Assignment of the alpha-1-microglobulin bikunin precursor (AMBP) gene to porcine chromosome 1 via a new PCR-based polymorphism.
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Animals↗
Biomedical subjects
R D Geisert
Publications and source records attributed to R D Geisert.
Endometrial responsiveness to oxytocin during diestrus and early pregnancy in pigs is not controlled solely by changes in oxytocin receptor population density.
These studies were performed to test the hypotheses that: 1) endometrial responsiveness to oxytocin (OT) in pig endometrium is associated with changes in OT receptor (OTr) population density resulting from corresponding regulation of OTr gene transcription, 2) endometrial responsiveness to OT is controlled solely through a mechanism involving changes in OTr population density, and 3) OTr population density and endometrial responsiveness to OT differ between diestrus and early pregnancy in pigs. In experiment 1, OTr population density and dissociation constant (Kd) in cyclic pigs were constant on Days 10-16 but increased (p < 0.05) between Days 10 and 12 of pregnancy before decreasing (p < 0.05) through Day 16. OT induced phosphoinositide (PI) hydrolysis and prostaglandin (PG) F2 alpha secretion in cyclic pigs only on Day 16 (p < 0.05), and during pregnancy only on Day 12 (p < 0.05). Activation of G protein by aluminum fluoride (AIF4-) treatment maximally stimulated (p < 0.05) PI hydrolysis and PGF2 alpha secretion in cyclic pigs on all days, indicating that downstream from the OTr, the PGF2 alpha secretory pathway was fully functional. During pregnancy, PI hydrolysis and PGF2 alpha secretion in response to AIF4- decreased (p < 0.01) on Days 14 compared to Days 10 and 12, and AIF4- did not stimulate PGF2 alpha release on Day 16. In experiment 2, abundance of OTr mRNA in cyclic pigs decreased between Days 0 and 5 before increasing between Days 5 and 12 (p < 0.05), but it was higher (p < 0.05) on Days 10-15 of pregnancy than on equivalent days in cyclic gilts. These results indicate that control of PGF2 alpha secretion in cyclic pigs appeared to occur primarily at the level of OTr coupling to G protein because changes in OTr number were not associated with increased sensitivity to OT or G-protein activation by AIF4-. During pregnancy, control was exerted at multiple levels, which included the OTr, G protein, phospholipase C, and subsequent aspects of the secretory pathway. The present study also indicated that endometrium was responsive to OT during luteolysis in cyclic pigs but not during corpus luteum maintenance in pregnant pigs.
Aluminum Compounds↗
Expression of an inter-alpha-trypsin inhibitor heavy chain-like protein in the pig endometrium during the oestrous cycle and early pregnancy.
In pigs, changes in an unidentified endometrial glycoprotein, pGP30, are temporally associated with rapid trophoblast elongation and initial placental attachment on day 12 of gestation. Identification of endometrial pGP30 was undertaken through protein purification, NH2-terminal amino acid sequencing and cDNA sequencing of products generated through reverse transcription-polymerase chain reaction. Sequencing of 35 amino acids from the NH2-terminal end of pGP30 revealed that the 30 kDa glycoprotein is a cleavage product from the C-terminal region of inter-alpha-trypsin inhibitor heavy chain 4 (I alpha IH4), previously known as inter-alpha-trypsin inhibitor heavy chain-like protein. I alpha IH4 is unique compared with the three other inter-alpha-trypsin inhibitor heavy chains as it does not contain a binding site for bikunin that has serine protease inhibitory activity and is sensitive to cleavage by kallikrein. Endometrial gene expression of I alpha IH4 was detected during the oestrous cycle (days 0-18) and early pregnancy (days 10-18). Gene expression of I alpha IH4 appeared to be enhanced during the midluteal phase (days 12 and 15) of the oestrous cycle and the period of trophoblast attachment (days 12-18). Expression of I alpha IH4 was not detected in day 12 conceptus tissue mRNA. Endometrial expression of I alpha IH4 in pigs may function as an acute phase protein for protection of the uterus from the inflammatory response induced by conceptus attachment to the uterine epithelium.
Amino Acid Sequence↗
Rapid communication: the porcine Inter-alpha trypsin inhibitor-heavy chain 4 (ITIH4) gene maps to chromosome 13.
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Animals↗
Ultrasonic evaluation for the time of ovulation in ewes treated with norgestomet and norgestomet followed by pregnant mare's serum gonadotropin.
Progestogens and follicular stimulants have proved reasonably successful for estrus synchronization, but time of ovulation relative to removal of the progestogen is not clearly established. We monitored time of ovulation in ewes following synchronized estrus. Ovaries of 40 Dorset and Rambouillet x Dorset ewes were evaluated during the spring and fall (20/replicate). Ewes were randomly assigned to one of two treatment groups (n = 20/group): implant-only (I) ewes received a norgestomet implant for 10 d; and implant + PMSG (PI) ewes received a norgestomet implant for 10 d with an i.m. injection of 500 IU of PMSG at implant removal. Time of estrus onset was detected with the HeatWatch Estrus Detection System. Following the initiation of estrus, ovaries were monitored via rectal ultrasonography every 6 h to determine the interval from implant removal to ovulation (OVUL), and onset of estrus to ovulation (INT). Four I and PI ewes that lost implants before scheduled removal on d 10 were removed from the study. Estrus was detected in 13 of 16 (81%) I and 14 of 16 (88%) PI ewes within 108 h after implant removal. The interval from implant removal to estrus (EST), INT, and OVUL were not affected by replicate. The INT was not different in I (33.8 h) and PI (35.0 h) ewes. Mean OVUL was greater (P < .01) in I (79.8 h) than in PI ewes (68.6 h), and mean EST was also greater (P < .01) for I (46.0 h) than for PI ewes (32.6 h). The present data indicate that ovulation occurs on average 70 to 80 h after implant removal in ewes treated with norgestomet, and PMSG reduces the interval from implant removal to ovulation.
Animals↗
GnRH antagonist inhibition of luteinizing hormone secretion and ovulation in the pig.
A GnRH antagonist, Ac-D-p-Cl-Phe1,2, D-Trp3, D-Arg6, D-Ala10 GnRHb (Organon), was utilized to determine the effective dosage and duration to inhibit LH secretion in the pig. In a preliminary trial, barrows received either 10, 50, or 250 micrograms/kg BW of the GnRH antagonist. Secretion of LH was inhibited within 30 min for a duration of 12 h with the 100 micrograms/kg dose but persisted for greater than 48 h with the 250 micrograms/kg treatment. A second study determined effectiveness of the antagonist for inhibiting ovulation in cyclic gilts. At first detection of standing estrus, cyclic gilts were treated with either saline (control), 100, or 200 micrograms/kg BW of the GnRH antagonist (GnRH1). A second group of GnRH antagonist gilts received 200 micrograms/kg BW of the GnRH antagonist approximately 8 h prior to standing estrus (GnRH2). The GnRH1-treatment failed to inhibit or delay ovulation. Ovulation was inhibited and estrous cycles lengthened in GnRH2-treated gilts. These preliminary results suggest that ovulation in the gilt can be inhibited if the GnRH antagonist is administered prior to the LH surge.
Animals↗
Characterization and proteolytic activity of a cathepsin L-like polypeptide in endometrium and uterine flushings of cycling, pregnant and steroid-treated ovariectomized gilts.
Cathepsin L has been proposed to be involved with the endothelial-chorial type of placentation in the cat. Little information concerning the presence and secretion of cathepsin L is available for a species with noninvasive epitheliochorial placentation such as the pig. Cathepsin L activity in uterine flushings and endometrium from gilts during different days of the oestrous cycle and early pregnancy was analysed through specific substrate metabolism and Western blot analyses with antiserum against cat endometrial cathepsin L. This antiserum was utilized to determine the cellular localization of the enzyme within porcine endometrium. Cathepsin L activity within uterine flushings was elevated on Day 15 of the oestrous cycle and early pregnancy, with activity declining on Day 18. Cat cathepsin L antiserum cross-reacted with a group of 46, 40 and 38 kDa uterine proteins and detected a product within the surface and glandular epithelium of the endometrium. The appearance of the 40 kDa protein was first detected on Day 10 of the oestrous cycle with the 38 kDa proteins appearing on Day 15 and 18 of pregnancy. The 40 and 38 kDa uterine proteins appear to be steroid regulated as 12 days of progesterone administration is necessary to detect the proteins and cathepsin L activity.
Animals↗
Detection of transcripts for retinoic acid receptors, retinol-binding protein, and transforming growth factors during rapid trophoblastic elongation in the porcine conceptus.
Early conceptus development (Days 10-12) in the pig is characterized by rapid remodeling and elongation of the trophoblastic membrane. Endometrial synthesis and secretion of retinol-binding protein (RBP; transporter of retinol) into the uterine lumen parallels the rapid morphological transformation of the conceptus. Hence, retinoic acid, a powerful morphogen, may serve a critical function in trophoblastic remodeling and conceptus elongation through the various isoforms of retinoic acid receptor (RAR). We hypothesize that various transcripts for RAR will be expressed, possibly differentially, in the early developing porcine conceptus on Days 10-12 of gestation. Conceptuses were collected from pregnant gilts and individually frozen in liquid nitrogen after classification for morphological size. A total of 36 conceptuses (4 groups of 9) representing spherical (2-9 mm), early tubular (10-12 mm), late tubular (13-60 mm), and filamentous (> 100 mm) morphologies were assembled for qualitative assessment of mRNA for RAR alpha, RARbeta, RARgamma, RBP, and transforming growth factor beta (TGFbeta)-2 and -3 with reverse transcription polymerase chain reaction (RT-PCR). RBP gene expression was affected by stage of conceptus development (p < 0.05) as RBP expression increased in a linear fashion from 2-mm to 8-mm spherical conceptuses, decreased slightly in the tubular stage, and increased dramatically in filamentous conceptuses. RAR alpha, RARbeta, and RARgamma gene expression was evident at all stages of conceptus development. RAR alpha gene expression increased (p < 0.05) in a linear fashion from 2- to 8-mm spherical conceptuses, and expression plateaued in the 9- to 12-mm conceptuses. Expression of RAR alpha declined slightly at the tubular stage but dramatically increased in filamentous conceptuses. RARbeta gene expression was present across all stages of conceptus development although in diminutive amounts. RARgamma gene expression was evident in all stages of conceptus development but was not affected (p > 0.05) by developmental stage. Gene expression for TGFbeta-2 was not detected by RT-PCR at any stage of conceptus development, whereas TGFbeta-3 expression increased (p < 0.05) from the 2- to 8-mm conceptuses and remained high throughout trophoblastic elongation into the filamentous stage. The presence of mRNA for the RAR alpha, RARbeta, and RARgamma in elongating porcine conceptus provides evidence that RAR, particularly RAR alpha, may function as an important acceptor for retinoic acid and possibly regulate gene expression prior to and during rapid trophoblastic elongation in the porcine conceptus.
Actins↗
Ontogeny of elongation and gene expression in the early developing porcine conceptus.
Early porcine conceptus development is characterized by rapid trophoblastic elongation between Days 11 and 12 of pregnancy, a period of embryonic loss in the pig. Growth factors and steroids secreted by the conceptus and uterus, as well as ligand receptors produced by the conceptus, are thought to regulate trophoblastic elongation. Therefore, the objectives of this study were to characterize conceptus gene expression for the steroidogenic enzymes 17alpha-hydroxylase and aromatase and the mesodermal marker brachyury, as well as the expression of receptors for fibronectin (integrin beta-1), progesterone, estrogen, oxytocin, prostaglandin F2alpha, and leukemia inhibitory factor (LIF), prior to and during trophoblastic elongation. Total RNA was extracted from individual conceptuses from Day 10 to Day 12 of pregnancy. Gene expression was determined by reverse transcription polymerase chain reaction on conceptuses having 2- to 4-, 5-, 6-, 7-, 8-, 9-, and 10- to 12-mm spherical, 13- to 25-mm tubular, and > 100-mm filamentous morphologies. There was a stage of development effect on both 17alpha-hydroxylase (p < 0.001) and aromatase (p < 0.003) gene expression. Initial 17alpha-hydroxylase gene expression was detected in early spherical conceptuses (2-4 mm), increasing abruptly through to 7-mm conceptuses. Aromatase gene expression increased dramatically in 6- to 7-mm conceptuses, with increased expression throughout development. Gene expression for LIF receptor (LIFR) (p < 0.02) was similar to that for 17alpha-hydroxylase, while brachyury gene expression began in 6-mm conceptuses and increased (p < 0.001) throughout development. Integrin beta-1 was expressed at all stages of development. Conceptus gene expression was not detected for progesterone, estrogen, oxytocin, and prostaglandin F2alpha receptors. Prior to elongation, dynamic changes in gene expression are occurring that appear to be associated with estrogen production and preparation of the conceptuses for elongation. LIFR expression is highly associated with steroidogenic enzyme production with an initial peak preceding rapid trophoblastic elongation, suggesting that LIF may be involved in early conceptus development in the pig.
Animals↗
Persistence of the dominant follicle during melengestrol acetate administration and its regression by exogenous estrogen treatment in beef cattle.
Experiments were conducted to test the hypothesis that estrogen treatment will regress a persistent dominant follicle developed during melengestrol acetate (MGA) treatment in the absence of a functional corpus luteum (CL) with normal fertility following development and ovulation of a newly recruited follicle. In Exp. 1, nonlactating beef cows (n = 31) were administered .5 mg.cow-1.d-1 of MGA (d 0) for 14 d with 25 mg of prostaglandin F2 alpha (PGF2 alpha) administered on d 6 and 8 to regress the CL. On d 11 of treatment, approximately half the MGA-treated cows received 5 mg of estradiol valerate (EV) i.m. (MGAEV, n = 14) and the remainder were maintained on MGA (n = 17). Ovaries were checked with ultrasound on d 8, 10, 12, and 14 of MGA treatment and every day until ovulation. A persistent dominant follicle developed in 90% of the MGA-treated cows by d 10 of treatment. Most of the MGA-treated cows ovulated the persistent dominant follicle (n = 13/17), whereas EV treatment regressed the persistent dominant follicle (n = 10/14) with the recruitment of a new follicle that ovulated (n = 8/10). Diameter of the ovulatory follicle was larger (P < .05) for the MGA (19.8 +/- .6 mm) than for the control (15.1 +/- .8 mm) and MGAEV (14.8 +/- .7 mm) cows. In Exp. 2, nonlactating, multiparous beef cows (n = 97) and yearling heifers (n = 38) were equally allotted to either a control, MGA alone, or MGA + estradiol-17 beta (MGAE) group with the same dose of MGA as administered in Exp. 1. The 1st d of MGA feeding was the 1st d of treatment. On d 10 of treatment half the MGA-treated animals were injected i.m. with 5 mg of estradiol-17 beta. In controls, behavioral estrus was detected and animals were artificially inseminated (AI) for 5 d (d 10 to 14 of experiment). All controls not exhibiting estrus by d 15 of experiment were injected with 25 mg of PGF2 alpha. The remaining controls and all MGA cows were observed for behavioral estrus and AI commenced for 7 d following withdrawal of MGA (d 15 to 21 of experiment). More (P < .05) controls (90.3%) than MGA (84.8%) or MGAE (63.6%) cows showed estrus within 7 d after MGA withdrawal. The percentage of animals conceiving to the synchronized estrus did not differ (P > .05) among treatments. The data support our hypothesis that a persistent dominant follicle developed and can be regressed with exogenous estrogen treatment followed by the recruitment and ovulation of a new follicle after MGA withdrawal and fertility of that estrus does not seem to be significantly compromised.
Animals↗
Regulation of conceptus development and attachment in pigs.
Implantation/placentation in domestic pigs is preceded by synthesis of oestrogen by the conceptus to maintain functional corpora lutea throughout pregnancy and a rapid morphological transformation of conceptuses from spherical to long filamentous thread-like structures. Initial conceptus expansion, reaching a metre in length, not only delineates the surface area for placental attachment, but also provides the mechanism for delivery of oestrogen to signal events necessary for placentation throughout the uterine horn. Timing for conceptus gene expression to induce trophoblast expansion and attachment in pigs is temporally associated with downregulation of progesterone receptors and increase in oestrogen receptors within the uterine epithelium. Within the confines of the uterine lumen, pig conceptuses normally do not erode or invade through the uterine epithelial surface. However, the pig conceptus possesses extensive proteolytic activity as it is highly invasive outside the uterine lumen of the pig. Initial release of oestrogen by the elongating pig conceptus induces endometrial release of cytokines and a variety of protease inhibitors. Recently, endometrial expression for the inter-trypsin inhibitor (I alpha I) family of protease inhibitors has been detected in the pig endometrium during conceptus elongation and attachment. It is possible that I alpha Is may function to inhibit trophoblast invasion and also serve as targets for adhesion molecules, such as integrins and heparin, to aid in placental attachment to the uterine epithelium.
Animals↗
Linkage mapping of the retinol-binding protein 4 (RBP4) gene to porcine chromosome 14.
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Alleles↗
Rescue of pregnancy and maintenance of corpora lutea in infertile transgenic mice expressing an ovine metallothionein 1a-ovine growth hormone fusion gene.
Transgenic female mice expressing a temporally regulatable ovine metallothionein 1a-ovine growth hormone (oMT1a-oGH) fusion gene can cycle, mate, and support early embryonic development; but they fail to maintain pregnancy or, in most cases, to exhibit signs of pseudopregnancy. The present study was designed to determine whether or not the infertility of oMT1a-oGH transgenic female mice is due to luteal insufficiency. A series of five experiments was conducted in which various hormonal therapies were utilized in an attempt to overcome this infertility. Provision of 1 mg progesterone to oMT1a-oGH females from Days 1 to 17 of gestation rescued pregnancy in 19 of 20 females with an increased mean litter size (p < 0.01) compared to that of wild-type control females. Supplementation with progesterone also appeared on the basis of gross anatomical appearance to maintain CI. Provision of 2 mg progesterone in combination with 25 ng estradiol-17 beta from Days 4 to 17 rescued pregnancy in 14 of 24 oMT1a-oGH females, while provision of 100 or 25 micrograms ovine prolactin from Days 3 to 17 failed to overcome oMT1a-oGH infertility. Transgenic females with rescued pregnancies lactated and raised heavier pups than wild-type females by Day 15 postnatal (p < 0.05). Results indicate that infertility of oMT1a-oGH transgenic mice is caused by luteal insufficiency. Supplementation of transgenic females with progesterone appeared to maintain morphologically normal CL and pregnancy with subsequently increased litter size and normal lactation.
Animals↗
Isolation and characterization of a 30-kDa endometrial glycoprotein synthesized during the estrous cycle and early pregnancy of the pig.
Endometrial polypeptide synthesis, which is regulated through ovarian steroid secretion and steroid production by the developing conceptus, not only provides the necessary secretory components vital to conceptus development but also presents the adhesive changes in the epithelial surface essential for conceptus attachment. In the present study, a 30-kDa, basic endometrial glycoprotein (pGP30) was isolated and characterized during the estrous cycle and early pregnancy of the pig. Uterine flushings and endometrial culture media were obtained from gilts on Days 0, 5, 10, 12, 15, and 18 of the estrous cycle and Days 10, 12, 15, and 18 of pregnancy. A polyclonal antibody was generated to pGP30 after isolation of medium from Day 15 pregnant endometrial cultures separated by gel filtration and PAGE. Western blot analysis indicated that the antiserum reacted with isoforms of pGP30 and cross-reacted with a 90-kDa component in serum that was not removed after cleavage of the oligosaccharide chains from the 90-kDa glycoprotein. Antiserum did not detect a 30-kDa band in media from cultures of kidney, fat, heart, muscle, liver, or serum; however, heart and muscle did contain bands of different molecular masses that cross-reacted with the antiserum. Multiple bands of higher molecular mass (35-40 kDa) were detected in the endometrial cultures from gilts on Days 0 through 10 of the estrous cycle. Treatment of ovariectomized gilts with estradiol-17 beta stimulated a similar response. During the mid- to late luteal phase of the estrous cycle (Days 12-18), the 30-kDa band as well as an additional 32-kDa band was present on Western blots. Administration of progesterone for 14 days stimulated the synthesis of both the 30- and 32-kDa products in ovariectomized gilts. However, only the pGP30 was detected on Days 12-18 of pregnancy. Immunocytochemical localization with antiserum to pGP30 indicated that the glycoprotein is present in the endometrial epithelium, with the surface epithelium demonstrating the strongest reaction product. Discrete changes in staining and cellular localization were observed during the early stages of the estrous cycle (Days 0-5) and the midluteal (Day 10) phase. A similar response was achieved with administration of steroids to ovariectomized gilts. Data indicate that discrete changes in epithelial synthesis of the endometrial glycoprotein occur at the time of conceptus trophoblastic elongation and placental attachment in the pig.
Animals↗
Sex identification in mammals with polymerase chain reaction and its use to examine sex effects on diameter of day-10 or -11 pig embryos.
The objectives of this study were to develop a rapid method for sex determination for several mammalian species using polymerase chain reaction (PCR) and to use this method to determine whether there is a significant developmental difference in spherical diameter between male and female d-10 or -11 porcine embryos. The PCR system was developed and verified using genomic DNA from pigs of known sex, then it was tested with genomic DNA from several other mammalian species. Sex is determined by amplification of two genes in a single reaction. The presence or absence of a region of the Sry (sex-determining region Y) gene determines sex, and amplification of the Zfy (male) or Zfx (female) genes acts as a positive control for PCR. Sex determination was successful for all animals tested, including pigs, cattle, sheep, goats, llamas, horses, humans, baboons, dogs, cats, rats, and mice. A total of 209 embryos were collected from 21 crossbred gilts on d 10 or 11 of gestation, and their diameters were measured. No significant difference in embryo diameter was detected between male and female embryos, indicating that sexual dimorphism in embryonic growth in pigs does not occur before the period of rapid embryo elongation. The present sexing technique using PCR is rapid (approximately 6 h from receipt of embryos to results), and it may be useful for examining the effects of sex on any trait of interest in early porcine embryos and embryos from several other mammals.
Animals↗
Immunocytochemical localization and changes in endometrial progestin receptor protein during the porcine oestrous cycle and early pregnancy.
Changes of progesterone receptor (PR) protein and cellular localization in the endometrium were evaluated during the oestrous cycle and early pregnancy of the gilt. During the oestrous cycle, the concentration of total PR protein within the endometrium was highest on Days 0-5 and decreased on Day 10. The endometrial concentration of PR reached a nadir on Day 12 and this level was maintained throughout the remainder of the oestrous cycle (Day 18). In pregnant gilts, the concentration of endometrial PR protein from Day 10 to Day 18 was similar to that in cyclic gilts. Western blot analysis with antiserum specific for the A and B isoforms of PR indicated that porcine endometrium expresses both isoforms of PR. Immunostaining was detectable for both the A and B isoforms of PR from Day 0 to Day 12 of the oestrous cycle. However, no staining was observed on Day 15 and Day 18 of the oestrous cycle or pregnancy Immunocytochemical localization of PR in the endometrium of cyclic gilts and pregnant gilts indicated that there was intense staining for PR in surface epithelium and glandular epithelium during oestrus (Day 0) and on Day 5. However, the staining was less intense on Day 7 and Day 10 of the oestrous cycle and no epithelial staining was observed after Day 12. PRs were present in the stroma and myometrium throughout the oestrous cycle and early pregnancy. The presence of conceptuses during pregnancy did not affect the loss of PR from the uterine epithelium after Day 10 of gestation. Down-regulation of epithelial PR might be one factor involved in the timing of luteolysis during the oestrous cycle as well as conceptus growth and placentation during early pregnancy.
Animals↗
Effect of ovarian antral follicle cauterization on the interestrus interval of the gilt.
The objective of the present study was to determine if destruction of ovarian antral follicles by laser-cauterization affects CL lifespan during the estrous cycle of the gilt. Cyclic gilts were randomly assigned to either SHAM, laser (L) or laser-estradiol (L-E2) treatment groups, with the L-E2 group receiving a 5-mg intramuscular (i.m.) injection of estradiol-17beta cypionate at the time of the first surgery. Ovarian antral follicles were laser-cauterized on either Days 12 and 14 (L12) or Days 14 and 17 (L14) of the estrous cycle. In the L12-E2 group, 3 of 4 gilts had extended mean interestrus intervals of more than 22 days compared with 0 of 4, 0 of 6, 0 of 7 and 1 of 5 gilts in the SHAM, L12, L14 and L14-E2 groups, respectively. The L12-E2 gilts had a longer (P<0.05) mean interestrus interval (23.5+/-1.3 days) than the L12 (20.0+/-1.1 days), L14 (20.7+/-1.0 days) and SHAM (20.5+/-1.3 days). The mean interestrus interval of L14-E2 gilts (21.8+/-1.2 days) did not differ from those of the L12-E2 group or the L12, L14 and SHAM group gilts. Six additional gilts were injected with 5 mg estradiol cypionate-17beta to serve as nonsurgical controls for E2 treatment. Gilts (3 of 3) given an E2 injection on Day 12 had extended mean interestrus interval (26.0+/-2.6 days), while 2 of 3 gilts injected with E2 on day 14 had extended mean interestrus intervals (27.7+/-2.1 days). These results indicate that in cyclic gilts destruction of ovarian follicles by laser-cauterization did not affect CL lifespan, and that luteolysis is not dependent on the presence of antral follicles.
Journal Article↗
Changes in oestrogen receptor protein, mRNA expression and localization in the endometrium of cyclic and pregnant gilts.
Conceptus secretion of oestrogen on Day 11 of gestation is involved with establishment of pregnancy in the pig. Changes in oestrogen receptor (ER) protein, mRNA and cellular localization in the endometrium were evaluated during the oestrous cycle and early pregnancy of the gilt. In nonpregnant gilts, concentration of nuclear ER in the endometrium increased from Days 0 to 12 followed by a decline on Day 15 of the oestrous cycle. In pregnant gilts, changes in endometrial nuclear ER during Days 10, 12, 15 and 18 were similar to that in cyclic pigs. Analysis of endometrial ER mRNA expression did not detect any difference between cyclic and pregnant pigs between Days 10 and 15 postoestrus. Expression of ER mRNA in endometrium of cyclic and pregnant gilts was greatest on Day 10 followed by a decline on Day 15. Endometrial ER mRNA increased on Day 18 of the oestrous cycle, but remained low during pregnancy. Immunocytochemical localization of ER in the endometria of cyclic and pregnant gilts indicated that there was intense staining for ER in stromal cells and moderate to strong staining in surface and glandular epithelial cells during oestrus (Day 0) and Day 18 of the oestrous cycle. However, stromal ER staining was absent from Days 5 to 15 of the oestrous cycle and continued to be suppressed on Day 18 of pregnancy. Immunocytochemical staining of ER in the surface and glandular epithelium was readily detectable from Days 0 to 12 of the oestrous cycle and during pregnancy. Intensity of staining for ER declined in surface epithelial cells on Day 15 in both cyclic and pregnant pigs whereas positive staining for ER in glandular epithelium was absent. Staining for ER on uterine surface epithelial cells increased during pro-estrus (Day 18) of cyclic gilts but remained similar to Day 15 in pregnant gilts. Changes in endometrial ER protein, mRNA and localization in surface epithelium are consistent with a physiological role for conceptus oestrogen secretion in uterine function and maternal recognition of pregnancy in the pig.
Animals↗