Monkey Uterus Research Articles

Sperm intrusion into the implantation-stage blastocyst and its potential biological significance.

The human embryo derives from fusion of oocyte and sperm, undergoes growth and differentiation, resulting in a blastocyst. To initiate implantation, the blastocyst hatches from the zona pellucida, allowing access from external inputs. Modelling of uterine sperm distribution indicates that 200-5000 sperm cells may reach the implantation-stage blastocyst following natural coitus. We show ultrastructural evidence of sperm cells intruding into trophectoderm cells of zona-free blastocysts obtained from the uterus of rhesus monkeys. Interaction between additional sperm and zona-free blastocyst could be an evolutionary feature yielding adaptive processes influencing the developmental fate of embryos. This process bears potential implications in pregnancy success, sperm competition and human health.

Long-term effects of repeated superovulation on the uterus and mammary gland in rhesus monkeys.

The aim of this study is to evaluate the effect of repeated controlled ovarian hyperstimulation (COH) on the structure and function of the uterus and mammary gland. Three adult female rhesus monkeys were superovulated up to four times, and three spontaneously ovulating monkeys were used as controls. After a 5-year period, the uterus and mammary gland tissue samples were collected for examination of their structure and function. Further, the expression of certain tumor markers was examined to assess the cancer risk for each organ. Expression of Wnt7a (associated with the functional/developmental status of the uterus) was significantly decreased in the uterus of superovulated monkeys, and decreased expression of proliferation marker PCNA was found in uterine cells. Meanwhile, abnormal Golgi-derived secretory vesicles with an irregular shape were observed in the mammary glands of the superovulated monkeys, and decreased PCNA expression together with increased expression of caspase-3 (an apoptosis marker) was indicated in the mammary cells. The expression of tumor molecular markers of the uterus and mammary gland was not significantly different between the two groups. Repeated COH affects the expression of the uterine development-related gene several years later, and uterine cells exhibited a low proliferation status. The ultrastructure of the mammary gland epithelial cells was abnormal, and the cells exhibited both low proliferation and high apoptosis status. Cancer risk for these organs was not observed. Given that primates are the closest relatives of humans, the results obtained from this study provide more intuitive information for optimization of clinical COH.

Receptors for thyrotropin-releasing hormone, thyroid-stimulating hormone, and thyroid hormones in the macaque uterus

Thyroid gland dysfunction is associated with menstrual cycle disturbances, infertility, and increased risk of miscarriage, but the mechanisms are poorly understood. However, little is known about the regulation of these receptors in the uterus. The aim of this study was to determine the effects of long-term treatment with steroid hormones on the expression, distribution, and regulation of the receptors for thyrotropin-releasing hormone (TRHR) and thyroid-stimulating hormone (TSHR), thyroid hormone receptor α1/α2 (THRα1/α2), and THRβ1 in the uterus of surgically menopausal monkeys. Eighty-eight cynomolgus macaques were ovariectomized and treated orally with conjugated equine estrogens (CEE; n = 20), a combination of CEE and medroxyprogesterone acetate (MPA; n = 20), or tibolone (n = 28) for 2 years. The control group (OvxC; n = 20) received no treatment. Immunohistochemistry was used to evaluate the protein expression and distribution of the receptors in luminal epithelium, glands, stroma, and myometrium of the uterus. Immunostaining of TRHR, TSHR, and THRs was detected in all uterine compartments. Epithelial immunostaining of TRHR was down-regulated in the CEE + MPA group, whereas in stroma, both TRHR and TSHR were increased by CEE + MPA treatment as compared with OvxC. TRHR immunoreactivity was up-regulated, but THRα and THRβ were down-regulated, in the myometrium of the CEE and CEE + MPA groups. The thyroid-stimulating hormone level was higher in the CEE and tibolone groups as compared with OvxC, but the level of free thyroxin did not differ between groups. All receptors involved in thyroid hormone function are expressed in monkey uterus, and they are all regulated by long-term steroid hormone treatment. These findings suggest that there is a possibility of direct actions of thyroid hormones, thyroid-stimulating hormone and thyrotropin-releasing hormone on uterine function.

In the fetal rhesus monkey uterus, in utero exposure to whom it may concern: bisphenol A (BPA) is associated with changes in gene expression and accelerated adenogenesis

In the fetal rhesus monkey uterus, in utero exposure to whom it may concern: bisphenol A (BPA) is associated with changes in gene expression and accelerated adenogenesis

Endometrial epithelial cell modifications in response to embryonic signals in bonnet monkeys (Macaca radiata)

The present investigation reports embryo-induced modifications in the epithelial cells of the endometrium in a primate species. In vivo, epithelial cell response to the embryonic signals was assessed at the embryo attachment stage in the gestational uterus of bonnet monkeys (Macaca radiata) and in vitro response was investigated by treating human endometrial epithelial cell line (Ishikawa) with human embryo conditioned media (CM). Endometrial epithelial (EE) cells at the embryo attachment stage in bonnet monkeys revealed higher proliferation accompanied by significant up regulation (p < 0.05) in the expression of estrogen receptor (ER)α and down regulation (p < 0.05) in ERβ expression. Further gestational EE cells showed higher (p < 0.001) expression of mucin-1, except in the embryo attachment site. Also, observed were significantly higher expression (p < 0.05) and altered cytoplasmic distribution of α(v) and β(3) integrins, when compared to non-pregnant animals. In pregnant animals, the embryo attachment zone showed differential expression of immunoreactive integrins as compared to the non-attachment zone. This suggested the role of embryo secreted factors in modulation of the epithelial cell profile. In vitro studies partially supported this assumption. Significantly higher proliferation (p < 0.05), as well as increased expression of ERα, integrin β(3) and mucin-1 (p < 0.05) were observed in Ishikawa cells, on stimulation with CM. Taken together, these results indicated the proliferation and modulation in the expression of estrogen receptors and cell adhesion molecules in the EE cells; at the embryo attachment stage in bonnet monkeys. Further it is likely that embryo secreted factors contribute to some of these modifications in EE cells. This report is the first account of discrete cellular events, which occur in the uterine epithelium, at the embryo attachment stage in a primate species.

Correlation between CD4+ T-cell loss and Gag-specific T cells in different intestinal sites of chronically SIV-infected rhesus monkeys

To determine the loss of CD4+ T cells and virus-specific cytotoxic T cells (CTL) in different mucosal sites of rhesus monkeys infected with simian immunodeficiency virus (SIV). A cross-sectional comparative investigation of seven different mucosal sites from chronically SIV-infected rhesus monkeys was performed by analyzing blood and mucosal lymphocytes. Mucosal lymphocytes were isolated from duodenum, jejunum, ileum and colon as well as from vagina, cervix and uterus of SIV-infected rhesus monkeys at necropsy. CD4+ T cells and SIV-Gag-specific CTL were determined in blood and mucosal samples by flow cytometry. A significant depletion of CD4+ T cells was observed in blood and all mucosal sites of SIV-infected rhesus monkeys compared to uninfected animals. But the mean percentage loss of CD4+ T cells varied between 66 and 95% between the different mucosal tissues. The frequency of CTL ranged between 0.4 and 2.4% with the highest proportions in vagina and cervix. Among the intestinal sites the mean levels of CTL correlated with mean percentage loss of CD4+ T cells. A discriminative pronounced loss of CD4+ T cells among the mucosal tissues confirmed that viral replication affects different mucosal sites in a distinct way. Despite high levels of CTL, especially in vagina and cervix, the severe loss of mucosal CD4+ T cells could not be prevented during chronic SIV infection. However, within the four sites of the intestine a high virus-specific cellular immune response correlated with better preservation of CD4+ T cells.

Ovarian and Uterine Ultrasonography in Aotus azarai infulatus

We evaluated the uterus and ovaries of owl monkeys (Aotus azarai infulatus) via gynecological ultrasound examination. We evaluated the subjects in 2 different time periods. The first period (P1) was characterized by the absence of mating, with daily examinations, during 4 mo (n = 10). At the end of P1, we paired the subjects for 30 d, but without ultrasonographic evaluation. The second period (P2) was characterized by the presence of mating, with examinations once a week, during 7 consecutive months (n = 9). We evaluated the uterus and ovaries in sagittal and transverse scans, using a 5–12 MHz linear array probe. The uterine volume (UV) was directly proportional to the number of previous parturitions. The right ovary volume (RtOV) is greater than the left (LtOV) in P1 and P2. There is a positive correlation (p < 0.05) between the females’ mass, RtOV (r = 0.28) and LtOV (r = 0.16).

Immunohistochemical evaluation of uteri from female rhesus monkeys (macaca mulatta) after cyclic intramuscular estrogen treatment

OBJECTIVE: We previously reported significant changes of estrogen receptor-α (ER) and progesterone receptor (PR) in the uteri of perimenopausal female rhesus monkeys that underwent ovariectomies (OVXs) and were then randomized to receive intramuscular 17-estradiol treatment (E). We now report the expression of oncogenic receptors HER2 and BCL2.DESIGN: A prospective, randomized double-blind study.MATERIALS AND METHODS: Monkeys underwent OVXs and were randomly assigned to age-matched OVX plus E or OVX plus placebo vehicle (V) treatment groups. After a post-OVX interval of 30 ± 1.7 weeks (mean ± SEM), 15 monkeys received E (100 g/ml) and 13 monkeys received V every 3 weeks for 2 to 3 years. Paraffin sections of the uterine corpus and lower uterine segment (LUS) were stained using standard techniques for HER2 and BCL2. Immunohistochemical evaluation of glandular (G), myometrium (M), and stromal (S) compartments were performed separately on uterine corpus and LUS regions. Scoring for HER2 and BCL2 were performed by multiplying the percentage of positive cells (0-25% =1, 26-50% =2, 51-75% =3, and 76-100% = 4) by the intensity of staining (graded 1-4). Mixed model analysis was used to account for correlation between repeated measures made within a given monkey.TableGroupNMeanMedianNMeanMedianWilcoxon P-valueHER2 Corpus G150.200131.5400.112HER2 Corpus M150.000130.000NAHER2 Corpus S150.000130.000NAHER2 LUS G150.200130.6900.235HER2 LUS M150.000130.000NAHER2 LUS S150.000130.000NABCL2 Corpus G152.671101.5000.218BCL2 Corpus M154.604122.6710.251BCL2 Corpus S152.201121.330.50.440BCL2 LUS G152.332102.700.50.597BCL2 LUS M154.003123.924.50.980BCL2 LUS S151.460121.5800.546 Open table in a new tab CONCLUSIONS: Although E administration in ovariectomized monkeys results in a decrease in ER and an increase in PR in the uterus, we now demonstrate that E administration does not significantly change the expression of HER2 and BCL2. OBJECTIVE: We previously reported significant changes of estrogen receptor-α (ER) and progesterone receptor (PR) in the uteri of perimenopausal female rhesus monkeys that underwent ovariectomies (OVXs) and were then randomized to receive intramuscular 17-estradiol treatment (E). We now report the expression of oncogenic receptors HER2 and BCL2. DESIGN: A prospective, randomized double-blind study. MATERIALS AND METHODS: Monkeys underwent OVXs and were randomly assigned to age-matched OVX plus E or OVX plus placebo vehicle (V) treatment groups. After a post-OVX interval of 30 ± 1.7 weeks (mean ± SEM), 15 monkeys received E (100 g/ml) and 13 monkeys received V every 3 weeks for 2 to 3 years. Paraffin sections of the uterine corpus and lower uterine segment (LUS) were stained using standard techniques for HER2 and BCL2. Immunohistochemical evaluation of glandular (G), myometrium (M), and stromal (S) compartments were performed separately on uterine corpus and LUS regions. Scoring for HER2 and BCL2 were performed by multiplying the percentage of positive cells (0-25% =1, 26-50% =2, 51-75% =3, and 76-100% = 4) by the intensity of staining (graded 1-4). Mixed model analysis was used to account for correlation between repeated measures made within a given monkey. CONCLUSIONS: Although E administration in ovariectomized monkeys results in a decrease in ER and an increase in PR in the uterus, we now demonstrate that E administration does not significantly change the expression of HER2 and BCL2.

Study of the relationships between electrical and mechanical activities in the pregnant monkey's uterus

The efficiency of uterine contraction is usually measured by the intra uterine pressure (IUP) clinically measured through the deformation of the woman's abdomen. This indirect measurement can be inaccurate sometimes. The objective of this research is to identify in the electrical bursts, generated by the uterus during contraction electrohysterogram (EHG), some parameters that can give direct information on the IUP. In this paper, we explore the correlation that can exist between temporal, energetic as well as frequency EHG parameter and the IUP curve. Preliminary results show the existence of strong correlations between several evaluated parameters and the IUP. It suggests that non invasive recording of the electrical activity of the uterus is a promising tool for pregnancy monitoring giving thus, non invasively, direct information on the mechanical activity of the uterus.

Immunohistochemical evaluation of uteri from female rhesus monkeys (MACACA MULATTA) after cyclic intramuscular estrogen treatment

To evaluate the immunohistochemical parameters of estrogen receptor-α (ERα) and progesterone receptor (PR) in the uteri of perimenopausal female rhesus monkeys that underwent ovariectomies (OVXs) and were then randomized to receive intramuscular 17β-estradiol treatment (E). A prospective, randomized double-blinded study. Monkeys underwent OVXs and were randomly assigned to age-matched OVX plus E or OVX plus placebo vehicle (V) treatment groups. After a post-OVX interval of 30 ± 1.7 weeks (mean ± SEM), 15 monkeys received E (100 μg/ml) and 13 monkeys received V every 3 weeks for 2 to 3 years. Paraffin sections of the uterine corpus and lower uterine segment (LUS) were stained using standard techniques for ERα and PR. Immunohistochemical evaluation of glandular (G), myometrium (M), and stromal (S) compartments were performed separately on uterine corpus and LUS regions. Scoring of immunopositivity for ERα and PR were performed by multiplying the percentage of positive cells (0–25% = 1, 26–50% = 2, 51–75% = 3, and 76–100% = 4) by the intensity of staining (graded 1–4). Repeated measures analysis via SAS's Proc Mixed was used on natural log transformed data. Geometric means and their ratios are reported and were obtained by exponentiating the mixed model estimates. The uterine histology from E and V treated monkeys revealed no evidence of hyperplasia or cancer. The interaction between treatment effect and the repeated measure effect of group and the P-value for interaction was significant: P=0.017. TableGroup EGroup VRatio of GeometricGeometric MeanGeometric MeanMeans (E/V)P-value95% CIERα Corpus S1.00002.33750.42780.00720.2352, 0.7782ERα LUS S1.00002.46550.40560.00350.2277, 0.7224PR Corpus M9.15772.55393.58580.00031.9160, 6.7110PR LUS G13.88058.29521.67330.03831.0304, 2.7174PR LUS M7.38352.18803.37460.00021.8789, 6.0610PR LUS S7.44172.84132.61920.01541.2207, 5.6197 Open table in a new tab E administration in ovariectomized monkeys results in a significant decrease in ERα in the S compartment of both the uterine corpus and LUS, and a significant increase in PR in the LUS in the G, M, and S compartments. These findings demonstrate E administration plays a significant role in uterine physiology and results in the differential expression of ERα and PR in the female rhesus monkey.

Expression of Smad Ubiquitin Regulatory Factor 2 (Smurf2) in Rhesus Monkey Endometrium and Placenta During Early Pregnancy

Smad ubiquitin regulatory factor 2 (Smurf2) is an E3 ubiquitin ligase that is involved in the Smad-mediated TGF-beta signaling. TGF-beta has been shown to play an important role during normal embryo implantation, but whether Smurf2 is involved in this process has not been reported. This study was first conducted to investigate the expression of Smurf2 transcript and protein in different compartments of the rhesus monkey uteri and placenta during early pregnancy. The results showed that both the cloned partial sequence of Smurf2 gene and the corresponding amino acid residues shared 99% identity with those of human homologs. On day 12 (D12) of pregnancy, strong signals of Smurf2 mRNA were found in basalis glandular epithelium and luminal epithelium, and moderate expressions were detected in functionalis glandular epithelium. During early villi stage and villi placental stage, Smurf2 mRNAs were mainly localized in the placenta villi, trophoblastic column, trophoblastic shell, and basalis glandular epithelium. There appeared strong staining signals in the arterioles on D26 of pregnancy, but faint staining signals on D18 of pregnancy. No specific staining of Smurf2 mRNA was observed in stromal cells and myometrium. The expression pattern of Smurf2 protein was generally similar to that of its mRNA. These results provide the first evidence that Smurf2 may play specific roles in glandular secretion, trophoblastic cell invasion, and placentation through mediating the expression of the related proteins of TGF-beta signaling pathway during early pregnancy.

Comparative Expression and Distribution ofc-fos, Estrogen Receptorα (ERα), and p38α in the Uterus of Rats, Monkeys, and Humans

The uterine cellular expression and distribution of c-fos, ERalpha and p38alpha was compared in humans, nonhuman primates, and rats using immunohistochemistry. ERalpha and c-fos were present in the glandular (GE) and luminal epithelial cells (LE) of humans and nonhuman primates, with differing expression patterns evident between proliferative and secretory cycle phases. In rats, the highest and lowest expression of c-fos was present during proestrus and estrus, respectively, in the LE and GE. The most intense ERalpha staining in rats was observed during proestrus in the GE, while the least intense staining was seen in the LE during proestrus. Strong LE and GE expression of p38alpha was present in rats in all stages of the estrous cycle and during the proliferative phase in both humans and nonhuman primates. No p38alpha expression was observed during the secretory phase in either humans or nonhuman primates. Our work suggests that c-fos, ERalpha and p38alpha (a) are primarily expressed during the proliferative phase, but not the secretory phase and exhibit interspecies expression variability, and (b) rats exhibit cyclic changes in the expression of c-fos and ERalpha.

Expression of adamalysin 19/ADAM19 in the endometrium and placenta of rhesus monkey (Macaca mulatta) during early pregnancy

A disintegrin and metalloproteinase (ADAM) 19 may contribute to multiple processes including proteolysis, adhesion and intracellular signalling. These processes are also critical for embryo implantation. The aim of this study was to investigate the spatio-temporal expression of the ADAM19 in rhesus monkey uteri on days 12, 18 and 26 of pregnancy. The results showed that in the cloned monkey 346 bp ADAM19 gene fragment and 114 amino acid residues were 98 and 100% identical to those of human homologues, respectively. In-situ hybridization confirmed that the ADAM19 mRNA was located in the luminal and glandular epithelium on day 12 of pregnancy. On day 18 of pregnancy, strong signals of the ADAM19 mRNA were detected in the placental villi, trophoblastic column and glandular epithelium near the myometrium. Moderate expression of the ADAM19 mRNA was seen in the trophoblastic shell and stromal cells. The placental villi and trophoblastic column expressed abundant ADAM19 mRNA, and ADAM19 transcripts were also detected in the trophoblastic shell and fetal-maternal border on day 26 of pregnancy. The expression pattern of the ADAM19 protein was similar to its transcript, but signals for the ADAM19 protein in the stromal cells and trophoblastic shell increased more than its mRNA on day 18 of pregnancy. Statistical analysis demonstrated that the expression level of ADAM19 significantly increased on day 18 of pregnancy. These data suggest that the ADAM19 may be involved in the key processes of glandular secretion, trophoblast invasion and degradation of extracellular matrix during early pregnancy.

Expression of proteasome subunits low molecular mass polypeptide (LMP) 2 and LMP7 in the endometrium and placenta of rhesus monkey (Macaca mulatta) during early pregnancy.

Previous studies have demonstrated that the ubiquitin-proteasome pathway plays an important role in embryo implantation. Low molecular mass polypeptide (LMP) 2 and LMP7 are the two subunits of 20S proteasome, which are critical for proteasome activity. To further elucidate the roles of LMP2 and LMP7 in embryo implantation during early pregnancy, we cloned partial sequences of the LMP2 and LMP7 genes and studied the spatiotemporal expression of LMP2 and LMP7 in rhesus monkey (Macaca mulatta) uteri on Days 12, 18, and 26 of pregnancy. The results showed that the 349-base pair (bp) LMP2 fragment and the 340-bp LMP7 fragment were 97% and 99% identical, respectively, to those of human homologues. From the statistical results of reverse transcription-polymerase chain reaction, in situ hybridization, and immunohistochemistry, we found that the expression levels of LMP2 and LMP7 significantly increased with the elongation of pregnancy. The LMP2 and LMP7 mRNAs were mainly expressed in the luminal and glandular epithelium on Day 12 of pregnancy. On Days 18 and 26 of pregnancy, strong signals of LMP2 and LMP7 mRNAs were detected in the placental villi, trophoblastic column, and arterial endothelial cells close to the implantation site, and moderate expressions were found in the trophoblastic shell and glandular epithelium. The LMP2 and LMP7 mRNAs were extensively distributed in the stroma on Day 26 of pregnancy. The expression patterns of LMP2 and LMP7 proteins were similar to those of their transcripts, but weak immunostaining of LMP2 and LMP7 proteins was detected in stroma at all stages of pregnancy. These results suggest that LMP2 and LMP7 may be involved in some key processes of trophoblastic invasion, angiogenesis, degradation of the extracellular matrix, immune tolerance, and glandular secretion.

Identification and specific expression of matrix metalloproteinase-26 in rhesus monkey endometrium during early pregnancy.

Matrix metalloproteinases (MMPs) and their tissue inhibitors may play important roles in tissue remodelling processes of the uterus. This study identified MMP-26 (endometase/matrilysin-2) in the endometrium of pregnant rhesus monkeys (Macaca mulatta) and monitored the spatial and temporal expression of the transcript and protein in the uteri on days 12, 18 and 26 of pregnancy. The partial monkey MMP-26 gene sequence of 289 nucleotides was 98% identical to that of its human homologue and its protein fragment contained a PHCGVPDGSD sequence in the prodomain identical to that in human MMP-26. RT-PCR analysis demonstrated that the average level of MMP-26 mRNA in the endometrium was high on day 12 of pregnancy, but significantly decreased on days 18 and 26 (P < 0.05). In-situ hybridization confirmed that MMP-26 mRNA is specifically localized in the endometrial compartments, with intense signals in the glandular epithelium on day 12 and in the walls of spiral arterioles adjacent to the implantation site on day 26. The hybridization signal for MMP-26 mRNA in the glandular epithelium decreased dramatically on day 18 and was undetectable on day 26. No MMP-26 mRNA transcripts were detected in the placental villi on days 18 and 26. Immunohistochemistry showed that the expression pattern of MMP-26 protein was similar to that of its mRNA. The restricted expression pattern of MMP-26 in the monkey uterus implies that this new MMP is involved in the highly regulated tissue remodelling processes of the glandular epithelium and spiral arteries during early pregnancy.

Calcitonin immunostaining in monkey uterus during menstrual cycle and early pregnancy.

Calcitonin has been shown to be a progesterone-regulated potential marker of the receptive endometrium in the rat and human. The present study was undertaken to immunohistochemically investigate the changes in calcitonin in the monkey uterus during the menstrual cycle and periimplantation period. Calcitonin immunostaining was primarily localized in the glandular epithelium on d 16, 20, and 25 of the menstrual cycle. During early pregnancy, calcitonin immunostaining was strongly observed in the glandular epithelium only on d 9 of pregnancy, the day before implantation. Since the high level of calcitonin immunostaining in the glandular epithelium during the luteal phase of the menstrual cycle and periimplantation period matched the high level of maternal progesterone during this period, the expression of calcitonin in monkey endometrium may be under the regulation of maternal progesterone.

Characterization of relaxin binding in the uterus of the marmoset monkey.

The ovarian peptide hormone relaxin (RLX) plays an important role in the regulation of the endometrium both during the cycle and in early pregnancy. RLX interacts with specific receptors on endometrial stromal cells causing these to decidualize. In order to characterize the molecules with which RLX interacts in the primate uterus, a methodology based on a fully bioactive preparation of biotinylated porcine RLX was applied to cryosections of the uterus of female marmoset monkeys. Specific RLX binding was weakly detected in the proliferative phase in isolated endometrial stromal cells. In the secretory phase, the positively reacting cells increased in staining intensity and in number and also included some epithelial cells. Further increases occurred in pregnancy, but RLX binding in the endometrium decreased at the end of the cycle if pregnancy did not occur. The myometrium showed weak staining which did not vary through the cycle, but increased in pregnancy. Electrophoretic analysis of the RLX-binding moieties in these tissue sections indicated that a protein of approximately 40 kDa was the principal RLX-binding molecule, while minor specific bands were detectable at approximately 100 and approximately 200 kDa. The binding of biotinylated RLX could be specifically suppressed by co-incubation with unlabelled RLX, but not by insulin, IGF-I or biotin. This technique therefore allows the detection and molecular characterization of specific RLX binding in the primate uterus. In the marmoset monkey, the pattern of specific binding closely reflects the RLX-dependent physiology during implantation and early pregnancy, implying the probable involvement of a specific RLX receptor.

Expression of matrix metalloproteinase-2, -9, -14, and tissue inhibitors of metalloproteinase-1, -2, -3 in the endometrium and placenta of rhesus monkey (Macaca mulatta) during early pregnancy.

The extracellular matrix proteolytic machinery has long been recognized as one of the most important mechanisms for regulating trophoblast invasion. Matrix metalloproteinases (MMPs) are a group of proteases involved in this process, and their activities are regulated by tissue inhibitors of MMPs (TIMPs). In this study, we collected rhesus monkey uteri on Days 12, 18, and 26 of pregnancy and examined the mRNA expression of MMP-2, -9, -14, and TIMP-1, -2, -3, as well as the activities of MMP-2 and -9 by using in situ hybridization and gelatin zymography, respectively. The results showed that MMP-2 and -9 were expressed earlier than MMP-14 and TIMPs in the pregnant endometrium. MMP-14 and TIMP-2 mRNAs appeared in perivascular decidual cells earlier than MMP-2 mRNA. On Day 26 of pregnancy, placental villi expressed little MMP-2, -14, and TIMP transcripts but abundant MMP-9 mRNA. Furthermore, MMP-2, -9, -14, and TIMP-1, -2, -3 were highly expressed on the fetal-maternal border but were absent in the myometrium. TIMP-3 mRNA in the endometrium was specifically localized to some cells lining the outer membrane of several groups of arterioles. Combined with the results obtained by gelatin zymography, we found that active MMP-2 existed in the endometrium throughout these three phases, while MMP-9 showed considerable activities only on Days 18 and 26 of pregnancy. The data suggest key roles for MMP-2 and -9 in invasion of trophoblast cells into the endometrium and the development of the placenta and might indicate that these processes are regulated by MMP-14 and TIMP-1, -2, and -3.

Leukemia inhibitory factor, leukemia inhibitory factor receptor, and glycoprotein 130 in rhesus monkey uterus during menstrual cycle and early pregnancy.

This goal of this study was to examine immunohistochemical distribution of leukemia inhibitory factor (LIF), LIF receptor (LIFR), and glycoprotein (gp) 130 in rhesus monkey uterus during the menstrual cycle and early pregnancy. Pregnancy rate was significantly reduced in the control group from 66.7% (12 of 18) to 22.2% (4 of 18) with an injection of goat anti-human recombinant LIF immunoglobulin G into the uterine lumen on Day 8 of pregnancy. LIF was mainly localized in glandular and luminal epithelium. LIF immunostaining during the luteal phase was stronger than it was during the proliferative phase. LIF staining gradually increased from Day 3 of pregnancy and reached its highest level on Day 9. LIFR was mainly localized in the glandular and luminal epithelium. LIFR staining during the luteal phase was stronger than it was during the proliferative phase. LIFR staining began to increase from Day 3 of pregnancy and reached a high level on Days 9 and 11. Gp130, a signal-transducing receptor component of LIF, was mainly localized in the glandular epithelium. A high level of gp130 was found on Days 16 and 20 of menstrual cycle, and from Days 5 to 11 of pregnancy. These results suggest that LIF may play an important role in monkey implantation, as it does in mice.

Photodynamic endometrial ablation in the nonhuman primate.

To test the effectiveness of 5-aminolevulinic acid (ALA)-induced photodynamic endometrial ablation in the rhesus monkey under varying conditions of light delivery (fractionated versus continuous) and steroid priming. Photodynamic endometrial ablation was carried out in 17 rhesus monkeys that were either postmenopausal or in the early proliferative phase. Four hours after intralumenal injection of ALA (250 mg in 1 mL hyskon), a quartz fiber with a diffusing tip was inserted. A KTP tunable dye laser delivered 300 mW of light (635 nm) for 60 minutes in either continuous or fractionated fashion (20 minutes on, 5 minutes off, and 40 minutes on). In some experiments, thermistors were used to monitor temperature in the lumen and myometrium during light treatment. Hysterectomy was performed 3 or 4 days after treatment, and endometrial damage was assessed histologically. Two additional monkeys (one rhesus and one cynomolgus monkey) were exposed to the same protocol, except hyskon was substituted for ALA to control for potential ablative effects due to light treatment alone. Endometrial ablation was evident in all ALA-photosensitized specimens. The degree of ablation around the light fiber ranged from moderate to complete. The depth of ablation ranged from 1.14 +/- 0.54 to 2.15 +/- 1.62 mm (mean +/- standard deviation). Ablation was most complete in uteri of menopausal monkeys. Light treatment after ALA increased lumenal temperature from 36 C to 50 C, whereas temperature was not significantly increased by light treatment in the controls. This is the first report of endometrial destruction in the primate using a photodynamic approach. Whereas clinical application of photodynamic therapy (PDT) requires complete endometrial ablation to produce long-lasting amenorrhea, our results suggest that PDT may offer a simple office-based approach to endometrial ablation.