Cellular and molecular deviations in bovine in vitro-produced embryos are related to the large offspring syndrome.
The large offspring syndrome (LOS) is observed in bovine and ovine offspring following transfer of in vitro-produced (IVP) or cloned embryos and is characterized by a multitude of pathologic changes, of which extended gestation length and increased birthweight are predominant features. In the present study, we used bovine blastocysts to analyze cellular parameters, i.e., the number of cells in Day 7 blastocysts and the size of Day 12 elongating blastocysts, and molecular parameters, i.e., the relative abundance of developmentally important genes: glucose transporter (Glut) 1, Glut-2, Glut-3, Glut-4, heat shock protein (Hsp) 70.1, Cu/Zn-superoxide dismutase (SOD), histone H4.1, basic fibroblast growth factor (bFGF), insulin-like growth factor (IGF) I receptor (R), and IGFII-R. Some blastocysts were produced by in vitro maturation and fertilization followed by in vitro culture in synthetic oviduct fluid medium supplemented with BSA or human serum or by in vivo culture in the sheep oviduct. Other blastocysts were derived in vivo from the uterine horns of superovulated donors. The findings made in the early embryos were related to a representative number of calves obtained from each production system and from artificial insemination (AI). In vitro culture of bovine embryos in the presence of high concentrations of serum or BSA significantly increased the number of cells in Day 7 blastocysts, the size of blastocysts on Day 12, and the relative abundance of the transcripts for Hsp70.1, Cu/Zn-SOD, Glut-3, Glut-4, bFGF, and IGFI-R when compared with embryos from the in vivo production groups. Birthweights of calves derived from IVP embryos were significantly higher than those of calves derived from sheep oviduct culture, superovulation, or AI. The results support the hypothesis that persistence of early deviations in development is causally involved in the incidence of LOS, in particular in increased birthweights. The cellular and molecular parameters analyzed in this study can be considered early markers of LOS in cattle.
Laboratorio di Tecnologie della Riproduzione, Cremona, Italy.
Giovanna
G
Lazzari
Christine
C
Wrenzycki
Doris
D
Herrmann
Roberto
R
Duchi
Theo
T
Kruip
Heiner
H
Niemann
Cesare
C
Galli
D000818:Animals; D001724:Birth Weight; D001755:Blastocyst / Q000737:chemistry* / Q000166:cytology*; D002417:Cattle / Q000196:embryology*; D002418:Cattle Diseases / Q000209:etiology*; D046508:Culture Techniques / Q000379:methods; D005314:Embryonic and Fetal Development; D005260:Female; D005307:Fertilization in Vitro / Q000009:adverse effects / Q000662:veterinary*; D016222:Fibroblast Growth Factor 2 / Q000235:genetics; D051272:Glucose Transporter Type 1; D051273:Glucose Transporter Type 2; D051274:Glucose Transporter Type 3; D051275:Glucose Transporter Type 4; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics; D006657:Histones / Q000235:genetics; D009004:Monosaccharide Transport Proteins / Q000235:genetics; D009124:Muscle Proteins; D009419:Nerve Tissue Proteins; D015800:Protozoan Proteins / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D017526:Receptor, IGF Type 1 / Q000235:genetics; D013482:Superoxide Dismutase / Q000235:genetics; D013577:Syndrome
D051272:Glucose Transporter Type 1; D051273:Glucose Transporter Type 2; D051274:Glucose Transporter Type 3; D051275:Glucose Transporter Type 4; D018840:HSP70 Heat-Shock Proteins; D006657:Histones; D009004:Monosaccharide Transport Proteins; D009124:Muscle Proteins; D009419:Nerve Tissue Proteins; D015800:Protozoan Proteins; D012333:RNA, Messenger; C081884:heat-shock protein 70.1; D016222:Fibroblast Growth Factor 2; D013482:Superoxide Dismutase; D017526:Receptor, IGF Type 1