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GATA4 and GATA5 are essential for heart and liver development in Xenopus embryos.

Haworth KE, Kotecha S, Mohun TJ, Latinkic BV - BMC Dev. Biol. (2008)

Bottom Line: In this study we show that in Xenopus embryos GATA5 is essential for early development of heart and liver precursors.In addition, we have found that in Xenopus embryos GATA4 is important for development of heart and liver primordia following their specification, and that in this role it might interact with GATA6.Our results suggest that GATA5 acts earlier than GATA4 to regulate development of heart and liver precursors, and indicate that one early direct target of GATA5 is homeobox gene Hex.

View Article: PubMed Central - HTML - PubMed

Affiliation: School of Biosciences, Cardiff University, Museum Avenue, Cardiff, CF10 3US, Wales, UK. haworthk@cardiff.ac.uk

ABSTRACT

Background: GATA factors 4/5/6 have been implicated in the development of the heart and endodermal derivatives in vertebrates. Work in zebrafish has indicated that GATA5 is required for normal development earlier than GATA4/6. However, the GATA5 knockout mouse has no apparent embryonic phenotype, thereby questioning the importance of the gene for vertebrate development.

Results: In this study we show that in Xenopus embryos GATA5 is essential for early development of heart and liver precursors. In addition, we have found that in Xenopus embryos GATA4 is important for development of heart and liver primordia following their specification, and that in this role it might interact with GATA6.

Conclusion: Our results suggest that GATA5 acts earlier than GATA4 to regulate development of heart and liver precursors, and indicate that one early direct target of GATA5 is homeobox gene Hex.

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Related in: MedlinePlus

Injection of G5 MO, but not of C1 and C2 control MOs, cause severe size reduction or loss of heart and liver in Xenopus embryos. A: Translation of injected GATA5-GR.HA mRNA, detected by Western blotting with anti-HA antibody, is blocked by G5 but not by other MOs indicated. G5UTR MO ("G5U") does not affect translation of GATA5-GR.HA, as this constructs lacks the 5'UTR sequence. 1 ng of mRNA was injected into 1- or 2-cell embryos and 10–15 minutes later the embryos were injected with 10 ng of G5, or 50 ng of G5UTR or G4 MOs. E-Uninjected embryos. B: Injection of 5 ng of G5 MO results in a loss (B1,2) or severe reduction (B3-5) of cardiac and liver precursors, as revealed by whole-mount in situ hybridisation for MLC2 (purple) and Hex (turquoise). Posterior injection of G5 MO (5ng) has no obvious effects (B6). Control MOs 1 or 2 (C1 or C2, 50 ng/embryo) have no effect on normal development of heart and liver precursors (B7-9). B1-4, 8,9: ventral view; B5-7: lateral view (anterior to the left). In B3-5 arrows point to the remnants of the heart. C: G5 MO (1 ng/embryo) causes heart defects in X. tropicalis (Xt) embryos (arrow points to the heart remnant), and C1 MO (10 ng) has no effect on normal heart development (arrowhead in C2). Complete bleaching of X. tropicalis embryos has removed the morphological landmarks, and cement gland (cg) and eyes (e) have been indicated to add visualisation.
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Figure 1: Injection of G5 MO, but not of C1 and C2 control MOs, cause severe size reduction or loss of heart and liver in Xenopus embryos. A: Translation of injected GATA5-GR.HA mRNA, detected by Western blotting with anti-HA antibody, is blocked by G5 but not by other MOs indicated. G5UTR MO ("G5U") does not affect translation of GATA5-GR.HA, as this constructs lacks the 5'UTR sequence. 1 ng of mRNA was injected into 1- or 2-cell embryos and 10–15 minutes later the embryos were injected with 10 ng of G5, or 50 ng of G5UTR or G4 MOs. E-Uninjected embryos. B: Injection of 5 ng of G5 MO results in a loss (B1,2) or severe reduction (B3-5) of cardiac and liver precursors, as revealed by whole-mount in situ hybridisation for MLC2 (purple) and Hex (turquoise). Posterior injection of G5 MO (5ng) has no obvious effects (B6). Control MOs 1 or 2 (C1 or C2, 50 ng/embryo) have no effect on normal development of heart and liver precursors (B7-9). B1-4, 8,9: ventral view; B5-7: lateral view (anterior to the left). In B3-5 arrows point to the remnants of the heart. C: G5 MO (1 ng/embryo) causes heart defects in X. tropicalis (Xt) embryos (arrow points to the heart remnant), and C1 MO (10 ng) has no effect on normal heart development (arrowhead in C2). Complete bleaching of X. tropicalis embryos has removed the morphological landmarks, and cement gland (cg) and eyes (e) have been indicated to add visualisation.

Mentions: To assess the role of GATA5 in Xenopus development we first used a translation-blocking MO, G5. This MO targets both GATA5 pseudoalleles in Xenopus laevis and GATA5 mRNA in diploid Xenopus tropicalis (see Additional File 1). Doses of 20 ng or greater per X. laevis embryo resulted in gastrulation defects and lethality by mid-neurula stages. Therefore, to investigate the role of GATA5 in heart and liver development doses of 5–10 ng/embryo were used. This dose of G5 MO was shown to specifically and effectively block translation of the injected tagged GATA5 mRNA (Fig. 1A).


GATA4 and GATA5 are essential for heart and liver development in Xenopus embryos.

Haworth KE, Kotecha S, Mohun TJ, Latinkic BV - BMC Dev. Biol. (2008)

Injection of G5 MO, but not of C1 and C2 control MOs, cause severe size reduction or loss of heart and liver in Xenopus embryos. A: Translation of injected GATA5-GR.HA mRNA, detected by Western blotting with anti-HA antibody, is blocked by G5 but not by other MOs indicated. G5UTR MO ("G5U") does not affect translation of GATA5-GR.HA, as this constructs lacks the 5'UTR sequence. 1 ng of mRNA was injected into 1- or 2-cell embryos and 10–15 minutes later the embryos were injected with 10 ng of G5, or 50 ng of G5UTR or G4 MOs. E-Uninjected embryos. B: Injection of 5 ng of G5 MO results in a loss (B1,2) or severe reduction (B3-5) of cardiac and liver precursors, as revealed by whole-mount in situ hybridisation for MLC2 (purple) and Hex (turquoise). Posterior injection of G5 MO (5ng) has no obvious effects (B6). Control MOs 1 or 2 (C1 or C2, 50 ng/embryo) have no effect on normal development of heart and liver precursors (B7-9). B1-4, 8,9: ventral view; B5-7: lateral view (anterior to the left). In B3-5 arrows point to the remnants of the heart. C: G5 MO (1 ng/embryo) causes heart defects in X. tropicalis (Xt) embryos (arrow points to the heart remnant), and C1 MO (10 ng) has no effect on normal heart development (arrowhead in C2). Complete bleaching of X. tropicalis embryos has removed the morphological landmarks, and cement gland (cg) and eyes (e) have been indicated to add visualisation.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2526999&req=5

Figure 1: Injection of G5 MO, but not of C1 and C2 control MOs, cause severe size reduction or loss of heart and liver in Xenopus embryos. A: Translation of injected GATA5-GR.HA mRNA, detected by Western blotting with anti-HA antibody, is blocked by G5 but not by other MOs indicated. G5UTR MO ("G5U") does not affect translation of GATA5-GR.HA, as this constructs lacks the 5'UTR sequence. 1 ng of mRNA was injected into 1- or 2-cell embryos and 10–15 minutes later the embryos were injected with 10 ng of G5, or 50 ng of G5UTR or G4 MOs. E-Uninjected embryos. B: Injection of 5 ng of G5 MO results in a loss (B1,2) or severe reduction (B3-5) of cardiac and liver precursors, as revealed by whole-mount in situ hybridisation for MLC2 (purple) and Hex (turquoise). Posterior injection of G5 MO (5ng) has no obvious effects (B6). Control MOs 1 or 2 (C1 or C2, 50 ng/embryo) have no effect on normal development of heart and liver precursors (B7-9). B1-4, 8,9: ventral view; B5-7: lateral view (anterior to the left). In B3-5 arrows point to the remnants of the heart. C: G5 MO (1 ng/embryo) causes heart defects in X. tropicalis (Xt) embryos (arrow points to the heart remnant), and C1 MO (10 ng) has no effect on normal heart development (arrowhead in C2). Complete bleaching of X. tropicalis embryos has removed the morphological landmarks, and cement gland (cg) and eyes (e) have been indicated to add visualisation.
Mentions: To assess the role of GATA5 in Xenopus development we first used a translation-blocking MO, G5. This MO targets both GATA5 pseudoalleles in Xenopus laevis and GATA5 mRNA in diploid Xenopus tropicalis (see Additional File 1). Doses of 20 ng or greater per X. laevis embryo resulted in gastrulation defects and lethality by mid-neurula stages. Therefore, to investigate the role of GATA5 in heart and liver development doses of 5–10 ng/embryo were used. This dose of G5 MO was shown to specifically and effectively block translation of the injected tagged GATA5 mRNA (Fig. 1A).

Bottom Line: In this study we show that in Xenopus embryos GATA5 is essential for early development of heart and liver precursors.In addition, we have found that in Xenopus embryos GATA4 is important for development of heart and liver primordia following their specification, and that in this role it might interact with GATA6.Our results suggest that GATA5 acts earlier than GATA4 to regulate development of heart and liver precursors, and indicate that one early direct target of GATA5 is homeobox gene Hex.

View Article: PubMed Central - HTML - PubMed

Affiliation: School of Biosciences, Cardiff University, Museum Avenue, Cardiff, CF10 3US, Wales, UK. haworthk@cardiff.ac.uk

ABSTRACT

Background: GATA factors 4/5/6 have been implicated in the development of the heart and endodermal derivatives in vertebrates. Work in zebrafish has indicated that GATA5 is required for normal development earlier than GATA4/6. However, the GATA5 knockout mouse has no apparent embryonic phenotype, thereby questioning the importance of the gene for vertebrate development.

Results: In this study we show that in Xenopus embryos GATA5 is essential for early development of heart and liver precursors. In addition, we have found that in Xenopus embryos GATA4 is important for development of heart and liver primordia following their specification, and that in this role it might interact with GATA6.

Conclusion: Our results suggest that GATA5 acts earlier than GATA4 to regulate development of heart and liver precursors, and indicate that one early direct target of GATA5 is homeobox gene Hex.

Show MeSH
Related in: MedlinePlus