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Belliveau MJ, Cepko CL Extrinsic and intrinsic factors control the genesis of amacrine and cone cells in the rat retina. Development. 126:1999;555-566. This paper explores the role of the early postnatal environment (i.e. at P0) on the cell fate decisions made by the progeny of E16 cells. The co-aggregate culture system of Akagawa et al. [9] and Watanabe and Raff [8] was used. Twentyfold more P0 cells than E16 cells were present in the co-aggregates. The results indicate that the P0 environment presented at least two signals that affected the fates of the E16 progeny. One signal was inhibition of amacrine cell production. This signal was found to originate from amacrine cells themselves and is thus a form of feedback inhibition. The other signal induced formation of cone photoreceptors. Interestingly, the postnatal environment did not induce the formation of cell types normally made postnatally, such as bipolar neurons, Müller glia, or a large number of rods. These data suggest that progenitor cells from different ages are intrinsically different, but responsive to extrinsic cues in a limited fashion. In addition to an analysis of the effects on cell fate, the time period for inhibition to the amacrine fate was examined, and found to end as the cell entered its terminal M phase.
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Postmitotic cells fated to become rod photoreceptors can be respecified by CNTF treatment of the retina
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Ezzeddine ZD, Yang X, DeChiara T, Yancopoulos G, Cepko CL Postmitotic cells fated to become rod photoreceptors can be respecified by CNTF treatment of the retina. Development. 124:1997;1055-1067. The effects of the cytokine family represented by CNTF on the development of the postnatal rat and mouse retina were examined. Application of CNTF to intact organ (explant) cultures resulted in almost a complete elimination of cells expressing opsin, a marker of rod photoreceptors. This was true for both rat and mouse cultures. In rat explants, but not in mouse, there was a five- to sevenfold increase in the number of bipolar cells, and these were shown to be the cells that would have become rods in the absence of CNTF. The plasticity of the cells in the rat explants was observed to extend into the period when the cells are postmitotic, and resistance to CNTF was not present until opsin expression was detectable. These data indicate that extrinsic cues can affect the specification of retinal cells. Experiments were also performed using explants from knockout mice in which a component of the CNTF receptor was eliminated (i.e. CNTF receptor-α knockout or LIF receptor-β knockout). These explants were cultured in the absence of added factors, and the expression of opsin and markers of other cell fates were examined. The number of cells expressing opsin was elevated by 25-30% in the knockout explants relative to wild-type or heterozygous littermates, suggesting that events triggered by these receptors play a role in normal retinal development.
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Waid DK, McLoon SC Ganglion cells influence the fate of dividing retinal cells in culture. Development. 125:1998;1059-1066. The generation of ganglion cells in vitro from E4 chicks was studied. When E4 cells were cultured adjacent to older explants, fewer ganglion cells were made by the E4 progenitors. This suggested that perhaps the ganglion cells in the older explants might be providing feedback inhibition. Ganglion cells were then eliminated from the older explants by section of the optic nerve, which results in ganglion cell death. This procedure eliminated the inhibitory activity of the older explants, demonstrating that the inhibitor was dependent on the presence of ganglion cells. When conditioned medium was prepared from older explants in which ganglion cells were present, inhibitory activity could be recovered from a fraction of <3 kDa. These studies suggest that the number of ganglion cells produced normally during retinal development is regulated by previously born ganglion cells through production of soluble factors.
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Bao ZZ, Cepko CL The expression and function of Notch pathway genes in the developing rat eye. J Neurosci. 17:1997;1425-1434.
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Müller-cell-derived leukemia inhibitory factor arrests rod photoreceptor differentiation at a postmitotic pre-rod stage of development
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Neophytou C, Vernallis AB, Smith A, Raff MC Müller-cell-derived leukemia inhibitory factor arrests rod photoreceptor differentiation at a postmitotic pre-rod stage of development. Development. 124:1997;2345-2354. Dissociated murine retinal cells were cultured in monolayers, and the expression of markers of rod photoreceptors (e.g. rhodopsin, recoverin, and arrestin) was examined. The authors found that serum was very inhibitory to the formation of rods, and that it stimulated the production of Müller glia. Müller glial cells were partially purified and shown to inhibit the formation of opsin-positive rods in a co-culture, as was conditioned medium prepared from Müller cell cultures (MCMs). The cells appeared to be arrested at a pre-rod stage, as indicated by the appearance of their nuclei, which was similar to that of mature rods. LIF and CNTF were found to mimic the effect of Müller cells and serum. Neutralizing antibodies directed against LIF were able to almost eliminate the inhibitory activity in the MCM, and a LIF receptor antagonist was able to block the inhibition of added CNTF, MCM, or serum. The inhibition by MCM or CNTF was found to be reversible in that addition of the receptor antagonist could reverse inhibition if added after 5 days in culture in the presence of MCM or CNTF.
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McFarlane S, Zuber ME, Holt CE A role for the fibroblast growth factor receptor in cell fate decisions in the developing vertebrate retina. Development. 125:1998;3967-3975. The role of FGFRs was examined in Xenopus retinal development. The expression of FGFR-1, FGFR-2, and FGFR-4 in mitotic cells was observed. In the maturing retina, FGFR-1 expression was the first to decrease; later, in the more mature retina, FGFR-1 expression was absent, but FGFR-4 persisted in developing photoreceptors, some inner nuclear layer cells, and in the ciliary margin zone. A dominant-negative form of FGFR (D48) was injected into blastomeres that produce the retina, and the stage 40 retina was examined for effects on clonal composition. Several changes in clonal composition were noted: a reduction in the number of rod photoreceptors and amacrine cells, and an increase in the number of cone photoreceptors, horizontal cells, and Müller glia. The effects did not appear to be attributable to selective death or premature differentiation of Müller cells. An additional construct encoding a form of the FGFR that does not affect FGF signaling (HAV0), but that does affect activation by non-FGF ligand(s), was tested. This receptor also affected clonal composition in the Xenopus retina, in a manner almost opposite to that of the dominant-negative FGFR, suggesting that a non-FGF ligand that signals through this receptor might also affect cell fate determination.
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Jensen AM, Raff MC Continuous observation of multipotential retinal progenitor cells in clonal density culture. Dev Biol. 188:1997;267-279. The behavior of E17 murine retinal progenitor cells plated at clonal density in vitro was observed. The cells were taken from a strain that expressed Bcl2 from the neuron-specific enolase promoter, which led to the suppression of at least some cell death. Culture conditions were determined to optimize survival; a rich medium that was supplemented with known growth factors (e.g. bFGF, EGF, NT3, BDNF and CNTF) and chick brain extract was thus chosen. Clone size and composition were examined at different time points, and in some cases, time-lapse video microscopy was used to monitor the clones. Proliferation was found to vary, with some cells dividing after 1 day in vitro, and others not until after 4 days in vitro. Overall, many fewer cell divisions were observed relative to in vivo. Specific retinal cell fates were not assessed, but generic fates were assessed by morphology and by staining with anti-MAP2 (for neurons) or anti-GFAP (for glia). Both types of cells could be seen in the clones. Size and clonal composition did not seem to be correlated, but neurons usually appeared before glia.
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The chick NeuroM gene, related to the Drosophila atonal gene, was isolated, and its expression pattern in the developing chick spinal cord, tectum, retina, and dorsal root ganglia was studied. In all cases, NeuroM appears to be in newly postmitotic, premigratory or migratory neurons, and it is also maintained in the older chick retina in the outer portion of the inner nuclear layer. Expression of NeuroD in the early stages of development was also examined and found to be complementary to that of NeuroM at the later stages, with expression persisting in the outer nuclear layer and ganglion cell layer.
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Roztocil T, Matter-Sadzinski L, Alliod C, Ballivet M, Matter JM NeuroM, a neural helix-loop-helix transcription factor, defines a new transition stage in neurogenesis. Development. 124:1997;3263-3272. The chick NeuroM gene, related to the Drosophila atonal gene, was isolated, and its expression pattern in the developing chick spinal cord, tectum, retina, and dorsal root ganglia was studied. In all cases, NeuroM appears to be in newly postmitotic, premigratory or migratory neurons, and it is also maintained in the older chick retina in the outer portion of the inner nuclear layer. Expression of NeuroD in the early stages of development was also examined and found to be complementary to that of NeuroM at the later stages, with expression persisting in the outer nuclear layer and ganglion cell layer.
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Morrow EM, Furukawa T, Lee JE, Cepko CL NeuroD regulates cell fate determination in the developing neural retina. Development. 126:1999;23-36. The expression of NeuroD in mouse and rat retina was examined. It was expressed in cells that appeared to be newly postmitotic and fated to be photoreceptors or amacrine cells. Later, expression appeared to be confined to a subset of differentiating amacrine cells; it was then extinguished in mature amacrine cells, but maintained in a subset of photoreceptor cells. The function of NeuroD was studied by forcing expression in developing and mature retinal cells of the rat using a retroviral vector. Several effects were seen: there was a slight reduction in clone size when introduced at P0; there was a complete lack of Müller glia in the marked clones; and there was an increase in amacrine cells and a decrease in bipolar cells. In order to rule out an indirect effect on clonal composition due to forcing cells to prematurely exit the cell cycle, the virus was introduced into the latest stages of development, P4 and P6. At these times, most cells are normally exiting the cell cycle, and the production of Müller glia and bipolar cells is at its peak. The same effects were seen on clone composition as were seen at P0, indicating that these effects were most probably directly on the cell fate choices. Retinal explant cultures were prepared from a NeuroD knockout and examined for effects on the cellular composition. In keeping with the viral misexpression data, there was a fourfold increase in Müller glia, and a twofold increase in the number of bipolar cells. Amacrine differentiation also appeared to have been affected. As Müller glia can divide and may also be more resistant to death in the culture, the change in Müller cell number could have been attributable to proliferation and/or selective survival. Both of these possibilities were examined and disproved. These data indicate that NeuroD has a role in cell fate determination, and most probably plays additional roles in proliferation and differentiation in the retina.
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Xath5 participates in a network of bHLH genes in the developing Xenopus retina
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From Xenopus retina, the authors isolated Xath5, a bHLH gene that is related to the Drosophila atonal gene. Xath5 is expressed in retinal progenitor cells and newly differentiating neurons. It is expressed in some of the same cells as another bHLH gene, NeuroD. Overexpression of Xath5 resulted in an increase in ganglion cells and a decrease in amacrine cells, bipolar cells, and Müller glial cells. Overexpression of NeuroD also resulted in fewer Müller glial cells, but resulted in more bipolar and amacrine cells and had very little effect on ganglion cells. Potential regulatory interactions between these two bHLH genes, and another bHLH gene, Xash3 were explored in the early Xenopus embryo in a primary neurogenesis assay. The authors found that introduction of Xath5 led to broad upregulation of NeuroD, and that introduction of NeuroD led to a more spatially restricted upregulation of Xath5.
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Kanekar S, Perron M, Dorsky R, Harris WA, Jan LY, Jan YN, Vetter ML Xath5 participates in a network of bHLH genes in the developing Xenopus retina. Neuron. 19:1997;981-994. From Xenopus retina, the authors isolated Xath5, a bHLH gene that is related to the Drosophila atonal gene. Xath5 is expressed in retinal progenitor cells and newly differentiating neurons. It is expressed in some of the same cells as another bHLH gene, NeuroD. Overexpression of Xath5 resulted in an increase in ganglion cells and a decrease in amacrine cells, bipolar cells, and Müller glial cells. Overexpression of NeuroD also resulted in fewer Müller glial cells, but resulted in more bipolar and amacrine cells and had very little effect on ganglion cells. Potential regulatory interactions between these two bHLH genes, and another bHLH gene, Xash3 were explored in the early Xenopus embryo in a primary neurogenesis assay. The authors found that introduction of Xath5 led to broad upregulation of NeuroD, and that introduction of NeuroD led to a more spatially restricted upregulation of Xath5.
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(1997)
Neuron
, vol.19
, pp. 981-994
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Kanekar, S.1
Perron, M.2
Dorsky, R.3
Harris, W.A.4
Jan, L.Y.5
Jan, Y.N.6
Vetter, M.L.7
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42
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0032528384
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The genetic sequence of retinal development in the ciliary margin of the Xenopus eye
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Note
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Perron M, Kanekar S, Vetter ML, Harris WA The genetic sequence of retinal development in the ciliary margin of the Xenopus eye. Dev Biol. 199:1998;185-200. The expression of transcription factors and Notch pathway genes was examined in the Xenopus retina. The authors report the order of expression of genes within cells in the central retina, where differentiation is occurring, and in the ciliary margin zone (CMZ), where the most immature, proliferative cells are located. Co-expression of XNotch-1, X-Delta-1, ESR1, ESR2, Xash3, Xash1, and Xath5 in cells just central to the most immature cells (referred to by the authors as stem cells) of the CMZ was observed. The domain of expression of Xash3 was more narrow, however, ending more peripherally than that of the others, whereas the domain of Xath5, ATH3, and X-MyT1 began more centrally. NeuroD appeared to begin later than the others and to be maintained in the outer portion of the inner nuclear layer (INL) and in the outer nuclear layer (ONL). ATH3 also was maintained in the outer portion of the INL. As predicted by their function in early eye development, XSix3 and Xrx1 were expressed in the most peripheral cells, along with a low level of Pax6, throughout the CMZ, whereas Xotx2 began to be expressed at the same level as NeuroD. Expression of some of these genes in mitotic cells was determined by performing in situ hybridization on BrdU-labeled material. X-Notch-1 and X-Delta-1 were found to be expressed in dividing cells, as shown previously, and Xath5 and NeuroD expression was found to begin in late-stage mitotic cells. Synthesis of all of the above patterns suggests that there is a particular order of expression of these genes, such that four zones of gene expression can be defined. In addition, different combinations of genes appear to be expressed in different layers of the retina: XSix3, Pax6, and Brn 3.0 in the GCL; XSix3, ATH3, Xotx2, NeuroD (and in some cells, Xrx1) in the outer portion of the INL; XSix3, X-MyT1, and Pax6 in the inner portion of the INL; and NeuroD and Xrx1 in the ONL.
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(1998)
Dev Biol
, vol.199
, pp. 185-200
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Perron, M.1
Kanekar, S.2
Vetter, M.L.3
Harris, W.A.4
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43
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0031695418
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NeuroD induces photoreceptor cell overproduction in vivo and de novo generation in vitro
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•]. Overexpression in the retina using a retroviral vector resulted in an approximately 50% increase in photoreceptor cells, without a concomitant decrease in any other cell population assayed. Infection of cultures of retinal pigmented epithelial cells resulted in an abundance of photoreceptor cells, and no increase in other retinal cell types.
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•]. Overexpression in the retina using a retroviral vector resulted in an approximately 50% increase in photoreceptor cells, without a concomitant decrease in any other cell population assayed. Infection of cultures of retinal pigmented epithelial cells resulted in an abundance of photoreceptor cells, and no increase in other retinal cell types.
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(1998)
J Neurobiol
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Yan, R.-T.1
Wang, S.-Z.2
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44
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Plasticity and differentiation of embryonic retinal cells after terminal mitosis
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Adler R, Hatlee M Plasticity and differentiation of embryonic retinal cells after terminal mitosis. Science. 243:1989;391-393.
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Science
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Adler, R.1
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0027371915
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Taurine promotes the differentiation of a vertebrate retinal cell type in vitro
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Altshuler D, LoTurco J Jr., Cepko CL Taurine promotes the differentiation of a vertebrate retinal cell type in vitro. Development. 119:1993;1317-1328.
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(1993)
Development
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Altshuler, D.1
Loturco J., Jr.2
Cepko, C.L.3
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46
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0025823975
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EGF and TGF-α stimulate retinal neuroepithelial cell proliferation in vitro
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Anchan RM, Reh TA, Angello J, Balliet AM, Walker M EGF and TGF-α stimulate retinal neuroepithelial cell proliferation in vitro. Neuron. 6:1991;923-936.
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Anchan, R.M.1
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Walker, M.5
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0026533318
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S-laminin expression in adult and developing retinae: A potential cue for photoreceptor morphogenesis
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Hunter DD, Murphy MD, Olsson CV, Brunken WJ S-laminin expression in adult and developing retinae: a potential cue for photoreceptor morphogenesis. Neuron. 8:1992;399-413.
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(1992)
Neuron
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Hunter, D.D.1
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Retinoic acid promotes differentiation of photoreceptors in vitro
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Kelley MW, Turner JK, Reh TA Retinoic acid promotes differentiation of photoreceptors in vitro. Development. 120:1994;2091-2102.
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(1994)
Development
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Kelley, M.W.1
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49
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0028825233
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Ligands of steroid/thyroid receptor induce cone photoreceptor in vertebrate retina
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Kelley MW, Turner JK, Reh TA Ligands of steroid/thyroid receptor induce cone photoreceptor in vertebrate retina. Development. 121:1995;3777-3785.
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(1995)
Development
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Kelley, M.W.1
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50
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Changes in retinal cell fate induced by overexpression of EGF receptor
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Lillien L Changes in retinal cell fate induced by overexpression of EGF receptor. Nature. 377:1995;158-162.
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(1995)
Nature
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Lillien, L.1
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Two phases of rod photoreceptor differentiation during rat retinal development
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The kinetics of opsin synthesis in rod photoreceptors was analyzed. Several questions were addressed. One was whether there is any synchrony in the kinetics among cells born on the same day, and whether there is a consistent lag between the terminal mitosis and the onset of opsin synthesis. It was found that cells born on or after E19 had, on average, a 6 day lag before turning on opsin. There was synchrony among cells born on the same day, in that the majority of cells turned on opsin over a 2 day period. In contrast, cells born before E19 had longer and more variable lags, extending from, on average, 7-14 days. All of the rod photoreceptors born prior to E19 turned on opsin beginning at about P2. The effect of the environment on this behavior was analyzed. It was found that the embryonically born cells did not alter their kinetics when cultured in a postnatal environment, as reported previously by Watanabe and Raff [8].
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Morrow EM, Belliveau MJ, Cepko CL Two phases of rod photoreceptor differentiation during rat retinal development. J Neurosci. 18:1998;3738-3748. The kinetics of opsin synthesis in rod photoreceptors was analyzed. Several questions were addressed. One was whether there is any synchrony in the kinetics among cells born on the same day, and whether there is a consistent lag between the terminal mitosis and the onset of opsin synthesis. It was found that cells born on or after E19 had, on average, a 6 day lag before turning on opsin. There was synchrony among cells born on the same day, in that the majority of cells turned on opsin over a 2 day period. In contrast, cells born before E19 had longer and more variable lags, extending from, on average, 7-14 days. All of the rod photoreceptors born prior to E19 turned on opsin beginning at about P2. The effect of the environment on this behavior was analyzed. It was found that the embryonically born cells did not alter their kinetics when cultured in a postnatal environment, as reported previously by Watanabe and Raff [8].
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(1998)
J Neurosci
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Morrow, E.M.1
Belliveau, M.J.2
Cepko, C.L.3
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Restriction of late cerebral cortical progenitors to an upper-layer fate
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Frantz GD, McConnell SK Restriction of late cerebral cortical progenitors to an upper-layer fate. Neuron. 17:1996;55-61.
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Neuron
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Fates of visual cortical neurons in the ferret after isochronic and heterochronic transplantation
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McConnell SK Fates of visual cortical neurons in the ferret after isochronic and heterochronic transplantation. J Neurosci. 8:1988;945-974.
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Diversity and pattern in the developing spinal cord
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Tanabe Y, Jessell TM Diversity and pattern in the developing spinal cord. Science. 274:1996;1115-1123.
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Science
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Tanabe, Y.1
Jessell, T.M.2
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