Regulation Of Reproductive Function Research Articles

Defining the Gonadotrophin‐Releasing Hormone Neuronal Network: Transgenic Approaches to Understanding Neurocircuitry

The gonadotrophin-releasing hormone (GnRH) neurones are the final downstream effector neurones driving the central regulation of reproductive function and fertility in all mammalian species. Although it is abundantly clear that successful fertility relies upon the communication of a variety signals regarding internal and external cues to the GnRH neuronal population, how this is achieved remains poorly understood. A range of technical limitations has posed significant hurdles to defining, with any certainty, the complexities of the synaptic neuronal network regulating GnRH neurones. However, recent advances in transgenic technology have opened up new avenues to reconsider questions aimed at understanding this critical network. This article addresses some of the latest advances that use transgenic mouse models as tools to understand the neuronal circuitry underpinning the regulation of the GnRH neurones. By incorporating standard morphological and viral tract tracing techniques with innovative transgenic tools, recent studies have uncovered previously unappreciated qualities of the GnRH neurone, including extensive dendritic lengths, numerous somal and dendritic spines and plasticity over pubertal development, along with beginning to define the primary and higher-order afferents that make up the GnRH neuronal network.

Direct stimulatory effect of ghrelin on pituitary release of LH through a nitric oxide-dependent mechanism that is modulated by estrogen

Ghrelin, a gut peptide with key actions on food intake and GH secretion, has been recently recognized as potential regulator of reproductive function. Thus, in adult female rats, ghrelin has been proven to modulate GnRH/LH secretion, with predominant inhibitory effects in vivo. We analyze herein potential direct pituitary effects of ghrelin on basal and GnRH-stimulated gonadotropin secretion in prepubertal female rats, and its interplay with ovarian inputs, nitric oxide (NO), and hypothalamic differentiation. In the experimental setting, pituitaries from intact and ovariectomized prepubertal female rats were challenged with ghrelin in vitro and LH secretion was monitored. Our results demonstrate that 1) ghrelin consistently stimulated in vitro pituitary LH secretion under different experimental conditions; 2) the sensitivity to ghrelin, expressed either as the minimal effective dose or the amplitude of the LH response, was modulated by ovarian inputs; 3) the blockade of estrogen action significantly augmented the stimulatory effect of ghrelin; 4) the stimulatory effect of ghrelin on LH secretion required proper NO synthesis; and 5) the ability of ghrelin to elicit LH secretion in vitro was preserved after alteration (masculinization) of brain sexual differentiation. Overall, our present data reinforce the concept that ghrelin participates in the control of LH secretion, with potential stimulatory actions at the pituitary level that require the presence of NO and are modulated by ovarian signals.

Current knowledge and future challenges in camelid reproduction

Reproductive biology research on camelids offers some interesting peculiarities and challenges to scientists and animal production specialists. The objective of this paper is to review camelid reproduction, advances in reproductive physiology and reproductive biotechnologies in camelids and discuss some areas for further research. In the female, the focus has been on understanding follicular dynamics. This has allowed development of synchronization and superovulation strategies to support embryo transfer technologies which are now commonly used in camels. Some advances have been achieved in preservation of embryos by vitrification. Fertilization, early embryo development and embryo signaling for maternal recognition of pregnancy are still not fully understood. New information on the interaction of the developing embryo and the endometrium may shed some light on this signaling as well as the mechanism of prevention of luteolysis. The presence of a seminal ovulation-inducing factor (OIF) was confirmed in llamas and alpacas. Chronology of oocytes maturation has been described. In vitro production of embryos has been achieved resulting in successful pregnancies and births in the dromedary. These techniques offer a new tool for the production and study of interspecies/cross-species embryos and their effect on pregnancy. Male reproductive function remains poorly studied. Semen preservation and artificial insemination still present many challenges and are not used in production at the moment. The involvement of climatic and nutritional conditions as well as the role of leptin in the regulation of reproductive function need to be evaluated.

Effects of congenital hydrocephalus on the hypothalamic gonadotrophin-releasing hormone system

Despite the investigations that have linked hydrocephalus to reproductive system abnormalities, no researchers have attempted to identify the pathophysiological mechanism of this relationship. Because the role of the hypothalamic gonadotrophin-releasing hormone (GnRH) system in the regulation of reproductive functions is well established, the authors used immunohistochemical and radioimmunoassay (RIA) techniques to determine the morphological and biochemical effects of hydrocephalus on the hypothalamic GnRH system. Hypothalamic GnRH levels, fiber density, and cell types were studied in 21- and 50-day-old control and congenitally hydrocephalic Texas rats. Results of RIA indicated a significant (8.4%) increase in GnRH in 21-day-old hydrocephalic rats (9.17 +/- 0.64 pg/ng total protein) compared with that in controls (0.97 +/- 0.74 pg/ng total protein). In addition, the 50-day-old hydrocephalic animals had a significantly higher level of GnRH compared with age-matched controls (20.4 pg/ng compared with 1.88 +/- 2.1 pg/ng total protein). This increase was accompanied by changes in the fiber appearance and a shift from low GnRH producing cells to high GnRH producing cells in the hydrocephalic animals; however, there was no significant difference in the fiber density between the control and hydrocephalic animals at 21 days. In addition, poor neurological scores correlated with the severity of hydrocephalus. These results demonstrated that hypothalamic GnRH levels are significantly affected by fetal-onset hydrocephalus and that the mechanisms responsible for these effects may take place at the cellular rather than the gross structural level. Furthermore, they suggest that impairments in the GnRH system may be protracted in neonates and infants with hydrocephalus, and thus may be overcome by relatively early treatment with ventricular diversion. However, the clinical implications of GnRH perturbations in shunt-dependent patients must await a forthcoming study in shunted animals.

Nutritional inputs into the reproductive neuroendocrine control system – a multidimensional perspective

Evolution has shaped regulatory systems to improve the chance of reproductive success in a somewhat unpredictable environment. One of the more powerful regulators of reproductive function in both sexes is metabolic status, defined as the availability of nutrients and energy to the tissues. Here, we briefly review the basics of the relationship between metabolic status and the activity of the system that controls pulsatile GnRH and LH secretion. We then reflect on these relationships within the framework of a model that comprises four interdependent 'dimensions': 1) genetic, 2) structural, 3) communicational, and 4) temporal. Using two major examples, the male sheep and the post-partum dairy cow, we illustrate aspects of each dimension that seemed to have evolved to limit the risks associated with 'the decision to reproduce'. The results of recent studies have also led us to include in our model the concepts of 'metabolic memory' and 'nutrient sensing' to help explain some aspects of the temporal dimension. Throughout the review, we propose directions for future research that could shed light on pathways that have evolved to ensure that animals are able to take the least risky 'decision'.

Role of Central Nervous System and Ovarian Insulin Receptor Substrate 2 Signaling in Female Reproductive Function in the Mouse1

Insulin receptor signaling regulates female reproductive function acting in the central nervous system and ovary. Female mice that globally lack insulin receptor substrate (IRS) 2, which is a key mediator of insulin receptor action, are infertile with defects in hypothalamic and ovarian functions. To unravel the tissue-specific roles of IRS2, we examined reproductive function in female mice that lack Irs2 only in the neurons. Surprisingly, these animals had minimal defects in pituitary and ovarian hormone levels, ovarian anatomy and function, and breeding performance, which indicates that the central nervous system IRS2 is not an obligatory signaling component for the regulation of reproductive function. Therefore, we undertook a detailed analysis of ovarian function in a novel Irs2 global null mouse line. Comparative morphometric analysis showed reduced follicle size, increased numbers of atretic follicles, as well as impaired oocyte growth and antral cavity development in Irs2 null ovaries. Granulosa cell proliferation was also defective in the Irs2 null ovaries. Furthermore, the insulin- and eCG-stimulated phosphoinositide-3-OH kinase signaling events, which included phosphorylation of Akt/protein kinase B and glycogen synthase kinase 3-beta, were impaired, whereas mitogen-activated protein kinase signaling was preserved in Irs2 null ovaries. These abnormalities were associated with reduced expression of cyclin D2 and increased CDKN1B levels, which indicates dysregulation of key components of the cell cycle apparatus implicated in ovarian function. Our data suggest that ovarian rather than central nervous system IRS2 signaling is important in the regulation of female reproductive function.

Inhibin: Regulation of reproductive function and practical use in females

ABSTRACTInhibins are gonadal glycoprotein hormones selectively and potently inhibiting follicle‐stimulating hormone (FSH) secretion from the pituitary gland. Inhibins are produced mainly by the ovary and are purified from follicular fluid. Inhibins were shown to be produced in two forms through dimeric assembly of an α‐subunit and one of two closely related β‐subunits to form inhibin A (α‐βA) and inhibin B (α‐βB). Although inhibin subunits are expressed in various tissues, the gonads are the major source of circulating inhibins. While inhibins may act as a paracrine or autocrine factor in some tissues, their best understood roles are as endocrine regulators of pituitary FSH. In this review we focus our attention on more recent developments in inhibin research. We describe patterns of inhibin A and B secretion during the estrous cycle. We also review the immunization against inhibin α subunit as a practical method for superovulation. Superovulation has been induced successfully by passive or active immunization against the inhibin α‐subunit in several species such as mice, rats, hamsters, guinea pigs, cows, mares, ewes and goats. Furthermore, several studies have shown that oocytes superovulated with immunization against inhibin α‐subunit have the ability to develop normally, suggesting that inhibin immunization could be used as a practical method for superovulation in a wide range of animal species.

Seasonal reproductive activity and innervation of vas deferens and accessory male genital glands in the water buffalo (Bubalus bubalis)

Autonomic nerves supplying mammalian male internal genital organs have an important role in the regulation of reproductive function. To find out the relationships between the neurochemical content of these nerves and the reproductive activity, we performed an immunohistochemical study in a species, the water buffalo, exhibiting a seasonal sexual behaviour. The distribution of noradrenergic and peptide-containing nerves was evaluated during the mating (autumn-winter) and non-mating (spring-summer) periods. During the mating period, a dense noradrenergic innervation was observed to supply the vas deferens as well as the accessory genital glands. Peptide-containing nerves were also observed but with a lower density. During the non-mating period noradrenergic nerves dramatically reduced. These results suggest that there is a neuro-endocrine interaction between androgen hormones and the autonomic nerve supply in the regulation of male water buffalo reproductive functions.

Role of cytokines in the regulation of reproductive function

The levels of 17 cytokines in the follicular fluid were measured by multiplex proteome analysis at the peak of stimulated ovulation during the in vitro fertilization cycle. In patients with ineffective folliculogenesis, the concentrations of IL-2, IL-4, IL-7, and granulocytic CSF in the follicular fluid were significantly lower than in women with greater mean number of collected oocytes. It was shown that multicomponent cytokine deficit was associated with lower production of high quality oocytes and lower efficiency of fertilization of the resultant oocytes in vitro. The absence of pregnancy after in vitro fertilization cycle is associated with lower levels of IL-2, IL-4, IL-7, granulocytic CSF, and macrophagic inflammatory protein 1b in combination with elevated contents of IL-8 and IL-13. The results attest to an important role of cytokines in the regulation of oogenesis and in preparation of the endometrium to implantation of the embryo.

Prenatal melatonin and its interaction with tachykinins in the hypothalamic - pituitary - gonadal axis

The pineal gland, through its hormone melatonin, influences the function of the hypothalamic-pituitary-gonadal axis. Tachykinins are bioactive peptides whose presence has been demonstrated in the pineal gland, hypothalamus, anterior pituitary gland and the gonads, in addition to other central and peripheral structures. Tachykinins have been demonstrated to influence the function of the hypothalamic-pituitary-gonadal axis, acting as paracrine factors at each of these levels. In the present review, we examine the available evidence supporting a role for melatonin in the regulation of reproductive functions, the possible role of tachykinins in pineal function and the possible interactions between melatonin and tachykinins in the hypothalamic-pituitary-gonadal axis. Evidence is presented showing that melatonin, given to pregnant rats, influences the developmental pattern of tachykinins in the hypothalamus and the anterior pituitary gland of the offspring during postnatal life. In the gonads, the effects of melatonin on the tachykinin developmental pattern were rather modest. In particular, in the present review, we have included a summary of our own work performed in the past few years on the effect of melatonin on tachykinin levels in the hypothalamic-pituitary-gonadal axis.

Effects of 3-Methyl-4-Nitrophenol in Diesel Exhaust Particles on the Regulation of Reproductive Function in Immature Female Japanese Quail (Coturnix japonica)

In a previous study, we found that 3-methyl-4-nitrophenol (PNMC), a component of diesel exhaust particles and also a degradation product of the insecticide fenitrothion, exhibits reproductive toxicity in the adult male Japanese quail. The present study investigated the toxicity of PNMC in the female Japanese quail and its ability to influence reproduction in immature females. The quail (21-day-old) were injected intramuscularly (im) with PNMC at doses 0.1, 1 or 10 mg/kg body weight daily for 3 days. There was no significant difference in body growth between the PNMC-administered and control birds. However, the weights of the oviducts were significantly lower in the PNMC-treated birds at all doses. Furthermore, the plasma concentrations of luteinizing hormone (LH) and estradiol-17 beta were significantly decreased with 1 and 10 mg/kg of PNMC. These findings suggest that PNMC might influence the hypothalamo-pituitary-gonadal axis with decreasing in secretion of GnRH, LH and ovarian steroid hormones and subsequently disturb growth of the reproductive organs of immature female quail. This study indicates that PNMC induces reproductive toxicity at the central level and disrupts reproductive function in the immature female quail.

Estradiol and progesterone synthesis in human placenta is stimulated by calcitriol

Calcitriol exerts a diverse range of biological actions including the control of growth and cell differentiation, modulation of hormone secretion, and regulation of reproductive function. The placenta synthesizes calcitriol through the expression of CYP27B1, but little is known about local actions of this hormone in the fetoplacental unit. The objective of this study was to investigate the effects of calcitriol upon progesterone (P4) and estradiol (E2) secretion in trophoblasts cultured from term human placenta. Cells were incubated in the presence of calcitriol for 18h and pregnenolone or androstenedione were subsequently added as substrates for the 3β-hydroxysteroid dehydrogenase (3β-HSD) or P450-aromatase (CYP19), respectively. Calcitriol stimulated in a dose-dependant manner E2 and P4 secretion. The use of a selective inhibitor of PKA prevented the effects of calcitriol upon E2 secretion, but not on P4. These results show that calcitriol is a physiological regulator of placental E2 and P4 production and suggest a novel role for calcitriol upon placental steroidogenesis.

Social and photoperiod effects on reproduction in five species of Peromyscus

At temperate latitudes, mammals and birds use changes in day length to time their reproductive activities to coincide with seasonal fluctuations in the environment. Close to the equator, however, conditions permissive of breeding do not track changes in day length as well, so other cues may be more important than photoperiod. In a variety of vertebrates, social interactions regulate breeding condition. We hypothesized that individuals of different species of Peromyscus mice found closer to the equator would respond more strongly to housing with an opposite sex conspecific than they would to photoperiod. To test this hypothesis, we compared the effects of long and short day lengths versus 8 days of pair housing with a female on reproductive tissue weights and testosterone (T) concentrations in five species of Peromyscus ( P. aztecus, P. eremicus, P. maniculatus, P. melanophrys, and P. polionotus). After 13 weeks of short days (8L:16D), P. maniculatus, P. melanophrys, and P. polionotus significantly reduced relative testes mass compared to long day (16L:8D) housed animals. Social housing, however, had no effect on tissue weights in any species. However, male P. polionotus paired with females for 8 days increased T concentrations compared to single-housed males, whereas paired P. maniculatus reduced T. These data suggest that mechanisms of photoperiodic and social regulation of reproductive function are mediated by different physiological mechanisms among closely-related species and that both phylogeny and environmental factors contribute to patterns of reproductive plasticity.

Decidualization and maintenance of a functional prostaglandin system in human endometrial cell lines following transformation with SV40 large T antigen

Prostaglandins (PGs) are key regulators of reproductive function and associated pathologies. We have established stable endometrial stromal and epithelial cell lines with SV40 large T antigen (TAG) as a model to study PG action in the human endometrium. Two clones for each cell type were selected for rapid growth, PG production and response to interleukin-1beta (IL-1beta). The resulting stromal (HIESC) and epithelial (HIEEC) cells retain their characteristics for at least 40 population doublings (PDs). The selected clones express progesterone (PR) and estrogen receptor-alpha (ER-alpha) at both mRNA and protein levels. By contrast, with the existing known human endometrial cell lines Ishikawa and KLE, HIESC and HIEEC increase their production of PGF2alpha and PGE2 and cyclooxygenase (COX)-2 protein expression in response to IL-1beta. The latter cells also express the main biosynthetic enzymes involved in PG production, cytosolic phospholipase A2 (PLA2), COX-1 and COX-2, PGF synthase and PGE synthase and the corresponding EP2, EP3, EP4 and FP receptors. The selective COX-2 inhibitor NS-398 completely inhibits the increased production of PGs induced by IL-1beta in both cell types, whereas dexamethasone (DEX) exerts a stronger inhibition in HIESC than in HIEEC. The latter observation may be related to the higher expression of COX-1 measured in HIEEC. On the basis of the present characterization and previous determination of corresponding primary cell cultures, HIESC and HIEEC appear appropriate to study the contribution of PGs in the regulation of human endometrium function and associated pathologies.

15-Hydroxyprostaglandin dehydrogenase in the bovine endometrium during the oestrous cycle and early pregnancy

Prostaglandins (PG) are primary regulators of reproductive function. In ruminants, the relative production of PGE2 and PGF2alpha determines the return to a new oestrous cycle or to the establishment of pregnancy in response to a viable embryo. PG action depends on biosynthesis, transport and interaction with their receptors, which are all expressed differentially during the oestrous cycle. PGs are, however, local mediators and thus the onsite degradation by enzymes such as 15-hydroxyprostaglandin dehydrogenase (HPGD), also known as 15-PGDH, is another factor to consider in the regulation of physiological action. Little information is available on PG catabolism in the endometrium during the oestrous cycle or early pregnancy. The purpose of this study was to clone the bovine 15-PGDH, produce the recombinant protein and generate a specific antibody to study its activity and its expression in the endometrium during the oestrous cycle. We have found that the bovine 15-PGDH is highly homologous to the rat and human isoforms. 15-PGDH is localized principally in the glandular epithelium and to a lesser extent in stromal and luminal epithelial cells. The enzyme expression is regulated during the oestrous cycle and it reaches its maximal level on days 16-18. Transient expression is observed in luminal epithelial and trophoblast cells on day 21 of pregnancy. The mRNA is expressed at a constant high level throughout the cycle. The activity of the recombinant 15-PGDH was also tested and was found comparable for PGF2alpha and PGE2. These data suggest that 15-PGDH contributes to the tight regulation of PG action in the endometrium especially at the critical period of recognition of pregnancy.

A novel post-transcriptional mechanism of regulation of luteinizing hormone receptor expression by an RNA binding protein from the ovary

Luteinizing hormone/human chorionic gonadotropin (LH/hCG) receptor, a member of the rhodopsin/β 2 adrenergic receptor subfamily of G-protein coupled receptors, is expressed primarily in the gonads and essential for the regulation of reproductive function. In the ovary, the expression of the receptor is post-transcriptionally regulated under physiological conditions. Studies from our laboratory showed that the ligand-induced down-regulation of the receptor occurs by accelerated degradation of the mRNA rather than by decreased transcription. We have identified a cytoplasmic LHR mRNA binding protein (LRBP) as a trans-acting factor in regulating LHR mRNA levels. LRBP binds to the coding region of LHR mRNA and causes accelerated degradation of mRNA. The RNA binding activity of LRBP was found to be inversely correlated to LH/hCG receptor mRNA levels. LRBP was purified to homogeneity and its identity was established as mevalonate kinase by N-terminal microsequencing and MALDI analysis. Mevalonate kinase, an enzyme involved in de novo synthesis of cholesterol, belongs to the GHMP family of kinases having a potential RNA binding fold. The expression of MVK mRNA and MVK protein levels were induced in response to hCG treatment prior to the down-regulation of LH/hCG receptor mRNA expression. A model for the post-transcriptional regulation of LH/hCG receptor in the ovary by mevalonate kinase is proposed.

Énergétique et régulation de la fonction de reproduction chez des femelles captives Babouins Olive (Papio anubis)i

The aim of this study was to assess energetic and social parameters of lactating female baboons, and to test the possible role of these factors on different reproductive variables such as postpartum amenorrhea duration or interbirth intervals. A 2-year follow-up was carried out for a sample of 23 multiparous and semi-free living lactating female baboons. There were no rank-related differences in mean energy intakes among the females. All females were in good physical condition and had positive energy balance. The energy costs of lactation were primarily covered by 2 behavioural adaptations: moderate energy intakes increase, particularly during the second and third quarter of lactation, and physical activity decreases. Compared with free-living female baboons, reduced energetic constraints (adequate energy intakes and moderate energetic expenditure) explain short postpartum amenorrhea durations and interbirth intervals in our sample. Fertility is strongly influenced by dominance rank, low-ranking females having longer delays before conception and longer inter-birth intervals. Moreover, mothers with heavier infants have shorter postpartum amenorrhea durations and need fewer cycles to conceive. High-ranking females or those in good physical condition appear likely to withstand relatively rapid infant growth which is independent of the infant’s sex, and have therefore higher reproductive rates. It is unlikely that nutritional mechanisms alone affect the relationship between dominance status and fertility, and our results suggest rather that social stress would also interfere with regulation of reproductive function.

Deoxyribonucleic Acid Microarray Analysis of Gene Expression Pattern in the Arcuate Nucleus/Ventromedial Nucleus of Hypothalamus during Lactation

Lactation is characterized by extreme hyperphagia and negative energy balance resulting from a large energy drain due to milk production and by a suppression of cyclic ovarian function. Increases in neuropeptide Y and agouti-related protein and a decrease in proopiomelanocortin expression in the arcuate nucleus of hypothalamus (ARH) may contribute to the hyperphagia to maintain energy balance and to the suppression of LH secretion associated with lactation. However, little is known about the full extent of neuroendocrine changes in the ARH that may contribute to the various adaptations occurring during lactation. To address this issue, we used Affymetrix microarray to acquire a reliable profile of the lactation-induced transcriptional changes in micropunches containing the ARH and a portion of the ventromedial nucleus of the hypothalamus. Using high stringency criteria, 12 genes were identified as being differentially regulated during lactation, and an additional 10 genes and three transcribed sequences were identified using moderate stringency criteria. Changes in neuropeptide Y, enkephalin, tyrosine hydroxylase, and dynorphin, genes previously shown to be differentially regulated during lactation, provide validation for the microarray analysis. New genes identified as being differentially expressed include those related to neurotransmission, growth factors, signal transduction, and structure remodeling. These data identify new genes in ARH/ventromedial nucleus of the hypothalamus that may play an important role in the adaptations of lactation related to hyperphagia, milk production, and the suppression of cyclic reproductive function and may contribute to elucidating a framework for integrating changes in energy intake with the regulation of reproductive function during lactation.

Kisspeptins: Regulators of Metastasis and the Hypothalamic‐Pituitary‐Gonadal Axis

The kisspeptins are the peptide products of the KiSS-1 gene and the endogenous agonists for the GPR54 receptor. Although KiSS-1 was initially discovered as a metastasis suppressor gene, recent evidence suggests the kisspeptin/GPR54 system is a key regulator of the reproductive system. Disrupted GPR54 signalling causes hypogonadotrophic hypogonadism in rodents and man. Central or peripheral administration of kisspeptin potently stimulates the hypothalamic-pituitary-gonadal axis, increasing circulating gonadotrophin concentrations in a number of animal models. These effects appear likely to be mediated via the hypothalamic gonadotrophin-releasing hormone system, although kisspeptins may have direct effects on the anterior pituitary gland. Hypothalamic KiSS-1 expression is regulated by circulating sex steroids. The precise physiological role of the kisspeptin system in the regulation of reproductive function remains to be elucidated.

Expression of Human Prostaglandin Transporter in the Human Endometrium across the Menstrual Cycle

Prostaglandins (PGs) are important regulators of reproductive function. The mechanism by which PGs are transported across the biological membrane is a new emerging field of investigation. Prostaglandin transporter (PGT) has been identified as a functional PG carrier. The aim of our study was to outline the expression of PGT in the human endometrium across the menstrual cycle. Quantitative RT-PCR showed human PGT (hPGT) expression to be strong in the proliferative and early secretory phases and low in the middle to late secretory phase. Northern blot analysis revealed hPGT mRNA transcript of 4 kb in the human endometrium. A peptide-directed polyclonal antibody was generated in rabbits against the 22 amino acids forming the C terminus of hPGT. Antibody specificity was demonstrated by Western blot. Immunoblots of endogenous hPGT in the human endometrium revealed a 70-kDa protein in endometrial cells. Endometrial biopsies collected across the menstrual cycle were used to assess hPGT protein expression by immunohistochemistry. hPGT was immunolocalized to luminal, glandular epithelial, and stromal cells. Because it was observed at the mRNA level, semiquantitative analysis showed a higher protein expression in proliferative and early secretory phases than in the mid-late secretory phase. In conclusion, our study revealed that hPGT expression is modulated in epithelial and stromal cells of the human endometrium at both mRNA and protein levels during the menstrual cycle. These findings support a role for hPGT as an important new player in the regulation of PG action in the human endometrium.