Impact of equine assisted reproductive technologies (standard embryo transfer or intracytoplasmic sperm injection (ICSI) with in vitro culture and embryo transfer) on placenta and foal morphometry and placental gene expression.

Assisted reproductive technologies (ARTs) such as intracytoplasmic sperm injection (ICSI), in vitro embryo culture and embryo transfer (ET) may be associated with alterations in fetal and placental development. In horses, ET has been used for decades. More recently, in vitro embryo production by ICSI and in vitro culture, followed by embryo transfer (ICSI-C) has become an accepted method for clinical foal production. However, no information is available on the effects of ICSI-C or even of standard ET itself on placental and neonatal parameters in horses. We therefore evaluated placental and neonatal morphology and placental gene expression in reining- and cutting-type American Quarter Horse foals produced using different technologies. Thirty foals and placentas (naturally conceived (NC), ET and ICSI-C; 10 in each group) were examined morphometrically. The only parameter that differed significantly between groups was the length of the foal upper hindlimb, which was longer in ET and ICSI-C than in NC foals. Evaluation of placental mRNA expression for 17 genes related to growth and vascularisation showed no difference in gene expression between groups. These data indicate that within this population, use of ARTs was not associated with meaningful changes in foal or placental morphometry or in expression of the placental genes evaluated.

Production of a mitochondrial-DNA identical cloned foal using oocytes recovered from immature follicles of selected mares.

Cloned animals possess mitochondria derived from the host ooplast, which typically differ genetically from those of the donor. This is of special concern to horse breeders, as maternal lines are prized and athletic performance is a key factor in genetic value. To evaluate the feasibility of producing mitochondrial-identical cloned foals, we collected oocytes from immature follicles of two mares, BL and SM, maternally related to the donor stallion. In vitro matured, enucleated oocytes were treated with roscovitine-synchronized donor cells and blastocysts were transferred transcervically to recipient mares. In Mare BL, 10 aspiration sessions yielded 45 oocytes, of which 12 matured and seven were successfully recombined. One blastocyst was produced, which did not yield a pregnancy. In Mare SM, three aspiration sessions yielded 53 oocytes, of which 27 successfully recombined. These were assigned to either Scriptaid or Scriptaid plus Vitamin C treatments for the first 12 to 16 hours of embryo culture. Two blastocysts were produced from each treatment. One pregnancy was established after transfer from the Scriptaid treatment. This resulted in a viable foal whose genomic DNA and mitochondrial DNA matched to those of the donor animal. These results indicate that production of mitochondrial-identical cloned foals can be achieved using oocyte recovery from a very small number of selected mares. Despite mitochondrial homogeneity, the results varied with mare; Mare BL yielded both significantly fewer oocytes per aspiration session (P < 0.001) and significantly fewer reconstructed oocytes per oocyte recovered ( P < 0.001) than did Mare SM.