Changes in Ribosomal Ribonucleic Acid Content Within in Vitro-produced Bovine Embryos1

Bilodeau‐Goeseels, Sylvie; Schultz, Gilbert A.

doi:10.1095/biolreprod56.5.1323

Cited by 51 publications

(24 citation statements)

References 33 publications

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“…To this end, total RNA did not change during meiotic maturation which is consistent with previous findings for bovine oocytes [44,67]. The amount of RNA obtained for GVstage and matured oocytes was in agreement with others [~2 ng/ oocyte; 67,68]. Finding similar amounts of total RNA in oocytes and 4 to 8-cell embryos but higher levels in blastocyst-stage embryos was also consistent with a recent report by Gilbert et al [69].…”

Section: Discussionsupporting

confidence: 92%

Impact of Heat Stress Exposure During Meiotic Maturation on Oocyte, Surrounding Cumulus Cell, and Embryo RNA Populations

et al. 2011

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Abstract. To determine if reductions in developmental competence related to heat stress exposure were correlated with perturbations in certain RNA populations, poly(A) RNA, total RNA, RNA size distribution, and the abundance of transcripts (cyclin B1, GDF9, BMP15, poly(A) polymerase, HSP70, 18S & 28S rRNA) were examined in oocytes matured at 38.5 or 41 C. Performing in vitro fertilization resulted in embryos for examining RNA. Relative to germinal vesiclestage oocytes, total amount of poly(A) RNA decreased similarly in oocytes matured at 38.5 or 41 C. Total RNA did not change during meiotic maturation or up through the 4 to 8-cell stage of embryonic development. Blastocyst-stage embryos had more total RNA; those originating from heat-stressed oocytes had more than those from nonheat-stressed oocytes. Oocytes and 4 to 8-cell embryos had similar RIN values and ratios for rRNA, 18S/fast region, and 18S/inter region. Values obtained for blastocyst-stage embryos were similar to those obtained for cumulus cell RNA, which did not change during maturation. Culture at 41 C for the first 12 h of meiotic maturation had no impact on RNA size distribution or transcripts examined from oocytes, surrounding cumulus or resultant 4 to 8-cell embryos. Interestingly, however, RNA from blastocysts originating from heat-stressed oocytes had lower 18S/fast region and 18S/inter region ratios compared to other developmental stages and cumulus cells. Although biological significance of these RNA changes is unclear, differences at the molecular level in embryos from heat-stressed oocytes emphasize the importance of minimizing stress exposure during meiotic maturation, if the intent is to obtain developmentally-competent embryos. Key words: Cumulus, Heat stress, Microcapillary electrophoresis, Oocyte, RNA (J. Reprod. Dev. 57: [481][482][483][484][485][486][487][488][489][490][491] 2011) griculturally-important females exposed to environmental heat stress experience reduced fertility [1][2][3][4]. In cattle, this is largely due to hyperthermia [5][6][7] whereby rectal temperatures may reach or exceed 41 C [5,[8][9][10][11][12]. Effects of environmental heat stress to reduce female fertility are pronounced during or near estrus [13][14][15]. This corresponds to the time period when the female is receptive to mating and the oocyte within the Graafian follicle resumes meiosis and progresses to metaphase II (MII; meiotic maturation) in preparation for fertilization. Hyperthermia during this time has been associated with reductions in early embryonic development [16] and increases in embryonic loss near the time of placental attachment [17].The negative impact of hyperthermia during estrus may be attributable to indirect effects on the maternal environment [reviewed by 18] and/or due to direct effects on the cumulus-oocyte complex. Direct exposure of the cumulus-oocyte complex to 41 C during meiotic maturation reduces early embryo development after fertilization by 30 to 65% [19][20][21][22][23][24][25] similar to consequences observed when e...

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Section: Discussionsupporting

confidence: 92%

Impact of Heat Stress Exposure During Meiotic Maturation on Oocyte, Surrounding Cumulus Cell, and Embryo RNA Populations

et al. 2011

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“…S3). BMP15, KIT, GDF9, STAT3, C-MOS and ZP2, 3, 4 are oocyte markers (2,18,(21)(22)(23)(24)(25). The array analysis of bovine embryos revealed expression of ZP2, 3, 4, GDF9, BMP15, cadherin (CDH3), but also OCT4 (POU5F1), CLDN, ZO3, and TP53 (Fig.…”

Section: Confirmation Of Array Data By Real Time Rt-pcrmentioning

confidence: 92%

Genome-wide expression profiling reveals distinct clusters of transcriptional regulation during bovine preimplantation development in vivo

Kues

Sudheer

Herrmann³

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

152

113

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Bovine embryos can be generated by in vitro fertilization or somatic nuclear transfer; however, these differ from their in vivo counterparts in many aspects and exhibit a higher proportion of developmental abnormalities. Here, we determined for the first time the transcriptomes of bovine metaphase II oocytes and all stages of preimplantation embryos developing in vivo up to the blastocyst using the Affymetrix GeneChip Bovine Genome Array which examines approximately 23,000 transcripts. The data show that bovine oocytes and embryos transcribed a significantly higher number of genes than somatic cells. Several hundred genes were transcribed well before the 8-cell stage, at which the major activation of the bovine genome expression occurs. Importantly, stage-specific expression patterns in 2-cell, 4-cell, and 8-cell stages, and in morulae and blastocysts, were detected, indicating dynamic changes in the embryonic transcriptome and in groups of transiently active genes. Pathway analysis revealed >120 biochemical pathways that are operative in early preimplantation bovine development. Significant differences were observed between the mRNA expression profiles of in vivo and in vitro matured oocytes, highlighting the need to include in vivo derived oocytes/embryos in studies evaluating assisted reproductive techniques. This study provides the first comprehensive analysis of gene expression and transcriptome dynamics of in vivo developing bovine embryos and will serve as a basis for improving assisted reproductive technology.

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“…The transcription of embryonic genome starts in bovines in the late 8-cell stage (8c; Camous et al 1986, King et al 1988, Kopecny et al 1989, Pavlok et al 1993) and this event is called EGA. Until EGA, all the mRNAs and proteins are of maternal origin (Bilodeau-Goeseels & Schultz 1997). Some authors suggest that there is also a so-called minor genome activation between 1-and 4c, which is followed by major genome activation in the 8c (Memili & First 2000, Jakobsen et al 2006.…”

Section: Introductionmentioning

confidence: 99%

Silencing CENPF in bovine preimplantation embryo induces arrest at 8-cell stage

Toralova¹,

Šušor²,

Němcová³

et al. 2009

Reproduction

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Identification of genes that are important for normal preimplantation development is essential for understanding the basics of early mammalian embryogenesis. In our previous study, we have shown that CENPF (mitosin) is differentially expressed during preimplantation development of bovine embryos. CENPF is a centromere-kinetochore complex protein that plays a crucial role in the cell division of somatic cells. To our best knowledge, no study has yet been done on either bovine model, or oocytes and preimplantation embryos. In this study, we focused on the fate of bovine embryos after injection of CENPF double-stranded RNA (dsRNA) into the zygotes. An average decrease of CENPF mRNA abundance by 94.9% or more and an extensive decline in immunofluorescence staining intensity was detected relative to controls. There was no disparity between individual groups in the developmental competence before the 8-cell stage. However, the developmental competence rapidly decreased then and only 28.1% of CENPF dsRNA injected 8-cell embryos were able to develop further (uninjected control: 71.8%; green fluorescent protein dsRNA injected control: 72.0%). In conclusion, these results show that depletion of CENPF mRNA in preimplantation bovine embryos leads to dramatic decrease of developmental competence after embryonic genome activation.

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Changes in Ribosomal Ribonucleic Acid Content Within in Vitro-produced Bovine Embryos1

Cited by 51 publications

References 33 publications

Impact of Heat Stress Exposure During Meiotic Maturation on Oocyte, Surrounding Cumulus Cell, and Embryo RNA Populations

Impact of Heat Stress Exposure During Meiotic Maturation on Oocyte, Surrounding Cumulus Cell, and Embryo RNA Populations

Genome-wide expression profiling reveals distinct clusters of transcriptional regulation during bovine preimplantation development in vivo

Silencing CENPF in bovine preimplantation embryo induces arrest at 8-cell stage

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