Mapping of the porcine alpha-fetoprotein (AFP) gene to swine chromosome 8.
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Animals↗
Biomedical subjects
R K Christenson
Publications and source records attributed to R K Christenson.
Linkage mapping of a single nucleotide polymorphism (SNP) in the porcine QDPR gene to chromosome 8.
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Animals↗
Mapping of the porcine AREG and EGF genes to SSC8.
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Alleles↗
Interrelationships among conceptus size, uterine protein secretion, fetal erythropoiesis, and uterine capacity.
The interrelationships among d-11 conceptus size, d-105 placental weight, placental efficiency (the ability of the placenta to support fetal growth and development), fetal erythropoiesis, and uterine capacity were examined in 1/2 Meishan, 1/2 White crossbred gilts that were unilaterally ovariohysterectomized at 90 to 100 d of age. In Exp. 1, gilts were mated after at least one normal estrous cycle and then slaughtered at 105 d of gestation and number of fetuses and CL, placental weights, fetal weights, hematocrits, fetal plasma iron, and fetal plasma folate were measured. In Exp. 2, gilts were mated and plasma progesterone was measured on d 2 and 3 of gestation. On d 11, the length of the remaining uterine horn was recorded and the uterine horn was flushed with minimal essential medium. Number of CL, conceptus number, conceptus diameters, and total uterine flush retinol-binding protein (tRBP), acid phosphatase (tAP), and folate-binding protein (tFBP) were measured. Gilts were mated again and slaughtered at 105 d of pregnancy and the same traits measured in Group 1 were recorded. Plasma progesterone concentrations on d 2 and 3 were correlated with average conceptus diameter on d 11 (r = 0.60, P < 0.01, for each day). In contrast, tRBP (r = 0.49, P < 0.01), tAP (r = 0.53, P < 0.01), and tFBP (r = 0.51, P < 0.01) in uterine flushings on d 11 were only correlated with d-3 plasma progesterone concentrations. No correlations between d-11 average conceptus diameter or d-11 uterine length with d-105 uterine capacity were observed. Uterine capacity was negatively correlated with placental weight, fetal weight and fetal hematocrit (r = -0.36, P < 0.01; r = -0.44, P < 0.01; r = -0.32, P < 0.01; respectively). Hematocrits were correlated with fetal plasma iron (r = 0.50, P < 0.01) and folates (r = 0.44, P < 0.01). Hematocrit, plasma iron, and plasma folate were each correlated with residual fetal weights after adjusting for placental weight (a measure of placental efficiency), and accounted for 11% of the variation in this trait. These data suggest that conceptus diameter and uterine protein secretion on d 11 may be influenced by the onset of progesterone secretion by the CL, but do not support an influence of conceptus growth during early pregnancy on uterine capacity. These results also suggest that reducing placental and fetal weights will likely result in increased uterine capacity.
Acid Phosphatase↗
Erythropoietin mRNA expression in pig embryos.
To address whether altered erythropoietin (EPO) synthesis might be involved in prenatal pig mortality, studies were conducted to measure porcine embryonic EPO mRNA expression during early gestation (days 24-40). Three pig models differing in embryonic survival from days 24-40 were investigated: intact white crossbred gilts (INT), white crossbred gilts that were unilaterally hysterectomized-ovariectomized before puberty and whose pregnant uterus constituted a crowded environment (UHO), and prolific, intact Meishan gilts (ME). A partial cDNA for porcine EPO, developed via reverse transcription and polymerase chain reaction procedures was used to generate a 32P-labeled probe for use in Northern analyses. In an initial study, embryonic liver EPO mRNA was greatest on day 24, decreased by day 30 (P<0.01), and was barely detectable by day 40. EPO mRNA expression was not influenced by pig model. Placental EPO mRNA expression was detectable in only 4 of 53 placentae examined. In a second study at day 35 of gestation, embryonic liver EPO mRNA expression was measured in the same three pig models and in two embryos of divergent weights from each gilt. Meishan embryos had lower (P<0.01) plasma immunoassayable EPO concentrations (P=0.04) and higher survival rates (87+/-2.7%) at day 35 than did white crossbred embryos (75+/-5%). Liver EPO mRNA expression did not differ among animal models, nor did plasma EPO or tissue EPO mRNA expression differ between large and small embryos. There was no apparent relationship between embryonic development, measured as embryonic and placental size, and plasma EPO concentrations or liver EPO mRNA expression. These results indicate that at the gestational ages examined, the embryonic liver is one source of plasma erythropoietin and suggest that at the ages sampled, EPO is not a limiting factor in embryonic development.
Actins↗
Enhancer regions of ovine interferon-tau gene that confer PMA response or cell type specific transcription.
Interferon-tau (IFNtau), produced by the trophectoderm of peri-implantation conceptuses in ruminant ungulates, attenuates the uterine production of a luteolytic factor, prostaglandin F(2alpha), resulting in the maintenance of corpus luteum function. However, molecular mechanisms regulating the temporal/spatial expression of IFNtau gene are not clearly understood. The 5'-upstream region of the sheep IFNtau (oIFNtau) gene was examined for its transcriptional regulation in two different cell types; JEG3 cells supported the transactivation of oIFNtau-reporter construct, but HeLa cells did not. In a heterologous SV40 enhancer-oIFNtau promoter or oIFNtau enhancer-SV40 promoter systems, elements required for such cell specific transactivation were localized between -654 and -555 bases, the enhancer, but not the basal promoter region of the oIFNtau gene. In these combinations, high degrees of transactivation were observed in JEG3 cells and the activity was further enhanced by the addition of phorbol 12-myristate 13-acetate (PMA), while those responses were absent in HeLa cells. To identify nucleotide sequences responsible for cell specific expression, transient transfection studies with sequential point mutations in the enhancer elements were executed. Transactivation of oIFNtau enhancer-reporter constructs was primarily regulated by three regions containing AP-1 site, GATA like sequence and site(s) unidentified. In gel mobility shift assays (GMSAs), the AP-1 site located in the enhancer region was recognized by nuclear extracts from both cell types. However, one of the GMSA probes containing GATA-like sequence exhibited different DNA-protein complex patterns in JEG3 and HeLa cells. Observations, in which the same upstream sequence behaved differently due possibly to kinds of nuclear factors available in these cell lines, suggest that such a sequence may be involved in cell specific transactivation of the oIFNtau gene. Furthermore, the same enhancer sequences were also recognized by nuclear extracts from sheep trophoblasts, suggesting that the enhancer sequences between -654 and -555 bases of oIFNtau gene may be functioning in vivo.
Animals↗
Characterization of uterine epidermal growth factor during early pregnancy in pigs.
Genomic research has identified a quantitative trait locus for uterine capacity, a component trait contributing to litter size, on porcine chromosome 8. The epidermal growth factor (EGF) gene, on porcine chromosome 8, may influence uterine capacity because of its growth-promoting activities. Using reverse transcription-polymerase chain reaction (PCR) and iterative screening of a porcine reproductive tissue cDNA library, 4932 bp cDNA sequence coding for porcine EGF precursor was obtained. The predicted protein sequence of the EGF precursor contained 1214 amino acids, similar to human EGF precursor (1207 amino acids, 81% identity). Curiously, the sequence of the mature peptide was less homologous between species than other regions of EGF precursor. The presence of conserved regions outside the mature peptide may suggest that these regions are functionally important. Expression of EGF mRNA in the endometrium of White crossbred gilts (n = 3 to 5 each) was determined by Northern blotting using 20 microg of total RNA from endometrium of D 10, 13, and 15 cyclic, and D 10, 13, 15, 20, 30, and 40 of pregnant gilts. A 3342 bp probe from EGF precursor was used. The bands corresponding to EGF mRNA were quantified by densitometry and results were analyzed by ANOVA. EGF mRNA expression decreased significantly from D 13 to 15 of the cycle and pregnancy (P = 0.04), and from D 30 to 40 of pregnancy (P = 0.01). These findings show that EGF mRNA expression is temporally regulated during the cycle and early pregnancy, and this pattern of gene expression may be important during early conceptus development.
3' Untranslated Regions↗
Level of dietary energy during prepubertal growth and reproductive development of gilts.
Development of gilts that conceive early and continue to produce offspring is a primary objective of swine production. The objective of this study was to determine the degree of feed restriction during development required to optimize reproductive performance and efficiency in gilts. The effects that various patterns of growth had on reproductive development and performance of gilts through d 30 of gestation were investigated. At 13 wk of age and 41 kg BW, 192 white crossbred gilts were penned individually and assigned to receive 87.5%, 75%, 62.5%, and 50% of predicted ad libitum energy intake. The study was replicated in two seasons. At 25 wk of age, gilts were moved to group pens and allowed ad libitum access to feed, and estrous detection was initiated. Gilts were inseminated at first observed estrus and those recycling were remated. Post-mating gilts were fed 1.5x maintenance in stalls. Gilts that did not return to estrus 17 to 30 d after mating were slaughtered at 30 d of gestation. Reproductive tracts were collected and numbers of corpora lutea and live embryos were recorded. Feed restriction during development resulted in differences in BW and backfat thickness at the start of the breeding period and differences in feed intake during breeding. Gilts subjected to the greatest feed restriction during development consumed the greatest quantity of feed during breeding. Feed intake during breeding was associated with BW and backfat gain during breeding. The treatment group that entered breeding lightest and leanest (50% of predicted ad libitum intake) had the least number of days to first estrus, followed by the fattest, heaviest group (87.5% of predicted ad libitum intake). Treatment groups did not differ (P > 0.38) in ovulation rate or live embryo numbers. Significant relationships between quantity of GE consumed during development and variables considered important in reproductive development and performance were evident, such as BW and fatness at start of breeding and first detected estrus, and ovulation rate. Variation in dietary energy during the development period impacted many aspects of reproductive development and performance. However, coupling restricted energy intake during development with ad libitum intake during breeding negated many of the effects of feed restriction during the development period.
Animal Feed↗
Intravenous infusion of iron and tetrahydrofolate does not influence intrauterine uteroferrin and secreted folate-binding protein content in swine.
The effect of exogenous iron and folate on reproductive performance in swine is equivocal. However, the effect of exogenous iron and folate on secretion of their respective uterine transport proteins has never been reported. Twenty gilts were infused (n = 5 per treatment) with either 1) saline, 2) alpha-tocopherol, 3) alpha-tocopherol plus iron citrate, or 4) alpha-tocopherol plus tetrahydrofolate on d 11 to 14 of pregnancy. Intravenous infusion of iron citrate and tetrahydrofolate increased (P < 0.05) plasma iron and folate, respectively, for 6 to 8 h after treatment. Treatments had no effect on uterine content of uteroferrin or secreted folate-binding protein in uterine flushings obtained on d 15 of pregnancy. These data suggest that uterine secretion of uteroferrin and secreted folate-binding protein are not influenced by plasma levels of iron and folate, respectively, and may provide an explanation for the equivocal effect of iron and folate treatment on reproductive performance in gilts.
Acid Phosphatase↗
Effect of prepubertal feeding regimen on reproductive development and performance of gilts through the first pregnancy.
Development of gilts that conceive early and continue to produce offspring is an objective of swine production. We investigated different patterns of growth on reproductive development and performance of gilts through the first farrowing. At 13 wk of age and 43 kg BW, 286 white crossbred gilts were penned individually and assigned to treatments: Ad lib, ad libitum intake from 13 to 25 wk of age; Control, ad libitum intake from 13 wk of age until 100 kg BW and then 90% of ad libitum intake until 25 wk of age; and Restricted, 74% of ad libitum intake from 13 wk to 25 wk of age. Feed was formulated to restrict energy intake. The study was replicated in three seasons. At 25 wk of age, gilts were moved by treatment to group pens, fed for ad libitum consumption, and estrus detection was initiated. Gilts were inseminated at first estrus, and those recycling were remated. Postmating gilts were fed 1.5x maintenance until 105 to 110 d of pregnancy. Gilts were moved either to the farrowing facility or the abattoir at 105 to 110 d of pregnancy. Those taken to the abattoir were slaughtered and number, weight, and condition of the fetuses were recorded. Gilts moved to the farrowing facility were allowed to farrow, and number, weight, and condition of the piglets were recorded. Daily feed intake during breeding was 3.4 kg/d by Restricted gilts, 2.9 by Control gilts, and 2.7 kg/d by Ad lib gilts. Increased feed intake by Restricted gilts during breeding resulted in compensatory gains that overcame the reduced reproductive performance that resulted from the reduced BW and backfat these gilts carried at the start of breeding. Days to first estrus and pregnancy were not influenced by development period treatment (P < 0.13). Percentage of Ad lib, Control, and Restricted gilts that successfully completed their pregnancies were 61, 74, and 66, respectively (P > 0.19). Total feed fed from 13 wk of age to end of the first pregnancy per gilt assigned did not differ among Ad lib (506 kg) and Control (498 kg) gilts but was less (P < 0.01) in Restricted gilts (451 kg). Number of piglets born per gilt assigned (P > 0.09) and piglets produced per kilogram of feed fed from 13 wk of age to term (P > 0.29) were 6.47 and 0.0134 in Ad lib gilts, 7.26 and 0.0150 in Control gilts, and 6.38 and 0.0149 in Restricted gilts, respectively. Moderate feed restriction, 74% of ad libitum intake, reduced feed consumed from 13 wk of age to end of the first pregnancy with no significant impact on efficiency of piglet production.
Animal Feed↗
Increased plasma follicle-stimulating hormone concentrations in prepubertal gilts from lines selected for increased number of corpora lutea.
Plasma follicle-stimulating hormone (FSH) was evaluated in gilts from two studies in which ovulation rate was increased through direct selection for number of corpora lutea (CL) to determine whether selection for ovulation rate affected FSH secretion during prepubertal development. In the first study, 76 control and 110 selected gilts of University of Nebraska gene pool lines were bled twice during prepubertal development. Plasma FSH concentrations were greater (P < 0.05) at 53 (13.5%) and 75 (21.3%) d of age in selected than in control gilts. In the second study, 254 control gilts, 261 gilts from a line selected for ovulation rate, and 256 gilts from a line selected for uterine capacity were bled at three prepubertal ages. Plasma FSH was greater (P < 0.05), relative to controls, on d 34 (> 24%), 55 (> 13%), and 85 (> 10%) in White Composite gilts selected for either increased ovulation rate or for greater uterine capacity. Unilateral ovariectomy and hysterectomy were performed at 160 d of age on random gilts in these three lines (n = 377); weights of these organs were evaluated to determine whether selection affected their development. Ovarian and uterine weights were less (P < 0.01) in the control than in the ovulation rate line. Subsequently, ovulation rate was determined during pregnancy (n > or = 130 gilts/line). Controls had fewer (P < 0.01) CL (14.6) than gilts of the ovulation rate line (17.7) but numbers similar (P > 0.10) to those of gilts of the uterine capacity line (14.7). Within each line, plasma FSH only on d 85 correlated positively with subsequent ovulation rate (P < 0.03, 0.001, and 0.08; r = 0.17, 0.30, and 0.15 for control, ovulation rate, and uterine capacity lines, respectively). Ovarian weight at 160 d of age also correlated with subsequent ovulation rate (P < 0.03 and 0.001; r = 0.23 and 0.38) in control and ovulation rate gilts but not in uterine capacity gilts (P > 0.10; r = 0.11). Gilts selected for increased number of CL, in two independent studies, had greater concentrations of FSH during prepubertal development than respective controls. The modest but significant, positive association of FSH at 85 d of age with subsequent ovulation rate provides additional support for using plasma FSH in prepubertal gilts to indirectly select for ovulation rate.
Animals↗
Antibody repertoire development in fetal and neonatal piglets. II. Characterization of heavy chain complementarity-determining region 3 diversity in the developing fetus.
Since the actual combinatorial diversity in the V(H) repertoire in fetal piglets represents <1% of the potential in mice and humans, we wondered whether 1) complementarity-determining region 3 (CDR3) diversity was also restricted; 2) CDR3 diversity changed with fetal age; and 3) to what extent CDR3 contributed to the preimmune VDJ repertoire. CDR3 spectratyping and sequence analyses of 213 CDR3s recovered from >30 fetal animals of different ages showed that >95% of VDJ diversity resulted from junctional diversity. Unlike sheep and cattle, somatic hypermutation does not contribute to the repertoire. These studies also revealed that 1) N region additions are as extensive in VDJ rearrangements recovered at 30 days as those in late term fetuses, suggesting that TdT is fully active at the onset of VDJ rearrangement; 2) nearly 90% of all rearrangement are in-frame until late gestation; 3) the oligoclonal CDR3 spectratype of 30-day fetal liver becomes polyclonal by 50 days, while this change occurs much later in spleen; 4) there is little evidence of individual variation in CDR3 spectratype or differences in spectratype among lymphoid tissues with the exception of the thymus; and 4) there is a tendency for usage of the most J(H) proximal D(H) segment (D(H)B) to decrease in older fetuses and for the longer D(H) segment to be trimmed to the same length as the shorter D(H) when used in CDR3. These findings suggest that in the fetal piglet, highly restricted combinatorial diversity and the lack of somatic mutation are compensated by early onset of TdT activity and other mechanisms that contribute to CDR3 junctional diversity.
Animals↗
Luteinizing hormone secretion as affected by hypophyseal stalk transection and estradiol-17beta in ovariectomized gilts.
The objectives were to determine hypothalamic regulation of pulsatile luteinizing hormone (LH) secretion in female pigs and the biphasic feedback actions of estradiol-17beta (E(2)-17beta). In the first study, the minimum effective dosage of E(2)-17beta that would induce estrus in ovariectomized gilts was determined to be 20microg/kg body weight. In the second study, ovariectomized gilts were assigned randomly on day 0 to treatments: (a) hypophyseal stalk transection (HST), (b) cranial sham-operated control (SOC), and (c) unoperated control (UOC). On day 3, gilts from each group received a single i.m. injection of either E(2)-17beta (20microg/kg body weight) or sesame oil. Blood was collected from an indwelling jugular cannula at 15min intervals for 3h before (day -2) and after treatment (day 2) from HST, SOC and UOC gilts. On day 3, blood was collected at 2h intervals for 12h after E(2)-17beta or sesame oil injection and at 4h intervals thereafter for 108h. Pulsatile LH secretion in all gilts 2 days after ovariectomy exhibited a frequency of 0.9+/-0.06peaks/h, amplitude of 1.3+/-0.13ng/ml, baseline of 0.8+/-0.07. Serum LH concentrations from SOC and UOC gilts were similar on day 2 and profiles did not differ from those on day -2. In HST gilts pulsatile LH release was abolished and mean LH concentration decreased compared with controls (0 versus 0.9+/-0. 06peaks/h and 0.77+/-0.03 versus 1.07+/-0.07ng/ml, respectively; P<0. 05). E(2)-17beta or sesame oil did not affect serum LH concentration in HST gilts, and LH remained constant throughout 120h (0.7+/-0. 07ng/ml). In SOC and UOC control gilts, E(2)-17beta induced a 60% decrease (P<0.05) in LH concentration within 12h, and LH remained low until 48h, then increased to peak values (P<0.05) by 72h, followed by a gradual decline to 120h. Although pituitary weight decreased 31% in HST gilts compared with controls (228 versus 332mg, P<0.05), an abundance of normal basophils was evident in coronal sections of the adenohypophysis of HST comparable to that seen in control gilts. The third and fourth studies determined that hourly i. v. infusions of LHRH (2microg) and a second injection of E(2)-17beta 48h after the first had no effect on the positive feedback action of estrogen in UOC. However, in HST gilts that received LHRH hourly, the first injection of E(2)-17beta decreased (P<0.05) plasma LH concentrations while the second injection of E(2)-17beta failed to induce a positive response to estrogen. These results indicate that both pulsatile LH secretion and the biphasic feedback action of E(2)-17beta on LH secretion depend on hypothalamic regulatory mechanisms in the gilts. The isolated pituitary of HST gilts is capable of autonomous secretion of LH; E(2)-17beta will elicit direct negative feedback action on the isolated pituitary gland if the gonadotropes are supported by exogenous LHRH, but E(2)-17beta at high concentrations will not induce positive feedback in isolated pituitaries. Thus, the direct effect of E(2)-17beta on the pituitary of monkeys cannot be mimicked in pigs.
Animals↗
Genetic differences in the frequency of the hinge variants of porcine IgA is breed dependent.
The distribution of the IgA(a) and IgA(b) alleles of porcine IgA in over 160 randomly-selected animals revealed an abundance of heterozygotes but only two b/b homozygotes. Since the IgA(b) allotype is a splice site mutant lacking two-thirds of the hinge, this study tests the hypothesis that pigs with this genotype may be at a selective disadvantage while heterozygous individuals may be at some advantage. This hypothesis was tested by collecting data on 374 animals of known breed and often parentage. We show here that when breed was not considered, young animals of known parentage had genotypic frequencies identical to that expected for Mendelian alleles but that a/b heterozygotes were overrepresented in adults. However, when analyzed with regard to breed, a very strong association between breed and the frequency of the IgA(a) and IgA(b) alleles was discovered. Meishan and NIH minipigs were homozygous for IgA while heterozygotes predominated in Berkshire, Chester White, Durocs, Hampshire and Landrace. Animals homozygous for IgA(b) were best represented in the White Cross line. We show here that this very strong breed dependency of IgA allotypy in swine can produce a sample bias that can explain why only two b/b homozygotes (1.3%) were found in the 160 randomly-selected samples since the original samples came from primarily Landrace and Yorkshire animals. The expected frequency of b/b homozygotes in these breeds would be <3%. Thus, the data presented here reject the hypothesis that swine homozygous for a trait that results in loss of two-thirds of the IgA hinge, are selected against and that heterozygotes are positively selected. Rather, the study shows that IgA(a) and IgA(b) appear to be simple, breed-dependent allotypic markers.
Alleles↗
Porcine erythropoietin receptor: molecular cloning and expression in embryonic and fetal liver.
The full coding sequence for porcine erythropoietin receptor (EPOR) was elucidated using reverse transcription polymerase chain reaction (PCR) (rtPCR) and 3' and 5' rapid amplification of cDNA ends (RACE) procedures. Total RNA collected from Day 30 fetal liver was used as starting material. A 1843 bp sequence was obtained from which could be inferred a 509 amino acid protein which was 79-85% identical to the amino acid sequence of erythropoietin receptor from other species. Total RNA samples collected from white crossbred intact, white crossbred UHO and Meishan gilts on Days 24, 30 and 40 of gestation were subjected to Northern blotting using porcine EPOR cDNA as probe. Results indicated that (1) a major and two minor forms of mRNA are present, (2) fetal liver mRNA concentrations for EPOR are low on Day 24 of gestation and increase dramatically by Day 30 and (3) mRNA concentrations for EPOR tended to be decreased by intrauterine crowding.
Amino Acid Sequence↗
Analysis of possible silencer elements of ovine interferon-tau gene.
During the peri-implantation period significant production of ovine interferon-tau (olFNtau) by the trophectoderm is detected in day 13-16 conceptuses, but its level rapidly declines thereafter. To understand molecular mechanisms by which oIFNtau gene expression is down-regulated, a variety of deletion constructs were prepared from upstream sequences of the oIFNtau gene and examined for possible silencer regions by using transient transfection into human choriocarcinoma, JEG3, cells. Two regions between -700 to -654 bases (distal region) and from -503 to -453 bases (proximal region) were found to be the possible negative regulatory regions. With probes prepared from these regions, gel mobility shift assay (GMSA) was then conducted. DNA-protein complexes were observed, but the gel shift pattern was different between nuclear extracts from days 14 (active oIFNtau production) and 20 (minute oIFNtau production) ovine trophoblasts. Day 20 nuclear extracts exhibited more band patterns than those of day 14; most notably the distal region between -692 and -668 bases exhibited the distinct band with nuclear extracts from day 20, but not from day 14 trophoblasts. In addition, the band patterns from day 20 trophoblast nuclear proteins were similar to those detected with JEG3 and HeLa cell nuclear extracts. Taken together, these observations suggest that the upstream sequences identified could serve as negative regulatory regions to which various nuclear factors bind, resulting in reduction of oIFNtau gene transcription.
Animals↗
Identification of a functional transcriptional factor AP-1 site in the sheep interferon tau gene that mediates a response to PMA in JEG3 cells.
To examine regulatory mechanisms of sheep interferon tau (oIFNtau) gene expression, potential enhancer/silencer elements of the oIFNtau gene were examined using a transient transfection system with oIFNtau gene-chloramphenicol acetyltransferase (oIFNtau-CAT) reporter constructs in human choriocarcinoma cells, JEG3. Experiments with 5'-deletion constructs revealed that the upstream regions from bases -654 to -607 and from bases -606 to -555 were essential for oIFNtau gene expression. In a heterologous transcriptional system in which the upstream regions of oIFNtau were inserted in front of simian virus 40 (SV40) promoter, the regions between bases -654 and -555 were determined as being the enhancer region required for oIFNtau-SV40-CAT transactivation. A subsequent study with the oIFNtau-CAT constructs lacking the upstream region between bases -542 and -124 revealed that, in addition to the further upstream region between bases -1000 and -654, the sequences from bases -543 to -452 seemed to act as silencer regions. The oIFNtau-CAT constructs with site-specific mutagenesis revealed that multiple enhancer elements existed between bases -654 and -555 of the oIFNtau gene. On the basis of nucleotide sequence analysis, there are numerous sites between bases -654 and -555 to which potential transcriptional factors, AP-1, GATA and GATA-related proteins, could bind. Furthermore, gel mobility-shift assays revealed that AP-1 or other nuclear factors could bind to these elements. In co-transfection studies, the expression of c-Jun plus c-Fos enhanced the transactivation of oIFNtau-CAT but the expression of GATA-1, GATA-2 or GATA-3 did not. Taken together, these results suggest that the upstream region between bases -654 and -555 could be considered as the enhancer region for oIFNtau gene transactivation.
Animals↗
Cortisol concentrations in early porcine embryos.
A study was conducted to determine the presence of cortisol in body tissue of porcine embryos at days 25 and 35 of gestation. Cortisol concentrations (ng/mg DNA) were low but measurable at day 25 and increased eightfold by day 35 during a time when body weight increased 6.4-fold. At day 35, there was a highly significant positive linear regression of body weight on cortisol concentrations. The source of this embryonic cortisol is not known, but its presence suggests the opportunity for cortisol to influence porcine embryonic development at these early gestational stages.
Animals↗