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Epigenetic regulation of histone modifications and Gata6 gene expression induced by maternal diet in mouse embryoid bodies in a model of developmental programming

Epigenetic regulation of histone modifications and Gata6 gene expression induced by maternal diet in mouse embryoid bodies in a model of developmental programming
Epigenetic regulation of histone modifications and Gata6 gene expression induced by maternal diet in mouse embryoid bodies in a model of developmental programming
Background: dietary interventions during pregnancy alter offspring fitness. We have shown mouse maternal low protein diet fed exclusively for the preimplantation period (Emb-LPD) before return to normal protein diet (NPD) for the rest of gestation, is sufficient to cause adult offspring cardiovascular and metabolic disease. Moreover, Emb-LPD blastocysts sense altered nutrition within the uterus and activate compensatory cellular responses including stimulated endocytosis within extra-embryonic trophectoderm and primitive endoderm (PE) lineages to protect fetal growth rate. However, these responses associate with later disease. Here, we investigate epigenetic mechanisms underlying nutritional programming of PE that may contribute to its altered phenotype, stabilised during subsequent development. We use embryonic stem (ES) cell lines established previously from Emb-LPD and NPD blastocysts that were differentiated into embryoid bodies (EBs) with outer PE-like layer.

Results: Emb-LPD EBs grow to a larger size than NPD EBs and express reduced Gata6 transcription factor (regulator of PE differentiation) at mRNA and protein levels, similar to Emb-LPD PE derivative visceral yolk sac tissue in vivo in later gestation. We analysed histone modifications at the Gata6 promoter in Emb-LPD EBs using chromatin immunoprecipitation assay. We found significant reduction in histone H3 and H4 acetylation and RNA polymerase II binding compared with NPD EBs, all markers of reduced transcription. Other histone modifications, H3K4Me2, H3K9Me3 and H3K27Me3, were unaltered. A similar but generally non-significant histone modification pattern was found on the Gata4 promoter. Consistent with these changes, histone deacetylase Hdac-1, but not Hdac-3, gene expression was upregulated in Emb-LPD EBs.

Conclusions: first, these data demonstrate ES cells and EBs retain and propagate nutritional programming adaptations in vitro, suitable for molecular analysis of mechanisms, reducing animal use. Second, they reveal maternal diet induces persistent changes in histone modifications to regulate Gata6 expression and PE growth and differentiation that may affect lifetime health.
maternal low protein diet, embryoid body, mouse blastocyst, histone epigenetics, metabolic disease, gata6, primitive endoderm, chromatin immunoprecipitation
1471-213X
1-10
Sun, Congshan
19650d2f-f387-444d-9874-1621763fcfd9
Denisenko, Oleg
ff4478dd-c2c4-4033-964f-47f39a0f825d
Sheth, Bhavwanti
2ca6ed58-a992-47b7-b3a5-3c5df82aada7
Cox, Andy
5580a193-f4dd-4762-86d3-f184805c6996
Lucas, Emma S
a74aaa8d-5d8d-4cea-b288-267b85b6ae61
Smyth, Neil R
0eba2a40-3b43-4d40-bb64-621bd7e9d505
Fleming, Tom P
2abf761a-e5a1-4fa7-a2c8-12e32d5d4c03
Sun, Congshan
19650d2f-f387-444d-9874-1621763fcfd9
Denisenko, Oleg
ff4478dd-c2c4-4033-964f-47f39a0f825d
Sheth, Bhavwanti
2ca6ed58-a992-47b7-b3a5-3c5df82aada7
Cox, Andy
5580a193-f4dd-4762-86d3-f184805c6996
Lucas, Emma S
a74aaa8d-5d8d-4cea-b288-267b85b6ae61
Smyth, Neil R
0eba2a40-3b43-4d40-bb64-621bd7e9d505
Fleming, Tom P
2abf761a-e5a1-4fa7-a2c8-12e32d5d4c03

Sun, Congshan, Denisenko, Oleg, Sheth, Bhavwanti, Cox, Andy, Lucas, Emma S, Smyth, Neil R and Fleming, Tom P (2015) Epigenetic regulation of histone modifications and Gata6 gene expression induced by maternal diet in mouse embryoid bodies in a model of developmental programming. BMC Developmental Biology, 15 (1), 1-10. (doi:10.1186/s12861-015-0053-1).

Record type: Article

Abstract

Background: dietary interventions during pregnancy alter offspring fitness. We have shown mouse maternal low protein diet fed exclusively for the preimplantation period (Emb-LPD) before return to normal protein diet (NPD) for the rest of gestation, is sufficient to cause adult offspring cardiovascular and metabolic disease. Moreover, Emb-LPD blastocysts sense altered nutrition within the uterus and activate compensatory cellular responses including stimulated endocytosis within extra-embryonic trophectoderm and primitive endoderm (PE) lineages to protect fetal growth rate. However, these responses associate with later disease. Here, we investigate epigenetic mechanisms underlying nutritional programming of PE that may contribute to its altered phenotype, stabilised during subsequent development. We use embryonic stem (ES) cell lines established previously from Emb-LPD and NPD blastocysts that were differentiated into embryoid bodies (EBs) with outer PE-like layer.

Results: Emb-LPD EBs grow to a larger size than NPD EBs and express reduced Gata6 transcription factor (regulator of PE differentiation) at mRNA and protein levels, similar to Emb-LPD PE derivative visceral yolk sac tissue in vivo in later gestation. We analysed histone modifications at the Gata6 promoter in Emb-LPD EBs using chromatin immunoprecipitation assay. We found significant reduction in histone H3 and H4 acetylation and RNA polymerase II binding compared with NPD EBs, all markers of reduced transcription. Other histone modifications, H3K4Me2, H3K9Me3 and H3K27Me3, were unaltered. A similar but generally non-significant histone modification pattern was found on the Gata4 promoter. Consistent with these changes, histone deacetylase Hdac-1, but not Hdac-3, gene expression was upregulated in Emb-LPD EBs.

Conclusions: first, these data demonstrate ES cells and EBs retain and propagate nutritional programming adaptations in vitro, suitable for molecular analysis of mechanisms, reducing animal use. Second, they reveal maternal diet induces persistent changes in histone modifications to regulate Gata6 expression and PE growth and differentiation that may affect lifetime health.

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Accepted/In Press date: 6 January 2015
Published date: 21 January 2015
Keywords: maternal low protein diet, embryoid body, mouse blastocyst, histone epigenetics, metabolic disease, gata6, primitive endoderm, chromatin immunoprecipitation
Organisations: Biomedicine

Identifiers

Local EPrints ID: 377496
URI: https://eprints.soton.ac.uk/id/eprint/377496
ISSN: 1471-213X
PURE UUID: 917a5fec-e9d2-40d8-851c-dbc78e3067b1

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Date deposited: 11 Jun 2015 13:17
Last modified: 28 Aug 2019 18:42

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