Vascular Endothelial Growth Factor A and VEGFR-1 Change during Preimplantation in Heifers

Abstract

Vascular endothelial growth factor A (VEGFA) plays a critical angiogenic role in the endometrium of placentalia during preimplantation. The role of VEGFA and its receptors is not fully characterised in bovine reproduction. We analysed the mRNA expression of VEGFA isoforms 121, 165 and 189, and VEGF receptors 1 and 2 in three experimental settings (A, B and C). We compared intercaruncular endometrium of cyclic to pregnant heifers at Days 12, 15 and 18 post insemination (Day 0), and between Day 15 and Day 18 conceptuses (A). We further compared caruncular versus intercaruncular endometrium at Day 15 (B), and endometrium of heifers carrying embryos originating from somatic cell nuclear transfer (SCNT) versus in vitro fertilisation (IVF) at Day 18 (C). Endometrial VEGFA protein was localised and quantified. Pregnant heifers displayed lower intercaruncular endometrial mRNA expression of VEGFA-121 (p = 0.045) and VEGFA-189 (p = 0.009) as well as lower VEGFA protein abundance (p < 0.001) at Day 15. The VEGFA protein was localised in intercaruncular luminal, glandular epithelium and in tunica muscularis of blood vessels. At Day 15, caruncular endometrium displayed higher VEGFA mRNA expression than intercaruncular endometrium (p < 0.05). Intercaruncular endometrial VEGFA protein at Day 18 was higher in abundance in SCNT than in IVF (p = 0.038). Therefore, during preimplantation in cattle, there may be a need for timely physiological reduction in intercaruncular endometrial VEGFA expression in favour of the caruncular area to facilitate a gradient towards the implantation sites. A higher expression of VEGFA in SCNT may predispose for later placentation abnormalities frequently observed following SCNT.

Luminal and Glandular Epithelial Cells from the Porcine Endometrium maintain Cell Type-Specific Marker Gene Expression in Air–Liquid Interface Culture

“…Quantitative real-time PCR (qPCR) was carried out using the KAPA SYBR FAST qPCR Kit (Kapa Biosystems, Wilmington, USA) on a CFX384 Real-Time PCR Detection System (Bio-Rad, Munich, Germany). The relative expression level (ΔCq) of each gene was generated by scaling the target gene Cq of each individual sample to the geometric mean of the Cq of three reference genes [H3 histone family member 3A (H3F3A), tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta ( YWHAZ ) and beta-actin ( ACTB )] as described previously by Chiumia et al [ 35 ]. To calculate ΔΔCq to visualize the RNA expression pattern, ΔCq values of PEEC-G or ALI-PEEC-G were subtracted from ΔCq values of PEEC-L or ALI-PEEC-L, respectively.…”

“…Media compositions applied for cell culture purposes are listed in Table 1. Phenol red-free DMEM/ Hams F12 1:1 (21,041,025), Ham's F12 (11,765,054), GlutaMAX (35,050,087), and Nu-serum growth medium supplement (11,563,600) were obtained from ThermoFisher Scientific, Waltham, USA. FBS (S0115, Lot 0742C), penicillin/streptomycin (A2212), amphotericin B (A2612), HEPES (L1613), and PBS (L1825), were purchased from Biochrom AG, Berlin, Germany.…”

Pluripotency and Growth Factors in Early Embryonic Development of Mammals: A Comparative Approach

“…For example, VEGF expression has been related to fetal weight increase in porcine embryos [ 55 ], in which VEGF expression was detected in TE cells at Day 14 of pregnancy [ 50 ]. VEGF , VEGFR-1 , and VEGFR-2 mRNA have been found in vitellin sacs and TEs of bovine embryos [ 49 , 56 , 57 ]. In humans, VEGF expression increases in the late luteal phase, while in bovine corpus luteum, VEGF mRNA is upregulated in the early luteal phase, and then is progressively downregulated until its levels increase again during pregnancy [ 58 , 59 ].…”

Manipulating the Epigenome in Nuclear Transfer Cloning: Where, When and How

“…Studies in bovine showed that the endometrium responds differently to cloned embryos as compared to embryos produced by in vitro fertilization (IVF), indicating abnormalities in embryo-maternal communication and pregnancy recognition signaling [ 77 , 78 ]. Subsequent studies revealed dysregulation of genes involved in cell signaling and placental development [ 79 , 80 ].…”