Search PubMed⌕ Search

Biomedical subjects

R B Heap

Publications and source records attributed to R B Heap.

At least 37 records · Page 2Linked to original sources
PubMed1992

Regulation of steroid synthesis and metabolism in isolated binucleate cells of the placenta in sheep and goats.

Binucleate cells of sheep and goat fetal placentae comprise about one-fifth of the trophectodermal layer at the feto-maternal interface. When isolated and incubated in vitro they produce the steroids that are synthesized by the placenta in vivo (progesterone in sheep, 5 beta-pregnane-3 alpha,20 alpha diol in goats). This study demonstrates that progesterone synthesis in binucleate cell preparations in sheep was increased by prostaglandin (PG) E-2, nordihydroguaiaracetic acid (NDGA) and methylisobutylxanthine, but reduced by indomethacin, whereas in goats only NDGA produced any effect (an increase). None of the other compounds tested (luteinizing hormone, follicle stimulating hormone, prolactin, dibutyryl cAMP, A23187 or phorbolmyristic acetate) had any effect. Sheep binucleate cells also produced PGE-2 from arachidonic acid. These results suggest that, in sheep, products of both the cyclooxygenase (producing PGE-2) and lipoxygenase (inhibited by NDGA) pathways of arachidonic acid metabolism have regulatory roles in placental steroid synthesis, but only the lipoxygenase pathway is relevant in goats.

1-Methyl-3-isobutylxanthine↗
PubMed1992

Abnormal maternal behaviour in mice previously immunized against progesterone.

Anti-progesterone immunization leads to reversible infertility in mice; this can be achieved by passive immunization with a monoclonal antibody to progesterone (DB3), or by active immunization with either a progesterone-protein (bovine serum albumin; BSA) conjugate or anti-idiotype directed against DB3. Recovery of fertility in treated females varied from 39.5 to 75.5 median days after passive or active (progesterone-BSA) immunization respectively. Litter size after the first pregnancy also differed from 8.6 +/- 0.8 to 5.0 +/- 0.6 (mean +/- S.E.M.) per mother after passive or active immunization respectively. When litter size was standardized to a maximum of four pups per litter, aberrant maternal responses were observed in the first 5 days after delivery in 40-70% of the nursing mothers. These responses took the forms of cannibalism and failure to retrieve or to nurse pups and resulted in a high incidence of pup rejection (up to 40%), compared with no rejection in control mothers. When mothers were allowed to keep entire litters, an even higher incidence of pup rejection occurred (51% compared with 8% in controls). There was an apparent relation between the degree of negative maternal behaviour and the progesterone antibody concentration in the circulation during the infertile period. Whereas aberrant behaviour occurred mainly within the first 5 days of lactation, it was significantly reduced thereafter. Aberrant behaviour of the mother towards pups may be a consequence of the presence of residual progesterone antibodies in the circulation which affects the process of progesterone withdrawal at parturition that is essential for the establishment of normal maternal responses to the neonate.

Animals↗
PubMed1991

Oestrogen production by the preimplantation donkey conceptus compared with that of the horse and the effect of between-species embryo transfer.

Aromatase distribution in membranes of preimplantation horse and donkey conceptuses was compared by measuring the incorporation of [3H]androstenedione into oestrone and oestradiol-17 beta. In the donkey conceptus, aromatase activity was similar in all the tissues examined (yolk sac, chorionic girdle and allantochorion), whereas in the horse it was generally lower and showed the relationship chorionic girdle greater than yolk sac greater than allantochorion. A higher proportion of labelled precursor was incorporated into oestradiol-17 beta by extra-embryonic tissues of the donkey compared with those of the horse. In contrast to previous results, aromatase in the chorionic girdle did not decline progressively before its migration into the endometrium on Day 36 to form the endometrial cups. The chorionic girdle of a donkey conceptus carried in the uterus of a mare failed to invade the surrogate horse endometrium and aromatase activity was still high in this tissue at Day 42. Aromatase distribution in 2 transferred donkey-in-horse conceptuses resembled that of the fetal, rather than the maternal, genotype indicating a lack of effect of the maternal environment.

Amnion↗
PubMed1991

Progesterone and 5 beta-pregnanediol production by isolated fetal placental binucleate cells from sheep and goats.

Enzymic dispersion and density gradient separation were used for the isolation of enriched populations (60-90%) of cells from the corpus luteum, placenta and peripheral blood of pregnant sheep and goats. Analysis of the steroids produced from radioactive pregnenolone demonstrated that placental binucleate cells can produce progesterone and 5 beta-pregnanediol whereas white blood cells were relatively inactive. Thus, sheep binucleate cells converted pregnenolone predominantly to progesterone as did sheep luteal cells. However, goat binucleate cells produced 5 beta-pregnanediol as the major metabolite, which is consistent with its production in vivo during pregnancy. Production of progesterone (sheep) or 5 beta-pregnanediol (goat) by binucleate cells was shown to be proportional to the number and viability of the cells. In contrast with the binucleate cells there was no evidence that trophectodermal uninucleate cells play a significant role in placental progesterone or 5 beta-pregnanediol synthesis in either species.

Animals↗
PubMed1991

Mechanism of secretion of plasma insulin-like growth factor-I into milk of lactating goats.

125I-Labelled insulin-like growth factor-I (IGF-I) was infused as the free form directly into the pudic artery supplying one gland of lactating goats (n = 6). The infusion was for 60 min and 0.4 +/- 0.09% (S.E.M.) of the infusate was secreted into milk from the infused gland during its first passage through that gland. A large proportion of the 125I-labelled IGF-I escaped into the systematic circulation and was secreted into milk of both glands. A total of 5.2 +/- 0.4% of infused radioactivity was recovered in milk from both glands from 0 to 720 min. Radioactivity consisted of trichloroacetic acid (TCA)-precipitable and -soluble counts which were shown by gel filtration to be authentic IGF-I and degraded products of the peptide. The amount and time course of TCA-soluble radioactivity in milk from both glands was similar, suggesting degradation of 125I-labelled IGF-I at extramammary sites. Maximum specific activity for 125I-labelled IGF-I in milk from the infused gland was reached 80-120 min after the start of infusion and was 2.5-fold greater than milk from the non-infused gland. The time course of appearance of 125I-labelled IGF-I in milk suggests that transfer was via the transcellular pathway and this was further supported by comparing the pattern of transfer of [14C]sucrose and [14C]amino acids. When excess unlabelled IGF-I was included in the infusate, specific activity in milk from the infused gland was reduced to that of the non-infused gland, indicating a competitive and saturable mechanism of secretion for 125I-labelled IGF-I.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1991

Production of 5 alpha-dihydroprogesterone during late pregnancy in the mare.

Changes in the progesterone metabolite 5 alpha-dihydroprogesterone (5 alpha-DHP) in maternal plasma in late gestation, and possible sites of production of this steroid were studied in pony and Thoroughbred mares by an enzyme-linked immunosorbant assay for 5 alpha-DHP. In Thoroughbred mares, plasma 5 alpha-DHP increased from 63.7 +/- 10.5 ng/ml (27 days pre-partum) to 161.7 +/- 30.8 ng/ml (1 day pre-partum) falling to 90.2 +/- 16.1 ng/ml on the day of parturition. In pony mares, values rose from 30.8 +/- 8.1 ng/ml (27 days pre-partum) to 79.1 +/- 30.8 ng/ml (3 days pre-partum) and then decreased to 28.2 +/- 7.1 ng/ml on the day of parturition. Concentrations of 5 alpha-DHP were greater in the umbilical and uterine veins than in the corresponding umbilical and uterine artery samples. Tissues incubated with isotopically-labelled substrates showed that the fetal placenta converted pregnenolone to progesterone (47.5 +/- 9.0%). Endometrium metabolised [3H]pregnenolone to progesterone and 5 alpha-DHP (11.0 +/- 3.0% and 7.8 +/- 2.4%, respectively), and [14C]progesterone to 5 alpha-DHP (9.3 +/- 2.5%). Deuterium-labelled substrates were used to confirm the identity of these products by gas chromatography-mass spectrometry. Negligible amounts of 5 alpha-DHP were formed by other fetal tissues (adrenal, liver, kidney and testis). The results show that 5 alpha-DHP production in late pregnancy is derived primarily from endometrial metabolism of pregnenolone and/or progesterone.

5-alpha-Dihydroprogesterone↗
PubMed1990

Purification and characterization of heparin-binding growth factors from porcine uterus.

Heparin-binding growth factors present in pig uterine tissue were purified by approx. 50,000-fold using a combination of ammonium sulphate precipitation, ion-exchange chromatography and heparin-affinity chromatography. Purification of the uterus-derived growth factors (UDGFs) was monitored by the stimulation of [3H]thymidine incorporation into Swiss 3T3 cells and by a radioreceptor assay using 125I-labelled epidermal growth factor (EGF) as the ligand. The latter was shown to be a novel, rapid and reliable assay for heparin-binding growth factors which utilizes their trans-modulation of EGF receptor affinity. UDGFs exhibit strong affinity for immobilized heparin and two forms, named alpha UDGF and beta UDGF, were distinguished by salt gradient elution from heparin-agarose affinity columns. beta UDGF activity was eluted from heparin-agarose between 1.5 M- and 1.8 M-NaCl, and was correlated with the elution of a protein doublet of 17.2 kDa and 17.7 kDa. Immunoblotting of heparin-purified beta UDGF indicated that the beta UDGF doublet is immunologically related to the 146-amino-acid form of bovine basic fibroblast growth factor (bFGF), and that the 17.2 kDa component is an N-terminally truncated form of the 17.7 kDa component. After purification by C4 reversed-phase h.p.l.c., this doublet was biologically active and greater than 95% pure as assessed by silver-stained SDS/PAGE. Amino acid composition and sequence analysis confirmed that these beta UDGF polypeptides were microheterogeneous forms of bFGF. Fractions containing alpha UDGF activity were eluted from heparin-agarose in 1.3 M-NaCl. These fractions contained a 16.5 kDa protein which co-migrated on SDS/polyacrylamide gels with recombinant human acidic FGF (aFGF) and which which cross-reacted with an antiserum raised against aFGF. The identification of heparin-binding growth factors in porcine uterus at the time of implantation raises the possibility that they function in the reproductive tract during early pregnancy.

Amino Acid Sequence↗
PubMed1990

Effect of passive immunization against progesterone on its distribution in circulation and tissues of mice.

Anti-progesterone monoclonal antibody, injected into mice 32 h after mating at a dose that blocks the establishment of pregnancy, produced a significant reduction in the concentration of progesterone in the ovary and uterus within 6 h after treatment. Uterine concentrations remained lower in treated compared with control animals for at least 24 h after injection. There was an associated transient increase in plasma LH and FSH concentrations, but there was no change in plasma prolactin values. The percentage of total progesterone in the circulation that was unbound was reduced after treatment, but the concentration of unbound progesterone was increased. Studies of antibody binding of steroid in the presence of uterine progesterone receptor protein showed that there was a stoichiometric relationship in the distribution of ligand between the two binders. The present findings suggest that the effects of passive immunization against progesterone are associated with perturbation of tissue concentrations of steroid in the target organ as a result of high antibody concentrations in the circulation.

Animals↗
PubMed1990

Progesterone and oestrogen concentrations in plasma of Barbary sheep (aoudad, Ammotragus lervia) compared with those of domestic sheep and goats during pregnancy.

Steroid hormone concentrations have been measured in the peripheral plasma of 3 Barbary sheep over 3 breeding seasons. During pregnancy mean progesterone values rose initially and after a small decline between Days 30 and 50, increased again and remained between 17 and 28 nmol/l until the last 2 days of pregnancy. Oestradiol-17 beta reached a peak of about 300 pmol/l during mid-pregnancy, increasing to over 400 pmol/l in the last 5 days of pregnancy. Oestrone sulphate began to increase in concentration from about Day 40 of pregnancy and reached a peak of about 19 nmol/l by Day 120. Following a slight decrease from Day 130, there was a further rise in values just before parturition. Values for these steroids in the Barbary sheep studied were between those expected for domestic sheep and goats.

Animals↗
PubMed1990

Pregnancy-blocking progesterone antibody targets specifically the uterus through its progesterone-binding sites.

Passive immunization with a mouse monoclonal antibody against progesterone, designated DB3, blocks pregnancy in several species. We have previously reported that DB3 localizes in the mouse uterine epithelium shortly before normal implantation. This phenomenon is pregnancy dependent and specific for the progesterone antibody. In this study we demonstrate that DB3 is present in the lumen of the uterus 36 h after an i.p. injection; this correlates with the time of maximum antibody reaction on the uterine epithelium. Incubation of DB3 with free progesterone, progesterone-hemisuccinate or progesterone-bovine serum albumin before administration prevented its localization on the epithelium, indicating that the localization requires free progesterone-binding sites and thus probably depends upon progesterone binding. In addition, studies in vitro show that DB3 can effectively bind to progesterone carried by high-affinity progesterone-binding protein purified from coypu plasma. We suggest that specific targeting of DB3 may be through progesterone associated with a progesterone-binding molecule on the membrane of the uterine epithelia. This may be an important part of the mechanism of antibody action against implantation.

Animals↗
PubMed1990

The antifertility effect of passive immunization of mice against progesterone is influenced by genetic factors other than H-2 haplotype.

The anti-fertility effect of passive immunization against progesterone is influenced by genotype in mice. In order to quantify this finding we determined the effective dose that blocks pregnancy in 50% of treated mice (ED50) for two different anti-progesterone monoclonal antibodies (DB3 and 11/32) in four different strains of mice (BALB/cJ, CBA/Ca, Tuck's no. 1 and F1C). Efficacy was greatest in the two inbred strains (BALB/cJ and CBA/Ca) with ED50 values of 0.46-1.4 nmol. The F1C hybrid mice were more resistant to antibody treatment (ED50 2.2-3.0 nmol), while the outbred Tuck's no. 1 strain required much higher doses (ED50 7.0-8.2 nmol). There were no intrastrain differences between the two monoclonal antibodies. We have examined the possible role of the H-2 haplotype on antibody efficacy in different strains and crosses. The antibody was highly effective in blocking implantation in three congenic BALB strains of different H-2 haplotype, in another inbred strain CBA/Ca, and in reciprocal BALB/cJ x CBA/Ca crosses. The F1 hybrid crosses were somewhat resistant, but the C57BL/10Sn and B10.BR congenic strains were most resistant to treatment. The results show that the pregnancy-blocking effect of the anti-progesterone antibody was not influenced by the H-2 haplotype, but rather by background genes.

Animals↗
PubMed1990

Increase in milk secretion and mammary blood flow by intra-arterial infusion of insulin-like growth factor-I into the mammary gland of the goat.

The close-arterial infusion of free insulin-like growth factor-I (IGF-I; 1.1 nmol/min) for 6 h into the pudic artery supplying one mammary gland of lactating goats caused a 25 +/- 6% (mean +/- S.E.M., n = 6) increase in the rate of milk secretion of that gland. The increase in the rate of milk secretion in the adjacent noninfused gland (14 +/- 4%) was not significantly different from that observed during saline infusion (4 +/- 5%). Blood flow to the infused gland was increased from 378 +/- 26 ml/min 1 h before to 487 +/- 56 ml/min approximately 5 h after the start of the infusion of IGF-I, declining to 420 +/- 44 ml/min approximately 2 h after the end of the infusion. The total concentration of IGF-I (free and bound) in milk of the infused gland was significantly higher than that of the non-infused gland. The concentrations of IGF-I in carotid arterial plasma samples increased during IGF-I infusion from a mean value of 32 +/- 2 nmol/l before to a maximum of 49 +/- 3 nmol/l 5 h after the infusion commenced. Circulating concentrations of total IGF-I declined slowly after the infusion with an estimated half-life of 5 h. Infusion of saline alone did not alter mammary blood flow or the concentration of total IGF-I in milk or plasma. The results indicate that the infusion of free IGF-I into the mammary arterial supply enhances milk secretion and mammary blood flow in intact, conscious goats.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1990

The role of trophoblastic binucleate cells in implantation in the goat: a morphological study.

In the goat conceptus individual intra-epithelial trophectodermal binucleate cells first appear 18 days post coitum and their incidence rapidly increases where the trophectoderm is apposed to the caruncular and intercaruncular sites of initial attachment to the uterine epithelium. Special staining techniques reveal that these cells, when mature, contain prominent Golgi bodies and numerous characteristic granules. Our evidence shows that at 19 days post coitum the binucleate cells migrate to the microvillar junction and fuse with individual uterine epithelial cells to form hybrid feto-maternal trinucleate cells. It is proposed that subsequent continued binucleate cell migration and fusion with trinucleate cells produce the syncytial plaques typical of the remainder of pregnancy. It is further suggested that the fusion is important in facilitating the delivery of the characteristic granules to the base of the uterine epithelial layer with subsequent exocytosis of their contents into maternal tissue.

Animals↗
PubMed1989

Cardiovascular responses and mammary substrate uptake in Jersey cows treated with pituitary-derived growth hormone during late lactation.

Pituitary-derived bovine growth hormone (bGH) was administered to Jersey cows during late lactation for 7 d. Milk yield increased significantly during treatment and by a maximum of 49.6% on d 7. The magnitude of the increase was similar to that of mammary plasma flow (47.8 +/- 18.3%) over the same period. By 15-21 d after treatment, both variables had returned to pretreatment values. With respect to milk composition, bGH had negligible effect on lactose and fat concentrations but there were significant decreases in protein, sodium and chloride. Arterial plasma concentrations of bGH increased substantially during treatment, but the associated rise in insulin was not statistically significant. Haematocrit decreased significantly, the lowest value being recorded 3 d after bGH treatment ceased. Mammary respiratory quotient fell progressively after the start of bGH treatment and reached the lowest recorded value 3 d after treatment ceased (62.2 +/- 7.3% of pretreatment value). Glucose and acetate uptake by the mammary gland increased significantly during treatment, increase in glucose uptake being due both to a greater arterio-venous difference and to mammary plasma flow. There was strong evidence that the acute response in increased milk yield was associated with multiple effects in terms of mammary plasma flow and metabolism, as well as haematocrit changes indicative of increased plasma volume.

Animals↗
PubMed1989

Blocking of pregnancy in mice by immunization with anti-idiotype directed against monoclonal anti-progesterone antibody.

Passive transfer of monoclonal anti-progesterone antibodies shortly after mating blocks the onset of pregnancy in different species (mouse, rat, and ferret). Here we report that BALB/c mice can be actively immunized against progesterone, and hence against pregnancy, by means of rabbit anti-idiotypic antibodies specific for a mouse monoclonal anti-progesterone antibody, DB3. Some of the anti-idiotypic antibodies reacted with the steroid-combining site on the DB3 molecule. In response to repeated anti-idiotypic immunization, mice produced serum anti-progesterone antibodies (up to 100 micrograms/ml) that resembled DB3 in idiotypy, affinity, and specificity for progesterone and other steroid ligands. Thus an anti-idiotype can mimic the antigenicity of a steroid hormone with a high degree of accuracy. Compared with immunization with a progesterone-bovine serum albumin conjugate, the anti-progesterone response to anti-idiotype was considerably lower and clonally restricted. When mated after completion of the immunization course, the fertility rate of anti-idiotype-immunized mice was reduced to 30% from a control level of 91%. The anti-fertility effect was correlated with the circulating anti-progesterone concentration in individual animals and persisted for 4 or 5 estrous cycles. Active immunization with progesterone-bovine serum albumin was a highly effective means of rendering mice infertile; it reduced the fertility rate to zero over 16 or 17 estrous cycles. Our results suggest that anti-idiotypes may form the basis of contraceptive vaccines.

Animals↗