Increasing expression of oxytocin and vasopressin receptors in the equine conceptus between Days 10 and 16 of pregnancy.
Oxytocin (OT) and arginine vasopressin (AVP) have been detected in the yolk sac of the pre-attachment equine conceptus. Therefore, we have assessed the presence of OT and AVP receptors in equine conceptuses between Days 10 and 16 of pregnancy by qualitative PCR, quantitative PCR and immunohistochemistry. Expression of OT receptor and of the AVP receptors V1aR and V2R could be verified after sequencing the RT-PCR products of the expected length. The size of conceptuses used for quantitative PCR significantly increased with day of pregnancy (P<0.01) as did their quantitative expression of OTR (P<0.01). Immunohistochemistry of OTR resulted in weak trophectodermal abundance on Day 10, increasing at Day 12. On Day 14, staining intensity increased in individual cells of the trophectoderm while it decreased in other cells; this trend became more apparent on Day 16. The endoderm of the trophoblast and surrounding subtrophoblastic compartments always showed moderate staining for OTR. On Day 10 immunoreactive V2R protein was localised in the trophectodermal apical membrane; on Day 12 it was also present in the basal membrane and weakly in the cytoplasm. On Day 14 only individual trophectodermal cells showed positive supranuclear cytoplasmic areas or V2R, whereas on Day 16 about one-third of the trophectodermal cells were stained entirely and intensely positive. These results suggest an involvement of OT and AVP action in the development and expansion of the early equine conceptus.
Centre for Artificial Insemination and Embryo Transfer, University of Veterinary Sciences, Vienna A-1210, Austria. sven.budik@vetmeduni.ac.at
Sven
S
Budik
Franziska
F
Palm
Ingrid
I
Walter
Magdalena
M
Helmreich
Christine
C
Aurich
D000818:Animals; D001483:Base Sequence; D004622:Embryo, Mammalian / Q000378:metabolism*; D005260:Female; D018507:Gene Expression Regulation, Developmental; D005865:Gestational Age; D006736:Horses / Q000196:embryology* / Q000235:genetics / Q000378:metabolism; D008297:Male; D008969:Molecular Sequence Data; D011247:Pregnancy; D011270:Pregnancy, Animal; D018045:Receptors, Oxytocin / Q000235:genetics* / Q000378:metabolism; D017483:Receptors, Vasopressin / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015854:Up-Regulation
D018045:Receptors, Oxytocin; D017483:Receptors, Vasopressin