FIGURE

Fig. 6

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ZDB-FIG-120810-55

Publication

Hutchinson et al., 2012 - Tbl3 regulates cell cycle length during zebrafish development

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Fig. 6

Prolonged proliferative phase in retina results from slowing of the cell cycle Proliferation is increased in cey mutant retinas. At 3 dpf there are more pH3⁺ (G2/M) cells in cey mutants in the central retina (white arrowheads; B) as compared to wild-type (A). Injection of both tbl3 MO and p53 MO also results in an increase in pH3⁺ cells in the central retina (white arrowheads) at 3 dpf (D) as compared to a p53MO only control (C). Quantification shows that the increase in pH3⁺ cells is significant in both larvae injected with tbl3MO alone or tbl3MO co-injected with p53MO at 3 dpf (E). Control larvae were injected with phenol red. Whole Z-stacks of dorsal view retinas were projected and the number of pH3⁺ cells were counted and then averaged to determine the average number of mitotic cells at 3 dpf (E, see the “Methods” section for more details on quantification). Tbl3 morphants also have an increased number of cells in S-phase as compared to wild-type (F) indicating there are more proliferating cells at 3 dpf. The average number of BrdU positive cells in five lateral view sections was calculated for each retina (F, see the “Methods” section for more details). The progression of cells from S-phase to G2/M is slower in Tbl3 morphants than control embryos as is indicated by the smaller percentage of cells co-labeled with BrdU and anti-pH3 over a 6 h period of BrdU recovery (G). At 1 hpBrdU N=4, 8, 9, 8 for control, p53MO, tbl3MO, and tbl3p53MO, respectively. At 2 hpBrdU N=4, 4, 5, 5. At 4 hpBrdU N=7, 4, 4, 5. At 6 hpBrdU N=9, 11, 8, 6. Error bars in (E–G) represent standard error of the mean. Scale Bars are 130μm. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Expression Data

Antibody:	Ab3-h3
Fish:	WT WT + MO1-tbl3 WT + MO2-tbl3 Df(Chr03:noxo1a,zgc:113100,sox8b,proza,tbl3)cz26/cz26
Knockdown Reagents:	MO1-tbl3 MO2-tbl3
Anatomical Term:	retina
Stage:	Protruding-mouth

Expression Detail

Antibody Labeling

Phenotype Data

Fish:	WT + MO1-tbl3 WT + MO2-tbl3 Df(Chr03:noxo1a,zgc:113100,sox8b,proza,tbl3)cz26/cz26
Knockdown Reagents:	MO1-tbl3 MO2-tbl3
Observed In:	cell population proliferation M phase retina morphogenesis in camera-type eye S phase
Stage:	Protruding-mouth

Phenotype Detail

Acknowledgments

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Reprinted from Developmental Biology, 368(2), Hutchinson, S.A., Tooke-Locke, E., Wang, J., Tsai, S., Katz, T., and Trede, N.S., Tbl3 regulates cell cycle length during zebrafish development, 261-272, Copyright (2012) with permission from Elsevier. Full text @ Dev. Biol.