232 DIFFERENTIAL EXPRESSION OF UTERINE CALCIUM-RELATED PROTEINS (NCKX3, NCX1, PMCA1, TRPV6, AND CABINDIN-D9k AND -D28k) DURING HUMAN MENSTRUAL CYCLE

2010 ◽  
Vol 22 (1) ◽  
pp. 274
Author(s):  
K. C. Choi ◽  
E. B. Jeung
Keyword(s):  
Menstrual Cycle ◽  
Medical Center ◽  
Expression Patterns ◽  
Embryo Implantation ◽  
Calcium Balance ◽  
Human Uterus ◽  
Secretory Phase ◽  
Total N ◽  
Proliferative Phase ◽  
Related Proteins

The endometrium is hostile to embryo implantation except during the window of receptivity. A change in endometrial gene expression is required for the development of receptivity. The uterine calcium balance is crucial for physiological functioning, including smooth muscle contraction and embryo implantation. The location of cytoplasmic calcium-related proteins (CRP) include the calcium transporters 1 (CaT1), calbindin-D9k/-D28k (CaBP- 9k/28k), plasma membrane Ca2+-ATPase 1b (PMCA1b), sodium/calcium exchangers (NCX1), and potassium-dependent Na+/Ca2+ exchanger (NCKX3). The expressions of these CRP and their potential roles in the uterus of human during the menstrual cycle remain to be clarified. Thus, in this current study, the expression patterns of CRP were examined for their roles in the human uterus during the menstrual cycle. Human endometrial tissues were collected by curettage from women undergoing hysteroscopy for investigation of tubal patency or tubal ligation. Approval was given by the Human Ethics Committee at SCH Medical Center, Bucheon, and signed consent was obtained in every case. Human uterus (total n = 51) were divided into 3 groups: menstrual, proliferative, and secretory phase. Reverse-transcription PCR and Western blot analysis were applied to measure the level of CRP mRNA and protein, respectively. During the menstrual cycle of human, the expression levels of CaT1 mRNA and protein were increased 5-fold at proliferative phase (Days 6 to 13) compared with secretory phase in the endometrium of uterus. The expression of CaBP-28k mRNA and protein was less 2-fold during the proliferative phase (Days 6 to 13) than during the secretory phase (Days 16 to 28). However, the expressions of NCX1, NCKX3, and PMCA1b mRNA and protein were not altered during cycle, whereas the expression of CaBP-9k was not observed in the uterus of human. In addition, spatial expression of CRP was detected by immunohistochemistry Uterine CRP was abundantly localized in the cytoplasm of the luminal and glandular epithelial cells during menstrual cycle. Taken together, these results indicate that uterine CRP is abundantly expressed in the uterus, suggesting that uterine expression of CRP might be involved in reproductive function during the menstrual cycle in human.

Polo-like kinase expression in normal human endometrium during the menstrual cycle

2000 ◽  
Vol 12 (2) ◽  
pp. 59 ◽  
Author(s):  
Noriyuki Takai ◽  
Tami Miyazaki ◽  
Isao Miyakawa ◽  
Ryoji Hamanaka
Keyword(s):  
Menstrual Cycle ◽  
Stromal Cells ◽  
Cellular Proliferation ◽  
Secretory Phase ◽  
Ki 67 ◽  
Normal Endometrium ◽  
Proliferative Phase ◽  
Normal Human ◽  
Late Secretory Phase

The enzyme, polo-like kinase (PLK), is a mammalian serine/threonine kinase involved in cell cycle regulation. A great deal of evidence regarding the role of PLK in the cell cycle has been obtained through studies of cultured cells, though little is known about its function or even expression in vivo. The endometrium undergoes rapid proliferation and differentiation under ovarian steroid hormone control during the 28-day cycle. Thus, normal endometrium provides an excellent model in which to study the hormone dependency of PLK expression. In the present study, we examined the features of PLK expression in 20 samples of normal human endometrium during the menstrual cycle. The expression of Ki-67 and proliferating cell nuclear antigen (PCNA) were also examined as markers of proliferation. Immunohistochemical studies showed that PLK staining was detected in the basement membrane of many endometrial glands, stromal cells, and some endothelial cells. The number of PLK-positive endometrial gland cells was significantly higher in the late proliferative phase (19.16% 4.98%) and the early secretory phase (19.28% 4.99%) than in the early proliferative phase (2.60% 2.33%) or the late secretory phase (5.76% 2.16%) (P<0.0001). PLK expression seemed to be correlated with the expression of Ki-67 and PCNA in many endometrial glands and stromal cells particularly in the late proliferative phase, reflecting a role of PLK in cellular proliferation. Nevertheless, in the early secretory phase, at which point the expression of Ki-67 and PCNA decreased in endometrial glands, PLK was strongly expressed. This finding suggests that PLK may have some post-mitotic functions in certain specialized cell types. Although the highest expression of PLK was observed in the late proliferative and the early secretory phases, the expression drastically decreased in the late secretory phase. These findings, taken together, indicate that the expression of PLK in normal endometrium fluctuates over the course of the menstrual cycle, suggesting in turn that PLK is associated with hormone-dependent cellular proliferation and that hormone functions may be involved in its regulation.

METABOLISM OF OESTRADIOL-17β AND OESTRONE IN THE HUMAN UTERUS

1975 ◽  
Vol 80 (4) ◽  
pp. 719-731 ◽  
Author(s):  
A. R. Krishnan ◽  
B. K. Bajaj ◽  
V. Hingorani ◽  
K. R. Laumas
Keyword(s):  
Metabolic Activity ◽  
Subcellular Fractions ◽  
Human Endometrium ◽  
Physiological Significance ◽  
Pyridine Nucleotides ◽  
Hormone Action ◽  
Human Uterus ◽  
Secretory Phase ◽  
Proliferative Phase ◽  
Metabolic Conversion

ABSTRACT A study of the metabolism of oestradiol in the human endometrium and myometrium of the proliferative and secretory phases of the cycle showed that the conversion of oestradiol to oestrone by endometrium in the proliferative phase was higher than that in the secretory phase. The decreased metabolic activity of the secretory phase endometrium was attributed to the influence of progesterone on the endometrium. The metabolic conversion of oestradiol to oestrone was enhanced when pyridine nucleotides were added to the system. The conversion of oestradiol to oestrone was maximum in the cytoplasmic and nuclear fractions of the endometrium. Furthermore, the conversion of oestradiol was low in all the subcellular fractions of the myometrium as compared with the endometrial subcellular fractions. The presence of co-factors increased the metabolic conversion of oestradiol to oestrone in the subcellular fractions of the endometrium. The presence of 17β-hydroxysteroid oxidoreductase was indicated in all the subcellular fractions. A correlation was found between the amount of oestradiol and oestrone bound to the receptors in the uterus and the rate of metabolism of oestradiol in the uterus. The physiological significance of metabolism of oestradiol and the hormone action are discussed.

scholarly journals The SARS-CoV-2 receptor, Angiotensin converting enzyme 2 (ACE2) is required for human endometrial stromal cell decidualization

2020 ◽  
Author(s):  
Sangappa B. Chadchan ◽  
Vineet K. Maurya ◽  
Pooja Popli ◽  
Ramakrishna Kommagani
Keyword(s):  
Menstrual Cycle ◽  
Stromal Cells ◽  
Stromal Cell ◽  
Converting Enzyme ◽  
Endometrial Stromal Cell ◽  
Secretory Phase ◽  
Endometrial Stromal Cells ◽  
Angiotensin Converting Enzyme 2 ◽  
Proliferative Phase ◽  
Human Endometrial Stromal Cell

AbstractSTUDY QUESTIONIs SARS-CoV-2 receptor, angiotensin-converting enzyme 2 (ACE 2) expressed in the human endometrium during the menstrual cycle, and does it participate in endometrial decidualization?SUMMARY ANSWERACE2 protein is highly expressed in human endometrial stromal cells during the secretory phase and is essential for human endometrial stromal cell decidualization.WHAT IS KNOWN ALREADYACE2 is expressed in numerous human tissues including the lungs, heart, intestine, kidneys and placenta. ACE2 is also the receptor by which SARS-CoV-2 enters human cells.STUDY DESIGN, SIZE, DURATIONProliferative (n = 9) and secretory (n = 6) phase endometrium biopsies from healthy reproductive-age women and primary human endometrial stromal cells from proliferative phase endometrium were used in the study.PARTICIPANTS/MATERIALS, SETTING, METHODSACE2 expression and localization were examined by qRT-PCR, Western blot, and immunofluorescence in both human endometrial samples and mouse uterine tissue. The effect of ACE2 knockdown on morphological and molecular changes of human endometrial stromal cell decidualization were assessed. Ovariectomized mice were treated with estrogen or progesterone to determine the effects of these hormones on ACE2 expression.MAIN RESULTS AND THE ROLE OF CHANCEIn human tissue, ACE2 protein is expressed in both endometrial epithelial and stromal cells in the proliferative phase of the menstrual cycle, and expression increases in stromal cells in the secretory phase. The ACE2 mRNA (P < 0.0001) and protein abundance increased during primary human endometrial stromal cell (HESC) decidualization. HESCs transfected with ACE2-targeting siRNA were less able to decidualize than controls, as evidenced by a lack of morphology change and lower expression of the decidualization markers PRL and IGFBP1 (P < 0.05). In mice during pregnancy, ACE2 protein was expressed in uterine epithelial and stromal cells increased through day six of pregnancy. Finally, progesterone induced expression of Ace2 mRNA in mouse uteri more than vehicle or estrogen (P < 0.05).LARGE SCALE DATAN/A.LIMITATIONS, REASONS FOR CAUTIONExperiments assessing the function of ACE2 in human endometrial stromal cell decidualization were in vitro. Whether SARS-CoV-2 can enter human endometrial stromal cells and affect decidualization have not been assessed.WIDER IMPLICATIONS OF THE FINDINGSExpression of ACE2 in the endometrium allow SARS-CoV-2 to enter endometrial epithelial and stromal cells, which could impair in vivo decidualization, embryo implantation, and placentation. If so, women with COVID-19 may be at increased risk of early pregnancy loss.STUDY FUNDINGS/COMPETING INTEREST(S)This study was supported by National Institutes of Health / National Institute of Child Health and Human Development grants R01HD065435 and R00HD080742 to RK and Washington University School of Medicine start-up funds to RK. The authors declare that they have no conflicts of interest.

scholarly journals Tripeptidyl peptidase I promotes human endometrial epithelial cell adhesive capacity implying a role in receptivity

2020 ◽  
Vol 18 (1) ◽  
Author(s):  
Leilani L. Santos ◽  
Cheuk Kwan Ling ◽  
Evdokia Dimitriadis
Keyword(s):  
Menstrual Cycle ◽  
Epithelial Cell ◽  
Stromal Cells ◽  
Embryo Implantation ◽  
Unexplained Infertility ◽  
Implantation Failure ◽  
Secretory Phase ◽  
Endometrial Epithelial Cell ◽  
Tripeptidyl Peptidase ◽  
Adhesive Capacity

AbstractThe endometrium undergoes cyclic remodelling throughout the menstrual cycle in preparation for embryo implantation which occurs in a short window during the mid-secretory phase. It is during this short ‘receptive window’ that the endometrial luminal epithelium acquires adhesive capacity permitting blastocysts firm adhesion to the endometrium to establish pregnancy. Dysregulation in any of these steps can compromise embryo implantation resulting in implantation failure and infertility. Many factors contribute to these processes including TGF-β, LIF, IL-11 and proteases. Tripeptidyl peptidase 1 (TPP1) is a is a lysosomal serine-type protease however the contribution of the TPP1 to receptivity is unknown. We aimed to investigate the role of TPP1 in receptivity in humans.In the current study, TPP1 was expressed in both epithelial and stromal compartments of the endometrium across the menstrual cycle. Expression was confined to the cytoplasm of luminal and glandular epithelial cells and stromal cells. Staining of mid-secretory endometrial tissues of women with normal fertility and primary unexplained infertility showed reduced immunostaining intensity of TPP1 in luminal epithelial cells of infertile tissues compared to fertile tissues. By contrast, TPP1 levels in glandular epithelial and stromal cells were comparable in both groups in the mid-secretory phase. Inhibition of TPP1 using siRNA compromised HTR8/SVneo (trophoblast cell line) spheroid adhesion on siRNA-transfected Ishikawa cells (endometrial epithelial cell line) in vitro. This impairment was associated with decreased sirtuin 1 (SIRT1), BCL2 and p53 mRNA and unaltered, CD44, CDH1, CDH2, ITGB3, VEGF A, OSTEOPONTIN, MDM2, CASP4, MCL1, MMP2, ARF6, SGK1, HOXA-10, LIF, and LIF receptor gene expression between treatment groups. siRNA knockdown of TPP1 in primary human endometrial stromal cells did not affect decidualization nor the expression of decidualization markers prolactin (PRL) and insulin-like growth factor-binding protein 1 (IGFBP1). Taken together, our data strongly suggests a role for TPP1 in endometrial receptivity via its effects on epithelial cell adhesion and suggests reduced levels associated with unexplained infertility may contribute to implantation failure.

182 CALBINDIN-D28k IS PREDOMINANTLY EXPRESSED AND REGULATED BY ESTROGEN IN HUMAN ENDOMETRIUM DURING MENSTRUAL CYCLE

2011 ◽  
Vol 23 (1) ◽  
pp. 192
Author(s):  
K.-C. Choi ◽  
H. Yang ◽  
E.-B. Jeung
Keyword(s):  
Menstrual Cycle ◽  
Calcium Binding ◽  
Potential Role ◽  
Embryo Implantation ◽  
Endometrial Receptivity ◽  
Human Endometrium ◽  
Ishikawa Cell ◽  
Proliferative Phase ◽  
Calbindin D28k ◽  
The Brain

The human endometrium resists embryo implantation except during the window of receptivity. A change in endometrial gene expression is required for the development of receptivity. Uterine calbindin-D28k (CaBP-28k) has been shown to be involved in the regulation of endometrial receptivity by intracellular Ca2+. Nowadays, this protein has been mainly linked to the brain, kidneys, and pancreas, but potential role(s) of CaBP-28k remain to be clarified in the uterus of humans during the menstrual cycle. Thus, we demonstrated in this study that the expression of CaBP-28k in the human endometrium in more divided in the menstrual phases. During the menstrual cycle of humans, uterine expression levels of CaBP-28k mRNA and protein increased at the proliferative phase and fluctuated in these tissues, compared with other phases. We assessed the effects of the sex-steroid hormones E2 and P4 on the expression of CaBP-28k in the Ishikawa cell line. A significant increase in the expression of CaBP-9k mRNA was observed at the concentration of 17β-oestradiol (E2; 10–9 to 10–7 M). In addition, spatial expression of CaBP-28k was detected by immunohistochemistry. CaBP-28k is abundantly localised in the cytoplasm of the luminal and glandular epithelial cells during the menstrual cycle. Taken together, these results indicate that CaBP-28k, a uterine calcium-binding protein, is abundantly expressed in the human uterus, suggesting that uterine expression of CaBP-28k may be involved in reproductive functions during the menstrual cycle of humans.

203 PLASMA MEMBRANE Ca2+-PUMPING ATPase 1 IS ABUNDANTLY EXPRESSED AND DISTINCTLY REGULATED BY ESTROGEN IN HUMAN ENDOMETRIUM DURING THE MENSTRUAL CYCLE

2011 ◽  
Vol 23 (1) ◽  
pp. 201
Author(s):  
H. Yang ◽  
E.-B. Jeung
Keyword(s):  
Plasma Membrane ◽  
Menstrual Cycle ◽  
Critical Role ◽  
Human Endometrium ◽  
Cycle Phase ◽  
Menstrual Phase ◽  
Menstrual Cycle Phase ◽  
Secretory Phase ◽  
Total N ◽  
High Level

Plasma membrane Ca2+-pumping ATPases (PMCA) play a critical role in maintaining cellular Ca2+ homeostasis. The PMCA mRNA are encoded on 4 genes, designated PMCA1 to PMCA4. In a previous study, we found that both PMCA1 and PMCA4 are expressed at similar levels in astrocytes and in neurons. Although PMCA1b is expressed in the uterus of rats during the oestrous cycle, the expression of PMCA1 and its potential roles has not been elucidated during the menstrual cycle in the human endometrium. Thus, in the current study, the expression pattern of PMCA1 was examined to predict its roles in the human endometrium during the menstrual cycle. Human uterine tissues (total n = 40) were separated into 3 groups according to menstrual cycle phase: menstrual phase, proliferative phase (early, mid, late), and secretory phase (early, mid, late). Using real-time PCR and Western blot analysis, uterine expression of PMCA1 mRNA and protein increased to 1.5-fold in the early-, mid- and late-proliferative phases in the endometrium of the human uterus, compared with other menstrual phases. In addition, uterine PMCA1 was abundantly localised in the cytoplasm of the luminal and glandular epithelial cells in the menstrual phases, indicating that this protein may participate in the uterine Ca balance of the human endometrium during the menstrual cycle. Taken together, these results suggest that a high level of uterine PMCA1 expression may be involved in reproductive functions during the menstrual cycle of humans.

THE METABOLISM OF PROSTAGLANDIN E2 BY TISSUES FROM THE HUMAN UTERUS AND FOETO-PLACENTAL UNIT

1978 ◽  
Vol 87 (3) ◽  
pp. 632-642 ◽  
Author(s):  
Eve A. Willman ◽  
W. P. Collins
Keyword(s):  
Endometrial Tissue ◽  
Prostaglandin E ◽  
Human Uterus ◽  
Slight Effect ◽  
High Turnover ◽  
Secretory Phase ◽  
Proliferative Phase ◽  
Catabolic Enzymes ◽  
Decidual Tissue

ABSTRACT The biosynthesis and metabolism of prostaglandin E2 was studied in cell-free homogenates of tissues from the uterus and foeto-placental unit using specific in vitro methods. The results show that the synthesis of prostaglandin E2 is greater in endometrial tissue from the secretory phase (53.07 ± 39.05; ng/100 mg wet tissue/h) than from the proliferative phase of the uterine cycle. Furthermore, endometrial tissue forms more of this compound than myometrium (P < 0.005). During early pregnancy, synthesis is decreased (25.10 ± 16.62); at term, myometrium produces more prostaglandin E2 than decidua. During labour, however, decidual tissue accumulates more of this compound (52.73 ± 18.04) than either myometrium (34.71 ± 13.19) or cord (28.63 ± 11.71). The catabolic enzymes are most active in the placenta and chorion, followed by the cord, myometrium, decidua and amnion, but the differences have only a slight effect on the amounts of prostaglandin E2 which remain at the end of the incubation. The results suggest a high turnover of prostaglandin E2 in the decidua, myometrium and cord.

UPTAKE AND METABOLISM OF TRITIATED OESTRADIOL AND OESTRONE BY HUMAN ENDOMETRIAL AND MYOMETRIAL TISSUE IN VITRO

1974 ◽  
Vol 62 (1) ◽  
pp. 109-123 ◽  
Author(s):  
R. G. GABB ◽  
G. M. STONE
Keyword(s):  
Hydroxysteroid Dehydrogenase ◽  
Endometrial Tissue ◽  
Human Uterus ◽  
Secretory Phase ◽  
Proliferative Phase ◽  
Significant Difference ◽  
Degree Of Oxidation ◽  
Tissue Homogenates ◽  
Uptake And Metabolism

SUMMARY Human endometrial and myometrial tissues were incubated in vitro with [3H]oestradiol-17β and [3H]oestrone to study the uptake and interconversion of these two steroids by the tissues. Endometrial tissue displayed a higher capacity for oestrogen interconversion than myometrial tissue and the oxidation of oestradiol-17β to oestrone was favoured, rather than the reverse reaction. A greater degree of oxidation was found in tissue taken from uteri in the secretory phase than in the proliferative phase. A study of the distribution of radiometabolites between 'soluble' and 'particulate' fractions of tissue homogenates showed that a greater proportion of the tissue radioactivity was associated with the 'particulate' fraction in the proliferative phase than in the secretory phase. After incubation of endometrial tissue from the secretory phase with tritiated oestrogens there was evidence of the formation of chloroform-insoluble radiometabolites and some of these were tentatively identified as oestrogen sulphates. In a second experiment, the uptake and metabolism of the same two oestrogens by tissue from leiomyomata uteri (fibroids) and normal myometrial tissue were compared. No significant difference between these tissues was found. The results suggest that the levels of 17β-hydroxysteroid dehydrogenase in the human uterus may be dependent upon the levels of oestrogens in the blood. The lack of a difference in the treatment of oestrogens by fibroid and normal myometrial tissue suggests that these tissues may have a similar mechanism for the uptake and retention of oestrogens.

scholarly journals Mitotic activity of smooth muscle cells of the myoma: Does hormonal stimulation have an effect on the number of mitoses?

2010 ◽  
Vol 62 (1) ◽  
pp. 39-45
Author(s):  
Tatjana Kastratovic ◽  
Irena Tanaskovic ◽  
Vesna Lackovic ◽  
Marija Sorak ◽  
Vesna Stankovic ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Menstrual Cycle ◽  
Smooth Muscle Cells ◽  
Mitotic Activity ◽  
Luteal Phase ◽  
Muscle Cells ◽  
Secretory Phase ◽  
Hormonal Stimulation ◽  
Proliferative Phase ◽  
Proliferating Activity

Myomas develop as a result of increased mitotic (proliferating) activity of smooth muscle cells. In this study we examined the pathohistological samples of 176 myomas and their endometria that were obtained after hysterectomy from patients in the proliferative (follicular) and secretory (luteal) phase of the menstrual cycle. We examined the mitotic activity of the myoma cells in both phases and established that the average number of mitoses in the proliferative phase was significantly larger compared to the secretory phase, and that in the proliferative phase of the cycle there exists a statistically significant convergent association of the number of mitoses in the endometrium and in myomas. The number of endometrial mitoses is significantly larger than in myomas in both phases of the cycle.

241. Changes in the expression of Annexin IV mRNA and protein in human endometrium

2005 ◽  
Vol 17 (9) ◽  
pp. 95
Author(s):  
A. P. Ponnampalam ◽  
P. A. W. Rogers
Keyword(s):  
Menstrual Cycle ◽  
Cellular Localization ◽  
Single Cells ◽  
Human Endometrium ◽  
Reproductive Age ◽  
Histological Evaluation ◽  
Secretory Phase ◽  
Proliferative Phase ◽  
Cyclic Changes ◽  
Late Secretory Phase

In a previous study investigating global gene expression throughout the menstrual cycle,1 Annexin 4 (ANXIV) was identified as having significant cyclic changes in human endometrium. ANXIV belongs to a ubiquitous family of Ca2+-dependent phospholipid and membrane-binding proteins. The aims of this study were to investigate the cellular localization and regulation of ANXIV mRNA and temporal expression of ANXIV protein in human endometrium during the menstrual cycle. mRNA Expression: The menstrual cycle was divided into seven stages by histological evaluation. Curettings of endometrium were collected from 60 cycling women. For cellular localization, tissues from eight endometrial curettings were dissociated with collagenase into single cells, separated into epithelial and stromal cell fractions and snap frozen. Total RNA was extracted and ANXIV mRNA was quantified by real-time PCR. Immunohistochemistry: Full thickness endometrial tissue was collected from 50 reproductive age women undergoing hysterectomy. Tissue sections were formalin-fixed and paraffin-embedded. Goat polyclonal ANXIV antibody was used to localize ANXIV protein. ANXIV mRNA was significantly upregulated in the whole tissue during mid-late secretory phase of the cycle, and was predominantly expressed in epithelial cells. ANXIV protein was detected in the luminal and glandular epithelium in high levels throughout the menstrual cycle except in early secretory (ES) phase. The intensity of immunostaining was stronger in the glands of the basalis compared to functionalis in early proliferative phase, however, by the late secretory phase the functionalis glands showed higher expression levels. ANXIV mRNA data are consistent with a role for progesterone in upregulating the expression of ANXIV, although protein levels remain high through menstruation and into the proliferative phase. ANXIV can indirectly inhibit prostaglandin production, which is important for implantation. Hence the low levels of ANXIV protein at ES phase may relate to processes involved in implantation. (1)Ponnampalam et al. (2004). Mol. Hum. Reprod. 10(12), 879–893.

Sign in / Sign up

Export Citation Format

Share Document