Prostaglandin receptors (EP2 and EP4) and angiotensin receptor (AGTR2) mRNA expression increases in the oviducts of Nelore cows submitted to ovarian superstimulation

Abstract

Many peptides are responsible for the coordination of muscle contraction, secretion and ciliary beating of the oviduct epithelium to allow the transport of gametes and embryos, including vascular endothelial growth factors (VEGF), prostaglandins (PGs), endotelin-1 (ET-1) and angiotensin II (Ang II). The effect of reproductive biotechnologies used to improve embryo yield on oviduct gene expression is poorly understood. Thus, the aim of the present study was to evaluate the effect of ovarian superstimulation on the mRNA expression of the genes encoding the major peptides involved in oviduct contraction in bovine. Therefore, Nelore cows were submitted to P-36 (n=5) or P-36/eCG (n=5) ovarian superstimulatory protocols and a control group of cows was not submitted to any superstimulatory protocol (n=5). The relative expression of VEGF (VEGF, Flk1, Flt1), Ang II (AGTR2, ACE1), ET1 (ET1, ECE1) and PG pathway members (PGES, EP2, EP4, COX1, COX2) was analyzed using real time RT-PCR in each of oviduct segment (infundibulum, ampulla and isthmus). All target genes were expressed in the three segments of the bovine oviduct; however, specific genes were regulated by ovarian superstimulation: EP2 and EP4 receptors mRNA was affected by P-36/eCG protocol, in the ampulla and infundibulum, respectively; and AGTR2 mRNA was up-regulated by both the P-36/eCG and P-36 protocols in the isthmus. The upregulation of EP2, EP4 and AGTR2 expression in the superstimulated cows suggests a suitable effect of FSH and eCG on bovine oviduct physiology, coordinating the contraction in Nelore cows.

Equine chorionic gonadotropin drives the transcriptional profile of immature cumulus‐oocyte complexes and in vitro‐produced blastocysts of superstimulated Nelore cows

“…The animals were maintained on pasture ( Urochloa brizantha ), with ad libitum access to water and a mineral supplement. Nelore ( Bos taurus indicus ) nonlactating multiparous cows of age 5–7 years, and with body condition scores ranging from 2.0 to 3.5, were submitted to P‐36 (FSH group; n = 10) or P‐36/eCG (FSH/eCG group; n = 10) ovarian superstimulation protocols as described previously (Castilho et al, ; Fontes et al, ; Santos et al, ). Briefly, at a random stage of the estrous cycle, all animals received progesterone‐releasing vaginal inserts (1.0 g; PRIMER®, Tecnopec, São Paulo, Brazil) and estradiol benzoate (2.5 mg, i.m., Estrogin®, Farmavet, São Paulo, Brazil) on Day 0.…”

“…The use of FSH alone activates genes involved in oxidative and endoplasmic reticulum stress response, apoptosis, disturbance of angiogenesis, matrix remodeling, and growth‐related genes in follicular granulosa cells from beef cows (Dias, Khan et al, ), and impacts blastocysts gene expression (Mundim et al, ). When FSH was combined with eCG, genes involved in oviduct contraction, and relaxation were upregulated (Fontes et al, ) along with mRNA‐encoding proteins involved in the LH receptor intracellular signaling in granulosa cells (Castilho et al, ). Furthermore, this hormonal combination was able to modify the lipid profile of follicular fluid and steroidogenic capacity, increase plasma estradiol, regulate the abundance of the VEGF system, and LHCGR mRNA, and suppress miR‐222 expression in bovine granulosa cells (Santos et al, ).…”

Composing the Early Embryonic Microenvironment: Physiology and Regulation of Oviductal Secretions

“…Stimulation with hCG/eCG before insemination in gilts decreased the synthesis of PGE 2 and increased those of PGF 2α in the oviductal ampullas and isthmus three days post-insemination [59]. In Nelore cows, superovulation treatment using FSH and eCG induced site-specific changes in the expression of receptors for prostaglandins (EP2 and EP4) and angiotensin II (AGTR2) in oviduct epithelial cells [94]. In Buffalo cows treated with repeated doses of FSH for superovulation, a decreased expression of progesterone receptor (PGR) and estrogen receptor 1 (ESR1), as well as vascular endothelial growth factor (VEGF) and its receptor vascular endothelial growth factor receptor 1 (FLT1), was observed in the oviduct [95].…”

Prostaglandin E2–Induced Immune Exhaustion and Enhancement of Antiviral Effects by Anti–PD-L1 Antibody Combined with COX-2 Inhibitor in Bovine Leukemia Virus Infection

“…cDNA was synthesized from the total RNA using PrimeScript Reverse Transcriptase (Takara Bio, Otsu, Japan) according to the manufacturer's instructions. To measure the mRNA expression levels of COX2, EP4, EP2 (25), HPGD (26), and IFN-g in total PBMCs and each subpopulation, qPCR was performed using a thermal cycler (LightCycler 480 System II; Roche Diagnostic, Mannheim, Germany) with SYBR Premix DimerEraser (Takara Bio), following the manufacturer's instructions. Internal control genes were b-actin (ACTB) and GAPDH.…”