Research articleTemporal analysis of prostaglandin F2α receptor, caspase 3, and cyclooxygenase 2 messenger RNA expression and prostaglandin F2α receptor and cyclooxygenase 2 protein expression in endometrial tissue from multiparous Nelore (Bos taurus indicus) cows treated with cloprostenol sodium during puerperium
Introduction
During the postpartum period, the uterus undergoes modifications that restore the environment required to sustain a new pregnancy. Ymer et al. [1] proposed that in cows, a 65-day puerperium period is optimal, and conception within 85 to 90 days is desirable. Therefore, the production of one calf per year is economically feasible.
The postpartum uterus produces prostaglandin F2α (PGF2α), which can assist in the process of uterine involution by stimulating myometrial contraction [2], [3]. Cloprostenol sodium is a synthetic analog of PGF2α that is used for estrous synchronization, disease treatment, and to reduce uterine involution during the postpartum period [4], [5], [6], [7].
Prostaglandin F2α binds to the G-protein–coupled prostaglandin F2α receptor (FP), which mediates the release of intracellular calcium (Ca+) to promote muscular contraction [6], [8]. The mature form of the FP is highly expressed in luteal [8], [9], [10], [11], [12] and follicular cells [13] and in fetal membranes and uterine tissue during gestation [14], [15].
During estrus, the mammalian endometrium undergoes a series of morphologic and functional changes [16]. Furthermore, during the postpartum period, the endometrial tissue undergoes a series of morphologic changes to restore the histologic features that were altered during pregnancy and delivery.
Apoptosis is physiological programmed cell death, which ensures the balance between cellular death and proliferation. Caspases are a group of cysteine proteases that degrade proteins by recognizing aspartic acid residues [17]. These proteases are involved in apoptosis, are synthesized as zymogens, and belong to two classes: the initiators (caspase 2, caspase 8, caspase 9, and caspase 10) and the executioners (caspase 3 [CASP-3], caspase 6, and caspase 7). The initiator caspases exist as monomers and are activated by dimerization. The executioner caspases exist as dimers and are activated by cleavage of their interdomain linkers by initiator caspases [17], [18], [19]. Caspase-3 function is present during the final step of apoptosis and can be activated by intrinsic (mitochondrial) or extrinsic (death receptor) pathways [18]. Also, CASP-3 mRNA expression has been observed in the bovine placenta and uterus during the estrous cycle and during gestation [16], [19].
Cyclooxygenase (COX) is the enzyme required for the biosynthesis of prostaglandins from arachidonic acid [20], and it exists in two forms, namely COX-1 and COX-2. Cyclooxygenase 1 is a constitutive enzyme that is present in most cells. Cyclooxygenase 2 gene expression is induced by stimuli or mediators [6]. Also, COX-2 is important for cell growth and during inflammation [20], and it may produce prostaglandin during inflammation [21]. The COX expression in different tissues differs among species [22], but COX-2 is expressed in bovine uterine tissue during the estrous cycle, gestation, and the postpartum period [22], [23], [24], [25].
This study is based on the hypothesis that cloprostenol sodium promotes modifications in FP, COX-2, and CASP-3 mRNA expression that may favor the process of postpartum uterine involution in multiparous Nelore (Bos taurus indicus) females. Additionally, we aimed to describe the presence and immunolocalization of the FP and the COX-2 protein in endometrial tissue at different postpartum time points in these females. We examined the effect of serial administration of cloprostenol sodium (Days 1 and 4, Day 0 = birth) during the puerperal period of multiparous Nelore cows. We used quantitative reverse transcriptase–polymerase chain reaction (qRT-PCR) to monitor mRNA expression (on Days 1, 7, 14, 28, and 42) and immunohistochemical analysis to monitor tissue-specific protein expression (on Days 1, 14, 28, and 42). The results of this study may provide a better understanding of the molecular mechanisms involved in the postpartum period and may direct future studies toward the more effective use of cloprostenol sodium in hormonal protocols during the postpartum period of Nelore females.
Section snippets
Ethical aspects
The study was performed according to the ethical guidelines recommended by the National Council for Control of Animal Experimentation and the College of Animal Experimentation, and it was approved by the institution's Animal Care and Experimentation Ethics Committee (protocol number 50/2012).
Animals
A total of 24 multiparous Nelore (B. taurus indicus) females with calves were used. The parity of the animals was between two and six (median = 3), their age was between 4 and 13 years (median = 9), and
Results
Differences (P ≤ 0.05) were observed between the patterns of mRNA expression in the PG and TG only for CASP-3 and for the FP. The FP, CASP-3, and COX-2 mRNA expression differed among all time points during the postpartum period in the endometrial samples from Nelore cows. Immunohistochemical analyses revealed cytoplasmic COX-2 and FP localization in the endometrial surface epithelium, stroma, and glandular epithelium.
Discussion
During the puerperium period, the uterus undergoes modifications that restore an environment able to maintain a new pregnancy. In cows, a postpartum period of 65 days is recommended, and new conception within 85 to 90 days is desired, according to a review [1].
Because most studies are performed during gestation or in the first few hours after delivery, it is difficult to compare our study with previous results. To our knowledge, this is the first study of the spatiotemporal expression patterns
Acknowledgments
We are grateful to Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) for a scholarship (Proc.2011/15330-0), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (Capes (Proc./096/2010) for financial support, and to Fazenda Marino Avaré.
References (34)
- et al.
Molecular cloning and expression of a cDNA of bovine prostaglandin F2α receptor
J Biol Chem
(1994) - et al.
Suppression of protein kinace C signaling by the novel isoform of bovine PGF2α receptor
Biochem Biophys Res Commun
(2001) - et al.
Expression of prostaglandin F2α (PGF2α) receptor and its isoforms in the bovine corpus luteum during the estrous cycle and PGF2α-induced luteolysis
Domest Anim Endocrinol
(2012) - et al.
In vivo intra-luteal implants of prostaglandin (PG) E1 or E2 (PGE1, PGE2) prevent luteolysis in cows. II: mRNA for PGF2alpha, EP1, EP2, EP3 (A–D), EP3A, EP3B, EP3C, EP3D, and EP4 prostanoid receptors in luteal tissue
Prostaglandins Other Lipid Mediat
(2012) - et al.
Regulation, action and transport of prostaglandins during the periovulatory period in cattle
Mol Cell Endocrinol
(2007) - et al.
Immunohistochemical demonstration of cyclooxigenase—2 (COX-2) and prostaglandin receptors EP2 and FP expression in bovine intercaruncular uterine wall around term
Anim Reprod Sci
(2008) - et al.
Remodeling of bovine endometrium throughout the estrous cycle
Anim Reprod Sci
(2013) - et al.
Mechanisms of caspase activation
Curr Opin Cell Biol
(2003) - et al.
Expressions of apoptosis-regulating factors in bovine retained placenta
Placenta
(2011) - et al.
Expression of cyclooxygenase-II (COX-2) and 20α-hydroxysteroid dehydrogenase (20α-HSD)/prostaglandin F-synthase (PGFS) in bovine placentomas: implications for the initiation of parturition in cattle
Placenta
(2006)
Messenger RNA expression of angiotensin-converting enzyme, endothelin, cyclooxigenase-2 and prostaglandin synthases in bovine placentomas during gestation and the postpartum period
Vet J
Ovarian activity in beef and dairy cows with prolonged postpartum period and heifers that fail to conceive
Trop Anim Health Prod
Source of F series prostaglandins during the early postpartum period in cattle
Biol Reprod
Avanços na utilização de prostaglandinas na reprodução de bovinos
Rev Bras Reprod Anim
Serum progesterone levels in post-partum dairy cows after repeated application of prostaglandin F2α analogue D (+) cloprostenol sodium
J Vet Med A
Postpartum plasma PGF metabolite profile in cows with dystocia and/or retained placenta, and effect of fenprostalene on uterine involution and reproductive performance
J Vet Med Sci
Farmacologia veterinária
Cited by (6)
In-vitro effect of heat stress on bovine monocytes lifespan and polarization
2020, ImmunobiologyN-arylmethylideneaminophthalimide: Design, synthesis and evaluation as analgesic and anti-inflammatory agents
2019, Mini-Reviews in Medicinal ChemistryThe prostaglandin E<inf>2</inf> receptor PTGER2 and prostaglandin F<inf>2α</inf> receptor PTGFR mediate oviductal glycoprotein 1 expression in bovine oviductal epithelial cells
2018, Journal of Reproduction and DevelopmentUterine immune response after single and double fresh sperm insemination in mares
2017, Kafkas Universitesi Veteriner Fakultesi DergisiCloprostenol administration in the first week postpartum reduces expression of oxytocin receptors in the endometrium in Holstein-Zebu cows
2017, Arquivo Brasileiro de Medicina Veterinaria e ZootecniaEffect of exogenous hormones in the expression level of OXTRs gene in cows using Rt PCR
2023, Iraqi Journal of Veterinary Sciences