Corpus luteum and endometrial function in ewes post partum: a study in vivo and in vitro

1992 ◽  
Vol 4 (1) ◽  
pp. 77 ◽  
Author(s):  
JM Wallace ◽  
CJ Ashworth ◽  
RP Aitken ◽  
MA Cheyne

Induction of ovulation post partum is associated with a high incidence of prematurely regressing corpora lutea. However, inadequate luteal function is not the sole reason for pregnancy failure, because ewes with normal corpus luteum function and successful fertilization also fail to establish pregnancies. The effects of suckling status and the interval from post partum to rebreeding on corpus luteum and endometrial function were examined in vivo and in vitro. Ewes were weaned early or allowed to lactate, induced to ovulate using a progesterone-impregnated controlled internal drug release device and an intramuscular injection of pregnant mare serum gonadotrophin, and inseminated (intrauterine) at either 21 or 35 days post partum (n = 10 per group). A further 10 standard ewes whose interval from parturition was in excess of 150 days were included for comparative purposes. On Day 10 after insemination the pregnancy rate was determined in four ewes from each of the post-partum groups and five standard ewes. These ewes were then ovariectomized and hysterectomized for studies in vitro. The incidence of premature luteal regression, as assessed by progesterone concentrations in peripheral blood was independent of the suckling stimulus but dependent on stage post partum (21 days post partum, 6 of 19 ewes; 35 days post partum, 0 of 19 ewes; P less than 0.05). Luteal function was normal in all standard ewes. Ovulation rate, corpus luteum weight, corpus luteum progesterone content and basal progesterone production in vitro were significantly less in 21-day than in 35-day post-partum ewes. Pregnancy rates as determined on Day 10 or at term were low in all post-partum groups (7 out of the 38 ewes inseminated) compared with standard ewes (8 of 10). Uterine function was assessed by culturing endometrial tissue from the tip and body of each uterine horn in the presence of [3H]leucine for 30 h at 37 degrees C. Incorporation of radiolabel into non-dialysable proteins synthesized and secreted by the endometrium in vitro was independent of uterine horn location and suckling status but was significantly lower (P less than 0.001) in media from 21-day than from 35-day post-partum ewes. Irrespective of treatment group, incorporation of radiolabel was positively correlated with mean plasma progesterone concentrations on Days 2-10 after insemination and with basal progesterone production in vitro. Secreted proteins were detected by two-dimensional-polyacrylamide-gel electrophoresis and fluorography.(ABSTRACT TRUNCATED AT 400 WORDS)

1993 ◽  
Vol 136 (1) ◽  
pp. 17-NP ◽  
Author(s):  
J. M. Wallace ◽  
M. G. Thompson ◽  
R. P. Aitken ◽  
M. A. Cheyne

ABSTRACT Induction of ovulation early post partum in sheep is associated with a high incidence (30–40%) of premature luteolysis. The present study was designed to characterize oxytocin receptor levels, oxytocin-stimulated inositol phosphate (IP) turnover (second messenger) and oxytocin-stimulated prostaglandin F2α (PGF2α) release in the endometrium of post-partum ewes induced to ovulate 21 days after parturition and expected to exhibit a range of corpus luteal functions subsequently. Ovulation was induced on day 21 post partum using a controlled internal drug release device and pregnant mare serum gonadotrophin, and uterine tissues were collected on days 5, 10 or 15 of the cycle (n = 4/day). A further 12 ewes whose interval from previous parturition exceeded 150 days were similarly treated and acted as controls. Measurement of daily peripheral progesterone concentrations revealed that while all control ewes exhibited normal luteal function, abnormal luteal function was evident in two, two and one post-partum ewes studied on days 5, 10 and 15 of the cycle respectively. Oxytocin receptor binding was detected (by receptor-binding assay and in-vitro autoradiography) in the endometrium and myometrium of post-partum ewes at all three stages of the oestrous cycle but only at day 15 in control ewes. To determine IP turnover, 100 mg caruncular endometrium was incubated in duplicate for 2·5 h with 10 μCi [3H]inositol and treated with 0 or 2 μmol oxytocin/l for 30 min, then [3H]inositol mono-, bis- and trisphosphates were quantified. Oxytocin stimulated total IPs in all day-5 and day-15 post-partum ewes, in three of four day-10 ewes and in all day-15 control ewes. Basal endometrial PGF2α release measured in triplicate (100 mg/well) during a 2 h incubation was higher in post-partum versus control ewes on days 5 and 10 but not on day 15 of the cycle. Similarly, oxytocin stimulated PGF2α release to varying levels at all stages of the cycle in post-partum ewes but only on day 15 in control ewes. Irrespective of the treatment group endometrial oxytocin receptor number was significantly (P < 0·001) correlated with oxytocin-stimulated IP turnover and PGF2α release. Thus the induction of ovulation and the subsequent luteal phase in post-partum ewes is against a back ground of high oxytocin receptor expression and enhanced PGF2α release which in some ewes may contribute to abnormal luteal function. Journal of Endocrinology (1993) 136, 17–25


1977 ◽  
Vol 84 (1) ◽  
pp. 142-154 ◽  
Author(s):  
F. E. Cole ◽  
P. C. Arquembourg ◽  
B. F. Rice

ABSTRACT Studies were performed to try to determine if gonadotrophins are altered during their interaction with tissue receptors. Immunologic, electrophoretic and binding properties of lactoperoxidase labelled [125I]HLH and [125I]HCG were examined before and after elution from mouse luteoma and human corpora lutea receptor preparations. The anti-HCG used in these studies at a 1:10 000 dilution precipitated 92% of a freshly iodinated [125I]HCG preparation. Receptor eluted [125I]HCG, derived from the same batch of labelled ligand, was virtually quantitatively precipitated by the same dilution of anti-HCG. [125I]HCG eluted from the human corpus luteum was electrophoretically more homogenous when compared to its heterogenous parent labelled preparation and migrated to a position similar to that of native HCG. In Ouchterlony double diffusion experiments against anti-HCG antiserum, corpus luteum eluted [125I]HCG and [125I]HLH showed immunologic identity with each other as well as with native HCG and HLH. Receptor eluted [125I]HCG from the mouse luteoma, following in vivo administration via tail vein injection or after incubation in vitro with labelled hormones, was immunologically indistinguishable from native HCG. The electrophoretic mobility of HCG was retarded when HCG was added to extracts of mouse luteoma, liver and kidney. Eluates of mouse luteoma, applied to Bio-Gel columns previously equilibrated with [125I]HCG showed the ability to concentrate [125I]HCG in the high molecular weight column fractions. Similar results were obtained with columns equilibrated with [125I]TSH and [125I]HGH. [125I]HCG eluted from the mouse luteoma was able to bind to fresh luteoma homogenate but, in contrast to an earlier report with [125I]HCG eluted from rat testis, no enhancement of binding of the eluted [125I]HCG was observed with fresh tissue. These results could be explained by the extraction of non-dialyzable intracellular component during the [125I]HCG elution procedure from the luteoma homogenate which combines with HCG to lower its binding and alter its electrophoretic mobility. This component could be extracted from other mouse tissues and combines with other labelled peptide hormones. Data in the present report support in part the hypothesis that gonadotrophins eluted from mouse luteoma and human corpus luteum are not altered by their interaction with tissue receptors.


1984 ◽  
Vol 103 (1) ◽  
pp. 107-110 ◽  
Author(s):  
M. G. Hunter

ABSTRACT Human luteal tissue recovered from varying stages of the luteal phase was minced and incubated for 3 h and the effect of human chorionic gonadotrophin (hCG), prolactin and hCG + prolactin on progesterone and oestradiol production measured. While hCG generally enhanced both progesterone and oestradiol synthesis, prolactin alone at either 20 or 200 μg/l had no significant effect on steroidogenesis. When prolactin was added along with hCG in four of six corpora lutea, however, progesterone production significantly increased and in three of six corpora lutea oestradiol production was increased above that induced by hCG alone. It is concluded that prolactin may play some role in the control of steroidogenesis by the human corpus luteum. J. Endocr. (1984) 103, 107–110


Reproduction ◽  
2015 ◽  
Vol 149 (1) ◽  
pp. 1-10 ◽  
Author(s):  
P Accialini ◽  
S F Hernández ◽  
D Bas ◽  
M C Pazos ◽  
G Irusta ◽  
...  

In this study, we investigated the interaction between the Notch pathway and progesterone to maintain the functionality of the corpus luteum (CL). When Notch signaling is activated, the γ-secretase complex releases the active intracellular domains (NICD) of their receptors, which exert survival effects. We designed studies to analyze whether thein vitroinhibition of Notch affects progesterone production, steroidogenic regulators, apoptotic parameters, and signaling transduction pathways in the cultures of CL isolated from pregnant and superovulated rats. We detected a decrease in progesterone production when corpora lutea (CL) were incubated withN-(N-(3,5-difluorophenacetyl-l-alanyl))-S-phenylglycine t-butyl ester (DAPT), a γ-secretase inhibitor. This effect could be in part due to the decrease detected in the CL protein levels of P450scc because STAR and 3β-hydroxysteroid dehydrogenase were not affected by Notch inhibition. Besides, the addition of aminoglutethimide to the CL culture medium decreased NICD of NOTCH1. We observed an increase in the expression of active CASPASE3 (CASP3) after inhibition by Notch, which was reversed by the presence of progesterone. The BAX:BCLXLratio was increased in CL treated with DAPT and the presence of progesterone reversed this effect. In addition, phosphorylation of AKT was inhibited in CL treated with DAPT, but had no effect on ERK activation. To demonstrate that the action of DAPT is specifically related with the inhibition of Notch, CLs were incubated with DLL4 antibody and a decrease in progesterone production was detected. These results suggest the existence of a novel link between progesterone and the Notch signaling pathway to maintain the functionality of the CL.


2015 ◽  
Vol 27 (2) ◽  
pp. 360 ◽  
Author(s):  
Rosemary A. Stewart ◽  
Adrienne E. Crosier ◽  
Katharine M. Pelican ◽  
Budhan S. Pukazhenthi ◽  
Brandon D. Sitzmann ◽  
...  

Exogenous gonadotrophins administered before AI can adversely alter endocrine dynamics and inhibit embryo development in felids. In the present study, we tested the hypothesis that priming the domestic cat ovary with progestin mitigates the negative influence of gonadotrophin therapy by normalising early embryogenesis and luteal function. Queens were given either: (1) progestin pretreatment plus chorionic gonadotrophins (n = 8; primed); or (2) gonadotrophins only (n = 8; unprimed). Ovulatory response was assessed laparoscopically, and cats with fresh corpora lutea (CL) were inseminated in utero. Ovariohysterectomy was performed 3 days later to recover intra-oviductal embryos for in vitro culture; one ovary was prepared for histology, and CL from the remaining ovary were excised and assessed for progesterone content and targeted gene expression. Of the six primed and seven unprimed queens inseminated, embryo(s) were recovered from five individuals per group. Embryos from progestin-primed donors more closely simulated normal stage in vivo development (P < 0.05). No 2- or 4-cell embryos from either group developed beyond 16-cells in vitro; however, 50% of unprimed and 66.7% of primed (P > 0.05) 5–16-cell embryos progressed to morulae or blastocysts by Day 4 of culture. Although histological characteristics were unaffected by progestin priming (P > 0.05), luteal progesterone was unusually high (P < 0.05) in unprimed compared with primed cats (72.4 ± 5.8 vs 52.2 ± 5.5 ng mg–1, respectively). Two genes associated with progesterone biosynthesis (luteinising hormone receptor and 3β-hydroxysteroid dehydrogenase) were upregulated in unprimed versus primed individuals (P = 0.05 and P < 0.05, respectively), indicating potential mechanistic pathways for the protective influence of pre-emptive progestin treatment. Building on earlier findings that progestin priming prevents spontaneous ovulation, increases ovarian sensitivity to gonadotrophins and ensures a normative endocrine environment, the present study demonstrates that pretreatment with this steroid also benefits embryo development and normalisation of early luteal function.


1994 ◽  
Vol 143 (3) ◽  
pp. 423-431 ◽  
Author(s):  
P L Keyes ◽  
J L Kostyo ◽  
R Towns

Abstract The rabbit corpus luteum possesses LH receptors that are coupled to adenylyl cyclase, but paradoxically it does not require LH as a luteotrophic factor for the maintenance of progesterone secretion. This suggests that rabbit luteal cells may not respond physiologically to LH. Therefore, the present study was undertaken to investigate the responsiveness of the rabbit corpus luteum of pseudopregnancy to human chorionic gonadotrophin (hCG) which acts on the same receptor as LH. Pseudopregnancy was induced by injection of 40 IU pregnant mare serum gonadotrophin followed 50 h later by an injection of 40 IU hCG (day 0). On days 7 and 11 of pseudopregnancy, corpora lutea were obtained and incubated for 2 or 5 h in the presence of either 0·1 or 1 μg/ml hCG or 1 mm monobutyryl cyclic AMP (bcAMP). Neither hCG nor bcAMP stimulated progesterone production by the isolated corpus luteum, despite a sustained high rate of progesterone production by the tissue throughout the incubation period. By contrast, Graafian follicles removed from the same ovaries and incubated under the same conditions responded both to hCG and bcAMP with large increases in progesterone production. To determine whether the cyclic AMP content of the corpus luteum was altered by in vitro exposure to hCG, day 7 and day 11 corpora lutea were incubated for 5 or 15 min with various concentrations of hCG, and cyclic AMP in the tissue was then measured. Even at the highest concentration of hCG tested (10 μg/ml), the cyclic AMP content of the corpus luteum was unaltered. Given this result, the acute effects of various concentrations of hCG on the adenylyl cyclase activity of homogenates of day 11 corpora lutea were examined. Consistent with previous reports of others, adenylyl cyclase activity was stimulated, but only at a high concentration of hCG (1 μg/ml), and the degree of stimulation of the enzyme (∼75%) was quite modest. By contrast, the adenylyl cyclase activity of homogenates of rabbit Graafian follicles was stimulated by even the lowest concentration of hCG tested (0·01 μg/ml). Thus, the adenylyl cyclase of the rabbit follicle is much more sensitive to hCG stimulation than the luteal form of the enzyme. Given the poor responsiveness of luteal adenylyl cyclase to hCG, the possibility was considered that cyclic AMP production in response to hCG might be obscured by luteal cell phosphodiesterase. When day 11 corpora lutea were incubated with hCG in the presence of the phosphodiesterase inhibitor, isobutyl methylxanthine (5 mm), there was a marked increase in the cyclic AMP content of the tissue. Despite this large increase in endogenous cyclic AMP, progesterone production by the corpora lutea was again unaffected. Thus, the rabbit corpus luteum is insensitive to stimulation in at least two major respects. The adenylyl cyclase coupled to the LH receptor is resistant to stimulation by LH (hCG), and steroidogenesis, as reflected by progesterone production in vitro, is not stimulated acutely by cyclic AMP. These results suggest that progesterone biosynthesis in the rabbit corpus luteum is not a regulated process, but rather a process that becomes autonomous as a result of the differentiation of granulosa cells into luteal cells. Therefore, the role of a luteotrophin, such as oestrogen in the rabbit, is to maintain the health and viability of the luteal cells, which have, as an intrinsic property, the capacity to produce progesterone at a high rate. Journal of Endocrinology (1994) 143, 423–431


Reproduction ◽  
2012 ◽  
Vol 143 (4) ◽  
pp. 491-500 ◽  
Author(s):  
Koumei Shirasuna ◽  
Ayumi Kobayashi ◽  
Akane Nitta ◽  
Sayo Nibuno ◽  
Kiemi Sasahara ◽  
...  

The development of the corpus luteum (CL), which secretes large amounts of progesterone to establish pregnancy, is accompanied by active angiogenesis, vascularization, and lymphangiogenesis. Negative feedback regulation is a critical physiological mechanism. Vasohibin-1 (VASH1) was recently discovered as a novel endothelium-derived negative feedback regulator of vascularization. We therefore investigated the expression of VASH1 in the bovine CL. Expression of VASH1 mRNA and protein was predominantly localized to luteal endothelial cells (LECs). VASH1 expression in the CL was constant through the early to late luteal phases and decreased during CL regression relating with the action of luteolytic prostaglandin F2α in vivo. To investigate the role of VASH1, we determined whether VASH1 treatment affects angiogenesis and/or lymphangiogenesis using LECs and lymphatic endothelial cells (LyECs) in vitro. Vascular endothelial growth factor A (VEGFA) stimulated the expression of VASH1 in LECs but not in LyECs, and VASH1 completely blocked VEGFA-induced formation of capillary-like tube structures of LECs and LyECs in vitro. In summary, VASH1 is predominantly located on LECs in the bovine CL and inhibits the angiogenic and lymphangiogenic actions of VEGFA. Bovine CL therefore has a VEGFA–VASH1 system that may be involved in regulation of luteal function, especially in the development of the CL. The results indicate that VASH1 has the potential to act as a negative feedback regulator of angiogenesis and lymphangiogenesis in the CL in cows.


1987 ◽  
Vol 112 (3) ◽  
pp. 449-457 ◽  
Author(s):  
G. E. Webley ◽  
J. P. Hearn

ABSTRACT The effect of human chorionic gonadotrophin (hCG) and melatonin on the local production of progesterone by the marmoset corpus luteum was investigated in vivo using a perfusion cannula system. Progesterone secretion was measured in 10-min fractions of buffer which had been perfused through the corpus luteum at a flow rate of 70 μl/min for a maximum of 3 h in anaesthetized animals. Two corpora lutea were cannulated in each animal; one for perfusion of test material and the other for perfusion with buffer alone as a control. Perfusion with hCG (25 i.u./ml), investigated as a positive control, produced a marked stimulation of progesterone secretion which increased 10–20 min from the start of perfusion and reached a peak after 30–60 min. A stimulation of progesterone was also observed after perfusion with melatonin (860 pmol/l). The response was evident within 10–30 min of the hormone reaching the corpus luteum and was similar in magnitude to that observed for hCG. The ability of melatonin to stimulate progesterone secretion supports previous in-vitro studies and suggests an ovarian action for melatonin in the primate. The local perfusion system described may have potential uses in studies of luteal function related to aspects of infertility or regulation of fertility. J. Endocr. (1987) 112, 449–457


1992 ◽  
Vol 127 (3) ◽  
pp. 246-251 ◽  
Author(s):  
Yasunori Yoshimura ◽  
Yukio Nakamura ◽  
Fumitaka Ichikawa ◽  
Takahisa Oda ◽  
Masao Jinno ◽  
...  

The present study was undertaken to assess the ability of human corpora lutea to produce leukotriene B4 (LTB4). The maximum capacity of luteal cells to secrete progesterone was attained on day 4, and both the basal production and the responsiveness to hCG decreased thereafter. In contrast, the production of LTB4 by cultured luteal cells was significantly reduced on day 4, but increased thereafter. The basal concentration of LTB4 produced by luteal cells varied from 75 to 590 pg/105 cells/2 days. LTB4 production appeared to decrease concomitantly with increased-progesterone production in cultured luteal cells. Exposure to hCG decreased significantly LTB4 production by cultured luteal cells on day 4. An inhibitor of the lipoxygenase pathway, nordihydroguaiaretic acid (NDGA), inhibited LTB4 production in a dose-dependent manner. However, NDGA did not affect basal progesterone production by the cultured luteal cells. A significant inverse relationship existed between the accumulation rates of progesterone and LTB4 in the luteal cells. Furthermore, the addition of LTB4 inhibited progesterone production in a dose-dependent manner in both the presence and absence of hCG. In conclusion, LTB4 could be synthesized by human corpora lutea in vitro, and correlated inversely with the secretion rates of progesterone. These data suggest that LTB4 produced locally in the corpus luteum may be an important regulator in human luteal regression.


1969 ◽  
Vol 43 (2) ◽  
pp. 301-307 ◽  
Author(s):  
R. M. MOOR ◽  
L. E. A. ROWSON ◽  
MARY F. HAY ◽  
B. V. CALDWELL

SUMMARY The relationship between the conceptus and the corpus luteum during the first 90 days of pregnancy was determined in sheep which were made unilaterally pregnant by egg transfer 4 or 5 days after oestrus. Additional corpora lutea were established approximately 24 hr. after an injection of human chorionic gonadotrophin given on day 20, 30, 50 or 70, and 18–24 days later the sheep were killed. When 17 unilaterally pregnant ewes, in which new corpora lutea had been induced on or before day 31, were killed, only the corpora lutea on the same side as the gravid horn were still functional; total regression of the corpora lutea had occurred in the ovary adjacent to the non-gravid horn. In 13 out of 14 sheep in which corpora lutea had been induced on day 51 or 71, the luteal tissue was still present in both ovaries at autopsy. Thus a local effect is exerted during the first one-third of pregnancy by conceptuses confined to one uterine horn, but this effect changes to a more general one about day 50. In five sheep, embryos were allowed to develop in an intact uterus and under these conditions no unilateral effect could be demonstrated on corpora lutea induced on day 31.


Sign in / Sign up

Export Citation Format

Share Document