Estrogens are steroid hormones that play a significant role in the reproductive physiology of horses. They are primarily produced by the ovaries, but can also be synthesized in smaller amounts by the adrenal glands and placenta. Estrogens are involved in the regulation of the estrous cycle, influencing reproductive behaviors, and preparing the reproductive tract for potential pregnancy. In mares, the most prominent estrogens include estradiol, estrone, and equilin. These hormones can also impact bone density, cardiovascular function, and metabolism. This page compiles peer-reviewed research studies and scholarly articles that explore the production, function, and clinical implications of estrogens in equine health and reproduction.
Bollwein H, Schulze JJ, Miyamoto A, Sieme H.The goal of this study was to investigate for the first time a possible association between plasma concentrations of testosterone and total estrogen and testicular blood flow in the stallion. Correlations between these variables were calculated before and after administration of human chorionic gonadotropin (hCG). Eight mature warmblood stallions received 5,000 IU hCG intravenously, and four stallions received solvent only. Testicular blood flow in the left and right testicular arteries was assessed using colour Doppler sonography by measuring blood flow volume (BFV) and pulsatility index (PI)...
Chang M, Overk CR, Kastrati I, Peng KW, Yao P, Qin ZH, Petukhov P, Bolton JL, Thatcher GR.Oxidative metabolism of estrogens has been associated with genotoxicity. O-methylation of catechol estrogens is considered as a protective mechanism. 4-Methoxyequilenin (4-MeOEN) is the O-methylated product of 4-hydroxyequilenin (4-OHEN). 4-OHEN, the major catechol metabolite of the equine estrogens present in the most widely prescribed hormone replacement therapeutics, causes DNA damage via quinone formation. In this study, estrogen receptor (ERa) binding of 4-MeOEN was compared with estradiol (E2) and equilenin derivatives including 4-BrEN using computer modeling, estrogen response element (...
Hejmej A, Bilińska B.Evidence collected over the years has demonstrated that cryptorchidism is associated with a defect in spermatogenesis and, as a consequence, with either reduced fertility or infertility. However, the effect of cryptorchidism on Leydig cell function is less clear. The aim of our study therefore was to investigate the regulation of steroid hormone biosynthesis and, additionally, intercellular communication in the cryptorchid equine testes. Methods: Testes of mature bilaterally cryptorchid horse and healthy stallions were used for this study. The expression of luteinising hormone receptor (LHR), ...
Roser JF.It is well established in many mammalian species, including the horse that normal testicular function is dependent upon a functional hypothalamic-pituitary-testicular (HPT) axis, which involves classic feedback mechanisms. The major HPT hormones involved in the stallion are gonadotropin-releasing hormone (GnRH), luteinizing hormone (LH), follicle stimulating hormone (FSH), testosterone (T), estrogens (Es) and inhibin (INH). Although prolactin (PRL) fluctuates with season in the stallion and both PRL and thyroid hormone (TH) affect reproduction in other male species, their effects on stallion r...
Volkmann D, Zent W, Little T, Riddle T, Durenberger J, Potenza K, Sibley L, Roser J.While searching for the cause of the Mare Reproductive Loss syndrome (MRLS), we postulated that 1 of 3 tissues in 40-120 D pregnant mares was the likely primary target of the noxious factor that caused early abortions: The corpora lutea (CL), the endometrium or the fetus and/or its membranes. At this stage of gestation, progesterone (P4) is solely produced by luteal tissue, eCG by endometrial cups in the endometrium and oestrogens by the feto-placental unit. We determined whether concentrations of P4, eCG and/or total conjugated oestrogens (CE) would indicate which tissue was targeted during t...
Rambags BP, van Tol HT, van den Eng MM, Colenbrander B, Stout TA.Progesterone and oestrogen play essential roles in the maintenance of pregnancy in eutherian mammals and are thought to exert their effects on the developing conceptus indirectly, via the endometrium. In some species, early embryos have themselves been shown to express steroid receptors, thereby suggesting that reproductive steroids may also influence embryonic development directly. The aim of this study was to determine whether early intrauterine equine conceptuses express either the classical intracellular progesterone (PR) and oestrogen receptors (ERalpha and ERbeta) or the more recently ch...
Weng Q, Tanaka Y, Taniyama H, Tsunoda N, Nambo Y, Watanabe G, Taya K.To elucidate the relationship between steroidogenic hormones and developing adrenal glands, we investigated the immunolocalization of steroidogenic enzymes in equine fetal adrenal glands during mid-late gestation. Fetal adrenal glands were obtained from three horses at 217, 225 and 235 days of gestation. Steroidogenic enzymes were immunolocalized using polyclonal antisera raised against bovine adrenal cholesterol side-chain cleavage cytochrome P450 (P450scc), human placental 3beta-hydroxysteroid dehydrogenase (3betaHSD), porcine testicular 17alpha-hydroxylase cytochrome P450 (P450c17) and huma...
Yasui M, Suzuki N, Liu X, Okamoto Y, Kim SY, Laxmi YR, Shibutani S.4-Hydroxyequilenin (4-OHEN)-dC is a major, potentially mutagenic DNA adduct induced by equine estrogens used for hormone replacement therapy. To study the miscoding property of 4-OHEN-dC and the involvement of Y-family human DNA polymerases (pols) eta, kappa and iota in that process, we incorporated 4-OHEN-dC into oligodeoxynucleotides and used them as templates in primer extension reactions catalyzed by pol eta, kappa and iota. Pol eta inserted dAMP opposite 4-OHEN-dC, accompanied by lesser amounts of dCMP and dTMP incorporation and base deletion. Pol kappa promoted base deletions as well as ...
Dehennin L, Petit E, Bonnaire Y, Bruyas JF, Le Bizec B, Plou P.Rules of horse racing stipulate that pregnant mares may compete under definite conditions of date, because early pregnant status may be misused for the sake of enhancing physical performance by putative anabolic steroid action. Screening for pregnancy is generally performed by plasma equine gonadotrophin (eCG) immunoassay, which covers the period between Days 40 and 120. In common screening for urinary anabolic steroids performed by gas chromatography-mass spectrometry, inclusion of two complementary criteria, i.e. the evaluation of total conjugates of 5(10)-estrene-3beta,17alpha-diol (EED) an...
Bilińska B, Wiszniewska B, Kosiniak-Kamysz K, Kotula-Balak M, Gancarczyk M, Hejmej A, Sadowska J, Marchlewicz M, Kolasa A, Wenda-Rózewicka L.The purpose of this article was to summarize our results on the role of androgens and estrogens in human, rodent and equine testes and epididymides, in both, physiological and patological conditions, obtained in the space of the Solicited Project (084/PO6/2002) financially supported by the State Committee for Scientific Research during the last three years. Testosterone produced by Leydig cells of the testes is clearly the major androgen in the circulation of men and adult males of most mammalian species. However, androgen metabolites make up a significant fraction of total circulating steroid...
Bleul U, Theiss F, Rütten M, Kähn W.This case report describes a three-year-old Friesian stallion with hypospadias. Physical examination revealed a ventrocaudal deviation of the shaft of the penis and an incomplete glans penis. The urethral opening was 4 cm in length, slit-like and had a mucous membrane covering. A short fold ran from the ventral aspect of the anus and ended with a non-pigmented hairless area of skin. A human choriongonadotropin (hCG)-stimulation test resulted in an increase in the plasma levels of estrogen sulfate and testosterone, indistinguishable from that of a normal stallion. The karyogram had a normal num...
Roberto da Costa RP, Ferreira-Dias G, Mateus L, Korzekwa A, Andronowska A, Platek R, Skarzynski DJ.Nitric oxide (NO) plays an important role in angiogenesis and in the regulation of the blood flow. This study was carried out to investigate (i) the effects of endogenous estrogens and progestins and exogenous progesterone (P(4)) (5 ng/ml or 1 microg/ml) or estradiol 17beta (E(2)beta) (50 pg/ml or 1 microg/ml) on in vitro endometrial NO synthesis; (ii) the presence of different isoforms of NO synthase; (iii) and their relationship to microvascular density in the equine endometrium during the estrous cycle. NOS expression was also evaluated in the myometrium. Expression of endothelial and induc...
Parlevliet JM, Pearl CA, Hess MF, Famula TR, Roser JF.The presence of steroids and their receptors throughout development, specifically androgen receptor (AR), estrogen receptor alpha (ERalpha) and estrogen receptor beta (ERbeta), in the epididymis of a high estrogen producing species like the stallion has not been determined. Epididymal and testicular samples were collected for analysis of testosterone and estradiol-17beta (E(2)) concentrations and for immunolocalization of AR, ERalpha and ERbeta. The concentration of testosterone in the testis and epididymis were not different among age groups (P>0.05). AR was localized in the principal cell...
Reepmeyer JC, Brower JF, Ye H.Equilin-3-sulfate and delta8,9-dehydroestrone-3-sulfate are two isomers found in equine conjugated estrogens that differ in structure only by the position of a double bond in the steroid B-ring. These geometric isomers were not resolved on a C18 column during the analysis of conjugated estrogen drug products by LC-MS using acetonitrile-ammonium acetate buffer as the mobile phase. While no separations of these two isomers were observed on C18 or other alkyl-bonded silica based phases using a variety of mobile phase conditions, partial separations were achieved on phenyl bonded silica phases wit...
Hartt LS, Carling SJ, Joyce MM, Johnson GA, Vanderwall DK, Ott TL.Uterine function is primarily controlled by the combined actions of oestrogen and progesterone working through their cognate nuclear receptors. The mechanism of establishment of pregnancy in the mare is of interest because it involves prolonged pre-attachment and conceptus migration phases, and both invasive and non-invasive placental cell types, and as such has been an important comparative model. This study characterised regulation of oestrogen (ER) and progesterone (PR) receptors in the endometrium of the mare during the oestrous cycle and early pregnancy. Endometrial tissues collected duri...
Vardy MD, Gardner TR, Cosman F, Scotti RJ, Mikhail MS, Preiss-Bloom AO, Williams JK, Cline JM, Lindsay R.The purpose of this study was to determine effects of ovariectomy (OVX) and conjugated equine estrogens plus medroxyprogesterone acetate (CEE/MPA), or ethinyl estradiol plus norethindrone acetate (EE/NA) on biomechanics of uterosacral (USL) and round (RL) ligaments in postmenopausal (PMP) monkeys. Methods: This was a randomized, triple blind, placebo-controlled study. OVX monkeys received 12 months no treatment (Pbo) (n = 19), CEE/MPA (n = 19), or EE/NA (n = 21). USL and RL step strains and stress-relaxation data were curve-fitted, giving strain-dependent tensile modulus (TM) from 0% to 30%. R...
Hejmej A, Gorazd M, Kosiniak-Kamysz K, Wiszniewska B, Sadowska J, Bilińska B.Androgen metabolism may proceed to amplify the action of testosterone by its aromatisation to oestradiol. Recently, a growing body of evidence suggests a role of oestrogens in the male reproductive tract via their specific oestrogen receptors (ERs). In order to check whether androgens are converted to oestrogens in the testis, epididymis and prostate of the stallion, the expression of aromatase was visualised by means of immunohistochemistry. Moreover, to show the cellular targets for oestrogens the presence of oestrogen receptor alpha (ERalpha) and oestrogen receptor beta (ERbeta) was demonst...
Register TC, Cann JA, Kaplan JR, Williams JK, Adams MR, Morgan TM, Anthony MS, Blair RM, Wagner JD, Clarkson TB.The effects of dietary soy isoflavones (IF) and conjugated equine estrogens (CEE) on circulating inflammatory markers were determined at the end of a 3-yr study of ovariectomized monkeys consuming a moderately atherogenic diet. Treatments were: 1) control, receiving alcohol-extracted soy-protein-based diet with low IF content (comparable to approximately 5 mg/d); 2) CEE, added to the control diet at a dose comparable to 0.625 mg/d; and 3) IF, consumed as a part of unextracted soy protein isolate at a dose comparable to 129 mg/d. Serum soluble vascular cell adhesion molecule-1 (sVCAM-1) was red...
Smith NL, Heckbert SR, Lemaitre RN, Reiner AP, Lumley T, Weiss NS, Larson EB, Rosendaal FR, Psaty BM.Clinical trial evidence indicates that estrogen therapy with or without progestins increases venous thrombotic risk. The findings from these trials, which used oral conjugated equine estrogens, may not be generalizable to other estrogen compounds. Objective: To compare risk of venous thrombosis among esterified estrogen users, conjugated equine estrogen users, and nonusers. Methods: This population-based, case-control study was conducted at a large health maintenance organization in Washington State. Cases were perimenopausal and postmenopausal women aged 30 to 89 years who sustained a first v...
Ousey JC.The endocrine profiles in the periparturient mares are dominated by increasing concentrations of progestagens and decreasing oestrogens. These hormones are produced by precursors from the foetus, metabolized by the placenta and act primarily on the maternal uterus. The circulating concentrations of hormones in maternal plasma, generally, represent a small proportion of those metabolized by the foetus and utero-placental tissues. There is clear evidence that the foetal hypothalamo-pituitary-adrenal (HPA) axis initiates the process of foetal maturation and the hormonal cascade which culminates i...
Briant C, Ottogalli M, Guillaume D.With the objective of controlling the day of ovulation, 40 mares were assigned to a control or three treated groups: A3d, A4d, and A5d. The treated groups received antarelix (Teverelix 0.01 mg/kg, i.v., twice a day) for 3, 4, or 5 days from the day the dominant follicle (F1) reached 28 mm (=D0), and one injection of hCG (1600 IU, i.v.) on D1, D2, or D3, respectively. Control mares received one injection of hCG when F1 reached 35 mm. Plasma LH, FSH, progesterone, and total estrogens were assayed. In the A3d, A4d, and A5d groups, 9 (90%), 6 (60%), and 5 (50%) out of 10 mares, respectively, ovula...
Raeside JI, Christie HL, Renaud RL, Waelchli RO, Betteridge KJ.Because estradiol (E(2)) production by the early equine conceptus is considered crucial to the establishment of pregnancy, the amounts of E(2), estrone (E(1)), and their sulfates (E(2)S, E(1)S) were measured by RIA in yolk-sac fluid of 63 conceptuses collected by transcervical lavage over the period of 11-26 days after ovulation. Amounts increased significantly with age of conceptus, especially for E(1)S. Then, the metabolism of E(2), which may be highly relevant for its action, was examined in the conceptus and endometrium over the period when the conceptus ceases to migrate within the uterus...
Li Y, Yao J, Chang M, Nikolic D, Yu L, Yager JD, Mesecar AD, van Breemen RB, Bolton JL.Catechol-O-methyltransferase (COMT) plays an important role in the inactivation of biologically active and toxic catechols. It has been shown that COMT is genetically polymorphic with a wild-type and variant form where a valine has been substituted with a methionine. Several, but not all, epidemiological studies have shown that women, homozygous with the variant form, have an increased risk of developing breast cancer. Previously, we showed that 4-hydroxyequilenin (4-OHEN), a cytotoxic/genotoxic equine catechol estrogen metabolite, is both a substrate of COMT and an irreversible inhibitor of t...
Brown KA, Boerboom D, Bouchard N, Doré M, Lussier JG, Sirois J.17Beta-hydroxysteroid dehydrogenase type 4 (17betaHSD4) has a unique multidomain structure, with one domain involved in 17beta-estradiol inactivation. The objective of the study was to investigate the regulation of 17betaHSD4 during human chorionic gonadotropin (hCG)-induced ovulation/luteinization. The equine 17betaHSD4 cDNA was cloned and was shown to encode a 735-amino acid protein that is highly conserved (81-87% identity) compared with other mammalian orthologs. RT-PCR/Southern blot analyses were performed to study the regulation of 17betaHSD4 transcripts in equine preovulatory follicles ...
Hess MF, Roser JF.The large amount of testicular estrogens produced by the stallion is unique compared to the amounts found in other domestic species. Although the cellular locale of the cytochrome P450 aromatase (P450arom) enzyme that converts C19 androgens to C18 estrogens has been identified in the Leydig cell of adult equine testis, the location in the immature equine testis is not known. The goal of this work was to localize the enzyme in colts and stallions during sexual development. Testes were obtained from prepubertal (n=7), pubertal (n=6), and postpubertal (n=8) colts and stallions during both the bre...
Guillaume D, Chavatte-Palmer P, Combarnous Y, Duchamp G, Martinat N, Nagy P, Daels PF.The aim of this study was to compare the effects of treatment with repeated injections of sulpiride (a dopamine D2 antagonist) on prolactin secretion and induced lactation in ovariectomized and intact adult mares and to verify if this induction was possible at the beginning and at the end of the birth season. Two experiments were carried out in September [experiment (expt) 1], and in March (expt 2), in France (48 degrees N). In expt 1, three groups of five mares were tested: intact-control, intact-treated and ovariectomized-treated mares. In expt 2, mares previously subjected to artificial pho...
Foster SJ, Marshall DE, Houghton E, Gower DB.In on-going studies of 'classical' and ring B-unsaturated oestrogens in equine pregnancy, the products of metabolism of [2,2,4,6,6-2H(5)]-testosterone and [16,16,17-2H(3)]-5,7-androstadiene-3 beta,17 beta-diol with equine placental subcellular preparations and allantochorionic villi have been identified. Using mixtures of unlabelled and [2H]-labelled steroid substrates has allowed the unequivocal identification of metabolites by twin-ion monitoring in gas chromatography-mass spectrometry (GC-MS). Two types of incubation were used: (i) static in vitro and (ii) dynamic in vitro. The latter invol...
Sipahutar H, Sourdaine P, Moslemi S, Plainfossé B, Séralini GE.High levels of plasma estrogens constitute an endocrine peculiarity of the adult stallion. This is mostly due to testicular cytochrome p450 aromatase, the only irreversible enzyme responsible for the bioconversion of androgens into estrogens. To identify more precisely the testicular aromatase synthesis sites in the stallion, testes from nine horses (2-5 years) were obtained during winter or spring. Paraplast-embedded sections were processed using rabbit anti-equine aromatase, followed by biotinylated goat anti-rabbit antibodies, and amplified with a streptavidin-peroxidase complex. Immunoreac...
Simon JA, Liu JH, Speroff L, Shumel BS, Symons JP.The purpose of this study was to compare the effects on vaginal bleeding patterns of continuous combined hormone replacement therapy with norethindrone acetate and ethinyl estradiol versus conjugated equine estrogens and medroxyprogesterone acetate. Methods: Three hundred fifty-seven postmenopausal women were selected randomly (in a blinded manner) to 12 months of treatment with 1 mg norethindrone acetate/5 microg ethinyl estradiol, placebo, or open-label 0.625 mg conjugated equine estrogens/2.5 mg medroxyprogesterone acetate (conjugated equine estrogens/medroxyprogesterone acetate [CEE/MPA]; ...
Lemazurier E, Séralini GE.Our previous work showed that stallion testis produces high amounts of estrogens which are subsequently found in the ejaculate. These estrogens are mainly synthesized by testicular aromatase, and the major estrogen produced is estrone sulfate (E1S). The objective of this study was to investigate the potential role of E1S as a source of estrogens in the male and female horse reproductive tracts by determining whether both estrone sulfatase (Sulf) and 17beta-hydroxysteroid dehydrogenase type I (17beta-HSD1) activities were present in equine testes, epididymis and uterus. We assessed E1S bioconve...
Kenney RM, Ganjam VK.Focal enlargements of the uterus are palpable through the rectal wall and have various aetiology. Aspects of their diagnosis, possible pathogenesis, significance and treatment are presented. Lymphatic lacunae appear to be quite common in older mares and may occasionally give rise to large endometrial cyst, but more frequently cause widespread change throughout the uterine horns. Three unusual ovarian abnormalities are presented, a large, oestrogen-producing granulosa cell tumour, an oestrogen-producing large follicle in a 4-month-old filly, and an ectopic progesterone-producing adrenal structu...
El-Sheikh Ali H, Legacki EL, Scoggin KE, Loux SC, Dini P, Esteller-Vico A, Conley AJ, Stanley SD, Ball BA.Equine placentitis is associated with alterations in maternal peripheral steroid concentrations, which could negatively affect pregnancy outcome. This study aimed to elucidate the molecular mechanisms related to steroidogenesis and steroid-receptor signaling in the equine placenta during acute placentitis. Chorioallantois (CA) and endometrial (EN) samples were collected from mares with experimentally induced placentitis (n = 4) and un-inoculated gestationally age-matched mares (control group; n = 4). The mRNA expression of genes coding for steroidogenic enzymes (3βHSD, CYP11A1, CYP17A1, CYP19...
Silva ESM, Ignácio FS, Fritsch SC, Zanoni DS, Pantoja JCF, Oliveira-Filho JP, Meira C.To test the hypothesis that the administration of 2.5 mg of estradiol benzoate (EB) followed by 1500 mg of long acting progesterone (LA P4) causes similar uterine changes and molecular dynamics in anovulatory mares to those observed in cyclic ones, we evaluated the changes of estrogen (ERα and ERβ) and progesterone receptors (PR) in anestrous, transitional and cyclic mares by RT-qPCR and immunohistochemistry. In addition, we evaluated uterine edema, tonus and estrogens and progesterone plasma profile. Endometrial biopsies were taken from anestrous and transitional mares immediately before ...
Almadhidi J, Moslemi S, Drosdowsky MA, Séralini GE.Aromatase (estrogen synthetase) is a steroidogenic enzyme complex which catalyzes the conversion of androgens to estrogens (termed aromatization). This enzyme was purified from adult equine testis to homogeneity by five chromatographic steps. The ability of purified and reconstituted equine aromatase to exhibit an estrogen 2-hydroxylase activity was tested and compared to testosterone aromatization. Enzymatic activities were assessed by tritiated water release from labelled estradiol and testosterone. Kinetic analysis of estradiol 2-hydroxylation showed an apparent K(m) of 23 microM and a V(ma...
Monfort SL, Arthur NP, Wildt DE.Immunoreactive urinary oestrogen conjugates were assessed in daily urine samples (approximately 5 samples/week) collected from 8 Przewalski's mares maintained under semi-free-ranging pasture conditions. The relative percentage contributions of immunoreactive urinary oestrogens during different reproductive stages (oestrus, luteal phase, early, mid- and late gestation) were determined using high-pressure liquid chromatography. In general, conjugated forms of oestrone (oestrone sulphate and oestrone glucuronide) were the major excreted immunoreactive oestrogens in nonpregnant and pregnant Przewa...
Koskinen E, Kuntsi H, Lindeberg H, Katila T.Follicular growth and hormone determinations were used to predict ovulation in the mare. Thirty Finnhorse mares were used for the investigation and 38 oestrous cycles were studied. The mares were examined by rectal palpation and ultrasonography every 6 hours during late oestrus. Daily blood samples were obtained for progesterone and oestrone sulphate determination. The preovulatory follicle grew 3 mm a day up to 2 days before ovulation. The size then remained constant, before diminishing by 2-3 mm during the last 12 hours. The maximal diameter of the follicle was 43 +/- 4 mm. In 89% of the fol...
Mlodawska W, Slomczynska M.Ovarian steroidogenesis from the neonatal to pubertal period in horses is poorly understood. This study was designed to immunolocalize cytochrome P450 aromatase in the ovarian follicles of slaughtered fillies ages approximately (I) 6-9 mo (5 mm in diameter. Staining intensity was dependent on the size and morphology of the follicle. In nonatretic follicles 5-10 mm in diameter, the reaction was weak and heterogeneous, while most intense staining was observed in preovulatory follicles. In follicles (diameter <20 mm) in the groups <10MF and 1YF, the reaction was less intense than in adult m...
Kalpokas I, Mattos RC, Cavestany D, Martínez MN, Perdigón F, Meikle A.The effect of side of corpus luteum on uterine gene expression and protein localization of estrogen receptor α (ERα) and progesterone receptor (PR) in healthy cyclic and pregnant mares 13 days after ovulation (day 0) was investigated. Transcervical biopsies were performed to collect endometrium ipsilateral and contralateral regarding the side of corpus luteum on day 13 post-ovulation in cyclic (n = 6) and pregnant (n = 6) mares. Blood samples were collected daily from day 0 until the day of biopsy for 17β-estradiol (E2) and progesterone (P4) determinations. Receptor expression was d...
Asa CS.Behavior during the estrous phase of the ovulatory cycle of the mare is analogous in most ways to that of estrous females of other species. Proceptive behaviors bring the mare into the proximity of the male and attract his attention. Positioning facilitates mounting, intromission, and ejaculation. Estrous signs appear to be more intense in the few days prior to ovulation than during the transition periods that separate the recurring estrous and diestrous phases. Sexual behavior is absent during diestrus. Detection of estrus in mares is problematic in that it requires the presence (or at least ...
Daels PF, DeMoraes JJ, Stabenfeldt GH, Hughes JP, Lasley BL.Thirty pregnant mares were assigned to 3 groups: Group 1 (n = 10) mares served as controls; Group 2 (n = 10) mares were treated with altrenogest (44 mg/day) from Day 16 to 80 and Group 3 (n = 10) mares were treated with a luteolytic dose of PGF2 alpha on Day 16 followed by altrenogest (44 mg/day) until Day 80. Concentrations of progesterone and chorionic gonadotrophin (CG) in plasma and oestrogen conjugate (OC) in urine were determined between Days 16 and 80 of gestation. In Group 3, complete luteolysis occurred in all 10 mares following administration of PGF2 alpha. Six of the 10 mares did no...
Simon JA, Liu JH, Speroff L, Shumel BS, Symons JP.The purpose of this study was to compare the effects on vaginal bleeding patterns of continuous combined hormone replacement therapy with norethindrone acetate and ethinyl estradiol versus conjugated equine estrogens and medroxyprogesterone acetate. Methods: Three hundred fifty-seven postmenopausal women were selected randomly (in a blinded manner) to 12 months of treatment with 1 mg norethindrone acetate/5 microg ethinyl estradiol, placebo, or open-label 0.625 mg conjugated equine estrogens/2.5 mg medroxyprogesterone acetate (conjugated equine estrogens/medroxyprogesterone acetate [CEE/MPA]; ...
Cox JE, Redhead PH, Dawson FE.The results of performing 1720 blood tests for equine cryptorchidism are described. Using the paired sample human chorionic gonadotrophin (hCG) stimulation test and measuring testosterone, 6.7 per cent of tests did not give a clear result. If only the testosterone concentration in the pre-hCG blood sample was used, this percentage rose to 14 per cent. The paired sample hCG stimulation test was 94.6 per cent accurate. A comparison was made between the paired hCG stimulation test and the measurement of conjugated oestrogen in a single sample. The latter did not give as many doubtfuls but gave fa...
Zhang N, Ding S, Kolbanovskiy A, Shastry A, Kuzmin VA, Bolton JL, Patel DJ, Broyde S, Geacintov NE.The equine estrogens equilin (EQ) and equilenin (EN) are the active components in the widely prescribed hormone replacement therapy formulation Premarin. Metabolic activation of EQ and EN generates the catechol 4-hydroxyequilenin (4-OHEN) that autoxidizes to the reactive o-quinone form in aerated aqueous solutions. The o-quinones react predominantly with C, and to a lesser extent with A and G, to form premutagenic cyclic covalent DNA adducts in vitro and in vivo. To obtain insights into the structural properties of these biologically important DNA lesions, we have synthesized site-specifically...
Almadhidi J, Seralini GE, Fresnel J, Silberzahn P, Gaillard JL.Estrogens are the major steroids produced by equine gonads. To identify the cells responsible for estrogen synthesis, an antiserum against purified equine testicular cytochrome P450 aromatase was produced in rabbits. The reactivity and specificity of the antiserum were assessed by ELISA, immunoblot analysis, and immunoneutralization studies. Immunofluorescence microscopy demonstrated that in the male gonad, cytochrome P450 aromatase (P450arom) was localized in the interstitial tissue, whereas, under the experimental conditions used, the Sertoli and germ cells did not show any specific staining...
Haffner JC, Fecteau KA, Eiler H, Tserendorj T, Hoffman RM, Oliver JW.Traditionally, analysis of blood cortisol alone has been used to evaluate adrenal function. Currently, multisteroid analyses are considered more informative than analysis of a single hormone to assess adrenal function. The objective of the present research was to create a database for steroid reference values for domestic Mongolian horses. Seven adrenal steroid levels were determined in the blood of 18 colts, 34 stallions, 25 geldings, 17 fillies, and 29 mares. Results were as follows (lowest and highest group median, in nanograms per milliliter): progesterone: <0.030 (fillies), 4.30 (mares), ...
Choi SJ, Anderson GB, Roser JF.In vitro production of free estrogens and estrogen conjugates by intact Day 12.5, 13.5 and 14.5 equine embryos was measured at 2-h intervals over a 24-h culture period. Production of free estrogens was higher for Day 14.5 than Day 12.5 embryos. Differences in production of conjugated estrogens were not significant, but a trend toward increased production with increased age of embryo was apparent. No trend toward increased free and conjugated estrogen production per cell was observed with age. Embryo diameter and number of cells increased with age but varied considerably within groups. The amou...
Barreto RSN, Romagnolli P, Mess AM, Rigoglio NN, Sasahara THC, Simões LS, Fratini P, Matias GSS, Jacob JCF, Gastal EL, Miglino MA.In horses, pregnancy is characterized by high levels of maternal estrogens that are produced largely by the interstitial tissue inside the gonads of the offspring, associated with a physiological gonadal hyperplasia, that is uncommon in other species. However, a detailed structural-functional understanding of the early stages of gonadal development and hyperplasia has remained elusive in horse pregnancy because of the lack of substantial data. The goal of this study was to describe the genital organs' development in 19 early horse embryos and fetuses (days 20-140 of gestation) of both sexes by...
Honnens A, Weisser S, Welter H, Einspanier R, Bollwein H.The objective of this study was to investigate the relationships between uterine perfusion and estrogen, progesterone and the uterine nitric oxide synthase (NOS) system in five trotter mares during the estrous cycle. Color Doppler sonography for measurement of uterine blood flow and collection of blood for determination of plasma estrogen and progesterone concentrations were performed on days 0 (= ovulation), 1, 5, 11 and 15 and daily during estrus (days -1 to -4) of one estrous cycle; endometrial biopsy collection for mRNA expression analysis of NOS and estrogen receptors was performed on day...
Li Y, Yao J, Chang M, Nikolic D, Yu L, Yager JD, Mesecar AD, van Breemen RB, Bolton JL.Catechol-O-methyltransferase (COMT) plays an important role in the inactivation of biologically active and toxic catechols. It has been shown that COMT is genetically polymorphic with a wild-type and variant form where a valine has been substituted with a methionine. Several, but not all, epidemiological studies have shown that women, homozygous with the variant form, have an increased risk of developing breast cancer. Previously, we showed that 4-hydroxyequilenin (4-OHEN), a cytotoxic/genotoxic equine catechol estrogen metabolite, is both a substrate of COMT and an irreversible inhibitor of t...
Zavy MT, Vernon MW, Sharp DC, Bazer FW.Comparisons of estrone, 17 beta-estradiol, and plasma progestin concentrations were made in uterine fluid and peripheral blood of nonpregnant and pregnant pony mares. Concentrations of these steroids were also measured within yolk sac fluid from blastocysts on days 12, 14, 16, and 18 of pregnancy to obtain more complete analyses of the uterine environment (uterine fluid plus yolk sac fluid) of early pregnancy. Thirty mares were randomly assigned to six treatment groups (n = 5/group), and uterine fluid and peripheral blood samples were obtained on days 8, 12, 14, 16, 18, and 20 postovulation. A...
Raś A, Rapacz A, Raś-Noryńska M, Janowski TE.Cryptorchidism is a partial or total failure of testicular descent. For a proper diagnosis different methods are required. The main aim of this study was to compare different diagnostic methods. Sixty two horses (15 stallions, 32 cryptorchids and 15 geldings) were used in the experiment. They were clinically examined and ultrasonography was used to locate the testes. Blood samples were taken from the animals to measure the plasma level of testosterone and total estrogen (RIA method). In 22 horses suspected of cryptorchidism, the hCG stimulation test was performed. Blood samples were taken ever...
Choi I, Collante WR, Simmen RC, Simmen FA.Pig blastocysts exhibit a transient period of estrogen production at periimplantation, with a second, more sustained period of estrogen synthesis occurring in endometrium and placenta at later pregnancy. Previously we reported the isolation of cDNA clones encoding a novel isoform of cytochrome P450 aromatase (the terminal enzyme in the estrogen biosynthetic pathway) from porcine periimplantation blastocysts. The present study investigated pregnancy-associated expression, in blastocysts and maternal reproductive tract tissues of this and an additional mRNA transcript encoding a distinct P450 ar...
Muesing RA, Miller VT, LaRosa JC, Stoy DB, Phillips EA.Administration of conjugated equine estrogen to 31 postmenopausal women for 3 months produced 14.6% and 9.4% decreases in low density lipoprotein cholesterol (LDL-C) and apolipoprotein-B (apoB), and 11.5%, 12.7%, and 9.6% increases in high density lipoprotein cholesterol (HDL-C), apoA-I and apoA-II, respectively. Phospholipids of HDL2 and HDL3 were increased 57.9% and 19.3%, respectively, while relatively small increases in cholesterol of the two subfractions were not significant. Compositions of LDL and HDL and its subfractions were altered substantially with estrogen treatment. The proportio...
Reiffsteck A, Dehennin L, Scholler R.Estrone, 2-methoxyestrone and estradiol-17 beta have been definitely identified in seminal plasma of man, bull, boar and stallion by high resolution gas chromatography associated with selective monitoring of characteristic ions of suitable derivatives. Quantitative estimations were performed by isotope dilution with deuterated analogues and by monitoring molecular ions of trimethylsilyl ethers of labelled and unlabelled compounds. Concentrations of unconjugated and total estrogens are reported together with the statistical evaluation of accuracy and precision.
Stabenfeldt GH, Daels PF, Munro CJ, Kindahl H, Hughes JP, Lasley B.A direct enzyme immunoassay was developed to measure conjugated oestrogens in the plasma of pregnant mares. The antibody was produced in rabbits using oestrone-3-glucuronide (E1G) conjugated to bovine serum albumin. The enzyme conjugate was E1G conjugated to horseradish peroxidase. A sharp increase in plasma E1G concentrations occurred between Days 35 and 40 of gestation. Values declined slightly to Day 45, remained relatively constant to around Day 70 and rose sharply thereafter. Fetal death before Day 35 had no effect on plasma concentrations of E1G. Fetal death after Day 35 in conjunction w...
Stickle RL, Erb RE, Fessler JF, Runnels LJ.Unilateral ovariectomy was performed on 3 mares affected with granulosa cell tumors. Tumor fluid in each mare was found to contain estrogen, testosterone, and progesterone. In 2 mares, preoperative blood plasma concentrations of these hormones were comparable to those of a series of clinically normal mares. The other mare, which had a history of aggressive, masculine behavior, had higher testosterone content in the tumor fluid and in the preoperative blood sample. After surgical removal of the tumors, each mare developed follicles and ovulated with the remaining ovary. Each was eventually bred...
Inoue J, Cerbito WA, Oguri N, Matsuzawa T, Sato K.The serum concentrations of testosterone and oestrogens were determined in stallions classified as geldings, normal (according to age) or infertile (azoospermic). There were significant differences in testosterone and oestrogen levels between the groups. Normal concentrations of testosterone and total oestrogens were attained after 16 months of age. Castrated and immature horses ( 4 years). The results suggest that serum levels of testosterone and total oestrogens may provide a sensitive index of the endocrine function of the testis in male horses.
