Expression of leukaemia inhibitory factor at the conceptus–maternal interface during preimplantation development and in the endometrium during the oestrous cycle in the mare

2016 ◽  
Vol 28 (10) ◽  
pp. 1642 ◽  
Author(s):  
M. de Ruijter-Villani ◽  
C. Deelen ◽  
T. A. E. Stout
Keyword(s):  
Early Pregnancy ◽  
Critical Role ◽  
Endometrial Receptivity ◽  
Oestrous Cycle ◽  
Inhibitory Factor ◽  
Leukaemia Inhibitory Factor ◽  
Luminal Epithelium ◽  
Signal Transducer ◽  
Blastocyst Development ◽  
Mrna And Protein Expression

Leukaemia inhibitory factor (LIF) plays a critical role in blastocyst development and implantation in several species. The present study investigated mRNA and protein expression for LIF, as well as the low-affinity LIF receptor (LIFR) and interleukin-6 signal transducer (IL6ST), in equine endometrium, trophoblast and histotroph during early pregnancy and in the endometrium during the oestrous cycle. Endometrial LIF mRNA expression was upregulated after Day 21 of pregnancy, whereas LIF immunoreactivity increased in the endometrium on Day 28. Expression of LIF mRNA in the yolk sac membrane increased from Day 21 of pregnancy, whereas LIF immunoreactivity increased from Day 28 in the trophoblast. LIFR and IL6ST mRNA was expressed in the endometrium during both the oestrous cycle and early pregnancy and, although LIFR and IL6ST protein were localised to the glandular epithelium during the cycle and first 14 days of pregnancy, from Day 21 they were located in the luminal epithelium. Trophoblast expression of LIFR and IL6ST increased as pregnancy proceeded. In conclusion, LIF expression increased at the conceptus–maternal interface during capsule attenuation. Because contemporaneous upregulation of both LIFR and IL6ST was also observed in the trophoblast, we propose that LIF plays an important role in the development of endometrial receptivity for trophoblast growth, apposition and adhesion in mares.

Oncostatin M and leukaemia inhibitory factor trigger signal transducer and activator of transcription 3 and extracellular signal-regulated kinase 1/2 pathways but result in heterogeneous cellular responses in trophoblast cells

2016 ◽  
Vol 28 (5) ◽  
pp. 608 ◽  
Author(s):  
Wittaya Chaiwangyen ◽  
Stephanie Ospina-Prieto ◽  
Diana M. Morales-Prieto ◽  
Francisco Lazaro Pereira de Sousa ◽  
Jana Pastuschek ◽  
...  
Keyword(s):  
Dna Binding ◽  
Cell Lines ◽  
Binding Capacity ◽  
Binding Activity ◽  
Oncostatin M ◽  
Inhibitory Factor ◽  
Therapeutic Interventions ◽  
Leukaemia Inhibitory Factor ◽  
Tetrazolium Salt ◽  
Signal Transducer

Leukaemia inhibitory factor (LIF) and oncostatin M (OSM) are pleiotropic cytokines present at the implantation site that are important for the normal development of human pregnancy. These cytokines share the cell membrane receptor subunit gp130, resulting in similar functions. The aim of this study was to compare the response to LIF and OSM in several trophoblast models with particular regard to intracellular mechanisms and invasion. Four trophoblast cell lines with different characteristics were used: HTR-8/SVneo, JEG-3, ACH-3P and AC1-M59 cells. Cells were incubated with LIF, OSM (both at 10 ng mL–1) and the signal transducer and activator of transcription (STAT) 3 inhibitor S3I-201 (200 µM). Expression and phosphorylation of STAT3 (tyr705) and extracellular regulated kinase (ERK) 1/2 (thr202/204) and the STAT3 DNA-binding capacity were analysed by Western blotting and DNA-binding assays, respectively. Cell viability and invasiveness were assessed by the methylthiazole tetrazolium salt (MTS) and Matrigel assays. Enzymatic activity of matrix metalloproteinase (MMP)-2 and MMP-9 was investigated by zymography. OSM and LIF triggered phosphorylation of STAT3 and ERK1/2, followed by a significant increase in STAT3 DNA-binding activity in all tested cell lines. Stimulation with LIF but not OSM significantly enhanced invasion of ACH-3P and JEG-3 cells, but not HTR-8/SVneo or AC1-M59 cells. Similarly, STAT3 inhibition significantly decreased the invasiveness of only ACH-3P and JEG-3 cells concomitant with decreases in secreted MMP-2 and MMP-9. OSM shares with LIF the capacity to activate ERK1/2 and STAT3 pathways in all cell lines tested, but their resulting effects are dependent on cell type. This suggests that LIF and OSM may partially substitute for each other in case of deficiencies or therapeutic interventions.

Expression of mRNA encoding leukaemia inhibitory factor (LIF) and its receptor (LIFRβ) in buffalo preimplantation embryos produced in vitro: markers of successful embryo implantation

Zygote ◽  
2012 ◽  
Vol 21 (2) ◽  
pp. 203-213 ◽  
Author(s):  
S. Eswari ◽  
G. Sai Kumar ◽  
G. Taru Sharma
Keyword(s):  
Culture Media ◽  
Total Cell ◽  
Inhibitory Factor ◽  
Preimplantation Embryos ◽  
Leukaemia Inhibitory Factor ◽  
Hatching Rate ◽  
Blastocyst Development ◽  
Cell Stage ◽  
Significant Difference

SummaryThe objective of this study was to evaluate the effect of supplementation of recombinant leukaemia inhibitory factor (LIF) in culture media on blastocyst development, total cell number and blastocyst hatching rates and the reverse transcription-polymerase chain reaction analysis of preimplantation buffalo embryos to determine whether they contain the LIF-encoding mRNA and its beta receptor (LIFRβ) genes in different stages of preimplantation buffalo embryos. Cumulus–oocyte complexes retrieved from slaughterhouse buffalo ovaries were matured in vitro and fertilized using frozen buffalo semen. After 18 h of co-incubation with sperm, the presumptive zygotes were cultured in modified synthetic oviductal fluid without (control) or with rhLIF (100 ng/ml). There was no significant difference in the overall cleavage rate up to morula stage however the development of blastocysts, hatching rate and total cell numbers were significantly higher in the LIF-treated group than control. Transcripts for LIFRβ were detected from immature, in vitro-matured oocytes and in the embryos up to blastocyst stage, while transcripts for the LIF were detected from 8–16-cell stage up to blastocyst, which indicated that embryo-derived LIF can act in an autocrine manner on differentiation process and blastocyst formation. This study indicated that the addition of LIF to the embryo culture medium improved development of blastocysts, functional (hatching) and morphological (number of cells) quality of the blastocysts produced in vitro. The stage-specific expression pattern of LIF and LIFRβ mRNA transcripts in buffalo embryos indicated that LIF might play an important role in the preimplantation development and subsequent implantation of buffalo embryos.

scholarly journals A potential molecular mechanism for regulating pre-mRNA splicing of implantation-related genes through unique uterine expression of splicing factor SC35 in women and rhesus monkeys

Reproduction ◽  
2002 ◽  
pp. 209-217 ◽  
Author(s):  
GY Nie ◽  
AL Hampton ◽  
GQ Fu ◽  
YX Liu ◽  
JK Findlay ◽  
...  
Keyword(s):  
Early Pregnancy ◽  
Rhesus Monkeys ◽  
Distinct Pattern ◽  
Oestrous Cycle ◽  
Splicing Factor ◽  
Trophoblast Invasion ◽  
Uterine Receptivity ◽  
Blastocyst Development ◽  
Specific Expression ◽  
Implantation Sites

Splicing factor SC35 is an essential component of the spliceosome, the cellular apparatus that removes introns from pre-mRNA to provide alternatively spliced isoforms. Many proteins associated with development of uterine receptivity and embryo implantation are present as isoforms, the tissue-specific expression of which may be regulated through alternative splicing. SC35 was identified as being increased at implantation sites during early pregnancy in mice. However, the present study has demonstrated that SC35 is present in human and rhesus monkey endometrium, that the protein is increased during the secretory phase of the oestrous cycle compared with the proliferative phase in both these primates and that it is present in a distinct pattern within the nucleus of both epithelial and stromal cells, as well as in cells of the vasculature. Both the intensity of immunoreactive protein and the proportion of cells that stain for SC35 alter with the phase of the oestrous cycle. A very precise expression pattern of SC35 (both protein and mRNA) was seen during early placentation in rhesus monkeys. At implantation sites between day 24 and day 35 of early pregnancy, SC35 was expressed strongly in cytotrophoblasts within the trophoblastic shell, in syncytiotrophoblast at the periphery of the cell column and in both cytotrophoblast and syncytiotrophoblast in the floating villi. In the adjacent maternal decidua, expression of SC35 was weak. These results indicate a role for SC35 in preparation of a receptive uterus, in the provision of secreted proteins to support blastocyst development and in trophoblast invasion.

scholarly journals The expression of the IGF system in the bovine uterus throughout the oestrous cycle and early pregnancy

2000 ◽  
Vol 165 (2) ◽  
pp. 231-243 ◽  
Author(s):  
RS Robinson ◽  
GE Mann ◽  
TS Gadd ◽  
GE Lamming ◽  
DC Wathes
Keyword(s):  
Cross Sections ◽  
Early Pregnancy ◽  
In Situ Hybridisation ◽  
Endometrial Biopsy ◽  
Oestrous Cycle ◽  
Luminal Epithelium ◽  
Mrna Levels ◽  
Igf Binding Proteins ◽  
Igf System ◽  
Igf I

The IGF system is expressed in the uterus during the oestrous cycle and early pregnancy and is likely to play an important role in regulating the development of the embryo and uterus. The IGF peptides (IGF-I and -II) mediate their effects through the type 1 IGF receptor (IGF-1R), while the IGF-binding proteins (IGFBP-1 to -6) modulate their interaction with the receptor. In this study, the expression of the IGF system in the bovine uterus was determined throughout the oestrous cycle and on day 16 of pregnancy. Endometrial biopsy samples were collected from four cows over three cycles such that there were samples for every 2 days from day 0 (oestrus) to day 14 and then every day until day 21. To assess the effect of pregnancy, uterine horn cross-sections were collected on day 16 from 15 pregnant (PREG), five inseminated non-pregnant (INP) and nine uninseminated cyclic controls (CONT). The expression of mRNA for the IGFs, IGF-1R and IGFBP-1 to -5 was determined by in situ hybridisation and the results were quantified by measuring the optical density units from autoradiographs. The main region of IGF-I mRNA expression was the sub-epithelial stroma underlying the luminal epithelium. The expression of IGF-I mRNA was highest at oestrus and lowest during the early and late luteal phases. On day 16, IGF-I mRNA levels were low in all groups, with pregnancy having no effect on the IGF-I mRNA concentrations. The strongest expression of IGF-II mRNA was in the caruncular stroma, with pregnancy having no significant effect in this region. IGF-1R mRNA was also present in the caruncles and was strongly expressed in all epithelial cells both throughout the oestrous cycle and during early pregnancy. The expression of IGFBP-1 mRNA was confined to the luminal epithelium, with the strongest expression seen on day 14 of the cycle. On day 16 the expression of IGFBP-1 mRNA was higher in the PREG group compared with the CONT group. The expression of IGFBP-2 mRNA was localised to the sub-epithelial stroma with more INP than PREG cows showing detectable levels of IGFBP-2. The strongest expression of IGFBP-3 mRNA was in the caruncular stroma; expression in the endometrial stroma was similarly decreased during early pregnancy. IGFBP-5 mRNA was mainly expressed in the inner ring of myometrium and was not affected by pregnancy on day 16. In conclusion, these results show that many components of the uterine IGF system are differentially regulated during the oestrous cycle and early pregnancy and suggest that modulation of the IGF system may influence uterine activity during this period.

scholarly journals Leukemia Inhibitory Factor: Roles in Embryo Implantation and in Nonhormonal Contraception

2014 ◽  
Vol 2014 ◽  
pp. 1-10 ◽  
Author(s):  
Naguib Salleh ◽  
Nelli Giribabu
Keyword(s):  
Leukemia Inhibitory Factor ◽  
Growth And Development ◽  
Immune Modulation ◽  
Embryo Implantation ◽  
Critical Role ◽  
Inhibitory Factor ◽  
Trophoblast Invasion ◽  
Leukaemia Inhibitory Factor ◽  
Multiple Processes

Leukaemia inhibitory factor (LIF) plays an indispensible role in embryo implantation. Aberrant LIF production is linked to implantation failure. LIF regulates multiple processes prior to and during implantation such as uterine transformation into a receptive state, decidualization, blastocyst growth and development, embryo-endometrial interaction, trophoblast invasion, and immune modulation. Due to its critical role, LIF has been a target for a nonhormonal contraception. In this review, we summarize up-to-date information on the role of LIF in implantation and its role in contraception.

109 EXPRESSION OF GENES RELATED TO ENDOMETRIAL RECEPTIVITY IN EQUINE ENDOMETRIUM DURING THE ESTROUS CYCLE AND EARLY PREGNANCY

2015 ◽  
Vol 27 (1) ◽  
pp. 147 ◽  
Author(s):  
M. Hitit ◽  
A. Guzeloglu ◽  
C. Ozel ◽  
M. O. Atli ◽  
E. Kurar ◽  
...  
Keyword(s):  
Early Pregnancy ◽  
Reproductive Tract ◽  
Cell Signalling ◽  
Endometrial Receptivity ◽  
Oestrous Cycle ◽  
Mrna Levels ◽  
Expression Levels ◽  
Expression Of Genes ◽  
Relative Mrna Expression ◽  
One Way Anova

A set of genes that display differential expression levels in the reproductive tract could serve as beneficial markers of endometrial receptivity. SERPINA14 is present in the uterus during pregnancy and suppresses lymphocyte accumulation. Osteopontin is the ligand of integrin β3 and enables trophoblast communication during implantation. Leukemia inhibitory factor (LIF) is involved in inflammatory cell signalling and contributes to implantation by regulating immune cells. The objective was to assess the expression of SERPINA14, osteopontin, and LIF mRNAs in the equine endometrium during the oestrous cycle and early pregnancy. Biopsies were obtained from mares on day of ovulation (d 0, n = 4), late diestrus (LD, n = 4, high progesterone [P4]), and after luteolysis at the beginning of oestrus phase (AL, n = 4, <1 ng mL–1 P4) of the cycle. Biopsies were also taken on days 14 (P14, n = 4), 18 (P18, n = 4), and 22 (P22; n = 4) of pregnancy. Relative mRNA expression levels of genes were quantified using real-time quantitative RT-qPCR in duplicate. Data were analysed using one-way ANOVA, and l.s.d. test was applied. Both the oestrous cycle and early pregnancy increased SERPINA14 mRNA levels compared to d0. Expression of LIF mRNA was not significantly regulated except for a decline at AL. Expression of osteopontin mRNA was up-regulated during the oestrous cycle at LD while early pregnancy inhibited this up-regulation. The results suggest that the genes studied related to endometrial receptivity are strictly regulated accordingly to the stage of oestrous cycle, probably by circulating ovarian steroids, specifically progesterone, and pregnancy-associated factors are also involved in this regulation. This project was partially funded by TUBITAK 107O035 to AG and DUBAP 14VF12 to MOA. MH was supported by OYP 2013-090.

scholarly journals Leukaemia inhibitory factor in implantation and uterine biology

Reproduction ◽  
2005 ◽  
Vol 130 (2) ◽  
pp. 131-145 ◽  
Author(s):  
Susan J Kimber
Keyword(s):  
Knockout Mouse ◽  
Reproductive Tract ◽  
Inhibitory Factor ◽  
Glandular Epithelium ◽  
Leukaemia Inhibitory Factor ◽  
Luminal Epithelium

Leukaemia inhibitory factor (LIF) is one of the most important cytokines in the reproductive tract. Without expression of LIF in the uterus, implantation of a blastocyst cannot begin. Yet, 13 years after publication of the phenotype of the LIF knockout mouse we are only just beginning to understand how LIF functions in the uterus. This review addresses our knowledge of the role of LIF in regulating implantation through its influence on the luminal epithelium and stromal decidualization, but also its influence on reproductive tract cells such as leukocytes and glandular epithelium, during the pre-implantation phase of pregnancy.

scholarly journals 227.An inhibitor of leukemia inhibitory factor signalling blocks embryo implantation in the mouse

2004 ◽  
Vol 16 (9) ◽  
pp. 227
Author(s):  
E. Dimitriadis ◽  
C. Stoikos ◽  
M. Baca ◽  
W. Fairlie ◽  
A. D. Uboldi ◽  
...  
Keyword(s):  
Early Pregnancy ◽  
Leukemia Inhibitory Factor ◽  
Embryo Implantation ◽  
Uterine Horn ◽  
Inhibitory Factor ◽  
Luminal Epithelium ◽  
Pregnant Uterus ◽  
Implantation Sites ◽  
Post Implantation

Embryo implantation is a critical step in the establishment of pregnancy. Endometrial leukemia inhibitory factor (LIF) is essential for embryo implantation in the mouse (1). Uterine LIF is expressed in the luminal epithelium on Day 3 of pregnancy (D3) (D0�=�day of plug detection) and signals via activation of signal transducer and activator of transcription (Stat) 3 (2). We examined the effect of a novel LIF signalling inhibitor on the phosphorylation (p) of Stat3 during early pregnancy and on embryo implantation in the mouse. We injected LIF inhibitor into one uterine horn and PBS into the other uterine horn of the mouse at D3 and examined the effect on pStat3 immunostaining in the luminal epithelium between 30 and 360�min later. We found no immunoreactive pStat3 in luminal epithelium following treatment with LIF inhibitor at 60 and 90�min but variable staining at other time points. The PBS-treated uterine horn showed intense immunostaining at all times. LIF inhibitor (1mg/kg body weight per day) or PBS was administered to mice (a) subcutaneously, (b) intraperitoneally, at 8-hourly intervals for 3�days from D2, or (c) continuously into the peritoneal cavity via Alzet pumps from D2. No effect was seen on implantation at D6. When LIF antagonist (3.5mg/kg/day) or PBS were administered by Alzet pumps from D2 together with ip injections, 4-hourly from D3 for 36�h, there were no implantation sites in the uteri of treated mice (n�=�5) while the control mice (n�=�4) had 3.6���0.5�sites (P�<�0.001). Histologically, the uteri of the treated mice resembled non-pregnant uterus, while the control uterus resembled post-implantation uterus. The results demonstrate that treatment of mice during early pregnancy with a novel LIF inhibitor blocks LIF action in vivo and embryo implantation. This knowledge is important for development of novel contraceptives. (1) Stewart, C. L., Kaspar, P., Brunet, L. J., Bhatt, H., Gadi, I., Kontgen, F., Abbondanzo, S. J. (1992) Nature 359, 76–79. (2) Cheng, J. G., Chen, J. R., Hernandez, L., Alvord, W. G., Stewart, C. L. (2001) Proc. Natl Acad. Sci. USA 98, 8680–8685.

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