title: Regulation of heat-inducible HSPA1A gene expression during maternal-to-embryo transition and in response to heat in in vitro-produced bovine embryos.
abstract: In in vitro-produced (IVP) bovine embryos, a burst in transcriptional activation of the embryonic genome (EGA) occurs at the 8-16-cell stage. To examine transcriptional regulation prior to EGA, notably in response to heat stress, we asked (1) whether the spontaneous expression of a luciferase transgene that is driven by the minimal mouse heat-shock protein 1b (hspa1b) gene promoter paralleled that of HSPA1A during EGA in IVP bovine embryo and (2) whether expression of the endogenous heat-inducible iHSPA group member HSPA1A gene and the hspa1b/luciferase transgene were induced by heat stress (HS) prior to EGA. Using two culture systems, we showed that luciferase activity levels rose during the 40-h long EGA-associated cell cycle. In contrast, iHSPA proteins were abundant in matured oocytes and in blastomeres from the two-cell to the 16-cell stages. However, normalised results detected a rise in the level of HSPA1A and luciferase mRNA during EGA, when transcription was required for their protein expression. Prior to EGA, HS-induced premature luciferase activity and transgene expression were clearly inhibited. We could not, however, establish whether this was also true for HSPA1A expression because of the decay of the abundant maternal transcripts prior to EGA. In bovine embryos, heat-induced expression of hspa1b/luciferase, and most likely of HSPA1A, was therefore strictly dependent on EGA. The level of the heat-shock transcription factor 1 molecules that were found in cell nuclei during embryonic development correlated better with the embryo's capacity for heat-shock response than with EGA-associated gene expression.
authors.affiliation: UMR BDR, INRA, ENVA, Université Paris Saclay, 78350 Jouy-en-Josas, France.
authors.forename: Jean-Marc
authors.initials: JM
authors.lastname: Lelièvre
authors.affiliation: UMR BDR, INRA, ENVA, Université Paris Saclay, 78350 Jouy-en-Josas, France.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Peynot
authors.affiliation: UMR BDR, INRA, ENVA, Université Paris Saclay, 78350 Jouy-en-Josas, France.
authors.forename: Sylvie
authors.initials: S
authors.lastname: Ruffini
authors.affiliation: UMR BDR, INRA, ENVA, Université Paris Saclay, 78350 Jouy-en-Josas, France.
authors.forename: Ludivine
authors.initials: L
authors.lastname: Laffont
authors.affiliation: UMR BDR, INRA, ENVA, Université Paris Saclay, 78350 Jouy-en-Josas, France.
authors.forename: Daniel
authors.initials: D
authors.lastname: Le Bourhis
authors.affiliation: Institut Curie, PSL Research University, CNRS UMR3347, INSERM U1021, 91405 Orsay, France.
authors.forename: Pierre-Marie
authors.initials: PM
authors.lastname: Girard
authors.affiliation: UMR BDR, INRA, ENVA, Université Paris Saclay, 78350 Jouy-en-Josas, France.
authors.forename: Véronique
authors.initials: V
authors.lastname: Duranthon
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000378:metabolism*; D002417:Cattle; D046149:Embryo Culture Techniques / Q000662:veterinary; D005260:Female; D018507:Gene Expression Regulation, Developmental; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics* / Q000378:metabolism; D018869:Heat-Shock Response / Q000235:genetics*; D006358:Hot Temperature; D011247:Pregnancy
chemical_list: D018840:HSP70 Heat-Shock Proteins