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T E Spencer

Publications and source records attributed to T E Spencer.

At least 19 recordsLinked to original sources
PubMed1998

The steroid receptor coactivator-1 contains multiple receptor interacting and activation domains that cooperatively enhance the activation function 1 (AF1) and AF2 domains of steroid receptors.

Steroid receptors are ligand-inducible transcription factors, and their association with steroid receptor coactivators (SRCs) upon binding to DNA is necessary for them to achieve full transcriptional potential. To understand the mechanism of SRC-1 action, its ability to interact and enhance the transcriptional activity of steroid receptors was analyzed. First, we show that SRC-1 is a modular coactivator that possesses intrinsic transcriptional activity when tethered to DNA and that it harbors two distinct activation domains, AD1 and AD2, needed for the maximum coactivation function of steroid receptors. We also demonstrate that SRC-1 interacts with both the amino-terminal A/B or AF1-containing domain and the carboxyl-terminal D/E or AF2-containing domain of the steroid receptors. These interactions are carried out by multiple regions of SRC-1, and they are relevant for transactivation. In addition to the inherent histone acetyltransferase activity of SRC-1, the presence of multiple receptor-coactivator interaction sites in SRC-1 and its ability to interact with components of the basic transcriptional machinery appears to be, at least in part, the mechanism by which the individual activation functions of the steroid receptors act cooperatively to achieve full transcriptional activity.

Binding Sites↗
PubMed1998

Ovary-independent estrogen receptor expression in neonatal porcine endometrium.

Effects of age and ovariectomy (OVX) at birth on uterine growth, endometrial development, and estrogen receptor (ER) expression were determined for intact and OVX gilts (n = 5 per day) hysterectomized on postnatal days (PND) 0, 15, 30, 60, 90, or 120. Uteri were evaluated histologically, and ER protein and mRNA expression were characterized immunohistochemically and by in situ hybridization. OVX did not affect uterine weight or endometrial thickness until after PND 60, when both increased more rapidly in intact gilts. Neither did it affect genesis of uterine glands, which were present and which proliferated after PND 0, or endometrial ER expression patterns in glandular epithelium (GE), luminal epithelium (LE), or stroma (S) between PND 0 and 120. Endometrium was ER negative at birth. On PND 15, the ER signal was strong in GE, weak in S, and effectively absent in LE. Thereafter, although the ER signal remained strong in GE and increased through PND 60 in S, it was not evident consistently until after PND 30 in LE. The data indicate that 1) porcine uterine growth and endometrial morphogenesis are ovary-independent processes before PND 60; 2) uterine gland genesis is associated temporally with development of ER-positive endometrial GE and S; and 3) regulation of endometrial ER expression is ovary independent between PND 0 and 120. The results establish the ER as a marker of GE differentiation and implicate this receptor in mechanisms regulating endometrial morphogenesis in the neonatal pig.

Aging↗
PubMed1998

Expression of interferon regulatory factors one and two in the ovine endometrium: effects of pregnancy and ovine interferon tau.

Availàble evidence suggests that interferon tau (IFNtau), the signal for pregnancy recognition in ruminants, suppresses transcription of the estrogen receptor (ER) gene in the endometrial lumenal epithelium (LE) and superficial glandular epithelium (sGE) to prevent oxytocin receptor (OTR) expression and pulsatile release of luteolytic prostaglandin F2alpha. The IFN regulatory factors one (IRF-1) and two (IRF-2) are transcription factors induced by type I IFNs that activate and silence gene expression, respectively. The objectives of these studies were to determine effects of pregnancy and IFNtau on expression of immunoreactive IRF-1 and IRF-2 proteins in the ovine endometrium. In study one, IRF-1 and IRF-2 were not detected in the LE or sGE of cyclic ewes. In pregnant ewes, IRF-1 expression was detected transiently in the LE and sGE only on Days 11 and 13, and IRF-2 was detected in these same epithelia on Days 13, 15, 17, and 20. In study two, 36 ewes were fitted with uterine catheters on Day 5 of the estrous cycle, and one uterine horn was double-ligated at the base. Uterine horns of each ewe received twice-daily injections of either recombinant ovine IFNtau or control proteins beginning on Day 11 until hysterectomy at 1, 3, 6, 12, 24, 48, 72, 96, or 120 h after initial injection. The IRF-1 was detected transiently in the endometrial LE and sGE only at 12 and 24 h in the uterine horn receiving IFNtau but not in those tissues receiving control proteins. The IRF-2 was expressed in the LE and sGE at 24 h and thereafter in the IFNtau-treated, but not control uterine horns. In control uterine horns, ER and OTR were first detected in the LE at 48 h and 72 h, respectively, and remained abundant thereafter. In horns receiving IFNtau, ER and OTR expression was not detected in the endometrial LE and sGE. Results suggest that IFNtau acts directly on the LE and sGE during pregnancy to sequentially induce IRF-1 and then IRF-2 gene expression, which is correlated temporally with an absence of ER and OTR. The ovine ER gene may contain an IFNtau-responsive element(s) that binds negative-acting, IFNtau-inducible transcription factors, such as IRF-2, which silences transcription of the ER gene in the endometrial epithelium during maternal recognition of pregnancy.

Animals↗
PubMed1998

Effects of the estrous cycle and early pregnancy on uterine expression of Mx protein in sheep (Ovis aries).

Conceptuses of ruminant ungulates produce large amounts of a type I interferon, interferon-tau (IFNtau), which is the signal for maternal recognition of pregnancy. Induction of cellular Mx proteins is an important component of the response to type I interferon in the immune system, but Mx regulation and function have not been studied in the uterus. This study examined temporal and spatial alterations in ovine uterine Mx expression during the cycle and early pregnancy using immunohistochemistry, in situ hybridization, and Northern and slot-blot analysis. Sheep uterine endometrium expressed a single approximately 2.5-kilobase Mx mRNA transcript that was detectable at all stages of the estrous cycle and early pregnancy examined. In cyclic ewes, mRNA abundance in endometrium increased from Day 1 to peak levels at Day 13 and then declined to Day 15. In pregnant ewes, steady-state levels of Mx mRNA were first detected above the level in cyclic ewes at Day 13 postmating, were greater than 10-fold higher at Day 15, and remained elevated at Day 19. Expression of Mx mRNA in the myometrium did not change during the estrous cycle but increased approximately 23-fold between Days 11 and 15 of pregnancy. Immunohistochemical and in situ hybridization analysis revealed a similar temporal pattern of Mx expression. In cyclic ewes, Mx protein and mRNA were initially localized to the luminal epithelium at Days 1 and 3, increased from Days 5 to 13, especially in the shallow uterine glands, and then declined at Day 15. Pregnancy resulted in up-regulation of Mx expression in the luminal and glandular epithelium, stroma, and myometrium. Punctate Mx immunostaining and Mx mRNA concentrations were greatest when progesterone production was maximal during the estrous cycle and were strongly up-regulated by the conceptus across the entire uterine wall. It is suggested that a cascade of induction of Mx gene expression proceeds from the luminal epithelium to the outer longitudinal myometrium and that transcriptional activation of the promoter may involve both soluble cytokines (i.e., IFNtau) and steroid hormones (i.e., progesterone).

Animals↗
PubMed1997

Steroid receptor coactivator-1 is a histone acetyltransferase.

Steroid receptors and coactivator proteins are thought to stimulate gene expression by facilitating the assembly of basal transcription factors into a stable preinitiation complex. What is not clear, however, is how these transcription factors gain access to transcriptionally repressed chromatin to modulate the transactivation of specific gene networks in vivo. The available evidence indicates that acetylation of chromatin in vivo is coupled to transcription and that specific histone acetyltransferases (HATs) target histones bound to DNA and overcome the inhibitory effect of chromatin on gene expression. The steroid-receptor coactivator SRC-1 is a coactivator for many members of the steroid-hormone receptor superfamily of ligand-inducible transcription factors. Here we show that SRC-1 possesses intrinsic histone acetyltransferase activity and that it also interacts with another HAT, p300/CBP-associated factor (PCAF). The HAT activity of SRC-1 maps to its carboxy-terminal region and is primarily specific for histones H3 and H4. Acetylation by SRC-1 and PCAF of histones bound at specific promoters may result from ligand binding to steroid receptors and could be a mechanism by which the activation functions of steroid receptors and associated coactivators enhance formation of a stable preinitiation complex, thereby increasing transcription of specific genes from transcriptionally repressed chromatin templates.

Acetylation↗
PubMed1997

Steroid receptor induction of gene transcription: a two-step model.

Coactivators, such as steroid receptor coactivator 1 (SRC-1A) and CREB (cAMP response element binding protein)-binding protein (CBP), are required for efficient steroid receptor transactivation. Using an in vitro transcription assay, we found that progesterone receptor (PR)-driven transcription is inhibited by a dominant negative PR ligand-binding domain-interacting region of SRC-1A, indicating that SRC-1A is required for actual transcriptional processes. In addition, these coactivators also possess intrinsic histone acetyltransferase (HAT) activity and bind to each other and another HAT, p300/CBP-associated factor. Here we show that the human PR also interacts with p300/CBP-associated factor in vitro. Recruitment of multiple HATs to target promoters suggests an important role for chromatin remodeling in transcriptional activation of genes by steroid receptors. In transient transfection assays, we found that addition of a histone deacetylase inhibitor, trichostatin A, strongly potentiated PR-driven transcription. In contrast, directing histone deacetylase-1 (HD1) to a promoter using the GAL4 DNA binding domain inhibited transcription. Furthermore, PR transactivation was repressed by recruiting HD1 into the PR-DNA complex by fusing HD1 to a PR ligand-binding domain-interacting portion of SRC-1. Collectively, these results suggest that targeted histone acetylation by recruited HAT cofactors and histone deacetylation are important factors affecting PR transactivation. Recruitment of coactivators and HATs by the liganded PR in vivo may result in (i) remodeling of transcriptionally repressed chromatin to facilitate assembly and (ii) enhanced stabilization of the preinitiation complex by the activation functions of coactivators and the liganded PR itself.

Chromatin↗
PubMed1997

Comparative study of uterine morphogenesis and protein secretion in neonatal White crossbred and Meishan gilts.

Thirty-five crossbred and 22 Meishan contemporary gilts were necropsied on Day 1, 14, 28, 42, or 56 of age (birth = Day 0). At necropsy, a cross section of one uterine horn was fixed for histomorphometric study, and minced uterine tissue was cultured with 50 muCi [3H]leucine. Secreted proteins were identified by two-dimensional PAGE, fluorography, and incorporation of radioactivity. Body weights at necropsy were similar for the two breeds and increased (p < 0.01) between Days 1 and 56 of age. Ovarian and uterine weights, as well as histomorphometric areas, were similar for the two breeds on Day 1 but increased markedly (p < 0.01) in Meishan gilts on Day 56. In gilts of both breeds, secretion of uterine proteins 1 (M(r) x 10(-3)/pI; 45/6.0), 2a and 2b (doublet, 25/6.2), and 3 (20/5.5) increased in association with endometrial gland development. A fourth protein (97/4.0) was observed in gilts of both breeds but was more abundant in Meishan; a fifth protein (13/6.0) was detected only in crossbred gilts on Day 56. Although specific regulatory roles for locally produced uterine proteins remain to be defined, the increase in specific uterine proteins and breed differences in uterine protein secretion suggest that uterine proteins may influence early uterine development.

Age Factors↗
PubMed1997

Effects of exogenous recombinant ovine interferon tau on circulating concentrations of progesterone, cortisol, luteinizing hormone, and antiviral activity; interestrous interval; rectal temperature; and uterine response to oxytocin in cyclic ewes.

Interferon tau (IFN tau) is the conceptus-produced antiluteolytic signal in ruminants. Three experiments examined the effects of s.c. administration of recombinant ovine (ro)IFN tau on interestrous interval (IEI), oxytocin (OT)-induced uterine prostaglandin F2alpha metabolite (PGFM) production, rectal temperature (RT), respiration rate (RR), and plasma concentrations of progesterone, cortisol, LH, and antiviral activity (AVA) in plasma and uterine flushings. In experiment I, 20 ewes were treated s.c. with either 0, 1, 2, or 4 mg/day roIFN tau (0.7 x 10(8) U/mg; 5 ewes/dosage) from Days 11 to 15 of the estrous cycle (estrus = Day 0) and were challenged with OT (30 IU) on Day 15. Jugular blood samples were collected at -10, 0, 10, 20, 30, 40, 50, and 60 min relative to the OT challenge and assayed for PGFM. Recombinant oIFN tau increased IEI (16.7, 18.7, and 22.6 +/- 0.6 days for 0, 2, and 4 mg roIFN tau, respectively, p < 0.01). Recombinant oIFN tau did not affect peak PGFM response to OT (2309 +/- 172 pg/ml; p > 0.1). However, the 4 mg/day dosage delayed the time to peak PGFM (32.4 vs. 47.5 +/- 3.4 min; p < 0.01, 0 vs. 4 mg) and resulted in approximately 200% higher concentrations of PGFM at 60 min post-OT (0 vs. 4 mg/day, p < 0.07). Experiment II was similar to experiment I, except that only the 0- and 4-mg/day dosages of roIFN tau were administered. Ewes were hysterectomized on Day 16, and assay of uterine flushes detected no AVA from ewes treated with either 0 or 4 mg/day roIFN tau. In experiment III, 20 ewes were treated s.c. with either 0, 2, 4, or 6 mg roIFN tau on Day 12. Blood samples, RT, and RR were obtained at frequent intervals for 24 h, and plasma was assayed for progesterone, cortisol, LH, and AVA. Plasma AVA, which increased in a dose-dependent manner, was detectable within 60 min and remained elevated at 24 h compared to control values. RT (elevated 0.5-1.0 degrees C), RR, and cortisol increased in response to all dosages of roIFN tau, with peak values occurring 150-180 min postinjection. For all dosages of roIFN tau, plasma progesterone declined from 120 to 360 min posttreatment and then returned to pretreatment values by 24 h (p < 0.01) as compared to controls. Overall, exogenous roIFN tau altered uterine PGFM response to OT from a pulse to a gradual and sustained elevation and extended IEI with only a transient decline in progesterone and mild hyperthermia, effects that are not expected to compromise pregnancy.

Animals↗
PubMed1997

Interferon tau: a novel pregnancy recognition signal.

PROBLEM: Trophectoderm of ruminant conceptuses (embryo and associated membranes) secretes tau interferons (IFNtau) as the pregnancy recognition signal. How does it act? METHOD: Review of current data. RESULTS: IFNtau acts on uterine epithelium to suppress transcription of the genes for estrogen receptor and oxytocin receptor. This blocks development of the uterine luteolytic mechanism and, therefore, release of luteolytic pulses of prostaglandin F2alpha, but it has no effect on expression of the progesterone receptor. Maintenance of progesterone secretion by the corpus luteum ensures establishment and maintenance of pregnancy. Secretion of IFNtau on days 12-15 for sheep and days 14-17 for cows and goats is essential for pregnancy recognition. CONCLUSION: We propose that IFNtau affects endometrial gene expression by activating the Jak/Stat pathway, which results in formation of the ISGF3alpha transcription factor complex. ISGF3alpha binds to interferon-stimulated response elements and activates transcription of interferon-responsive genes such as interferon regulatory factor-1 (IRF-1) which, in turn, activates expression of the negative-acting transcription factor IRF-2. Pregnancy (or intrauterine injection of roIFNtau) results in a transient increase in endometrial IRF-1 expression followed 36-48 hr later by a sustained increase in IRF-2. We propose that IRF-2, or an IFNtau-induced negative regulatory factor like IRF-2, suppresses expression of the estrogen receptor gene and directly or indirectly blocks expression of the gene for oxytocin receptor to abrogate the uterine luteolytic mechanism and ensure the establishment of pregnancy.

Animals↗
PubMed1997

Role of co-activators and co-repressors in the mechanism of steroid/thyroid receptor action.

Steroid/thyroid hormone receptors are ligand-dependent transcription factors that regulate diverse aspects of growth, development, and homeostasis by binding as homodimers or heterodimers to their cognate DNA response elements to modulate transcription of target genes. Transactivation by steroid/ thyroid hormone receptors involves a conserved AF-2 domain located in the distal carboxy-terminus of the receptors. The existence of co-factors, termed co-activators or adapters, was first suggested by transcriptional squelching between progesterone receptors and estrogen receptors. Co-repressors were also postulated to contribute to the silencing function of unliganded thyroid hormone receptor (TR). The yeast two-hybrid system and Far-Western blotting have been used to identify several proteins that interact with members of the steroid/thyroid hormone receptor superfamily in a ligand-sensitive manner. Our laboratory cloned the first functional co-activator, termed steroid receptor co-activator-one (SRC-1), that appears to be a general co-activator for all steroid receptors tested and enhances transactivation of steroid hormone-dependent target genes. Subsequently, many more putative co-activators have been reported, including the SRC-1 related proteins, TIF2 and GRIP1, and other putative and unrelated co-activators such as ARA70, Trip1, RIP140, and TIF1. In addition, another co-activator, CREB-binding protein (CBP), has been shown to enhance steroid receptor-dependent target gene transcription. CBP and SRC-1 interact and synergistically enhance transcriptional activation by the ER and PR. Therefore, a ternary complex-consisting of CBP, SRC-1, and liganded steroid receptors-may form to increase the rate of hormone-responsive gene transcription. Similarly, co-repressors, such as SMRT and N-CoR, for TR and retinoic acid receptors (RAR) have been identified. The unliganded TR and RAR have been shown to inhibit basal promoter activity; this silencing of target gene transcription by unliganded receptors is mediated by these co-repressors. Collectively, available evidence supports the following model of steroid-responsive gene transcription. Upon binding of agonist the receptor changes its conformation in the ligand-binding domain that enables recruitment of co-activators, which allows the receptor to interact with the basal transcriptional machinery more efficiently and to activate transcription. In contrast, binding of antagonists induces a different conformational change in the receptor. Although some antagonist-bound receptor can dimerize and bind to its cognate DNA element, it fails to dislodge the associated co-repressors, which results in a nonproductive interaction with the basal transcriptional machinery. Similarly, the TR and RAR associate with co-repressors in the absence of ligand, thereby resulting in a negative interaction with the transcriptional machinery that silences target gene expression. In the case of mixed agonist/antagonists, such as 4-hydroxytamoxifen, activation of gene transcription may depend on the relative ratio of co-activators and co-repressors in the cell or cell-specific factors that determine the relative agonistic or antagonistic potential of different compounds. These co-activators and co-repressors appear to act as an accelerator and/or a brake that modulates transcriptional regulation of hormone-responsive target gene expression. Thus, the recent discovery of co-activators and co-repressors expands our knowledge of the mechanisms of steroid receptor action.

Animals↗
PubMed1996

Effects of interferon-tau and progesterone on oestrogen-stimulated expression of receptors for oestrogen, progesterone and oxytocin in the endometrium of ovariectomized ewes.

The effects of recombinant ovine interferon-tau (IFN-tau) and progesterone on oestrogen-stimulated expression of endometrial receptors for oestrogen (ER), progesterone (PR) and oxytocin (OTR) were determined in ovariectomized ewes. Cyclic ewes (n = 16) were ovariectomized and fitted with uterine catheters on Day 4 of the oestrous cycle (Day O, oestrous) and assigned randomly in 2 x 2-factorial arrangement to receive daily intrauterine injections of either recombinant ovine IFN-tau (roIFN-tau; 2 x 10(7) anti-viral units) or control proteins from Day 11 to Day 15 and 50 mg progesterone from either Day 4 to Day 10 (E-P) or Day 4 to Day 15 (E+P). All ewes received 50 micrograms oestradiol-17 beta on Days 13, 14 and 15 and were hysterectomized on Day 16. In control ewes, endometrial ER mRNA, PR protein and OTR density were greater in E-P- than E+P- treated ewes. In E-P ewes, roIFN-tau decreased oestrogen-stimulated increases in ER and OTR, but not PR expression compared with control ewes. In E+P ewes, endometrial ER mRNA and protein, PR mRNA and protein, and OTR levels were lower in roIFN-tau-treated ewes than control ewes. Immunoreactive ER and PR were absent in the endometrial luminal and superficial glandular epithelium of roIFN-tau compared with control ewes, but were present in the deep glandular epithelium and stroma regardless of steroid or protein treatment. These results indicate that progesterone affects oestrogen-induced increases in endometrial ER, PR and OTR expression in the PR+ deep glandular epithelium and stroma, whereas IFN-tau suppresses oestrogen-induced increases ER, PR and OTR expression in the PR- luminal and superficial glandular epithelium. These combined actions of IFN-tau and progesterone to suppress oestrogen-induced increases in endometrial OTR formation would prevent pulsatile production of luteolytic prostaglandin F2 alpha by the endometrium during early pregnancy.

Animals↗
PubMed1996

Estrogen enhances endometrial estrogen receptor gene expression by a posttranscriptional mechanism in the ovariectomized ewe.

Prior influence of estrogen is required for many physiological effects of steroid hormones. This study addresses positive autoregulation of estrogen receptor gene expression in endometrium. Groups of ovariectomized ewes (n = 6) were treated with a single i.m. injection of 50 micrograms 17 beta-estradiol for 6, 12, 24, or 48 h or of vehicle for 24 h (control) prior to collection of endometrium. Three ewes received a regimen of estradiol-progesterone-estradiol (EPE) designed to mimic the estrous cycle. Northern analysis of endometrial RNA using an ovine estrogen receptor complementary RNA probe indicated that estradiol increased (p < 0.0001) estrogen receptor messenger RNA abundance over time to be 5-fold greater at 24 h postinjection. This effect also occurred after a period of progesterone dominance in EPE ewes (p < 0.05). Ribonuclease protection assays with a complementary RNA probe for the ovine progesterone receptor demonstrated that estradiol treatment increased progesterone receptor messenger RNA abundance at 48 h (p < 0.005). Nuclear runoff analyses indicated that whereas estradiol enhanced the transcription rates of progesterone receptor (p < 0.1) and 28S ribosomal RNA genes (p < 0.002), activity of the estrogen receptor gene was unchanged (p > 0.25). These results suggest that a physiological dose of estradiol, similar to the preovulatory surge of estrogen, up-regulates endometrial estrogen receptor gene expression by a posttranscriptional mechanism.

Amino Acid Sequence↗
PubMed1996

Placental interferons.

Trophectoderm of ruminant conceptuses (embryo and associated membranes) secrete tau interferons (IFN tau) as the pregnancy recognition signal. Secretion of IFN tau on gestational days 12-13 for sheep and gestation days 14-17 for cows and goats is critical for pregnancy recognition. IFN tau acts on uterine epithelium to suppress estrogen receptor and oxytocin receptor gene expression, which prevents uterine release of luteolytic pulses of prostaglandin F2 alpha (PGF). Expression of the progesterone receptor (PR) gene in uterine endometrium is not affected by oIFN tau. Maintenance of progesterone secretion by the corpus luteum (CL) ensures establishment of pregnancy. Pig conceptuses secrete both IFN alpha and IFN gamma between days 15-21 of gestation, but their role(s) in early pregnancy is unknown. Estrogen secreted by pig trophoblast between gestational days 11-13 and 15-25 increases endometrial receptors for prolactin and causes exocrine secretion of PGF into the uterine lumen to prevent luteolysis. Shared cell-signaling mechanisms by IFNs and lactogenic hormones through Janus kinases (JAK) 1 and 2 may provide a common pathway to abrogate luteolytic mechanisms to ensure establishment of pregnancy. The role(s) of IFNs produced by human and rodent placentae is not known.

Animals↗
PubMed1996

Ovine interferon tau suppresses transcription of the estrogen receptor and oxytocin receptor genes in the ovine endometrium.

Tau interferons (IFNtau) are a unique subclass of the omega interferons that are transiently produced by the trophectoderm of the conceptus (embryo and associated membranes) during early pregnancy in ruminants. IFNtau acts as an antiestrogen on the endometrium to suppress increases in estrogen receptor (ER) and oxytocin receptor (OTR) gene expression which prevents pulsatile production of prostaglandin F2alpha and regression of the corpus luteum or luteolysis. In this study, steady-state levels of ER mRNA and transcription rates of the ER and OTR genes were two-fold lower in the endometrium of pregnant as compared to cyclic ewes on day 15. Levels of ER mRNA and ER and OTR gene transcription were also two-fold lower in the endometrium of day 15 cyclic ewes receiving intrauterine injections of recombinant ovine IFNtau from day 11 to day 14 compared to control ewes. Results suggest that the antiluteolytic action of IFNtau is to suppress transcription of the ER gene by a negative-acting transcriptional mechanism which prevents estrogen-induced increases in OTR gene expression in the endometrium. The novel antiestrogenic effects of IFNtau, combined with its Type I IFN characteristics and low cytotoxicity, suggest that this trophoblast IFN may have potential value as a chemotherapeutic agent for the treatment of infertility, viral disease and estrogen-dependent malignant disorders.

Animals↗
PubMed1995

Maternal treatment with somatotropin alters embryonic development and early postnatal growth of pigs.

A possible management strategy to alter fetal development and enhance sow productivity and progeny performance was examined by maternal administration of porcine somatotropin during early gestation. Eighteen crossbred gilts were bred naturally to boars of similar genetics, and pregnancy was confirmed between Days 21 and 24 of gestation by ultrasound. All animals were allowed ad libitum consumption of a 16% CP gestation diet through Day 21 of gestation and 3.0 kg/d for the remainder of gestation. Gilts were injected twice daily with 0 (n = 10) or 15 micrograms/kg body weight (BW) (n = 10; total, 30 micrograms/kg BW per d) pituitary-derived porcine somatotropin (pST) during Days 28 to 40 of gestation. Data were collected postmortem during embryonic, neonatal, and market-weight phases. At 41 d of gestation, pST treatment increased embryonic survival (87.9 versus 77.0%; P < 0.05) and embryo crown rump lengths (77.96 versus 65.14 mm; P < 0.01), but embryo weight was not altered (10.15 and 9.03 g; P > 0.10). Pigs from pST-treated gilts had increased (P < 0.01) crown rump lengths at birth (31.5 versus 30.4 cm) and 21 d (50.9 versus 48.4 cm). However, no differences were observed in birth or 21-d weights as a result of pST treatment (P > 0.10). Neonatal carcasses of progeny (20 kg BW) from the pST-treated gilts had heavier semitendinosus muscles (76.1 versus 66.0 g; P < 0.10), larger longissimus muscle cross-sectional area (10.1 versus 8.2 cm2; P < 0.05), longer sides (51.2 versus 47.9 cm; P < 0.001), and decreased 10th rib backfat (6.67 versus 8.64 mm; P < 0.001) compared with those of controls. Carcasses of market-weight progeny (100 kg BW) from pST-treated gilts had larger longissimus muscle cross-sectional area (P < 0.10), heavier trimmed loins (P < 0.10), and longer carcass sides (P < 0.05). Data are supportive of a hypothesis that mechanisms during early embryonic development are sensitive to manipulation through selected management strategies of the sow and that modifications of this strategy may serve as a model for the examination of molecular and cellular events controlling early embryonic growth.

Animals↗
PubMed1995

Sulfated glycoprotein-1 (SGP-1) expression in ovine endometrium during the oestrous cycle and early pregnancy.

This study determined effects of day of oestrous cycle and early pregnancy on sulfated glycoprotein-1 (SGP-1) expression in ovine endometrium. A 364-bp clone of the ovine SGP-1 mRNA was amplified from reverse transcribed Day-15 cyclic endometrial mRNA using the polymerase chain reaction (PCR) and primers specific for the rat SGP-1 mRNA sequence. Nucleotide sequence of the ovine SGP-1 cDNA shared significant identity with rat SGP-1 and human prosaposin. Ewes (n = 40) were hysterectomized on either Day 1, 6, 11, 13 or 15 of the oestrous cycle or on Day 11, 13, 15, 17 or 25 of early pregnancy. Total cellular RNA was isolated from endometrium and subjected to Northern and slot blot hybridization analyses using an antisense cRNA probe transcribed from the ovine SGP-1 cDNA clone. A single 2.6-kb mRNA transcript was detected by Northern hybridization analyses. Slot blot hybridization analyses indicated that steady-state levels of endometrial SGP-1 mRNA varied during the oestrous cycle (cubic, P < 0.02) and increased between Day 11 and Day 25 of early pregnancy (linear, P < 0.01). On Days 11, 13 and 15, endometrial SGP-1 mRNA levels were greater in pregnant ewes than in cyclic ewes (day x pregnancy status, P < 0.01). Immunohistochemical localization of SGP-1 in uterine tissues with rabbit anti-rat SGP-1 antibody revealed intense immunoreactivity associated primarily with the endometrial epithelium. These results indicate that the ovine endometrium expresses SGP-1, a prosaposin, and that SGP-1 expression varies during the oestrous cycle and is enhanced by the conceptus. The presence of SGP-1 in the endometrium suggests intracellular and extracellular roles for this protein in glycosphingolipid metabolism or transport in the uterine environment.

Animals↗
PubMed1995

Ovine interferon-tau regulates expression of endometrial receptors for estrogen and oxytocin but not progesterone.

Ovine interferon-tau (oIFN-tau) may stabilize endometrial progesterone receptor (PR) and/or inhibit estrogen receptor (ER) gene expression during pregnancy recognition to suppress endometrial oxytocin receptor (OTR) formation and production of luteolytic prostaglandin (PG) F2 alpha pulses. This study determined whether or not oIFN-tau stabilized PR expression in the endometrium during PR down-regulation by continuous exposure to progesterone. Twenty cyclic ewes were bilaterally ovariectomized and fitted with uterine catheters on Day 2 of the estrous cycle (Day 0 = estrus). Ewes were then assigned randomly to be treated, in a 2 x 2 factorial arrangement, with recombinant oIFN-tau (roIFN-tau; 2 x 10(7) antiviral units per ewe per day) or control proteins (6 mg/day) by intrauterine injection from Days 10 to 14, and with daily i.m. injections of 20 mg progesterone from Days 2 to 14 (P) or progesterone from Days 2 to 14 plus 50 micrograms estradiol-17 beta from Days 12 to 14 (P+E). All ewes were hysterectomized on Day 15. Endometrial PR mRNA (p < 0.01) and protein (p < 0.03) were higher in ewes receiving P+E than in those receiving P alone. However, the increase in PR mRNA and protein was not as great in the endometrium of roIFN-tau-treated ewes as compared to controls (p < 0.08, treatment x steroid). In ewes receiving P alone, PR mRNA and immunoreactive PR were localized to stroma and deep glandular epithelium and were not present in endometrial luminal and shallow glandular epithelium. Values for endometrial ER mRNA (p < 0.02) and ER protein (p < 0.01) were greater in controls than in roIFN-tau-treated ewes regardless of steroid treatment. Among controls, ER mRNA and immunoreactive ER protein were present in the luminal and glandular epithelium and were increased in the epithelium and stroma in ewes receiving estrogen. In contrast, endometrial ER mRNA and immunoreactive ER protein were very low or absent in the endometrium of roIFN-tau-treated ewes and were not increased by estrogen. Among controls, endometrial OTR density was greater (p < 0.09) in ewes treated with P+E than in those treated with P alone. In roIFN-tau-treated ewes, endometrial OTR density was lower (p < 0.01) than in the controls. Results indicate that roIFN-tau did not stabilize or prevent autologous down-regulation of PR mRNA or protein expression in the endometrium. However, roIFN-tau did suppress endometrial ER expression and OTR formation in ewes regardless of steroid treatment. The results support the hypothesis that the antiluteolytic effects of oIFN-tau are to suppress endometrial ER gene expression in the endometrial epithelium, thereby inhibiting formation of OTR and production of luteolytic PGF2 alpha pulses.

Animals↗