Control Of Luteal Function Research Articles

Control of luteal function during early pregnancy in sheep

Prolactin and LH are required for the maintenance of normal luteal structures during early pregnancy in the ewe, whilst luteolysis, induced by prostaglandin F-2 alpha is inhibited by the presence of the conceptus. The trophoblast also secretes a local anti-luteolytic factor, trophoblastin, which is most probably a protein because it is thermolabile and inactivated by pronase. It is soluble at pH 9.6 and can be separated on Ultrogel. Trophoblastin is present for a short time (12th-22nd day of pregnancy) but its effect may last several months.

Computational interrogation of cis-regulatory elements of genes that are common targets of luteotropin and luteolysin in the primate corpus luteum

The rapid recent increase in microarray-based gene expression studies in the corpus luteum (CL) utilizing macaque models gathered increasing volume of data in publically accessible microarray expression databases. Examining gene pathways in different functional states of CL may help to understand the factors that control luteal function and hence human fertility. Co-regulation of genes in microarray experiments may imply common transcriptional regulation by sequence-specific DNA-binding transcriptional factors. We have computationally analyzed the transcription factor binding sites (TFBS) in a previously reported macaque luteal microarray gene set (n=15) that are common targets of luteotropin (luteinizing hormone (LH) and human chorionic gonadotropin (hCG)) and luteolysin (prostaglandin (PG) F2α). This in silico approach can reveal transcriptional networks that control these important genes which are representative of the interplay between luteotropic and luteolytic factors in the control of luteal function. Our computational analyses revealed 6 matrix families whose binding sites are significantly over-represented in promoters of these genes. The roles of these factors are discussed, which might help to understand the transcriptional regulatory network in the control of luteal function. These factors might be promising experimental targets for investigation of human luteal insufficiency.

Involvement of Src family of kinases and cAMP phosphodiesterase in the luteinizing hormone/chorionic gonadotropin receptor-mediated signaling in the corpus luteum of monkey

BackgroundIn higher primates, during non-pregnant cycles, it is indisputable that circulating LH is essential for maintenance of corpus luteum (CL) function. On the other hand, during pregnancy, CL function gets rescued by the LH analogue, chorionic gonadotropin (CG). The molecular mechanisms involved in the control of luteal function during spontaneous luteolysis and rescue processes are not completely understood. Emerging evidence suggests that LH/CGR activation triggers proliferation and transformation of target cells by various signaling molecules as evident from studies demonstrating participation of Src family of tyrosine kinases (SFKs) and MAP kinases in hCG-mediated actions in Leydig cells. Since circulating LH concentration does not vary during luteal regression, it was hypothesized that decreased responsiveness of luteal cells to LH might occur due to changes in LH/CGR expression dynamics, modulation of SFKs or interference with steroid biosynthesis.MethodsSince, maintenance of structure and function of CL is dependent on the presence of functional LH/CGR its expression dynamics as well as mRNA and protein expressions of SFKs were determined throughout the luteal phase. Employing well characterized luteolysis and CL rescue animal models, activities of SFKs, cAMP phosphodiesterase (cAMP-PDE) and expression of SR-B1 (a membrane receptor associated with trafficking of cholesterol ester) were examined. Also, studies were carried out to investigate the mechanisms responsible for decline in progesterone biosynthesis in CL during the latter part of the non-pregnant cycle.Results and discussionThe decreased responsiveness of CL to LH during late luteal phase could not be accounted for by changes in LH/CGR mRNA levels, its transcript variants or protein. Results obtained employing model systems depicting different functional states of CL revealed increased activity of SFKs [pSrc (Y-416)] and PDE as well as decreased expression of SR-B1correlating with initiation of spontaneous luteolysis. However, CG, by virtue of its heroic efforts, perhaps by inhibition of SFKs and PDE activation, prevents CL from undergoing regression during pregnancy.ConclusionsThe results indicated participation of activated Src and increased activity of cAMP-PDE in the control of luteal function in vivo. That the exogenous hCG treatment caused decreased activation of Src and cAMP-PDE activity with increased circulating progesterone might explain the transient CL rescue that occurs during early pregnancy.

Immunohistochemical Detection of CD4-, CD8- and MHC II-Expressing Immune Cells and Endoglin in the Canine Corpus Luteum at Different Stages of Dioestrus

Specific immunohistochemical methods were applied to detect the presence of CD4-, CD8- and major histocompatibility complex II (MHC II)-expressing immune cells and of endoglin in the canine corpus luteum between days 15 and 75, after ovulation. Corpora lutea were obtained from groups of three clinically healthy beagle bitches, ovariohysterectomized at the respective days. For all four parameters, the effect of time was highly significant. Quantitative evaluation yielded high values on day 15, followed by a decrease on day 30 (CD4, CD8 and endoglin) and day 45 (MHC II). While there were no further changes for cells staining positive for CD4 and endoglin, CD8-positive immune cells increased from day 45 to day 60 to drop again on day 75; MHC II-positive staining increased from day 45 to days 60-75. These data suggest an involvement of the immune system in control of luteal function also in the dog that may have both stimulatory and inhibiting effects.

Corpus luteum (CL) function: local control mechanisms

LH and PGF 2α are the principal luteotrophic and luteolytic hormones in domestic animals, however, it is becoming increasingly apparent that intra-ovarian factors can modulate luteal function. For example, the insulin-like growth factors (IGF-I and -II) can regulate ovarian function, and have direct effects on ovarian cells. An important role for the IGFs in regulating ovarian function is suggested by the multiple effects of IGFs on both follicular and luteal steroidogenesis. Expression of mRNA encoding IGF-I, IGF-II and the type 1 IGF receptor has also been detected in the ruminant CL and is suggestive of autocrine/paracrine roles for both IGF-I and -II in the regulation of luteal function. The actions of the IGFs are further modulated by their association with specific binding proteins (IGFBPs), which regulate the transport of IGFs and their presentation to specific receptors. IGFBPs have been detected in the CL of domestic animals, and inhibitory effects on IGF-I-stimulated progesterone production have been demonstrated. The rapid cyclical changes in luteal growth and regression are associated with rapid changes in vasculature. The principle angiogenic factors include the fibroblast growth factors (FGFs), vascular endothelial growth factor (VEGF) and the angiopoietins (Ang). Other locally produced factors include cytokines such as TNF-α and IL-1β. One such factor is monocyte chemoattractant protein (MCP-1), which increases after exogenous PGF 2α. An influx of macrophages takes place in the CL around luteolysis, possibly in response to MCP-1 release, but these changes are not observed in cattle when luteolysis is inhibited. In conclusion locally produced factors are important in the control of luteal function, although their roles have yet to fully elucidated.

Roles of reactive oxygen species in the regulation of luteal function.

Ephemerality and prolongation of luteal function have been matters of great concern in reproduction for many years. However, their control mechanisms are very complex and differ among mammals. Recently, evidence has indicated that reactive oxygen species may play important roles in the regulation of luteal function. Reactive oxygen species are present in most somatic cells and are involved in apoptosis, or 'physiological cell death'. In the corpus luteum, reactive oxygen species also exert luteolytic effects as well as some paradoxical luteotrophic effects. This paper discusses the possible roles of reactive oxygen species in the control of luteal function.

Roles of reactive oxygen species in the regulation of luteal function

Ephemerality and prolongation of luteal function have been matters of great concern in reproduction for many years. However, their control mechanisms are very complex and differ among mammals. Recently, evidence has indicated that reactive oxygen species may play important roles in the regulation of luteal function. Reactive oxygen species are present in most somatic cells and are involved in apoptosis, or 'physiological cell death'. In the corpus luteum, reactive oxygen species also exert luteolytic effects as well as some paradoxical luteotrophic effects. This paper discusses the possible roles of reactive oxygen species in the control of luteal function.

Corpus Luteum and Its Control by Intra-Luteal Regulators

It is well documented that the corpus luteum is required for a successful pregnancy. The primary function of the corpus luteum is to secrete progesterone, which has several biological effects on target tissues in the reproductive system to prepare them for support of pregnancy or to provide nourishment to the conceptus. In most domestic species including cattle, three organ systems regulate the function of the corpus luteum. The anterior pituitary gland secretes LH, which is the primary hormone responsible for stimulating the secretion of progesterone. The uterus has a luteolytic effect during the late luteal phase of the estrous cycle. This luteolytic effect of the uterus is mediated via secretion of prostaglandin F2α. Finally, the conceptus has either direct luteotropic or antiluteolytic effects. However, there is increasing evidence for the existence of substances in luteal tissue which are able to act locally, either alone or by modulating the action of the gonadotropins, thus modifying the functions of corpus luteum. The intra-luteal regulators consist of peptides, eicosanoids, growth factors, cytokines, and neurotransmitters. Available data suggest an important role for these substances in the control of luteal function during not only pregnancy but the estrous cycle as well.

Application of knowledge about corpus luteum function in control of estrus and ovulation in cattle

Understanding corpus luteum (CL) function has led to development of methods of estrus synchronization in cattle that either extend the estrous cycle by administration of exogenous progestins or shorten the cycle by induction of luteolysis. Both methods have limitations, which have been reduced or overcome through sequential or combined treatment with progestin and luteolytic drugs. Future improvements in estrus synchronization methodology are most likely to come from achieving more synchrony between the development of a highly fertile ovulatory follicle and control of luteal function.

Differential gene expression within the ovine corpus luteum: identification of secreted protein acidic and rich in cysteine as a major secretory product of small but not large luteal cells.

Limited information is available regarding secretory proteins of the corpus luteum (CL), and the potential local role these proteins may play in control of luteal function. An ovine small luteal cell complementary DNA library was immunologically screened with a polyclonal antiserum generated against small cell secretory proteins. A relatively abundant complementary DNA (approximately 2.1 kb) encoding a calcium binding glycoprotein Secreted Protein Acidic and Rich in Cysteine (SPARC) was isolated. Production of SPARC protein by ovine luteal cells was confirmed by immunoprecipitating it from labeled culture medium. Expression of SPARC messenger RNA (mRNA) was examined within CL collected on days 3, 7, 10, 13, and 16 post estrus (n = 4, 4, 4, 3, and 4 respectively), and within pools of purified small (n = 3) and large (n = 4) luteal cells by Northern and dot blot analysis. Amounts of SPARC mRNA increased during the early luteal phase, peaked by day 7 (P < 0.05) and subsequently declined on days 10 and 13 (P < 0.05). SPARC mRNA content was significantly higher in the small than in the large cells (P < 0.003). In situ hybridization showed that SPARC mRNA was localized to the thecal layer of Graafian follicles and to day 3 and day 10 CL. Within CL, immunohistochemistry indicated that SPARC protein was associated with small luteal cells (spindle shaped, avg = 17 microM in diameter) but not with large cells. This specific localization to small cells was confirmed by colocalization of SPARC with 3 beta-hydroxysteroid dehydrogenase. We conclude that SPARC is a major secretory product of small steroidogenic luteal cells of the ovine CL. As SPARC is known to modulate many aspects of tissue reorganization, expression by small luteal cells may play a role in regulation of CL maturation.

Development and Control of Luteal Function in Farm Animals

Development and Control of Luteal Function in Farm Animals

Effects of an antiprogesterone (RU486) on the hypothalamic-hypophyseal-ovarian-endometrial axis during the luteal phase of the menstrual cycle.

The impact of the antiprogesterone RU486 [17 beta-hydroxy-11 beta-(4-dimethylaminophenyl) 17 alpha-(1-propynyl)estra- 4,9-dien-3-one] on the hypothalamic-pituitary-ovarian-endometrial axis was examined in normal cycling women during the mid (MLP)- and late (LLP) luteal phases. During the MLP, 10 women received 3 mg/kg RU486 for 3 days. During the LLP, a single dose of 600 mg RU486 was administered to 4 women, and in another 4 women a single dose of 3 mg/kg was given during corpus luteum rescue by hCG. Longitudinal studies with daily and frequent blood samples (every 10 min for 10 h) were conducted during 3 consecutive cycles (control-treatment-recovery). During the MLP, RU486-induced uterine bleeding occurred in all 10 women 36-72 h after the first dose. No histological evidence of endometrial breakdown was found in endometrial biopsies taken 12-24 h before the onset of bleeding. Significant decreases in LH secretion (P less than 0.001) and LH pulse amplitude (P less than 0.006) and blunted pituitary responses to GnRH (P less than 0.01) were evident by the last treatment day, but LH pulse frequency did not change. Complete luteolysis occurred in 2 of the 10 women. Incomplete luteolysis occurred in 8 women and was associated with an initial decline of serum estradiol (P less than 0.001), but not progesterone levels, followed by rebound increases (P less than 0.001) in LH, estradiol, and progesterone levels 3 days later, which may have reversed the luteolytic processes and prolonged corpus luteum function. Spontaneous luteolysis ensued 3-5 days later with the onset of second episodes of uterine bleeding. For serum FSH, an early rise occurred during the luteal phase in advance of the onset of the second episodes of uterine bleeding. This rise may have resulted in early follicle recruitment and accounted for the shorter duration of the follicular phase during recovery cycles. During the LLP, the single RU486 dose resulted in significant decreases in LH pulse amplitude (P less than 0.03), frequency (P less than 0.05), and secretion (not significant) within 12 h. The recovery cycle was entirely normal. Corpus luteum rescue with incremental doses of hCG did not prevent uterine bleeding after RU486 treatment. These findings indicate that RU486 operates at multiple sites and implies that progesterone is important in the control of luteal function. Further, our data provide a basis for exploring the potential use of RU486 as a once a month birth control agent.

Prolactin and luteinizing hormone receptors in marsupial corpora lutea: relationship to control of luteal function.

Prolactin and LH receptor concentrations in tammar wallaby corpora lutea (CL) have been examined and related to the control of luteal function in this and other marsupial species. During embryonic diapause, quiescent CL contained high concentrations of prolactin receptors. This was consistent with an earlier suggestion that prolactin may act directly on the CL to maintain its quiescent state. However, despite an apparent seasonal change in the mechanism by which the CL is maintained in quiescence, the number of luteal prolactin receptors remained constant throughout the year. Reactivation of quiescent CL led to an approximate halving of prolactin receptors on a per cell basis, but pregnancy had no effect. None of the wallaby CL showed any significant degree of LH binding, although wallaby follicles and testicular tissue did show binding. This lack of LH binding was consistent with previous findings that quiescent wallaby CL reactivate in response to hypophysectomy and are refractory to LH when cultured in vitro. Parturition, which occurs approximately 2 days before ovulation in this species, had no effect on numbers of either prolactin or LH receptors despite the occurrence of a peak of plasma prolactin at this time. The red kangaroo, which also exhibits embryonic diapause, showed a similar pattern of luteal receptor numbers with high prolactin and negligible LH binding, whereas the brush-tailed possum, which does not exhibit embryonic diapause, gave a pattern more like that seen in Eutheria with many more LH than prolactin receptors.

Control of luteal function and implantation in the mink by prolactin.

Mink (Mustela vison) were treated during the period of embryonic diapause with prolactin or ergocryptine (CB-154). Prolactin advanced implantation time and hastened onset of luteal phase progesterone secretion. Duration of gestation in prolactin-treated adult mink was shorter than that of control mink. Ergocryptine had the opposite effects, prolonging gestation and inhibiting onset of luteal phase progesterone secretion. Prolactin is suggested to be the luteotrophin necessary for termination of embryonic diapause in mink.

Effect of ergot alkaloids upon luteal function in the ferret.

SUMMARY Since ergocornine is believed to block prolactin secretion in several species, the drug was given to pseudopregnant ferrets in an attempt to define the rôle of the hormone in the control of luteal function. The effects of ergocornine, and of the more potent analogue 2-bromo-α-ergocryptine, were followed by assay of the progestin content of the blood and histological examination of the ovaries. Ergocornine was given at the time of induction of pseudo-pregnancy by an injection of luteinizing hormone (day 0), on day 1, days 0 and 1, days 0, 2, 4 and 6, or day 7, and 2-bromo-α-ergocryptine on days 0, 1, 2 and 3, or 7, 8, 9 and 10. A lasting depression of plasma progestin content occurred after the injection of ergocornine on days 0 and 1, perhaps because of interference with the formation of the luteal bodies. Other injection régimes had little effect on progestin secretion.

Local control of luteal function by the uterus of the guinea-pig.

THERE is now no doubt that distension of the uterus of a number of species of mammal results in characteristic changes in the oestrous cycle which stem from a shortening of the life span of the corpora lutea in the ovaries1,2. Thus in the guinea-pig the presence of two glass beads in each horn of the uterus, when inserted during the first four days after oestrus, shortens the length of the cycle in progress at the time of operation and that of succeeding cycles3. The degree to which the cycle is shortened appears to be governed by the number of beads in the uterus, for when one bead is present in each horn the cycle in progress is shortened from a normal length of 16 days to 14 days, whilst when two beads are present in each horn the cycle is reduced to 12 days4. Although one bead located in one horn of the uterus of the sheep exerts a definite shortening effect on the oestrous cycle5, four beads in one horn of the guinea-pig uterus do not exert as marked an effect as when they are shared between both horns. The mechanism underlying this effect is not understood, although it has been suggested that the pituitary gland is involved in exercising a luteolytic function6. The present work has been concerned with the examination of this quantitative effect at the level of the ovaries, and attention directed to the morphological changes taking place in the corpora lutea, rather than to changes in the oestrous cycle over a longer period.