The effects of feeding sows at onset of farrowing supplemental energy (blend of carbohydrates and glycerol) on farrowing kinetics and piglet vitality.

Delivering piglets is one of the most energy-demanding activities sows undergo in their lifetime. Sows can have myometrial contractions from 2 to 12 h before the first piglet is expelled as well as a nest-building behavior. Thus, when the first piglet is delivered, the female has already used part of her energy supply. When the sow gets exhausted due to lack of energy, the farrowing process can be interrupted, causing damage to the viability and vitality of the piglets. In the present study, we evaluated the effects of feeding sows an energy supplement at the onset of farrowing on farrowing kinetics and piglet vitality. The energy supplement consisted of a blend of carbohydrates and glycerol which provides 439 kJ of metabolizable energy per kg of metabolic weight. A total of 180 sows were used. At the onset of farrowing, sows were assigned to one of the following treatments: sows that were not supplied energy at the onset of farrowing, serving as controls (CON, n = 85); sows fed the energy supplement at the onset of farrowing (ESP, n = 95). Farrowing kinetics, blood glucose concentration, and piglet vitality were recorded for each sow. Blood glucose concentration was assessed by puncturing the auricular vein and using a portable glucometer at four different time points: after the birth of the 1st piglet (T0), and at 20 (T20), 40 (T40), 80 (T80), and 180 (T180) min after the birth of the 1st piglet. The vitality of the 1st, 6th, 12th, 17th, and 20th piglet born was evaluated using the Apgar score. Piglet birth weight and average colostrum intake were measured. The farrowing duration was 20 min shorter (P < 0.05) for ESP sows in comparison with CON sows. Sows from ESP treatment had higher (P ≤ 0.05) blood glucose concentration at T20 and T40 compared to the CON sows. The inter-piglet birth interval was shortened (P < 0.05) by 14 min between the 1st and 2nd piglet for the ESP treatment. The 17th and 20th piglets born from ESP sows had higher (P < 0.05) Apgar score compared to piglets of the same birth order from CON sows. Colostrum intake was higher (P < 0.01) for piglets born from ESP sows. Litter growth performance did not differ (P > 0.05). In conclusion, feeding a blend of carbohydrates and glycerol as an energy supplement for farrowing sows improved farrowing kinetics and piglet vitality score.

Supplemental progesterone during early pregnancy exerts divergent responses on embryonic characteristics in sows and gilts.

Progesterone (P4) plays a key role in pregnancy establishment and maintenance; during early pregnancy, P4 stimulates the production and release of uterine secretions necessary for conceptus growth prior to implantation; therefore, exogenous P4 supplementation may improve embryo development. This study evaluated the effects of supplementation during early pregnancy with long-acting injectable progesterone or altrenogest on embryonic characteristics of sows and gilts. Thus, a total of 32 sows and 16 gilts were used. On day 6 of pregnancy sows and gilts were allocated to one of the following groups: non-supplemented; supplemented with 20 mg of altrenogest, orally, from days 6 to 12 of pregnancy; supplemented with 2.15 mg/kg of long-acting injectable progesterone on day 6 of pregnancy. Animals were killed on day 28 of pregnancy, and ovulation rate, embryo survival, embryo weight, crown-to-rump length, uterine glandular epithelium and endometrial vascularization were assessed. Treatments had no effect on pregnancy rate, embryo survival or endometrial vascular density (P > 0.05). Non-supplemented gilts presented larger and heavier embryos compared to gilts from supplemented groups (P < 0.05). Sows in the altrenogest group presented larger and heavier embryos compared to non-supplemented sows and sows supplemented with long-acting injectable progesterone. In conclusion, supplementation of sows and gilts with progestagen from day 6 of pregnancy can be used as a means to improve embryo survival without deleterious effects.

Persistence and Retention of Porcine Reproductive and Respiratory Syndrome Virus in Stable Flies (Diptera: Muscidae).

We investigated the acquisition of porcine reproductive and respiratory syndrome (PRRS) virus by the stable fly (Diptera: Muscidae; Stomoxys calcitrans (L.)) through a bloodmeal, and virus persistence in the digestive organs of the fly using virus isolation and quantitative reverse-transcription PCR (qRT-PCR). Stable flies were fed blood containing live virus, modified live vaccine virus, chemically inactivated virus, or no virus. Stable flies acquired PRRSV from the bloodmeal and the amount of virus in the flies declined with time, indicating virus did not replicate in fly digestive tissues. Virus RNA was recovered from the flies fed live virus up to 24 h postfeeding using virus isolation techniques and 96 h using qRT-PCR. We further examined the fate of PRRSV in the hemolymph of the flies following intrathoracic injection to bypass the midgut barrier. PRRSV was detected in intrathoracically inoculated adult stable flies for 10 d using qRT-PCR. In contrast to what we observed in the digestive tract, detectable virus quantities in the intrathoracically inoculated stable flies followed an exponential decay curve. The amount of virus decreased fourfold in the first 3 d and remained stable thereafter, up to 10 d.

Effects of stress associated with weaning on the adaptive immune system in pigs.

This study was designed to investigate the effects of weaning age on specific components of the adaptive immune system in pigs. Twenty-three crossbred pigs were randomly assigned to 1 of 3 treatments: weaning at 14 (14D, n = 8), 21 (21D, n = 7), or 28 (28D, n = 8) d of age. Peripheral blood samples, obtained when pigs were 13, 15, 20, 22, 27, 29, and 35 d of age, were analyzed for peripheral blood cell percentages and concentrations of neutrophils, lymphocytes, T cell subsets, mature B cells, and plasma cortisol concentrations. For each of the 3 groups, weaning increased plasma cortisol concentrations (P < 0.001) and reduced BW percentage change (P < 0.017). Lymphocyte concentrations displayed a treatment effect for the 14D (P = 0.074) and 28D (P = 0.014) groups. Albeit inconsistent, lymphocyte concentrations were less in weaned pigs on the day after weaning than in pigs remaining on the sow or weaned at a younger age. Specifically, mature B cells (CD21(+)) and CD4(+)CD8(+) cells decreased (P < 0.05) after weaning at 28 d of age. Other differences occurred among treatments; however, the differences apparently were not associated with weaning. Based upon the immunological measures used in the present study, there was not an explicit benefit to the adaptive immune system for any weaning age. Early weaning did not negatively affect the adaptive immunological competence of pigs as determined by changes in populations of immune cells.

Assessment of Stomoxys calcitrans (Diptera: Muscidae) as a vector of porcine reproductive and respiratory syndrome virus.

Porcine Reproductive and respiratory syndrome (PRRS) is a globally significant swine disease caused by an arterivirus. The virus replicates in alveolar macrophages of infected pigs, resulting in pneumonia in growing pigs and late-term abortions in sows. Outbreaks occur on disparate farms within an area despite biosecurity measures, suggesting mechanical transport by arthropods. We investigated the vector potential of stable flies, Stomoxys calcitrans (L.) (Diptera: Muscidae), in the transmission of porcine reproductive and respiratory syndrome virus (family Arteriviridae, genus Arterivirus, PRRSV) under laboratory conditions. Stable flies were collected around PRRS-negative boar stud barns in North Carolina and tested for presence of the virus. Stable flies were collected on alsynite traps placed near the exhaust fan of the close-sided tunnel-ventilated buildings, suggesting blood seeking flies are attracted by olfactory cues. No flies were positive for PRRSV. We assessed transmission of the virus through an infective bite by feeding laboratory reared stable flies on blood containing virus and transferring them to naive pigs for subsequent bloodmeals. Transmission of the virus to naive pigs by infective bites failed in all attempts. The volume of blood contained within the closed mouthparts of the stable fly seems to be insufficient to deliver an infective dose of the virus. Stable flies are unlikely to transmit PRRSV from one pig to another while blood feeding. The fate of the virus after a bloodmeal remains to be determined.

Pharmacology of tetracycline water medication in swine.

Medicating drinking water with tetracycline is commonly used in swine production systems to treat and prevent disease outbreaks. However, little information is known of the pharmacokinetics of this medication in water formulations. Twenty-four barrows, divided into 1 control group (of nontreated animals) and 3 equally sized treatments groups (n = 6/group), were treated with tetracycline water medication for 5 d at 125, 250, and 500 mg/L. Blood samples were collected at 0 (prestudy), 4, 8, 12, 24, 32, 48, 56, 72, 80, 96, and 104 h after exposure. Data analyses consisted of a noncompartmental pharmacokinetic analysis and statistical analysis of steady state concentrations with repeated measures ANOVA and multiple-comparison testing to determine whether plasma concentrations differed among groups. Derived pharmacokinetic parameters were consistent with previously published feed and intravenous data. Plasma tetracycline concentrations at steady state were 0, 0.33, 0.47, and 0.77 microg/mL for 0-, 125-, 250-, and 500-mg/L exposures, respectively. Treatment group steady-state plasma concentrations were significantly different from plasma concentrations in control animals (P < 0.0001); however, whereas the 125- and 250-mg/L groups were significantly different from the 500-mg/L group (P < 0.0001), their mean plasma tetracycline concentrations did not differ from one another. Furthermore, the study showed that tetracycline oral bioavailability is very small. The dose response curve also shows that concentrations of plasma tetracycline increase linearly, yet not in a 1 to 1 ratio, to the direct increase in water medication dose.

Expression and localization of vascular endothelial growth factor and its receptors in pig corpora lutea during the oestrous cycle.

Expression and localization of mRNAs for vascular endothelial growth factor (VEGF), VEGF receptor 1 (Flt) and VEGF receptor 2 (KDR) (VEGFR-1 and VEGFR-2, respectively) were investigated in pig corpora lutea. Northern blot analysis of total RNA indicated hybridization of pig VEGF, VEGFR-1 and VEGFR-2 cDNA probes to mRNA transcripts of approximately 3.9, 7.0 and 5.0 kb, respectively. The expression of mRNAs for VEGF and its receptors during the luteal phase (days 4, 7, 10, 13 and 15 after the onset of oestrus) were assessed by northern blot analysis, and hybridization signals were normalized to expression of pig 18S rRNA. Relative hybridization signals of expression of VEGF mRNA appeared to be constant; however, expression of VEGFR-1 mRNA was low on day 4, increased on day 7, and was higher on days 10, 13 and 15 (P<0.05, compared with day 4). In contrast, no changes in expression of mRNA for VEGFR-2 were evident on days 4-13, but a decrease was detected (P<0.05) at day 15. In situ hybridization revealed that VEGF mRNA was localized predominantly in large luteal cells, whereas both VEGFR-1 and VEGFR-2 were localized to small cells. These data indicate that the VEGF system may be involved in the regulation of luteal vasculature throughout the lifespan of the corpus luteum. Although the expression of VEGF mRNA was unchanged during the luteal phase, variations in the expression of VEGFR-1 and VEGFR-2 mRNAs indicate that differential regulation of expression of the VEGF receptors may play a role in the control of VEGF-mediated vascular growth at different phases of development and maturation of the pig corpus luteum.

Cloning of pig prostaglandin F2alphaFP receptor cDNA and expression of its mRNA in the corpora lutea.

Changes in the expression and localization of luteal mRNA for PGF(2alpha) (FP) receptors may be critical in determining the luteolytic action of PGF(2alpha) in pig corpora lutea. In this study, a full-length FP receptor (FPr) cDNA was isolated and cloned from pig corpora lutea. This isolate (GenBank accession no. U91520) contains an open reading frame of 1086 bases coding for a protein of 362 amino acids with seven potential transmembrane domains. The predicted amino acid sequence of this isolate was 83% identical to the FPr amino acid sequence of other species including sheep, cattle and humans. Northern blot analysis showed the presence of an FPr message of about 5 kb in mRNA from pig corpora lutea. Relatively weak FPr mRNA expression was detected on day 4 and day 7 of the oestrous cycle. The expression was greater (P < 0.05) on days 10, 13 and 15 than on days 4 and 7. In situ hybridization analysis revealed that mRNA for FPr was expressed predominantly in the steroidogenic large luteal subtype of cell, although there was some expression in small luteal cells, with histological appearance of steroidogenic small cells. Localization of hybridization signals of FPr was observed in luteal tissue at all stages examined. These data demonstrate that FPr is expressed in pig corpora lutea throughout the oestrous cycle and that upregulation of the FPr mRNA occurs when the corpora lutea becomes sensitive to PGF(2alpha). Direct luteal targets of PGF(2alpha) appear to be primarily large steroidogenic cells in this species.

Effects of feed physical form and buffering solutes on water disappearance and proximal stomach pH in swine.

The effects of the physical form of feed on water disappearance and the effects of buffered water on proximal stomach pH in swine were determined in two experiments. In Exp. 1, 32 barrows were used to evaluate the water disappearance in pigs fed a finely ground and pelleted diet vs those fed a coarsely ground and mashed diet for ad libitum consumption over a 2-wk interval. There were four replicates with eight pigs per replicate. Average daily water and feed disappearance did not differ (P = 0.06 and P = 0.10, respectively). However, average daily water to feed ratio was higher for pigs on the pelleted diet (4.21+/-0.31 L/kg vs 3.04+/-0.33 L/kg; P = 0.02). The higher ratio for the pelleted diet indicated that this may be the cause of a more fluid digesta allowing reflux of irritants from the distal stomach to damage the pars esophageal region of the proximal stomach. In Exp. 2, four barrows (25+/-2 kg) had gastric cannulas surgically implanted into the proximal region of the stomach. Pigs were given ad libitum access to a finely ground and pelleted diet. The experimental design was a Latin square. Water treatments included water (control), 200 mOsm NaHCO3, 250 mOsm NaHCO3, and 250 mOsm mono-dibasic sodium phosphate. Pigs were given a 4-d adjustment period, and pH measurements began on the morning of the 5th d and continued for 24 h under normal feeding conditions. Feed was removed and measurements were continued for 16 h. Buffered water raised the pH of the proximal region of the stomach compared to the control (P < 0.001). Average pH while consuming the water treatments was 3.65+/-0.11 (n = 4) for water control, 4.86+/-0.11 (n = 4) for the 200 mOsm NaHCO3, 4.63+/-0.11 (n = 4) for the 250 mOsm NaHCO3, and 4.59+/-0.14 (n = 3) for the 250 mOsm mono-dibasic sodium phosphate. Buffers also raised the pH of the proximal region of the stomach for the fed (P < 0.001) and the feed restriction (P < 0.01) phases of the trial. Water disappearance rates in pigs given NaHCO3 were higher than in the control (P < 0.01). Average daily water disappearance for the treatments was 9.13+/-0.74 L for the control, 13.56+/-0.74 L for 200 mOsm NaHCO3, 13.77+/-0.74 L for the 250 mOsm NaHCO3, and 10.33+/-0.95 L for the phosphate buffer. The proximal pH of the stomach was increased by adding buffers to the water supply. Addition of NaHCO3 buffers also caused increased water disappearance.

Experimental Salmonella typhi infection in the domestic pig, Sus scrofa domestica.

The domestic pig, Sus scrofa domestica, was examined as a model for typhoid fever, a severe and systemic disease of humans caused by Salmonella typhi. Six pigs were inoculated 1 week post-weaning with approximately 10(10)colony forming units (cfu) of wild type Salmonella typhi strain ISP1820 intranasally and observed for 3 weeks. S. typhi was cultured from the tonsils of 50% of the pigs at necropsy. Cultures from all other organs analysed (ileum, colon, spleen and liver) were negative. No clinical or histopathological signs of disease were observed. Pigs inoculated in parallel with swine-virulent S. choleraesuis all exhibited signs of systemic salmonellosis indicating that the parameters of the experimental infection with S. typhi (e.g. route) were appropriate. Whereas the pig has a gastrointestinal tract that is very similar to humans, our results indicated that the unique features of host and microbe interaction needed to produce typhoid fever were not mimicked in swine. Nevertheless, our observation of tonsillar involvement was consistent with former observations of S. choleraesuis and S. typhimurium infections in swine and supports a role for the tonsil in all porcine salmonella infections.

Effects of immune challenge on concentrations of serum insulin-like growth factor-I and growth performance in pigs.

This study was designed to determine the long-term effects of repeated endotoxin treatment or immunization against human serum albumin on concentrations of serum insulin-like growth factor-I (IGF-I) and other indicators of growth performance in growing pigs. Thirty gilts (38.5 +/- 0.9 kg) were randomly assigned to 5 treatment groups (n = 6 animals/group): 1) lipopolysaccharide injections, 2) lipopolysaccharide pair-fed, 3) human serum albumin immunization, 4) human serum albumin pair-fed, and 5) control. Pigs in the lipopolysaccharide group were treated intramuscularly with lipopolysaccharide on Days 0-3. The pigs in the human serum albumin group were immunized with human serum albumin emulsified in Freund's adjuvant on Day 0 and administered a booster on Day 28. The lipopolysaccharide pair-fed pigs were matched by body weight and pair-wise fed with pigs treated with lipopolysaccharide. Similarly, human serum albumin pair-fed pigs were matched to human serum albumin immunized pigs. Serum IGF-I concentrations did not differ between or within groups. There was no difference in feed disappearance between groups prior to the initiation of treatments. The lipopolysaccharide group had a decrease (P = 0.013) in feed disappearance on Day 0 compared with control and human serum albumin groups. On Day 1, both lipopolysaccharide and human serum albumin groups differed (P < 0.05) from control. Average daily gain and total weight gain did not differ between groups; however, feed efficiency differed (P < 0.05) between lipopolysaccharide and control groups. Long-term effects of repeated endotoxin challenge or immunization on IGF-I concentrations and growth were not evident in the present study. This failure presumably was due to the development of endotoxin tolerance and a relatively innocuous vaccination against human serum albumin.

PGE receptor characteristics on porcine luteal cells during the estrous cycle and early pregnancy.

This study examined the affinities and concentrations of prostaglandin E (PGE) receptors on porcine luteal cells during the estrous cycle and early pregnancy. Corpora lutea (CL) were obtained from nonpregnant gilts at days 9 (n = 4), 12 (n = 3), and 14 (n = 6); three gilts possessed red, vascular CL and three gilts had white nonvascular CL) of the estrous cycle, and days 9 (n = 4), 12 (n = 3), 14 (n = 5), and 30 (n = 5) of pregnancy. The CL were dissociated enzymatically to disperse single cells and the red blood cells were removed by elutriation. The luteal cells were assayed for specific PGE binding by displacement analysis with use of [3H] PGE2 and varying concentrations of unlabeled PGE2. The specific binding of [3H] PGE2 to luteal cells decreased (p < 0.05) from days 9 to 14 of the estrous cycle, but only decreased (p < 0.05) from days 9 to 12 of pregnancy. Specific binding was higher (p < 0.05) on day 14 of pregnancy than the comparable stage of the estrous cycle. The affinities of PGE receptors decreased (p < 0.05) only on the luteal cells dissociated from red, vascular CL of day 14 nonpregnant gilts compared with those of other days of the estrous cycle and pregnancy. The number of PGE receptors on porcine luteal cells was similar (p > 0.05) in pregnant and nonpregnant gilts, but decreased (p < 0.05) on days 12-14 postestrus. During early pregnancy, it was evident that high affinity PGE receptors are sustained on porcine luteal cells; however, the role of the PGE receptors in maternal recognition of pregnancy remains speculative.

Shigella infection as observed in the experimentally inoculated domestic pig, Sus scrofa domestica.

The domestic pig, Sus scrofa domestica, was investigated as a potential animal model for shigellosis. We examined the effects of pig age, pig breed and antibiotic pretreatment upon Shigella infection. Shigella dysenteriae, and Shigella flexneri (both virulent and avirulent strains) were utilized. Our results indicated that young (4-week-old), conventionally re ared, domestic pigs were routinely, but briefly, colonized (average=3.5+/-2.5 days) following oral or gavage administration ofS. flexneri, as determined by direct rectal cultures. The duration of S. dysenteriae colonization was significantly shorter. Inoculation of younger (2 days) or older (9 weeks) pigs with S. flexneri had no significant effect on infection duration. Similarly, infection of 4-week-old pigs with virulent and avirulent strains of S. flexneri had no effect upon the duration of infection, nor did the use of a swine-passaged S. flexneri isolate. Marked clinical, histopathological (gross and microscopic) and immunoIhistopathological signs of disease were absent in all infections. However, in instances where microscopic histopathological evidence was used to correctly identify infected pigs, tonsillar lesions were the consistently noted criteria. The tonsils are believed to be an important portal of entry for Salmonella choleraesuis, another member of the Enterobacteriaceae and a prevalent pig pathogen. Taken altogether, our results indicate that the domestic pig is unsuitable as a model for shigellosis.

Effects of LH, prostaglandin E2, 8-bromo-cyclic AMP and forskolin on progesterone secretion by pig luteal cells.

The present study examined the effects of LH, prostaglandin E2 (PGE2), 8-bromo-cyclic AMP (cAMP) and forskolin on progesterone secretion by small and large pig luteal cells. Corpora lutea were isolated from gilts (n > or = 3 per day) on days 9, 12 and 14 of the oestrous cycle and days 9, 12, 14 and 30 of pregnancy. After enzymatic dissociation of the corpora lutea, small and large luteal cells were obtained by elutriation. Culture plates (24-well) were then seeded with 150,000 small luteal cells or 30,000 large luteal cells per well in 1 ml M199 medium in the absence or presence of LH, PGE2, LH plus PGE2, 8-bromo-cAMP or forskolin. After 12 h of incubation, culture plates were centrifuged, and the supernatant collected and frozen for subsequent assay of progesterone. Differences within day were not detected between cyclic and pregnant gilts, and thus, results were combined for days 9, 12 and 14. Basal progesterone secretion by small luteal cells was less (P < 0.05) on days 14 and 30 than days 9 and 12. Treatment with LH, PGE2, 8-bromo-cAMP or forskolin increased (P < 0.05) progesterone secretion by small luteal cells on days 9 and 12; however, treatments had no effect on days 14 and 30. Basal progesterone production by large luteal cells was less (P < 0.05) on day 30 compared with other days. PGE2 stimulated (P < 0.001) progesterone production by large luteal cells at all days. In contrast, 8-bromo-cAMP and forskolin inhibited progesterone production by large luteal cells on day 12 (P < 0.05), and day 14 (P < 0.001). These data show that pregnancy status does not alter luteal cell response to the aforementioned secretagogues. However, regulation of progesterone secretion differs between small and large luteal cells, and the age of the corpora lutea. Also, it is unlikely that the stimulatory actions of PGE2 involve increased cAMP production in pig large luteal cells.

Effects of acute endotoxemia on serum somatotropin and insulin-like growth factor I concentrations in prepubertal gilts.

OBJECTIVE: To evaluate effects of endotoxemia on serum somatotropin (ST) and insulin-like growth factor I (IGF-I) concentrations in finishing pigs. ANIMALS: Eight female pigs (98 +/- 2 kg) randomly assigned to IV administration (time 0) of saline solution (n = 4) or Escherichia coli lipopolysaccharide (LPS; 5 micrograms/kg of body weight; n = 4). PROCEDURE: Serum ST concentration was determined in serum samples obtained at 20-minute intervals for 6 hours after treatment. Serum IGF-I concentration was determined in samples collected at 1-hour intervals for 6 hours and at 12, 15, 18, 24, 48, 72, and 96 hours after treatment. RESULTS: One distinct pulse of ST (peak 10.5 +/- 0.5 ng/ml) was observed at 40 minutes in each pig after administration of LPS. Control pigs had 2.25 +/- 0.48 ST pulses during the 6 hours of frequent sample collection; however, magnitude of the ST pulses was similar between gilts given LPS and control gilts. A temporal association between ST pulses and saline administration was not evident. Serum IGF-I concentration was similar between gilts of the LPS and control groups prior to treatment. The IGF-I concentration was similar between gilts of the LPS and control groups prior to treatment. The IGF-I concentration was lower (P < 0.01) in gilts of the LPS group (44 +/- 5 ng/ml) than in gilts of the control group (157 +/- 4 ng/ml) at 24 hours. The difference in IGF-I concentrations between groups was evident for 96 hours. CONCLUSIONS: Immediate release of ST was attributed to stress associated with acute endotoxemia and stimulation of the pituitary gland; immune stimulation by LPS may have contributed to the changes in IGF-I concentration. Because feed consumption was similar between the 2 groups of pigs, suppression of IGF-I concentration for 96 hours after administration of LPS was attributable to factors in addition to transient feed restriction. Thus, acute endotoxemia altered the positive association between ST and IGF-I, and provided evidence for a potential mechanism of impaired growth in endotoxemic animals.

Research applications using pigs.

The pig is an excellent research model for numerous human conditions and diseases. As a consequence, valuable information has been generated that has direct applications for human medicine. Research with an applied or agriculture-based emphasis also is essential to commercial pork production. Information must be exchanged between researchers using the pig as a biomedical model and investigators conducting applied research. The numerous research applications using the pig illustrate that the pig is a valuable resource for both biomedical and applied research.

Identification and distribution of prostaglandin E receptors on porcine luteal cells.

This study examined whether pig luteal cells contain specific prostaglandin (PG) E receptors. Corpora lutea, from PG 600R -induced cycling prepubertal gilts or naturally cycling gilts, were dissociated enzymatically, and large, small, and mixed cells were obtained by elutriation. Preliminary experiments demonstrated that optimal [3H]PGE2 binding to mixed cells was obtained after incubation at 22 degrees C for 2 h in binding buffer containing 1% BSA at pH 7.38. In addition, [3H]PGE2 binding was displaced by prostaglandins in the order of PGE2 approximately or = to PGE1 > PGF2a > > PGI2. The mixed, large, and small cells were assayed for specific PGE2 binding by displacement analysis with [3H]PGE2 and varying concentrations of unlabeled PGE2. Each gilt was assigned to group A (n = 6) or group B (n = 3) on the basis of whether the serum progesterone concentrations in that pig had increased or decreased, respectively, from 2 days before ovary collection to the time of ovary collection. Luteal cells from PG 600R -induced cycling prepubertal gilts had similar (p > 0.1) affinities and numbers of PGE receptors. In group A, the mixed cells contained a PGE binding site with a Kd of 21.1 +/- 1.1 nM and a concentration of 5.0 x 10(5) sites per large cell. Affinities of PGE receptors were similar (p > 0.1) regardless of cell types and functional status. PGE receptor concentrations were higher on large cells than on small cells (p > 0.05) and in group A than in group B (p = 0.006). These data demonstrated that PGE receptors primarily exist on porcine large luteal cells and that the concentration of PGE receptors appeared to be related to corpus luteum functional status.

Identification and distribution of tumor necrosis factor alpha receptors in pig corpora lutea.

This study examined whether the pig CL contains specific tumor necrosis factor alpha (TNF alpha) receptors and compared the binding affinities and capacities of small and large cell membranes. Aliquots of membranes, isolated from intact or dispersed luteal tissue, were homogenized, and membrane protein content was quantified. Luteal membranes were assayed for specific TNF alpha binding by displacement analysis, with use of [125I]TNF alpha and varying concentrations of unlabeled TNF alpha. Preliminary experiments demonstrated that TNF alpha binding was maximal after incubation at 22 degrees C for 180 min. In addition, [125I]TNF alpha binding was displaced by TNF alpha, but not by other cytokines. Small cell membranes contained a TNF alpha binding site with an affinity (Kd = 11.6-19 nM) different (p < 0.05) from that of the binding site on large cell membranes (Kd = 56.2-99.6 nM). TNF alpha binding capacities were similar in small and large cell membranes. These data demonstrate that pig CL contain specific, saturable TNF alpha binding sites. The higher-affinity binding sites were localized in the small cell population, which contains predominantly endothelial cells and small luteal cells, suggesting that TNF alpha acts primarily on one or both of these cell types within the CL.

Tumour necrosis factor-alpha differentially alters progesterone and prostaglandin F2 alpha production by porcine luteal cells.

This study examined the effects of tumour necrosis factor-alpha (TNF alpha) on basal and stimulated progesterone secretion, as well as prostaglandin F2 alpha production, by small, large and mixed porcine luteal cells and assessed the action of TNF alpha in the presence and absence of indomethacin. Corpora lutea were isolated from gilts on days 8-9 of the oestrous cycle and enzymatically dissociated. Luteal cells were either subjected to elutriation to isolate small and large cells or were separated from erythrocytes by a polysucrose gradient to serve as the mixed luteal cell group. Then 24-well culture plates were seeded with 150,000 small, 30,000 large and mixed (30,000 large+100,000-250,000 small) luteal cells suspended in 1 ml medium 199 media supplemented with 5 micrograms insulin/ml, 40 ng cortisol/ml and 50 micrograms low-density lipoproteins/ml. Cells were cultured for up to 24 h in a humidified incubator at 37 degrees C with 5% CO2 in air. TNF alpha time- and dose-dependently suppressed (P < 0.05) LH-induced, but not basal, progesterone secretion by small luteal cells. Moreover, TNF alpha inhibited (P < 0.05) forskolin-mediated, but not cyclic AMP-mediated, progesterone secretion by small luteal cells. The LH-stimulated progesterone secretion by small luteal cells was not affected by TNF alpha in the presence of indomethacin. Progesterone secretion by large and mixed luteal cells was not affected by TNF alpha. Prostaglandin F2 alpha production by small and mixed, but not large, luteal cells was enhanced (P < 0.05) by TNF alpha.(ABSTRACT TRUNCATED AT 250 WORDS)

Differential effects of LH and PGE2 on progesterone secretion by small and large porcine luteal cells.

This study examined the effects of LH and PGE2 on progesterone secretion by small and large porcine luteal cells with or without low-density lipoproteins. Corpora lutea were isolated from gilts 13-14 days after administering gonadotrophins; enzymatically dissociated and small and large cells were isolated by elutriation. Culture plates, 24-well, were then seeded with 150,000 small or 30,000 large luteal cells suspended in 1 ml M199 medium supplemented with 5 micrograms insulin ml-1, 40 ng hydrocortisone ml-1 and with or without low-density lipoproteins (50 micrograms cholesterol ml-1) or PGE2. Cells were cultured for up to 24 h in a humidified incubator at 37 degrees C under 5% CO2 in air. The low-density lipoproteins stimulated (P < 0.05) progesterone secretion by large, but not small, luteal cells. Prostaglandin E2 stimulated (P < 0.05) progesterone production by large luteal cells in a dose-dependent manner, and the stimulatory effects of PGE2 were greater (P < 0.05) in the presence than in the absence of low-density lipoproteins. Progesterone secretion by small luteal cells was not significantly affected by PGE2. Progesterone production by small luteal cells was enhanced (P < 0.05) by LH, and the stimulatory effects of LH were greater (P < 0.05) in the presence than in the absence of low-density lipoproteins. In the absence of these lipoproteins, LH had no effect on progesterone secretion by large luteal cells; however, in the presence of low-density lipoproteins, LH increased (P < 0.05) progesterone secretion by large cells, though to a lesser (P < 0.05) extent than the effect of LH on small cells. These data demonstrate that progesterone secretion by porcine luteal cells is stimulated differentially by LH and PGE2 and that small luteal cells are more responsive to LH and PGE2 acts primarily on large luteal cells.