Biomedical subjects

D Schams

Publications and source records attributed to D Schams.

At least 37 records · Page 2Linked to original sources

PubMed2002

Steroids as local regulators of ovarian activity in domestic animals.

The presented overview gives clear evidence for steroids as local regulators of follicular and luteal activity. In the follicle, estrogen receptor-alpha (ERalpha) and ERbeta expression are demonstrated in cow, ewe and pig. Besides species specific effects in general, there is evidence that estradiol-17beta (E(2)) exerts a dose-dependent inhibition on the secretion of progesterone (P(4)) by both theca interna cells (TI) and granulosa cells (GC). GC enhance the ability of the TI to produce androstendione by supplying them with progestin precursor. Androgen produced by TI enhances the ability of the GC to make E(2), and high concentrations of E(2) in the preovulatory follicle inhibit 3beta-HSD in both TI and GC and thus, may promote the use of the pathway Delta(5) for TI androgen production. The authors suggest that E(2) acts within the follicle to exert positive feedback on androgen and E(2) production, and exerts mitotic and anti-atretic or anti-apoptotic effects on follicular cells. Parts of the E(2)-mediated local action are regulated by stimulating effects on hormone receptors (LH, FSH, oxytocin). Gap junctions permit transfer of nutrients and cytokines to and from the avascular GC and oocyte, and formation is stimulated by estrogens. In bovine corpus luteum (CL) there is evidence that P(4) may directly regulate the production of P(4), oxytocin and prostaglandins (PGs) in a cycle dependent fashion. In most of domestic animal species, there is clear evidence for CL production of E(2) with clear stimulatory and luteotropic effects on P(4), and an intraluteal circuit that involves paracrine effects of E(2), oxytocin and PGF(2alpha) (especially in pigs). In contrast, there are species (ruminants, mares) in which the evidence for important local effects of E(2) is less clear, although expression of ERalpha, ERbeta and progesterone receptor (PR) is documented. Progesterone is very important for the regulation of CL lifetime by effects on the endometrium and release of the luteolytic signal PGF(2alpha). In conclusion, steroids as local regulators of ovarian activity are now documented and may stimulate further research in this field.

Animals↗

PubMed2002

Effect of dietary beta-carotene on the early embryonic development and uterine fluid composition of gilts.

The role of beta-carotene in reproduction was investigated in gilts assigned to three dietary supplementations: VA (4000 IU vitamin A); VA + VA (4000 IU + 8300 IU); VA + BC (4000 IU + 100 mg beta-carotene) per kg diet for 14 weeks. Gilts were slaughtered at day 12 of gestation. In the VA + BC group, number of corpora lutea was lowest, but the number of embryos was greatest, resulting in a non-significant decreased prenatal mortality (p<0.07). The proportion of less developed spherical and tubular embryos compared with filamentous was greatest in the VA + BC group (p<0.01). No differences were observed for vitamin A and retinal binding protein (RBP) in the uterine fluid. When animals were grouped according to the development of blastocysts, vitamin A and RBP levels were higher in the VA + BC group with only filamentous embryos (p<0.01). This indicates that the supplementation of beta-carotene to gilts might affect embryonic losses possibly because of slower alterations in the uterine environment, resulting in a higher and less variable number of embryos, despite an apparently more heterogeneous development.

Animal Feed↗

PubMed2001

The effects of conditioning to suckling, milking and of calf presence on the release of oxytocin in dairy cows.

The aim of the study was to record the oxytocin (OT) release during milking (M) without or with calf presence, suckling (S) and finally calf removal just before the next milking in cows during postpartum or early lactation periods. Furthermore, the release of OT was examined during S and M in unknown surroundings (parlour). A total of 20 Brown Swiss cows kept under loose housing environment were used in our experiment. In both periods, the cows were milked twice daily at 07.00 and 18.00h and suckled three times daily at 09.00, 14.00 and 20.00h in the stall (tie housing). In the postpartum period, 13 cows were suckled and milked in the presence of their calves in the stall for the first 5 days of postpartum. Five from seven primiparous cows were additionally suckled by their own calves in the parlour on day 5 at 20.00h. On day 6, calves were separated and moved from mothers to the calf barn 10min before morning M. After evening milking cows were relocated to the herd within the same stable and milked in the parlour for a period of 4 weeks without contact to their calf. For control, additional seven primiparous cows without calf presence (not suckled) were also milked in the tie housing. In the early lactation period, suckled cows were moved back to the tie housing 2 days before the start of two consecutive days of S by their own calves and milking. Afterwards, 10min before M calves were separated again. Before S, two consecutive M were considered as controls. Results: The S stimulus during postpartum resulted in a higher OT release as compared with M in the calf presence and M after calf separation but not during M of not suckled cows. S in parlour reduced OT release. However, when not suckled primiparous cows were first milked in parlour, OT release was more reduced and in some cows total inhibition was observed. In early lactation during the first S, release of OT was lower than during control M, but increased gradually with repeated S and reached a maximum already on the second day. After two S, during evening M, the M related OT release was reduced as compared with controls. Calf removal 10min before M reduced OT release as compared with control M or M in calf presence. In conclusion, the calf presence and its removal can negatively influence OT secretion during M. Conditioning to machine milking caused a short-term reduction of OT release during first suckling, which normalised within 1 day.

Journal Article↗

PubMed2001

Naloxone cannot abolish the lack of oxytocin release during unexperienced suckling of dairy cows.

To evaluate the role of opioids for the regulation of oxytocin release in response to teat stimulation, 10 brown-Swiss dairy cows were randomized to two experiments during mid of lactation. In the first experiment, four cows without previous suckling experience were suckled by an alien calf between two normal milkings. Before and during milking or suckling, frequent blood samples were collected via a jugular cannula for determination of oxytocin and beta-endorphin. In the second experiment, six cows were treated with naloxone or saline, 10min before the start of the first or second suckling, respectively. The collected blood samples were assayed for oxytocin.In the first experiment, the plasma levels of beta-endorphin were elevated during and after the unexperienced suckling in three cows, but not in the fourth cow, and the release of oxytocin during suckling was markedly reduced, suggesting no release of alveolar milk. In the second experiment, the release of oxytocin during suckling was again significantly reduced. Pretreatment with naloxone before suckling did not completely abolish the adverse effect of suckling and the oxytocin plasma level did not increase to levels comparable with control milking.In emotional stress situations, the release of oxytocin from the pituitary is inhibited with simultaneously elevated beta-endorphin plasma levels. Although there is some evidence for a regulatory role of opioids for the release of oxytocin, other mediators are suggested to be more potent in regulating oxytocin under stress conditions.

Journal Article↗

PubMed2001

Growth hormone, but not luteinizing hormone, acts with luteal peptides on prostaglandin F2alpha and progesterone secretion by bovine corpora lutea in vitro.

Prostaglandin F2alpha (PGF2alpha) is a major physiological luteolysin in the cow. However, injection of PGF2alpha before day 5 (day 0 = estrus) of the estrous cycle dose not induce luteolysis. On the other hand, the early corpus luteum (CL) actively produces PGF2alpha. This indicates that luteal PGF2alpha may play a key role in the refractoriness to PGF2alpha injected during the early luteal phase when angiogenesis is active in the CL. Thus, this study aimed to investigate the possible interaction between pituitary hormones and local factors (luteal peptides) on secretion of PGF2alpha and progesterone (P) by the early bovine CL, and to evaluate the effect of growth hormone (GH) as well as its interactions on production of PGF2alpha in the developing CL. A RT-PCR analysis revealed that mRNA for GH receptor in CL was fully expressed from early in the luteal phase throughout the estrous cycle, while luteinizing hormone (LH) receptor mRNA was expressed less by the early and regressing CL than those at mid or late luteal phases (P < 0.05). For the stimulation test, an in vitro microdialysis system (MDS) was used as a model. Each bovine early CL (days 3-4) was implanted with the MDS, and maintained in an organ culture chamber. The infusion of GH, insulin-like growth factor-I (IGF-I) and oxytocin (OT) increased (P < 0.05) PGF2alpha and P release. In contrast, LH had no effect (P > 0.05) on PGF2alpha secretion and little effect on P release. Unexpectedly, there was no distinct interaction between pituitary hormones and luteal peptides on secretion of PGF2alpha and P. These results indicate that GH is a more powerful stimulator of PGF2alpha and P production in the early bovine CL than LH and suggest that GH and luteal peptides, IGF-1 and OT, contribute to maintenance of elevated PGF2alpha production in the developing bovine CL.

Animals↗

PubMed2001

Inhibition of oxytocin release and milk let-down in postpartum primiparous cows is not abolished by naloxone.

About 10% of primiparous cows have no milk ejection during the first milkings after delivery. Therefore, 17 Brown Swiss dairy cows in their first lactation were used to evaluate the extent of disturbed milk let-down and the corresponding oxytocin (OT) plasma values in the 1st 5 days after delivery. The first milking was 9-22 h after parturition and served for classification of the cows to groups with inhibited (INH), bimodal (BIMO), or normal (NOR) milk let-down. The OT plasma levels before the start of manual teat stimulation and machine milking were comparably high during the first milking especially in NOR and BIMO cows. Ten minutes before the second milking (M2), 300 mg of the opioid antagonist naloxone was injected to test whether the disturbance was affected by the action of endogenous opioids on the neurohypophysis. The milk yield was not influenced by the naloxone treatment, and the INH cows had milk ejection only after a vaginal stimulation. Afterwards, the cows were milked twice every day, until the milk let-down and the OT release were unaffected (equal to control milking). Then, at the next milking, the cows were injected with 300 mg morphine 10 min before milking. The central OT release in response to manual teat stimulation and machine milking was completely blocked in all cows, but a vaginal stimulation was able to abolish this block, at least partially, in 16 cows. Thus, morphine produced a milk let-down characteristic as in the INH cows during the first three milkings. For the following milking, the cows were pre-treated with 300 mg naloxone (-15 min) plus 300 mg morphine (-10 min) before milking. The OT release and the milk yields were unaffected when compared with the control milking. This experiment demonstrates that exogenous opioids can affect the central release of OT in a naloxone-reversible manner even very soon after parturition. However, endogenous opioids are probably not the main mediators of disturbed central OT release and alveolar milk ejection in post-partum primiparous cows.

Animals↗

PubMed2001

Uterine responses to exogenous oxytocin before and after pre-partum luteolysis in the cow.

The aim of this study was to test the functional status of uterine oxytocin receptors in cows in vivo around parturition. The animals received consecutive, intra-arterial injections of 800, 1,600 and 3,200 mU of oxytocin at three different stages: during late gestation (days 260-274), at 12 h and at 24 h after intramuscular injection of a prostaglandin F2alpha analogue at day 275 to induce parturition. Cows (n = 6) had been provided with myometrial electrodes and a catheter had been installed in the aorta and in a branch of the uterine vein (UV). Regular blood samples were obtained from the UV from 5 min before until 45 min after each oxytocin injection to measure plasma levels of prostaglandin F2alpha (PGF2alpha) and oxytocin. Uterine electromyographic (EMG) activity was registered continuously during each experiment. The increase of oxytocin levels in UV plasma after intra-arterial injections was dose dependent (p < 0.02). Pre- and post treatment oxytocin levels at 24 h after induction of parturition were significantly increased (p = 0.0313). Both during late pregnancy and at 12 h after induction of parturition, oxytocin caused a significant increase in EMG activity (p = 0.022). After the 3,200 mU dose the increase was significantly higher than with the other 2 doses (p = 0.004). After each dose, EMG activity returned to baseline levels within some 15 min. At 24 h after induction of parturition, the pre-treatment level of EMG activity had increased. Doses of 800 mU and 1,600 mU of oxytocin produced a significant (p = 0.022) increment of EMG activity, which was of the same magnitude as during the preceding stages; after 3,200 mU of oxytocin the response was significantly higher than before (p = 0.008). No significant increases of PGF2alpha levels in UV plasma could be measured after oxytocin injections at any of the three stages. It is concluded that the myometrium of the pregnant cow responds in vivo to physiological doses of oxytocin. At 24 h after induction of parturition, when luteolysis has occurred and a parturient pattern of parturient myometrial activity has already started to develop, the response is enhanced. Physiological doses of oxytocin did not evoke a spurt release of PGF2alpha in uterine venous blood during the peripartal period.

Animals↗

PubMed2001

Stimulatory and synergistic effects of luteinising hormone and insulin like growth factor 1 on the secretion of vascular endothelial growth factor and progesterone of cultured bovine granulosa cells.

Vascular endothelial growth factor (VEGF) is the most important factor in the regulation of angiogenesis. Associated with luteinisation and formation of corpus luteum (CL) are alterations in luteal vascularity. The aim of the study was to test under in vitro conditions the stimulation of VEGF and progesterone (P) secretion of bovine granulosa cells by LH, IGF1 (insulin like growth factor) or by factors known to be produced by luteinised granulosa cells or in the early CL. Localisation of VEGF protein in preovulatory follicle and early CL were achieved by immunohistochemistry. LH and IGF1 stimulated dose dependently and significantly P and VEGF when tested alone. Both hormones added simultaneously had clear additive and even more interesting far greater (synergistic) effects on P with LH (0.1 ng/ml) plus 5 or 10 ng IGF1. In contrast, VEGF was stimulated only additively with 0.1 ng/ml of LH plus 5 or 10 ng IGF1. But with the higher dose of LH (1 ng/ml) additionally to the additive effect a tendency for a synergistic action (which was significant with 1 ng LH plus 5 ng IGF1/ml) was observed. Endothelin, oxytocin, progesterone, atrial natiuretic peptide, angiotensin II, prostaglandin F2 alpha alpha, prostaglandin E2, cortisol, fibroblast growth factor 1 and 2 and growth hormone showed no effect neither on P nor on VEGF. Tumour necrosis factor alpha (TNF alpha) stimulated (P < 0.05) VEGF with 10 or 100 ng/ml but not P. TPA (12-0 tetra decaenoyl-phorbol-13-acetate) or Ca2+ ionophore did not show a stimulatory effect in contrast to forskolin which increased P and VEGF secretion dose dependently. The VEGF protein was localised in follicle (granulosa cells, theca cells and some endothelial cells) and early (about 24 h after ovulation) CL (granulosa-lutein cells and endothelial cells). The same signalling pathway by stimulation of cAMP production and proteinkinase A activation for luteinisation and neo-vascularisation demonstrates a close temporal and spatial relationship of these normal physiological processes.

Animals↗

PubMed2001

Estradiol-17beta is produced in bovine corpus luteum.

The aim of this study was to investigate the expression of cytochrome P450 aromatase (aromatase) mRNA, its activity, and estradiol-17beta (estradiol) secretion in bovine corpus luteum (CL) during the estrous cycle. Expression of aromatase mRNA was examined in CL at the early, mid, late, and regressed luteal stages by using a reverse transcription-polymerase chain reaction. Aromatase mRNA was detected in all luteal stages examined, although aromatase expression was significantly lower during the early and regressed luteal phases compared to the mid and late luteal phases. Moreover, cultured midluteal cells clearly converted exogenous [(3)H]androstenedione into estradiol, and an aromatase inhibitor significantly inhibited this conversion. To characterize the local release of estradiol within the CL during the estrous cycle, an in vitro microdialysis system (MDS) of CL was conducted. Estradiol in MDS perfusate was confirmed by a reverse-phase high-performance liquid chromatography in combination with enzyme immunoassays. Basal release of estradiol from microdialyzed CL did not change during the estrous cycle. Additionally, when freshly prepared midluteal cells were exposed to estradiol (10(-14) to 10(-9) M), estradiol stimulated prostaglandin (PG) F(2alpha) secretion (P < 0.05), although it did not affect progesterone and oxytocin secretion. The overall results indicate that estradiol is produced locally in bovine CL throughout the estrous cycle, and they suggest that estradiol plays a role in regulating PGF(2alpha) production in CL as an autocrine/paracrine factor.

Androstenedione↗

PubMed2001

The expression of the IGF family and GH receptor in the bovine mammary gland.

To study the involvement of the IGFs in mammary development and lactation of the cow, the temporal expressions of IGF-I and -II, its receptor type 1 (IGFR-1), IGF-binding proteins (IGFBPs)-1 to -6 and GH receptor (GHR) mRNA were examined. This was carried out for different stages of mammogenesis, lactogenesis, galactopoiesis and involution in the bovine mammary gland of 26 animals. Furthermore, IGF-I was localised by immunohistochemistry. The highest mRNA concentrations for IGF-I were detected in the mammary tissue of late pregnant heifers (days 255-272) and significantly lower expression was detected during lactogenesis and galactopoiesis. Immunohistochemistry of IGF-I revealed only a weak staining in the epithelium of the ducts during mammogenesis. The epithelium of the alveoli were negative during mammogenesis, lactogenesis and galactopoiesis but displayed distinct IGF-I activity during involution. In the stroma a distinct staining of the cytoplasm of adipocytes and of vascular smooth muscle cells was observed. A certain percentage of fibroblasts (usually 20-30%) were also immunopositive. In contrast, highest expression for IGFR-1 was detected during galactopoiesis and involution. The lowest mRNA concentration for IGFR-1 was found during pregnancy (days 194-213). In general, the expression of IGF-II was not regulated during mammogenesis and lactation, but decreased during involution. The mRNA for the six binding proteins was detected in the bovine mammary gland. The dominant binding proteins were IGFBP-3 and -5. The highest expression of IGFBP-3 was observed during mid-pregnancy and the lowest during late lactation, involution and in non-pregnant heifers. The mRNA for IGFBP-5 increased during late mammogenesis and lactogenesis followed by a decrease thereafter. In general, the mRNA concentrations for IGFBP-2, -4 and -6 were barely detectable during all stages. In contrast, the expression for IGFBP-1 was upregulated in the mammary gland of virgin heifers and increased around the onset of lactation. mRNA for GHR was found during all stages examined without outstanding fluctuations. In conclusion, locally produced IGF-I and -II may mediate mammogenesis. The high mammary IGFR-1 mRNA during lactation suggests a role for peripheral IGF-I in maintenance of lactation. The role of IGFBPs in the mammary gland needs further evaluation.

Animals↗

PubMed2001

Production and localisation of angiotensin II in the bovine early corpus luteum: a possible interaction with luteal angiogenic factors and prostaglandin F2 alpha.

The newly formed corpus luteum (CL) rapidly develops after ovulation and has the features of active vascularisation and mitosis of steroidogenic cells. These stage-specific mechanisms also may contribute to gain the function of prostaglandin F2 alpha (PGF2 alpha)-resistant CL at this stage. Recent studies suggest that the vasoactive peptide angiotensin II (Ang II) regulates luteal function. Thus, this study aimed to investigate (i) the expression of angiotensin-converting enzyme (ACE) mRNA by RT-PCR and the ACE protein expression by immunohistochemistry, (ii) the effects of angiogenic growth factors, basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF), on the secretion of Ang II, PGF2 alpha, progesterone and oxytocin (OT), and (iii) the effects of luteal vasoactive peptides (Ang II and endothelin-1 (ET-1)) or OT on the secretion of PGF2 alpha, progesterone and OT from bovine early CL (days 3--4 of the oestrous cycle), and evaluate a possible interaction of these substances with PGF2 alpha. The expression of mRNA for ACE was found in theca interna of mature follicle, early CL and endothelial cells from developing CL as well as pituitary and kidney, but granulosa cells of mature follicle were negative. The immunohistochemical analysis revealed that blood capillaries (endothelial cells) were stained for ACE, but luteal cells were negative in early CL. To examine the effects of substances on the secretory function of the CL, an in vitro microdialysis system was used as a model. The infusion of bFGF and VEGF stimulated Ang II and PGF2 alpha secretion as well as progesterone, but not OT secretion in early CL. The infusion of Ang II after PGF2 alpha infusion continued the stimulatory effect on progesterone and OT release within early CL until 3 h thereafter. However, the infusion of ET-1 alone had no effect on progesterone or OT release. The infusion of luteal peptides such as Ang II and OT stimulated PGF2 alpha secretion, whereas the infusion of ET-1 did not. In conclusion, the overall results of this study indicate that a functional angiotensin system exists on the endothelial cells of early CL, and that angiogenic factors bFGF and VEGF upregulate luteal Ang II and PGF2 alpha secretion, which fundamentally supports the mechanism of progesterone secretion in bovine early CL. This idea supports the concept that the local regulatory mechanism involved in active angiogenesis ensures the progesterone secretion in the developing CL in vivo.

Angiotensin II↗

PubMed2000

Expression and localization of growth factors during mammary gland development.

Growth and differentiation of the mammary gland during development and lactation are controlled by complex hormonal mechanisms. Additionally growth factors are supposed to act as local mediators of the hormonally controlled developmental processes. Mammary tissue for this study was obtained from non pregnant control heifers, primigravid heifers (second part of pregnancy), around parturition, during lactation (early and late) and from dry cows. Using RT-PCR and ribonuclease protections assay (RPA) the expression of the following growth factors was studied in the different phases bovine mammary gland development: Insulin-like growth factor I (IGF-I), insulin-like growth factor II (IGF-II), fibroblast growth factor 1 (FGF-I), fibroblast growth factor 2 (FGF-2), transforming growth factor alpha (TGF-alpha). Additionally the expression of fibroblast growth factor receptor (FGFR) and growth hormone receptor (GHR) was investigated. The cellular distribution pattern of several of these growth factors and GHR was obtained using Immunocytochemical techniques. The detailed expression and localization pattern of these growth factors are presented and their role in the local regulation of the bovine mammary gland is briefly discussed.

Animals↗

PubMed2000

Cortisol and ACTH release in dairy cows in response to machine milking after pretreatment with morphine and naloxone.

The aim ot the study was to examine the effect of morphine and naloxone pretreatment on cortisol and ACTH concentrations in response to machine milking in dairy cows. In the first part of the experiment, the effects of i.v. morphine doses (0, 21, 70 and 210 mg, one dose each day) 10 min before morning milking were studied in six Brown Swiss dairy cows. In the second part, four cows were treated 1 d after the control milking with 210 mg morphine at 10 mm before milking and the next day with 210 mg naloxone at 15 min before milking followed by 210 mg morphine at 10 min before milking. In addition, four other cows were treated 1 d after the control milking with 210 mg naloxone at 10 min before milking. Pretreatment with morphine significantly suppressed the machine milking-induced increase of cortisol in blood plasma as compared with controls. Naloxone pretreatment overcame the inhibitory effect of morphine and elevated milking-induced cortisol concentrations. Naloxone administration alone significantly increased cortisol concentration resulting from milking as compared with controls. However, ACTH concentrations did not change in either control or treated animals, suggesting an ACTH-independent release mechanism for cortisol during milking. We conclude that the release of cortisol in response to machine milking seems to be modulated by emidogenous opioids at the adrenal level and does not appear to be under the control of ACTH.

Adrenocorticotropic Hormone↗

PubMed2000

Effects of morphine and naloxone on the release oxytocin and on milk ejection in dairy cows.

The aim of this study was to investigate the action of opioids (the mu receptor agonist morphine) and the antagonist naloxone on inhibition of oxytocin release and milk let-down in response to milking in dairy cows. In the first experiment, cows were injected with 0, 21, 70 and 210 mg morphine 10 min before milking on four successive days. Plasma oxytocin levels after 1 min manual stimulation of the udder were reduced by 70 and 210 mg morphine, and milk let-down was inhibited at the latter dose. In the second experiment, cows were injected after a control milking with 210 mg morphine (or 350 mg at 10 min before milking the following day if not effective) to inhibit milk flow. On the following day the inhibiting dose of morphine was given with 210 mg naloxone. Naloxone injection given before morphine had no effect on plasma oxytocin concentrations, but abolished the inhibition of oxytocin release by morphine and potentiated oxytocin release in response to milking. Naloxone alone injected the day after control milking increased oxytocin levels during milking, suggesting involvement of the opioid system in milking. A model has been developed for the control of opioid effects during milking. Morphine suppressed oxytocin release during milking in a dose-dependent manner and the effect was reversible by naloxone.

Animals↗

PubMed2000

Regulation of angiotensin II production and angiotensin receptors in microvascular endothelial cells from bovine corpus luteum.

Recent findings suggest that the ovarian renin-angiotensin system regulates ovarian function through the paracrine/autocrine actions of angiotensin (Ang) II. The aims of this study were to investigate 1) the endothelial cell capacity to convert Ang I to Ang II, 2) the effects of endocrine and paracrine/autocrine factors on Ang II production in microvascular endothelial cells (MVE) derived from the developing corpora lutea (CL), and 3) the relationship between Ang II peptide concentration and expression of mRNA for angiotensin type 1 and 2 receptors (ATR1 and AT2R) in the bovine CL at different stages of the estrous cycle. When Ang I was added to the MVE at a concentration of 10(-9) M, it was converted to Ang II (21%). The production of Ang II from Ang I time-dependently rose for 24 h. Addition of captopril (an inhibitor of Ang-converting enzyme [ACE]) to the MVE cultures significantly inhibited Ang II production from 6 h to 24 h (P < 0.05). Addition of estradiol-17beta (E(2)) + vascular endothelial growth factor and E(2) + basic fibroblast growth factor to MVE cultures increased Ang II production, whereas E(2) or growth factors alone had no effect. Specific transcription for AT1R and AT2R was detected in bovine CL and MVE. There were no significant changes in Ang II tissue concentration or AT1R mRNA expression using reverse transcription-polymerase chain reaction during the estrous cycle. In contrast, AT2R mRNA expression decreased during the midluteal phase (P < 0.05) and increased to the highest level during the late luteal phase (P < 0.05). Results demonstrated that Ang II is generated from Ang I in MVE isolated from the developing bovine CL, indicating that MVE have ACE activity. In addition, mRNA expression for Ang II receptors was detected in the bovine CL and the luteal MVE. These results suggest that Ang II is produced by actions of the local renin-angiotensin system, at least in part, on MVE in the bovine CL, and that this peptide may be involved in the regulation of luteal function during early development and luteolysis.

Angiotensin I↗

PubMed2000

Tumor necrosis factor-alpha and its receptor in bovine corpus luteum throughout the estrous cycle.

The objective of this study was to investigate tumor necrosis factor alpha (TNF-alpha) expression, the presence of functional TNF-alpha receptors, and expression of TNF receptor type I (TNF-RI) mRNA in the bovine corpus luteum (CL) during different stages of the estrous cycle. Reverse transcription (RT)-polymerase chain reaction (PCR) showed no difference in TNF-alpha mRNA expression during the estrous cycle. Concentrations of TNF-alpha in the CL tissue increased significantly from the mid to the late luteal stage and decreased thereafter (P < 0.05). An RT-PCR analysis showed higher levels of TNF-RI mRNA in CL of Days 3-7 than of other stages (P < 0.05). (125)I-TNF-alpha binding to the membranes of bovine CL was maximal after incubation at 38 degrees C for 48 h. The binding was much greater for TNF-alpha than for related peptides. A Scatchard analysis revealed the presence of a high-affinity binding site in the CL membranes collected at each phase of the estrous cycle (dissociation constant: 3.60 +/- 0.58-5.79 +/- 0.19 nM). In contrast to TNF-RI mRNA expression, the levels of receptor protein were similar at each stage of the estrous cycle. When cultured cells of all luteal stages were exposed to TNF-alpha (1-100 ng/ml), TNF-alpha stimulated prostaglandin F(2alpha) and prostaglandin E(2) secretion by the cells in a dose-dependent fashion (P < 0.01), especially during the early luteal phase, although it did not affect progesterone secretion. These results indicate the local production of TNF-alpha and the presence of functional TNF-RI in bovine CL throughout the estrous cycle, and suggest that TNF-alpha plays some roles in regulating bovine CL function throughout the estrous cycle.

Animals↗

PubMed2000

Expression and tissue concentration of vascular endothelial growth factor, its receptors, and localization in the bovine corpus luteum during estrous cycle and pregnancy.

The presence of vascular endothelial growth factor (VEGF) in the ovary has been reported in a number of species. The objective of the present study was to demonstrate the expression of VEGF, VEGF receptor (R)-1, and VEGFR-2 in detail by different methodological approaches in bovine corpora lutea (CL) obtained from different stages of the estrous cycle and during pregnancy. VEGF and VEGF receptor transcripts were analyzed by reverse transcription-polymerase chain reaction (RT-PCR) and ribonuclease protection assay. All components of the VEGF system were found in the bovine CL during the estrous cycle and pregnancy. Analysis of VEGF transcript by RT-PCR shows that CL tissues expressed predominantly the smallest isoforms (VEGF(121) and VEGF(165)). The highest mRNA expression for VEGF and VEGFR-2 mRNA was detected during the early luteal phase, followed by a significant decrease of expression during the mid and late luteal phase and a further decrease of VEGF mRNA after regression. During pregnancy, high levels of expression were always present. In contrast, no significant change in VEGFR-1 mRNA expression during the estrous cycle and pregnancy was found. The VEGF protein concentration in CL tissue was significantly higher (20.9-23.4 ng/g wet weight) during the early luteal phase (Days 1-7), followed by a decrease at the late luteal phase (14.3-18.7 ng/g wet weight) and, especially, after CL regression (2.8 ng/g wet weight). However, relatively high levels were found during pregnancy (10.1 ng/g wet weight). As achieved by immunohistochemistry, VEGF protein was localized predominantly in luteal cells. High VEGF protein and transcript concentrations and increased VEGFR-2 expression during the early luteal phase coincided with luteal vascularization. These results suggest an important role of VEGF in angiogenesis of the newly formed CL. The high VEGF mRNA expression and protein levels during matured vasculature in the mid-stage CL and pregnancy also suggest also a survival function for endothelial cells.

Animals↗

PubMed2000

Periovulatory changes in the local release of vasoactive peptides, prostaglandin f(2alpha), and steroid hormones from bovine mature follicles in vivo.

We previously proposed that an endothelin-angiotensin-atrial natriuretic peptide system may contribute to inducing ovulation of mature bovine follicles by modulating follicular secretion of steroids and prostaglandins (PGs). Thus, this study aimed to determine the real-time changes in the local release of angiotensin II (Ang II), endothelin (ET), atrial natriuretic peptide (ANP), PGF(2alpha), and steroid hormones from bovine mature follicles during the periovulatory period in vivo. Seven cows were treated for superovulation using FSH and PGF(2alpha) injections. Two dialysis capillary membranes per follicle were surgically implanted into the theca layer of mature follicles and connected to a microdialysis system (MDS). Fractions of the perfusate were collected from Day -1 (Day 0 = LH surge) to Day 3. Five out of seven treated cows were normally ovulated, and the newly formed corpora lutea were observed at the end of the experiment. In these five ovulated cows, the release of estradiol, androstenedione, and progesterone in the theca layer increased (P < 0.05) synchronously with the LH surge. Acute increases in PGF(2alpha) and Ang II concentrations in the ovarian venous plasma (OVP) were observed at 24-48 h after the peak of the LH surge, when multiple ovulations were expected to occur. The follicular Ang II release was low during the pre-LH surge period and rose (P < 0.05) at the beginning of the increase in the LH surge. On the other hand, ET-1 release dropped (P < 0.05) when plasma LH started to increase. However, no clear changes in ANP concentration in the MDS perfusate and plasma were observed. The above local changes in Ang II, PGF(2alpha), as well as steroid hormones were not observed in cows (n = 2) that did not show an LH surge and ovulation. The present results demonstrate for the first time the local release of Ang II, ET-1, and ANP from the bovine mature follicle in real-time in vivo and show that Ang II and PGF(2alpha) concentrations in the OVP acutely increase around the time of ovulation. The overall results support the concept of a local functional ET-Ang-ANP system in the bovine mature follicle that may be involved in the ovulatory process.

Angiotensin II↗