been to explant embryonic retina at an age when the antigens are not yet expressed in vivo, the appearance of the antigens in culture represented de novo expression. In contrast, neural retinal cells in dissociated cultures did not exhibit de novo expression of differentiated molecular properties. Collectively, these results indicate that the isolated neural retina is capable of cell birth, commitment and differentiation when the cells are maintained together in explant culture, whereas the environment of monolayer cultures is apparently unable to promote the generation of mature neuronal phenotypes.

AB - The differentiation of presumptive neural retina following its isolation from rat embryos and growth in explant and monolayer culture has been studied to obtain information regarding the extent to which factors extrinsic and intrinsic to the retina participate in determining molecular and cytological differentiation. Explanted retinal epithelium retained the capacity for mitosis, as shown by [3H]thymidine incorporation, and from the undifferentiated neuroepithelium, retinal cell-types emerged and acquired a laminar organization resembling that in vivo. Characterization of rod photoreceptor cells at both the light and electron microscopic level showed that these cells exhibit differentiated structural features including inner segments, connecting cilia and membranous expansions suggestive of forming outer segments. Immunofluorescent labeling with an antibody to a synaptic vesicle protein, and electron microscopic identification of synaptic elements showed formation of synapses by the photoreceptor cells within the explant. Neurites extending from the explants exhibited growth on laminin, fibronectin and collagen substrates. Since the neurites immunolabeled with antibodies to the 140 kDa subunit of neurofilament and with antibodies to Thy-1, they could be identified as axons of ganglion cells. Antibodies to a variety of cell-type specific antigens showed that the cells expressed molecules associated with the fully differentiated cell. Furthermore, since our approach has been to explant embryonic retina at an age when the antigens are not yet expressed in vivo, the appearance of the antigens in culture represented de novo expression. In contrast, neural retinal cells in dissociated cultures did not exhibit de novo expression of differentiated molecular properties. Collectively, these results indicate that the isolated neural retina is capable of cell birth, commitment and differentiation when the cells are maintained together in explant culture, whereas the environment of monolayer cultures is apparently unable to promote the generation of mature neuronal phenotypes.

UR - http://www.scopus.com/inward/record.url?scp=0025014708&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025014708&partnerID=8YFLogxK

U2 - 10.1016/0165-3806(90)90259-2

DO - 10.1016/0165-3806(90)90259-2

M3 - Article

VL - 51

SP - 69

EP - 84

JO - Developmental Brain Research

JF - Developmental Brain Research

SN - 0165-3806

IS - 1

ER -