Effects of oviductal fluid on the development, quality, and gene expression of porcine blastocysts produced in vitro.
In mammals, fertilization and early pre-implantation development occur in the oviduct. Previous results obtained in our laboratory have identified specific molecules in the oviduct that affect porcine sperm-egg interactions. The aim of the present study was to determine whether the contact between oocytes and oviductal fluid also affect embryo development, quality, and gene expression. In vitro matured porcine oocytes were exposed to bovine oviductal fluid (bOF) for 30 min prior to fertilization. Cleavage and blastocyst development rates were significantly higher from bOF-treated oocytes than from untreated oocytes. Blastocysts obtained from bOF-treated oocytes had significantly greater total cell numbers than those obtained from untreated oocytes. Using real-time PCR, grade 1 (very good morphological quality) and grade 2 (good morphological quality) blastocysts were analyzed for gene transcripts related to apoptosis (BAX, BCL2L1), mitochondrial DNA (mtDNA) transcription/replication (POLG, POLG2, and TFAM), blastomere connection and morula compaction (GJA1), and blastocyst formation and pluripotency (POU5F1). We found that the entire set of genes analyzed was differentially expressed between grade 1 and 2 blastocysts. Furthermore, bOF treatment reduced the ratio of BAX to BCL2L1 transcripts and enhanced the abundance of TFAM transcripts in grade 2 blastocysts. Not only do these findings demonstrate that factors within the bOF act on porcine oocytes both quickly and positively, but they also suggest that such factors could promote embryo development and quality by protecting them against adverse impacts on mtDNA transcription/replication and apoptosis induced by the culture environment.
Lloyd
Rhiannon E
RE
Institute of Zoology, Zoological Society of London, Regent's Park, London, UK.
Romar
Raquel
R
Matás
Carmen
C
Gutiérrez-Adán
Alfonso
A
Holt
William V
WV
Coy
Pilar
P
D000818:Animals; D001755:Blastocyst / Q000187:drug effects* / Q000378:metabolism; D012634:Bodily Secretions; D002417:Cattle; D004268:DNA-Binding Proteins / Q000378:metabolism; D047108:Embryonic Development / Q000187:drug effects*; D005307:Fertilization in Vitro; D015870:Gene Expression / Q000187:drug effects; D006023:Glycoproteins / Q000378:metabolism; D024101:Mitochondrial Proteins / Q000378:metabolism; D013552:Swine / Q000196:embryology*; D014157:Transcription Factors / Q000378:metabolism; D015044:Zona Pellucida / Q000378:metabolism; D051028:bcl-2-Associated X Protein / Q000378:metabolism; D051020:bcl-X Protein / Q000378:metabolism
D004268:DNA-Binding Proteins; D006023:Glycoproteins; D024101:Mitochondrial Proteins; C087523:OVGP1 protein, Bos taurus; D014157:Transcription Factors; D051028:bcl-2-Associated X Protein; D051020:bcl-X Protein; C422562:mitochondrial transcription factor A