Developmental kinetics and gene expression in male and female bovine embryos produced in vitro with sex-sorted spermatozoa.
Using bovine embryos generated in vitro from IVF with X-sorted, Y-sorted and unsorted spermatozoa, we compared the kinetics of male and female embryo development and gene expression between male and female blastocysts. Bovine in vitro-matured oocytes (n = 8858) were fertilised with spermatozoa from each of three different bulls (X-sorted, Y-sorted or unsorted spermatozoa depending on the experiment). The cleavage rate was assessed 24, 27, 30, 33, 36, 40, 44 and 48 h post insemination (h.p.i.) and blastocyst development was recorded on Days 6-9. The relative mRNA abundance of nine genes (GSTM3, DNTM3A, PGRMC1, TP53, BAX, COX2, IGF2R, AKR1B1 and PLAC8) was analysed in male and female Day 7 blastocysts produced with sorted and unsorted spermatozoa from one bull. Cumulative cleavage rate and blastocyst yield were significantly higher in the unsorted group compared with the X- or Y-sorted group from the same bull (P < or = 0.05). Although differences existed between bulls in terms of cleavage rate, no differences were observed in cleavage rate between X- and Y-sorted spermatozoa within a bull. The blastocyst yield was significantly higher only for Bull 3 when the Y-sorted spermatozoa were used (27.1+2.8 v. 19.1+1.4 for Y- and X-sorted spermatozoa, respectively; P < 0.05). There were no differences in the mRNA abundance of the nine genes analysed between embryos of the same sex produced with sorted or unsorted spermatozoa. However, significant differences in polyA mRNA abundance were observed between male and female blastocysts for three genes (GSTM3, DNMT3A and PGRMC1; P < or = 0.05). In conclusion, the use of sorted rather than unsorted spermatozoa in IVF significantly delays the onset of first cleavage. Differences were noted between bulls, but not between X- and Y-sorted spermatozoa, and although no differences were found in terms of the mRNA abundance of the nine genes tested between sorted and unsorted spermatozoa, sex-related differences were found in the case of three genes.
Bermejo-Alvarez
Pablo
P
Departamento de Reproducción Animal y Conservaciónde Recursos Zoogenéticos, INIA, Carretera De la Coruña Km 5.9, Madrid 28040, Spain.
Lonergan
Patrick
P
Rath
Detlef
D
Gutiérrez-Adan
Alfonso
A
Rizos
Dimitrios
D
D000818:Animals; D001483:Base Sequence; D001755:Blastocyst / Q000378:metabolism; D002417:Cattle / Q000196:embryology* / Q000235:genetics*; D002469:Cell Separation / Q000379:methods / Q000662:veterinary; D002970:Cleavage Stage, Ovum / Q000378:metabolism; D017931:DNA Primers / Q000235:genetics; D047108:Embryonic Development / Q000235:genetics*; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods / Q000662:veterinary*; D018507:Gene Expression Regulation, Developmental; D066298:In Vitro Techniques; D007700:Kinetics; D008297:Male; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013094:Spermatozoa
D017931:DNA Primers; D012333:RNA, Messenger