Relationship of plasma beta-carotene and vitamin A to luteal function in postpartum cattle

The influence of plasma concentrations of beta-carotene and vitamin A on in vivo progesterone production by bovine corpora lutea after gonadotropin-releasing hormone-induced LH release was assessed in 39 postpartum dairy cows. Thirty Holsteins and nine Jerseys were given 100 micrograms gonadotropin-releasing hormone on d 12 of an estrous cycle, which began from 30 to 49 d postpartum. Concentrations of beta-carotene and vitamin A in plasma and progesterone and LH in serum were determined prior to gonadotropin-releasing hormone injection (0 h); serum progesterone and LH concentrations were also determined 1, 2, and 3 h after injection of gonadotropin-releasing hormone. Serum concentrations of progesterone and LH were increased by gonadotropin-releasing hormone. Incremental progesterone production in an analysis of covariance was influenced by breed as well as the interactions of breed with vitamin A, of season with beta-carotene, and of season with vitamin A. The regression coefficients were positive for beta-carotene and negative for vitamin A in all cases. In conclusion, luteal function in the postpartum cow appears to be related to plasma concentrations of beta-carotene and vitamin A.     

Vitamin A and beta-carotene in bovine and porcine plasma, liver, corpora lutea, and follicular fluid

Bovine and porcine blood plasma, liver, corpora lutea, and follicular fluid were obtained from local abattoirs for study of distribution of vitamin A and beta-carotene. Retinol, retinyl esters, and beta-carotene were separated on alumina columns and subjected also to thin-layer chromatography. Retinol and retinyl esters were in corpora lutea and follicular fluid of both species. Concentrations of beta-carotene were high in bovine plasma, corpus luteum, and follicular fluid. In contrast, beta-carotene was lower in porcine tissues. Retinol, retinyl esters, and beta-carotene were closely correlated in bovine follicular fluid and blood plasma; however, correlations between bovine plasma and corpora lutea were not significant except for retinol. Only porcine retinol was closely correlated with plasma and follicular fluid, whereas correlations were nonsignificant between plasma and corpora lutea retinol, retinyl esters, and beta-carotene. Further studies, therefore, are needed to elucidate the physiological role of vitamin A and beta-carotene in regulating ovarian functions.     

Effect of supplemental beta-carotene on luteinizing hormone released in response to gonadotropin-releasing hormone challenge in ovariectomized Holstein cows

Thirteen lactating Holstein cows were assigned randomly to either a control or beta-carotene (600 mg/d) treatment group to determine the effect of long-term beta-carotene supplementation on releasable luteinizing hormone in response to gonadotropin-releasing hormone challenge. The experimental period was 28 wk. Milking was terminated at wk 16, ovariectomy performed at wk 20, and response to gonadotropin-releasing hormone determined at wk 27. Serum beta-carotene concentrations reflected dietary intake and were higher in the cows fed beta-carotene after 2 wk of supplementation and remained higher for the duration of the trial. Feeding supplemental beta-carotene had no effect on circulating progesterone concentration, corpora lutea size or corpora lutea progesterone concentration, or basal concentrations of luteinizing hormone, frequency, and amplitude of luteinizing hormone pulses, or the release of luteinizing hormone in response to gonadotropin-releasing hormone. Thus, pituitary responsiveness to exogenous gonadotropin-releasing hormone was not affected by feeding supplemental beta-carotene.     

Stimulation of luteal cell progesterone production by lipoproteins from cows fed control or fat-supplemented diets.

Plasma lipoproteins from lactating dairy cows fed 0 or 7% supplemental fat were examined for their composition and ability to stimulate luteal cell progesterone production in vitro. Ultracentrifugation was utilized to isolate blood lipoproteins, and heparin affinity chromatography allowed separation of lipoprotein fractions based on the presence (low density lipoproteins) or absence of apolipoprotein B (high density lipoproteins). A portion of high density lipoproteins was fractionated by size, utilizing gel filtration chromatography. Slaughterhouse corpora lutea were dissociated, and plasma lipoproteins were added to the luteal cells on d 3 of culture and incubated for 48 h. In Experiment 1, blood was collected from heifers fed a diet that was not supplemented with fat. The addition of cholesterol from large, high density lipoproteins with a high cholesterol to protein ratio to luteal cultures increased progesterone production by an average of 17% compared with the addition of cholesterol from small, high density lipoproteins with a low cholesterol to protein ratio. In Experiment 2, electrophoretic mobility, apolipoprotein composition, and size of lipoproteins from control and fat-supplemented cows were similar. Lipoproteins from cows assigned to either a control or fat-supplemented diet showed no difference in their ability to stimulate progesterone production. Increased plasma progesterone concentration in lactating dairy cows fed supplemental fat does not appear to be mediated by alterations in lipoprotein composition.     

The effect of human chorionic gonadotrophin on the expression of progesterone receptors in human luteal cells in vivo and in vitro

The human corpus luteum expresses genomic progesterone receptors (PRs) suggesting that progesterone may have an autocrine or paracrine role in luteal function. We hypothesised that the reduction in luteal PR reported in the late-luteal phase augmented progesterone withdrawal and had a role in luteolysis. We therefore tested the hypothesis that luteal rescue with human chorionic gonadotrophin (hCG) would maintain PR expression. PR was immunolocalised to different cell types in human corpora lutea (n = 35) from different stages of the luteal phase and after luteal rescue with exogenous hCG. There was no change in the staining intensity of theca-lutein cell or stromal cell PR throughout the luteal phase or after luteal rescue. In the late-luteal phase, granulosa-lutein cell PR immunostaining was reduced (P < 0.05) but the trend to reduction was also seen after luteal rescue with hCG (P = 0.055). To further investigate the effect of hCG on granulosa-lutein cell PR expression, an in vitro model system of cultured human luteinised granulosa cells was studied. Cells were cultured for 12-13 days exposed to different patterns of hCG and aminoglutethamide to manipulate progesterone secretion (P < 0.0001). Expression of PR A/B and PR B isoforms was examined by quantitative real-time RT-PCR. PR A/B mRNA was lower (P < 0.05) after 11-13 days of culture than after 7 days of culture. This reduction could not be prevented by hCG in the presence (P < 0.05) or absence (P < 0.05) of stimulated progesterone secretion. The expression of PR B mRNA showed a similar pattern (P = 0.054). Simulated early pregnancy in vivo and hCG treatment of luteinised granulosa cells in vitro did not appear to prevent the down-regulation of PR seen during luteolysis.     

Steroidogenic responses of pig corpora lutea to insulin-like growth factor I (IGF-I) throughout the oestrous cycle

This study was designed to investigate the roles of insulin-like growth factor I (IGF-I), IGF-type I receptor (IGF-IR) and IGF-binding proteins (IGFBPs) in regulating progesterone secretion by pig corpora lutea during the oestrous cycle, and the signal transduction pathways involved in mediating the steroidogenic actions of IGF-I. Corpora lutea were collected on days 4, 7, 10, 13 and 15 or 16 of the oestrous cycle, enzyme dissociated and the luteal cells were cultured for 24 h in Medium 199 with IGF-I (0-100 ng ml(-1)), long R(3)-IGF-I (0-100 ng ml(-1)), anti-IGF-I (Sm 1.2B; 0-10 microg ml(-1)), anti-IGF-IR (alphaIR3; 0-2 microg ml(-1)), or IGF-I signal transduction pathway inhibitors (phosphatidylinositol (PI)-3-kinase: 100 nmol Wortmannin l(-1) and 10 micromol LY 294002 l(-1); MAP kinase: 50 micromol PD 98059 l(-1)) to investigate their effects on IGF-I (100 ng ml(-1)) stimulated progesterone secretion. Pig luteal cells displayed dose-dependent responses to IGF-I and long R(3)-IGF-I on days 4 and 7 of the oestrous cycle, but not on days 10-16. There was no difference in the ED(50) or V(max) (maximal response) values between IGF-I and long R(3)-IGF-I. Neither anti-IGF-I nor anti-IGF-IR had significant effects on progesterone secretion, at any dose or day. Wortmannin and LY 294002 blocked IGF-I stimulated progesterone secretion, but PD 98059 was without effect. Finally, IGF-I (6 microg) infused into the ovary on day 7 in vivo significantly increased progesterone secretion within 45 min of infusion. The conclusions of this study are: (1) IGF-I has steroidogenic actions only on 'young' (days 4-7) pig corpora lutea; (2) endogenous IGF-I and IGFBP are insufficient to modulate progesterone secretion in vitro; and (3) the steroidogenic actions of IGF-I are mediated via PI-3-kinase.     

Ovarian function in ewes after treatment with mifepristone early during the oestrous cycle

The aim of this study was to determine whether endogenous progesterone regulates synthesis and secretion of luteal oxytocin. In Expt 1, mature ewes (n = 5 per group) were assigned randomly to control or mifepristone (RU486) treatment groups. Ewes were injected s.c. twice a day with vehicle or 10 mg RU486 on days 5-7 of the oestrous cycle (oestrus = day 0). On day 8, after an i.v. injection with prostaglandin F(2alpha) (250 microg cloprostenol), venous blood samples were collected at frequent intervals to determine plasma oxytocin concentrations. Plasma oxytocin concentrations of RU486-treated ewes were not significantly different from those of control ewes. In Expt 2, ewes were injected s.c. each day with vehicle or 175 mg RU486 on days 2-5 of the oestrous cycle followed by administration of prostaglandin F(2alpha) on day 6. Four of five RU486-treated ewes showed 'split-oestrus' (oestrous behaviour for 36 h and then again at 84-108 h after the onset of initial oestrus). There was no significant difference in mean plasma oxytocin or progesterone concentrations between treatment groups. The mean masses of mature corpora lutea from control and RU486-treated ewes on day 6 of the oestrous cycle did not differ significantly (394.8 +/- 28.8 versus 319.5 +/- 48.3 mg). RU486-treated ewes contained mature corpora lutea, new corpora lutea (two of four ewes) and preovulatory follicles (>or= 10 mm, two of four ewes). The average interoestrous interval for RU486-treated ewes was 9 days more than that for control animals (26.2 +/- 2.9 versus 17 +/- 0.5 days; P < 0.025).     

Administration of vascular endothelial growth factor Trap during the 'post-angiogenic' period of the luteal phase causes rapid functional luteolysis and selective endothelial cell death in the marmoset

The intense angiogenesis characteristic of early corpus luteum development is dependent upon vascular endothelial growth factor (VEGF) as inhibitors of VEGF administered at the peri-ovulatory period suppress endothelial cell proliferation and progesterone secretion. We now report that administration of VEGF Trap, a soluble decoy receptor-based inhibitor, at the mid- or the late luteal phase in the marmoset results in a rapid decline in plasma progesterone. Since vascularisation of the corpus luteum is largely complete by the mid-luteal phase, it suggested that this functional luteolysis involved mechanisms other than inhibition of angiogenesis. A second experiment investigated the role of VEGF in maintaining the integrity of the luteal vasculature and hormone-producing cells. VEGF Trap was administered to marmosets in the mid-luteal phase and ovaries were obtained 1, 2, 4 or 8 days later for localisation of activated caspase-3 staining in the corpus luteum and compared with those obtained 2, 4 and 8 days after administration of control protein. The number of cells with activated caspase-3 staining was significantly increased after administration of VEGF Trap. Dual staining of activated caspase-3 with the endothelial cell marker CD31 showed that at 1 day post-treatment, more than 90% caspase-3-stained cells were vascular endothelium, prior to detection of an increasing incidence in death of hormone-producing cells on days 2 and 4. Staining with CD31 showed that the endothelial cell area was decreased after treatment. By 8 days after treatment, corpora lutea had regressed to varying degrees, while all control corpora lutea remained healthy. These results show that VEGF inhibition in the mid- or the late luteal phase induces functional luteolysis in the marmoset that is associated with premature and selective death of endothelial cells.     

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.     

Potential role for peroxisome proliferator-activated receptor gamma in regulating luteal lifespan in the rat

Peroxisome proliferator-activated receptor gamma (PPARgamma) has been shown to stimulate progesterone production by bovine luteal cells. We previously reported higher expression of PPARgamma in old compared with new luteal tissue in the rat. The following studies were conducted to determine the role of PPARgamma in rat corpora lutea (CL) and test the hypothesis that PPARgamma plays a role in the metabolism of progesterone and/or luteal lifespan. Ovaries were removed from naturally cycling rats throughout the estrous cycle, and pseudopregnant rats. mRNA for PPARgamma and P450 side-chain cleavage (SCC) was localized in luteal tissue by in situ hybridization, and protein corresponding to PPARgamma and macrophages identified by immunohistochemistry. Luteal tissue was cultured with agonists (ciglitazone, prostaglandin J2) or an antagonist (GW-9662) of PPARgamma. Progesterone was measured in media by RIA and levels of mRNA for 20alpha-hydroxysteriod dehydrogenase (HSD) and bcl-2 were measured in luteal tissue after culture by RT-PCR. An inverse relationship existed between the expression of mRNA for SCC and PPARgamma. There was no effect of PPARgamma agonists or the antagonist on luteal progesterone production in vitro, or levels of mRNA for 20alpha-HSD. PPARgamma protein was localized to the nuclei of luteal cells and did not correspond with the presence of macrophages. In new CL, ciglitazone decreased mRNA for bcl-2 on proestrus, estrus, and metestrus. Interestingly, GW-9662 also decreased mRNA for bcl-2 on proestrus and diestrus in old and new CL, and on metestrus in new CL. These data indicate that PPARgamma is not a major player in luteal progesterone production or metabolism but may be involved in regulating luteal lifespan.     

Peripartum changes of plasma and milk vitamin A and beta-carotene among dairy cows with or without mastitis

Over 12 mo we studied the relationship between peripartum concentrations of vitamin A and beta-carotene in blood plasma and milk of 93 Holsteins with or without subsequent mastitis. Blood was sampled daily from 7 days prepartum through 7 days postpartum and on alternate weeks through wk 10 of lactation. Milk samples were collected daily for 7 days postpartum and then biweekly for 10 wk. Somatic cell counts were on biweekly milk samples. Vitamin A and beta-carotene of blood plasma decreased rapidly prepartum to reach minimum concentrations at calving (vitamin A) or on day 4 to 6 postpartum (beta-carotene). Thereafter, both vitamin A and beta-carotene increased rapidly through 10 wk postpartum. Concentrations of vitamin A and beta-carotene in colostrum were higher than concentrations in milk. Cows with mastitis (somatic cells greater than 500,000 cells/ml milk) had lower vitamin A in blood plasma during days 0 to 7 and wk 2 and 4 postpartum than cows without mastitis. When data were analyzed with loge of somatic cell count as an independent regression variable, results were similar. In contrast to vitamin A, peripartum beta-carotene in blood plasma was higher among mastitic cows and was related to higher loge of somatic cell count. No significant difference was observed between mastitic and non-mastitic cows for vitamin A and beta-carotene in milk. Lower concentrations of plasma vitamin A and higher concentrations of beta-carotene during the immediate postpartum period were associated with higher milk somatic cell counts among dairy cows during lactation.     

Early postpartum reproductive profiles in Holstein cows with retained placenta and uterine discharges

Sixty Holstein cows were allocated to three groups. Twenty cows had retained placenta. The remaining cows were examined on d 14 postpartum and those with purulent discharges (n = 22) were assigned to one group and the remaining (n = 18) to a control group. Within each group, cows were given randomly either gonadotropin-releasing hormone (i.m., 200 micrograms) or saline on d 15 postpartum to evaluate the effect on changes in ovarian structures and plasma progesterone through 50 d postpartum and fertility. Corpora lutea were found in control cows by d 21, cows with uterine discharge by d 28, and cows with retained placenta by d 27. Maximum progesterone production during the first luteal phase was higher in control cows than in cows with purulent discharge or retained placenta (4.66 ng/ml compared with 3.23 and 3.34 ng/ml, respectively). Duration of the first corpus luteum was affected by clinical condition. Only 6.2% of cows with retained placenta had corpora lutea of normal duration (12 to 16 d), whereas 43.8 and 50.0% of cows with uterine discharge and control cows had normal postpartum luteal phases. Measures of fertility were not affected by gonadotropin-releasing hormone. Control cows had less days to conception (97) and fewer services per conception (1.6) than cows with retained placenta (134 and 2.5, respectively). Clinical group affected reproduction more than gonadotropin-releasing hormone did, possibly by altering ovarian function.     

Corpora lutea induced by gonadotrophin-releasing hormone treatment of anoestrous Welsh Mountain ewes: reduced sensitivity to luteinizing hormone in vivo and to chorionic gonadotrophin in vitro

Seasonally anoestrous Welsh Mountain ewes received 250 ng gonadotrophin-releasing hormone (GnRH) every 2 h, with (Group 1; n=13) or without (Group 2; n=14) progesterone priming for 48 h. Fourteen control ewes (Group 3) were studied during the luteal phase in the breeding season. Animals in Group 4 (n=12) received progesterone priming followed by 250 ng GnRH at increasing frequency for 72 h, while ewes in Group 5 (n=13) were given three bolus injections of 30 microg GnRH at 90-min intervals. All treatment regimens induced ovulation. However, only corpora lutea (CL) from ewes in Group 3 (breeding season) or Group 4 exhibited normal luteal function. Luteal luteinizing hormone (LH) receptor levels were significantly higher on day 12 than day 4, and CL from groups with adequate CL (3 and 4) had significantly higher (125)I-human chorionic gonadotrophin (hCG)-binding levels than the three groups with inadequate CL on day 12. LH-binding affinity was unchanged. Exogenous ovine LH (10 microg) in vivo on days 3 or 11 after ovulation induced a pulse of progesterone in ewes with adequate CL: however, ewes in Groups 1, 2 and 5 showed no significant response. Basal progesterone secretion in vitro was significantly greater on day 4 than on day 12. Maximal steroidogenic responses of adequate and inadequate CL to hCG and to dibutyryl cyclic-3',5'-AMP were similar at both stages of the luteal phase. However, the EC50 for hCG on days 4 and 12 was 10-fold lower for groups with an adequate CL (0.1 IU hCG/ml) than for inadequate-CL groups (1 IU hCG/ml; P <0.05). Thus, in addition to the well-characterized premature sensitivity of GnRH-induced inadequate CL to endometrial luteolysin, we have shown (1) a marked decrease in total number of cells in the CL, a profound reduction in vascular surface area, and a decrease in mean large luteal cell volume (with no change in large luteal cell numbers), (2) decreased luteal LH receptor and progesterone content compared with adequate CL and (3) that CL that were becoming, or were destined to become, inadequate failed to respond to ovine LH in vivo and were 10-fold less sensitive to hCG in terms of luteal progesterone secretion in vitro.     

Seminal plasma regulates ovarian progesterone production, leukocyte recruitment and follicular cell responses in the pig

Seminal plasma (SP) acts to influence the uterine endometrium after mating, activating synthesis of embryotrophic cytokines and inflammatory changes that condition the tract for embryo implantation and establishing pregnancy. The objective of this study was to investigate in pigs whether the ovary might also be responsive to SP exposure. Prepubertal gilts were synchronised with exogenous gonadotrophins and received transcervical treatment with pooled boar SP or PBS; then the ovarian tissue was recovered at 34 h (preovulation) and on days 5 and 9 after treatment. The ovarian response was assessed by measuring ovulation rate, number and size of corpora lutea, ovarian leukocyte populations, progesterone production in vivo, as well as responses of retrieved granulosa cells cultured in vitro. In SP-treated gilts, leukocyte recruitment into the ovarian tissues was increased fourfold at 34 h, with macrophages comprising the most abundant cell lineage. There was no effect of SP on the number of oocytes ovulated; however, the weight of corpora lutea was increased in SP-treated gilts. SP also induced an increase in plasma progesterone content seen from day 5 to at least day 9 after treatment. In addition, granulosa cells and thecal tissue retrieved from preovulatory follicles of SP-treated gilts were more responsive in vitro to growth factor- and gonadotrophin-stimulated cell proliferation and progesterone synthesis. These results suggest that uterine exposure to SP influences immune cell trafficking in the ovary and enhances steroidogenesis in early pregnancy. The effects of SP on ovarian function potentially contribute to reproductive success in the pig.     

The human corpus luteum: which cells have progesterone receptors?

Studies comparing the regressing corpus luteum with the rescued corpus luteum have demonstrated that human chorionic gonadotrophin (hCG) has effects on cell types that do not express hCG receptors. As progesterone synthesis is hCG dependent and the corpus luteum has been shown to express genomic progesterone receptors, progesterone is a candidate molecule for these paracrine effects. This study aimed to define the cellular localisation of progesterone receptors in the human corpus luteum using dual-staining immunohistochemistry for genomic progesterone receptors and specific cellular markers. Well-characterised corpora lutea (n = 12) from different stages of the luteal phase were studied. The same distribution was observed in all corpora lutea examined. The steroidogenic cells (3beta-hydroxysteroid dehydrogenase positive) and both theca-lutein (17alpha-hydroxylase positive) and granulosa-lutein (aromatase positive) express progesterone receptors, as do stromal fibroblasts (vimentin positive, fibroblast antigen positive). Vascular endothelial cells (CD31 positive), pericytes (alpha-smooth muscle actin positive), macrophages (CD68 positive) and fibroblasts within the central clot do not express nuclear progesterone receptors. Progesterone is a candidate messenger molecule for the effects of hCG on the matrix metalloproteinase-producing stromal fibroblasts. Some of the effects of hCG on steroidogenic cells may be mediated by progesterone, but its effects on blood vessels and macrophages require alternate paracrine signalling mechanisms. In addition, there appears to be at least two fibroblast populations in the corpus luteum.     

Immunolocalization of steroidogenic enzymes in the corpus luteum and placenta of the Japanese black bear, Ursus thibetanus japonicus, during pregnancy

The Japanese black bear, Ursus thibetanus japonicus, is a seasonal breeder and shows delayed implantation for several months during pregnancy. The objective of this study was to clarify the steroidogenic capability of the corpus luteum and placenta during pregnancy, including both delayed implantation and fetal development, by immunolocalization of steroidogenic enzymes in these organs of the Japanese black bear. Ovaries and placentae from 15 wild Japanese black bears, which had been killed legally by hunters and were thought to be pregnant, were used in an immunocytochemical study to localize the cholesterol side chain cleavage cytochrome P450 (P450scc), 3beta-hydroxysteroid dehydrogenase (3betaHSD), 17alpha-hydroxylase cytochrome P450 (P450c17) and aromatase cytochrome P450 (P450arom) by the avidin-biotin-peroxidase complex method using polyclonal antisera raised in mammals against P450scc, 3betaHSD, P450c17 and P450arom. P450scc and 3betaHSD were localized in all luteal cells throughout pregnancy. P450c17 was present in a few luteal cells, especially in the outer area of the corpus luteum throughout pregnancy, but the number of positively immunostained cells decreased during the post-implantation period. Cells positively immunostained for P450c17 were significantly smaller than negatively immunostained cells (P < 0.01). P450arom was present sporadically in a few luteal cells throughout pregnancy, but the number of positively immunostained cells decreased during the post-implantation period. The size of cells positively immunostained for P450arom was not significantly different from that of negatively immunostained cells. The whole placenta was negatively immunostained for P450scc, 3betaHSD and P450c17, but P450arom was present in the syncytiotrophoblasts and endothelial cells of maternal blood vessels. These results indicate that, in the Japanese black bear, corpora lutea are a source of progesterone which may play an important role in the maintenance of delayed implantation and fetal development during pregnancy. Corpora lutea have a minimum capability to synthesize androgen in small luteal cells and oestrogen in normal-sized luteal cells during pregnancy, and placentae have the ability to synthesize oestrogen during late pregnancy.     

Effect of nitric oxide on the expression of insulin-like growth factors and the insulin-like growth factor binding proteins throughout the lifespan of the human corpus luteum

The presence of insulin-like growth factors (IGF), IGF binding proteins (IGFBP) and IGF receptor type 1 (IGF-IR) in the human corpus luteum was investigated by examining the expression and production of related proteins throughout the lifespan of the corpus luteum and the action of nitric oxide upon their production. The expression of proteins in corpora lutea from the early, mid-and late luteal phases was assessed by immunohisto-chemistry, evaluated by a semi-quantitative analysis and the functional study was performed in corpus luteum explants incubated with nitric oxide donors. IGF-I and -II and IGFBP-1 and -3 were measured in the culture media by specific immunoassays. The results showed that IGF-I and -II, IGFBP-1 to -6 and IGF-IR were detected in the human corpus luteum throughout the luteal phase. Moreover, the expression and production of IGF-I and IGFBP-1 increased progressively from corpora lutea from the early to late luteal phases (P < 0.05), whereas the expression and production of IGFBP-2, -4 and -5 were significantly higher in corpora lutea from the mid-luteal phase (P < 0.05). No differences were observed in the expression of IGF-II, IGFBP-3 and -6 and IGF-IR throughout the lifespan of the corpus luteum. However, functional studies showed that nitric oxide donors elicited a stimulatory action on production of IGF-I in corpora lutea from the early luteal phase (80%) and on production of IGFBP-1 in corpora lutea from the late luteal phase (50%) (P < 0.05), whereas production of IGF-II and IGFBP-3 was not affected by nitric oxide. In conclusion, the components of the IGF-IGFBP system are expressed in the human corpus luteum throughout its lifespan. Nitric oxide regulates IGF-I and IGFBP-1 production, indicating that the growth factors may serve, at least in part, as mediators of the action of nitric oxide in the human corpus luteum.     

Effect of presence or absence of corpora lutea on milk production in East Friesian dairy ewes

The potential luteal effects on milk production were examined in dairy ewes that were not superovulated in contrast to studies using superovulated ewes. Lactating East Friesian crossbred ewes (n = 24) were synchronized using intravaginal progesterone (controlled intravaginal drug-releasing device), PGF2alpha, and gonadotropins. After ovulation, corpora lutea (CL) were counted via laparoscopy on d 4 and 11. On d 5, ewes received either saline (CLYES, n = 12) or PGF2alpha (CLNO, n = 12) to allow CL persistence (2.4 +/- 0.3 CL on d 11) or regression (0 CL on d 11), respectively. Each ewe received two CIDR d 5 to 18 to maintain high concentrations of plasma progesterone (P4) and to suppress estradiol (E2). Each ewe received PGF2alpha on d 18. Data were collected during three periods (pretreatment: d 0 to 5; treatment: d 6 to 18; posttreatment: d 19 to 25). Milk yield and milking time were recorded daily, milk samples were obtained for analyses of fat and protein, and blood samples were collected for P4 and E2 immunoassay. During treatment, CLYES ewes had higher milk yield (1.56 vs. 1.44 +/- 0.01 kg/d), milk fat (92.2 vs. 81.1 +/- 1.3 g/d), and milk protein (83.7 vs. 77.5 +/- 0.8 g/d) compared with CLNO ewes, respectively. Differences were maintained posttreatment, despite luteolysis in CLYES ewes. Estradiol concentrations did not differ between treatments and were low after d 5. Milk production was increased in East Friesian ewes in the presence of an average of 2.4 corpora lutea, an effect independent of estradiol.     

Use of concentrations of progesterone and estradiol-17 beta in milk in monitoring postpartum ovarian function in dairy cows

Data from artificial insemination, rectal palpation, and hormone assays were used to characterize postpartum reproductive activity in 54 dairy cows. Progesterone and estradiol-17 beta were measured in milk samples collected for 120 d (Trial 1) or 65 d (Trial 2). Progesterone was higher and estradiol was lower in milk than in serum. Values for both hormones in milk were highly correlated with those in serum. Most cows (64%) had short first luteal phases (less than or equal to 12 d). First rise (28 d) in progesterone was later (33.4 vs. 24.9 d) for cows having short rather than normal (greater than 12 d) luteal phases. Cows were classified as having a short luteal phase followed by a normal luteal phase or as having normal luteal phases for the first two estrous cycles. Estradiol for the 6 d prior to each luteal phase was higher preceding the second phase than the short phase or those preceding both phases of cows with normal phases. Follicular function prior to ovulation, as measured by estradiol, was not responsible for short-lived corpora lutea. Concentrations of progesterone in milk in the late luteal phase prior to insemination were related to fertility.