Human Reproduction, Vol. 18, No. 1, 130-136,
January 2003
© 2003 European Society of Human Reproduction and Embryology
Retinoic acid decreases the viability of mouse blastocysts in vitro
1 Department of Obstetrics and Gynecology, Chang Gung Memorial Hospital, Kaohsiung, Taiwan, 2 Chang Gung University School of Medicine, 3 Department of Nutrition and Health Science, Fooyin Institute of Technology and 4 Department of Obstetrics and Gynecology, UCLA School of Medicine, California, USA 5 To whom correspondence should be addressed at: Department of Obstetrics and Gynecology, Chang Gung Memorial Hospital, 123, Ta-Pei Rd., Niao-Sung Hsiang, Kaohsiung County, Taiwan. e-mail: s10d1{at}mail.st1917.com.tw
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Abstract |
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BACKGROUND: This study was designed to examine the cytotoxic effect of retinoic acid on the blastocyst stage of mouse embryos and on subsequent early postimplantation embryo development in vitro. METHODS AND RESULTS: Mouse blastocysts were exposed for 24 h to doses of 0, 0.1 µmol/l and 10 µmol/l all-trans retinoic acid and observed for their capacity to implant and develop during the early postimplantation period in vitro. When retinoic acid-pretreated blastocysts were allowed to implant in vitro, significantly fewer embryos were able to reach a later stage of embryo development. Compared with the findings for the control blastocysts, exposure to retinoic acid resulted in a significant reduction in the average number of total cells in blastocysts and the trophectoderm/inner cell mass lineage. The effect was associated with a significant increase in the frequency of cells identified as being engaged in apoptosis by means of the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling and Annexin V techniques. CONCLUSIONS: This is the first evidence that retinoic acid induces cell death (apoptosis) and inhibits cell proliferation in mouse blastocysts. This results in the retardation of early postimplantation blastocyst development and subsequent blastocyst death.
Key words: apoptosis/blastocyst/implantation/postimplantation/retinoic acid
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Introduction |
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Vitamin A and its physiological metabolites, retinoic acid (RA) and other retinoids have profound effects on embryo morphogenensis, cell growth and differentiation, vision, and reproduction (DeLuca, 1991
; Gudas, 1992; Eskild and Hansson, 1994; Hofmann and Eichele, 1994). In addition, RA is a valuable compound for the therapy of cystic acne and numerous other dermatological disorders (Bollag et al., 1983; Becherel et al., 1996; Duell et al., 1996) and may have potential for combination biotherapy for the treatment of malignant disease including cancers of ovary, breast, lung, liver and blood (Solary et al., 1994; Kim et al., 1996; Guzey et al., 1998; Mangiarotti et al., 1998; Srivastava et al., 1999). While normal embryo development requires retinoids, too much or too little at the wrong stage or wrong time can adversely affect the developing embryo (Sporn et al., 1991). Several animal studies have demonstrated a potential adverse effect of RA when administered during mid and late pregnancy in mice. Administration of RA on day 7 or day 8 of gestation causes retardation of general development, abnormal differentiation of the cranial neural plate and abnormal development of the hindbrain (Morriss-Kay et al., 1991). Administration of RA on day 9 of gestation induces dysmorphogenensis of the inner ear in mice (Frenz et al., 1996). Abnormalities of limb and neural plate development have been induced when RA is administered between day 10 and day 16 of gestation (Kochhar, 1973; Kochhar et al., 1984; Stafford et al., 1995). Previous studies have shown the occurrence of mRNA expression of all three RA receptors (RAR) 
, ß and 
in mouse and bovine blastocysts (Wu et al., 1992; Mohan et al., 2001). These findings suggest that RA may be involved in the developmental process of preimplantation mammalian embryos. It has been demonstrated that in-utero exposure to excess RA during the preimplantation period (early cleavage stage) does not result in any adverse effect on preimplantation embryo development (blastocyst formation) (Huang and Lin, 2001). However, in-vitro exposure to excess RA during the peri-implantation (blastocyst stage) and early postimplantation period does result in early growth retardation and subsequent embryo death for postimplantation embryos (Huang et al., 2001). Some teratogenic effects of retinoids may be due to their ability to induce apoptosis. Cellular responses to RA in vitro range from cell death to differentiation. RA-induced cell death with characteristics of apoptosis has been observed in several murine embryonic studies (Kochhar et al., 1993; Lee et al., 1994; Clifford et al., 1996; Okazawa et al., 1996; Chiba et al., 1997; Finley et al., 1999; Lu and Dong, 1999; Shum et al., 1999; Stewart et al., 2000; Suwa et al., 2001). However, there are no reports that demonstrate the RA-induced effects on blastocysts. Because the blastocyst is composed of the inner cell mass (ICM), blastocoele and trophectoderm (TE), the effects of RA on blastocysts include: (i) the action of RA in the ICM; (ii) the action of RA in the TE; (iii) the interaction between ICM and TE. In order to study further the relevant mechanism by which RA affects the viability of blastocysts, this study was designed to examine the cell-killing effect of RA on the blastocyst stage of mouse embryos and on subsequent early postimplantation embryo development in vitro. |
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Collection of mouse blastocysts
ICR mice (6–8 weeks old) were induced to superovulate by injecting 5 IU pregnant mare’s serum gonadotrophin (PMSG; Sigma Chemical Co., St Louis, MO, USA) followed 48 h later by injecting 5 IU hCG (Serono, NV Organon Oss, The Netherlands). They were then mated overnight with a single fertile male of the same strain. Mating was confirmed by the presence of a copulatory plug the following day. The ICR virgin albino mice, male mice and pregnant mice were maintained on breeder chow and kept under a 12 h day/12 h night regimen, with food and water available ad libitum. All animals received humane animal care as outlined in the Guidelines for Care and Use of Experimental Animals (Canadian Council on Animal Care, Ottawa, 1984). The morning after an overnight mating period, female mice with vaginal plugs were separated and used experimentally. The day a vaginal plug was found was defined as day 0 of pregnancy. Embryos were obtained by flushing the uterine horn and Fallopian tubes on the afternoon of day 3 of gestation with CMRL-1066 culture medium (Gibco Life Technologies, Grand Island, NY, USA) containing 1 mmol/l glutamine and 1 mmol/l sodium pyruvate (Sigma). The embryos were collected in uncoated plastic 35 mm Falcon culture dishes and washed a minimum of three times. Expanded blastocysts from different females were pooled and randomly selected for the various experiments.
Embryo culture
Blastocysts were cultured for assessment of implantation in vitro and further embryonic differentiation according to a modification of an established method (Huang, 2001). The embryos were cultured in 1 ml of culture medium in 4-well Multidishes (Nunc, Roskilde, Denmark) at 37°C under 5% CO2 in air. For group culture, five embryos were cultured per well. CMRL-1066 was used as the basic culture medium. It was supplemented with 1 mmol/l glutamine and 1 mmol/l sodium pyruvate plus 50 IU/ml penicillin and 50 mg/ml streptomycin (Gibco). Following the 24 h culture using different dose of retinoic acid without serum supplementation, all groups of embryos were cultured in the same condition (RA-free) for 7 days. During the first 3 days the culture medium was supplemented with 20% fetal calf serum (Gibco) and thereafter with 20% heated-inactivated human placental cord serum. Embryos were inspected daily under a dissecting microscope and classified according to an established method (Witschi, 1972). Developmental parameters, such as hatching through the zona pellucida, attachment to the culture dishes, trophoblastic outgrowth and differentiation of the embryo proper into early or late egg cylinders (germ layer stage) or primitive streak to early somite stage were recorded daily throughout the 8 day culture period. Embryonic development was observed through a phase-contrast microscope (Olympus IMT-2, Tokyo, Japan). To decrease observer bias, all the data were analysed using the following criteria. Implanted blastocyst was defined as the attachment and outgrowth of the blastocyst to the culture dish. An early egg cylinder (EEC) embryo was defined as an embryo that had reached stages 9 or 10 by day 4. A late egg cylinder (LEC) embryo was defined as an embryo that reached stages 11, 12 or 13 by day 6 of culture. An early somite (ES) embryo was defined as an embryo that had reached stages 14 or 15 by day 8.
Experimental design (24 h exposure to all-trans retinoic acid at the blastocyst stage in all experiments)
Experiment 1 (morphological development in vitro)
Expanded blastocysts were exposed to 0 (n = 150), 0.1 (n = 145) or 10 (n = 165) µmol/l all-trans RA without serum supplementation for 24 h. This covers concentrations expected in embryos after oral administration (Kraft et al., 1989) in the treated group. They were then washed in RA-free medium and further cultured in CMRL-1066 medium in order to assess their further development as outlined above. All-trans RA (Sigma) was prepared in an aqueous solution of 0.01% dimethyl sulphoxide (DMSO) and a similar solution of DMSO alone was used as the control. Briefly, 0, 0.1 or 10 µmol/l of RA was added on the first day of culture to the treated group. Over the following 7 days of culture, the culture medium was changed to CMRL-1066 without RA in both the treatment and control groups.
Experiment 2 (effect of RA on cell proliferation in blastocysts)
A total of 208 blastocysts was incubated in 0, 0.1 or 10 µmol/l RA for 24 h and washed in RA-free medium following the incubation. The proliferation of blastocysts was evaluated by counting separately the number of ICM and TE cells identified by dual differential staining (Pampfer et al., 1990). This method is based on immunosurgery, considering the impermeability of the TE layer, which protects the ICM cells from exposure to the antibody and the complement reaction. The two cell lineages can be distinguished following fluorochrome staining (Figure 1). According to the protocol, the zona pellucida was removed by incubating the blastocysts in 0.4% pronase in M2 medium supplemented with 0.1% bovine serum albumin (M2-BSA). The denuded blastocysts were then exposed to 1 mmol/l of trinitrobenzenesulphonic acid (TNBS) in BSA-free M2 medium (M2) containing 0.1% PVP at 4°C for 30 min (Hardy et al., 1989) and washed with M2. Further, the blastocysts were treated with 30 µg/ml anti-dinitrophenol-BSA complex antibody in M2-BSA at 37°C for 30 min. After rinsing, the blastocysts were incubated in M2 supplemented with 10% whole guinea-pig serum as a source of complement, 20 µg/ml bisbenzimide and 10 µg/ml propidium iodide at 37°C for 30 min. The immunolysed blastocysts were gently transferred onto a slide and protected from light before observation. Under appropriate UV light excitation, the ICM cells, which take up bisbenzimide but exclude propidium iodide (PI) were stained blue, whereas the TE cells, which are labelled with both fluorochromes, were stained orange–red. Because multinucleated cells have been shown to be infrequent in preimplantation embryos (Gardner and Davies, 1993), the number of nuclei was considered as an accurate measure of the number of cells.
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Experiment 3 (effect of RA on cell death in blastocysts)
TUNEL labelling: A total of 284 blastocysts was incubated to 0, 0.1 or 10 µmol/l of RA for 24 h and washed in RA-free medium following the incubation. The blastocysts following the 24 h treatment were used in this study. After fixation, permeabilization, blocking and extensive washing, the embryos were subjected to TUNEL labelling involving dUTP biotin nick-end labelling by using an in-situ cell death detection kit (Roche Molecular Biochemicals, Mannheim, Germany) according to the company’s instructions. Briefly, each group of embryos was incubated with 20 µl of the TUNEL reaction mixture containing 2 µl of enzyme solution and 18 µl of labelling solution (containing fluorescein-conjugated nucleotides) for 30 min at 37°C. After incubation, embryos were washed extensively with phosphate-buffered saline (PBS) buffer plus 0.3% BSA. Converted-POD (20 µl) was added to embryos and incubated for 30 min at 37°C. After incubation, the embryos were washed extensively with PBS buffer. Finally, 20 µl of DAB-substrate solution was added to embryos and incubated for 2 min at room temperature. Photographic images (Figure 1A–C) were taken using a fluorescence microscope (Zeiss, Axioskop, Germany) under bright light.
Annexin staining: A total of 183 blastocysts was incubated in 0, 0.1 or 10 µmol/l of RA for 24 h and washed in RA-free medium following the incubation. The blastocysts following the 24 h treatment were used in this study. An Annexin V–FLUOS staining kit (Roche) was used to stain these embryos according to the manufacturer’s instructions. According to the protocol, the zona pellucida was removed by incubating the blastocysts in M2-BSA. Embryos were washed well with PBS plus 0.3% BSA. They were incubated with a mixture of 100 µl binding buffer, 1 µl fluorescein isothiocyanate (FITC) conjugated Annexin V and 1 µl PI for 60 min. After incubation, the embryos were washed well before photographic images were taken under a fluorescence microscope (Zeiss) with bright light and under UV light with DAPI/FITC/Ron Daman triple filters. This kit contains PI to determine the integrity of the plasma membrane and Annexin V to detect the translocation of phosphatidylserine (PS) from the inner face to the outer surface of the plasma membrane. In apoptotic cells, the cells translocate PS from the inner face of the plasma membrane to the cell surface while maintaining membrane integrity. Once on the cell surface, PS can easily be detected by staining with FITC-conjugated Annexin V, a protein that will specifically bind to PS. However, in necrotic cells, the broken plasma membrane allows PI, a DNA binding dye, to enter the cell through broken membranes to bind to the cellular DNA. Thus, apoptotic, necrotic and viable cells can be distinguished. Apoptotic cells stain positive for Annexin V (green) and necrotic cells stain positive for PI (red), while viable cells with an intact plasma membrane and no translocated PS are negative for both Annexin V and PI (colourless). The illustration is shown in Figure 1D–H.
Statistical analysis
2-test and one-way ANOVA were used as appropriate. Differences of P < 0.05 were considered to be significant.
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Effect of RA on blastocyst development in vitro
As shown in Table I, 165 blastocysts treated with 10 µmol/l RA, 145 blastocysts treated with 0.1 µmol/l and 158 control blastocysts were used for implantation in vitro. Implantation was similar in both treatment and control groups. However, in the treatment group, the formation of a 2-layer ICM and the formation of an ectoplacental cone were significantly reduced. In the treatment group, fewer embryos developed to the advanced egg cylinder stages (LEC and ES stages) when compared with the control group (logistic regression: effect of concentration, P < 0.001; effect of development, P < 0.0001; interaction, P = 0.02).
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Effect of RA on cell proliferation
Blastocysts that had been exposed to 0.1 or 10 µmol/l of RA developed less well and were found to contain fewer cells after 24 h than the control blastocysts. The differential staining techniques revealed that the effect of RA on cell number was more pronounced in the TE than in the ICM. A 17% cell deficiency was observed in the TE lineage following exposure to 10 µmol/l of RA (P = 0.007; Figure 2A).
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Effect of RA on cell death
The results of the culture experiments were confirmed in that cell death, as detected by TUNEL, increased 2–3-fold in blastocysts exposed to 10 µmol/l RA. The effect of RA on cell apoptosis was more pronounced in the ICM than in the TE (Figure 2B). Cell death was also detected by Annexin V staining kit. The percentage of Annexin V positive embryos in each group was analysed. There was a higher rate of Annexin V positive ICM in blastocysts treated with RA than in the control blastocysts (Figure 2C). However, there was a similar rate of Annexin V positive TE in blastocysts treated with RA than in the control blastocysts (Figure 2C). Furthermore, the mean number of necrotic cells in the blastocysts was similar between the control and treatment groups (Figure 2D).
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Discussion |
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The results of this study demonstrate that cell death in the mouse blastocysts that are transiently (24 h) exposed to RA and cultured in vitro is affected by excess RA. Blastocysts exposed to 10 µmol/l RA contain three times as many dead cells as blastocysts not exposed to RA as measured by the TUNEL staining method. Cell death was also detected by the Annexin V staining method and the percentage of apoptosis was significantly higher in the ICM lineage of the treated blastocysts than the control blastocysts. In contrast, the incidence of cell necrosis in RA-exposed blastocysts was similar to that of the control blastocysts. After 24 h exposure to RA, the blastocysts revealed retarded postimplantation development leading to embryonic death even though they could implant onto the cultured dishes. This suggests that excess RA can decrease the viability of mammalian blastocysts and can regulate cell death in the blastocysts.
Embryonic development is a complex process that is extremely sensitive to vitamin A as both deficiency and excess can lead to abortion and embryonic malformation. It is reasonable to interpret these results as suggesting that an excess of RA disrupts the normal developmental programme and leads to a serious retardation as demonstrated in the present study. In previous studies, the adverse effects of RA were found to be dose-dependent at the different stages of embryo development. Mouse embryos exposed to excess RA (12 mg/kg) on day 7
of development showed retardation of general development, abnormal differentiation of the cranial neural plate and abnormal development of the hindbrain (Morriss-Kay et al., 1991). The morphological features of embryos from treated mice were characterized by a reduced somite number, reductions in the pharyngeal arch size and number, a rostrally displaced otocyst, delayed closure of the anterior neuropore, as well as retardation of heart development. These effects of RA were related to the expression of the Hox-2.9 and Krox-20 genes, which in turn reduced the expression of the TGF-ß1 and TGF-ß2 proteins. Maternal exposure to RA (20 mg/kg) on day 9 of gestation has been found to induce dysmorphogenesis of the inner ear in mouse embryos (Frenz et al., 1996). Similarly, a congenital limb anomaly can be induced following exposure of pregnant Swiss–Webster mice to non-physiological levels of RA (120 mg/kg) on days 10 and 11 of gestation (Kochhar, 1973; Kochhar et al., 1984). Additionally, elevated RA doses (30 mg/kg) adversely affected early postimplantation embryogenesis on days 3 or 4 of gestation (F.J.Huang et al., unpublished results). However, 50 mg/kg of RA does not affect early or late preimplantation embryos adversely and even higher doses of RA (100 mg/kg) did not adversely affect the development of late preimplantation embryos (Huang and Lin, 2001). In contrast, RA in vitro (10 µmol/l) does affect germ layer and subsequent neurula development from day 3 to day 8 of gestation (Huang et al., 2001). These findings suggest that embryos at different stages have different tolerances for increases in RA concentration. It has been shown that RA suppresses mesodermal gene expression in mouse embryonic stem cells (Bain et al., 1996) and induces endodermal gene expression in F9 embryonal carcinoma cells (Cho et al., 1999). These findings suggest that the teratogenic effects of RA on early postimplantation embryos may be mediated by disruption of germ layer-specific gene activities.
Embryonic cells in embryo development are often poised between proliferation and apoptosis. It has been shown that RA-induced apoptosis is involved in embryonic development. For example, RA inhibited the proliferation and induced apoptosis of ectomesenchymal stem cells. These influences may contribute to the formation of cleft palate and other orofacial congenital malformation (Lu et al., 1999; Suwa et al., 2001). RA has an important role in digit separation with the increase of apoptotic cell death, as shown in limb development (Stewart et al., 2000; Kochhar et al., 1993; Lee et al., 1994; Tamagawa et al., 1995). RA also induced down-regulation of Wnt-3a, involving induction of widespread apoptosis in the tail bud cells of mouse embryo (Shum et al., 1999). In the present study, excess RA inhibited the proliferation (reduced cell number) and induced apoptosis in the ICM of mouse blastocysts. This may result in the postimplantation embryo retardation in vitro.
The TE is a sphere of epithelial cells surrounding the ICM and the blastocoel. Trophoblast cells arising at the blastocyst stage as the TE are required for development in the mammalian conceptus and contribute to the placenta (Cross et al., 1994), which is critical for the survival of mammalian embryos. Trophoblast stem cells treated with RA were compromised in their ability to proliferate and exhibited properties of differentiation into giant cells (Yan et al., 2001). This may implicate a function for RA in giant-cell formation during placentation. Furthermore, in-vitro studies performed with human trophoblastic cells demonstrated that the secretion of hCG and trophoblast differentiation are stimulated by retinoids (Tarrade et al., 2001). Although RA inhibited the proliferation (reduced cell number) and induced apoptosis in the TE of mouse blastocyst in the present study, this did not inhibit the embryo implantation in vitro. However, giant-cell formation and differentiation were difficult to demonstrate in the present in-vitro model.
The molecular mechanisms underlying the induction of developmental apoptosis by RA are beginning to be understood. A stress protein of the HSP 100/Clp family (HSP 110) could represent a biochemical event of apoptotic cell death induced by RA, administered to embryos at day 9 post coitum (Evrard et al., 2000). This effect resulted in craniofacial malformation similar to those of mandibulofacial dysostosis in man. Retinoic acid and bone morphogenetic protein 4 (BMP4) together induce p27 protein leading to retinoblastoma (Rb) protein activation and ultimately apoptosis in P19 embryonal carcinoma cells (Glozak and Rogers, 2001). Apoptosis induced by RA may operate by a mechanism activated by reactive oxygen species in embryonic stem cell death (Castro-Obregón and Covarrubias 1996).
There are several reports regarding the effects of RA on different stages of postimplantation embryos, embryonal stem cell, embryonal carcinoma cells, or trophoblast stem cells. As a follow-up to the previous studies (Huang and Lin, 2001; Huang et al., 2001), this study is the first to show that RA induced cell death (apoptosis) in blastocysts, which can result in subsequent retardation of postimplantation developmental and/or death. However, the molecular mechanism of apoptosis by RA in blastocysts and the interaction between ICM and TE need to be determined in further studies.
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Acknowledgements |
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We thank Mrs Zong-Xian Lin and Yu-Cheng Hsu, who have assisted in this study in the animal laboratory. We are also grateful to Hsueh-Wen Chang, PhD, who is at the Department of Biological Science, National Sun Yat-sen University, for his assistance in statistical analysis. This work was supported by grant NSC 89-2314-B-182A-105 from the National Science Council of the Republic of China. The preliminary data were presented as a poster at the 17th World Congress of the International Federation of Fertility Societies, Melbourne, Australia, November 25–30, 2001.
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References |
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Bain, G., Ray, W.J., Yao, M. and Gottlieb, D.I. (1996) Retinoic acid promotes neural and represses mesodermal gene expression in mouse embryonic stem cells in culture. Biochem. Biophys. Res. Commun., 223, 691–694.[CrossRef][Web of Science][Medline]
Becherel, P.A., Le Goff, L., Ktorza, S., Chosidow, O., Frances, C., Issaly, F., Mencia-Huerta, J.M., Debre, P., Mossalayi, M.D. and Arock, M. (1996) CD23-mediated nitric oxide synthase pathway induction in human keratinocytes is inhibited by retinoic acid derivatives. J. Invest. Dermatol., 106, 1182–1186.[CrossRef][Web of Science][Medline]
Bollag, W. (1983) Vitamins and retinoids: from nutrition to pharmacotherapy in dermatology and oncology. Lancet, 1, 860–863.[Web of Science][Medline]
Castro-Obregón, S. and Covarrubias, L. (1996) Role of retinoic acid and oxidative stress in embryonic stem cell death and neuronal differentiation. FEBS Lett., 381, 93–97.[CrossRef][Web of Science][Medline]
Chiba, H., Clifford, J., Metzger, D. and Chambon, P. (1997) Specific and redundant functions of retinoids X Receptor/Retinoic acid receptor heterodimers in differentiation, proliferation, and apoptosis of F9 embryonal carcinoma cells. J. Cell Biol. 139, 735–747.
Cho, S.Y., Lee, S.H. and Park, S.S. (1999) Differential expression of mouse Disabled 2 gene in retinoic acid-treated F9 embryonal carcinoma cells and early mouse embryos. Mol. Cells, 30, 179–184.
Clifford, J., Chiba, H., Sobieszczuk, D., Metzger, D. and Chambon, P. (1996) RXRalpha-null F9 embryonal carcinoma cells are resistant to the differentiation, anti-proliferative and apoptotic effects of retinoids. EMBO J. 16, 4142–4155.
Cross, J.C., Werb, Z. and Fisher, S.J. (1994) Implantation and the placenta: key pieces of the development puzzle. Science, 266, 1508–1518.
DeLuca, L.M. (1991) Retinoids and their receptors in differentiation, embryogenesis, and neoplasia. FASEB J., 5, 2924–2933.[Abstract]
Duell, E.A., Kang, S. and Voorhees, J.J. (1996) Retinoic acid isomers applied to human skin in vivo each induce a 4-hydroxylase that inactivates only trans retinoic acid. J. Invest. Dermatol., 106, 316–320.[CrossRef][Web of Science][Medline]
Eskild, W. and Hansson, V. (1994) Vitamin A functions in the reproductive organs. In Blomhoff R. (ed.) Vitamin A in health and disease. Marcel Dekker, New York, pp. 531–559.
Evrard, L., Vanmuylder, N., Dourov, N., Hermans, C., Biermans, J., Werry-Huet, A., Rooze, M. and Louryan, S. (2000) Correlation of HSP 110 expression with all-trans retinoic acid-induced apoptosis. J. Craniofac. Genet. Dev. Biol., 20, 183–192.[Web of Science][Medline]
Finley, M.F., Devata, S. and Huettner, J.E. (1999) BMP-4 inhibits neural differentiation of murine embryonic stem cells. J. Neurobiol., 40, 271–287.[CrossRef][Web of Science][Medline]
Frenz, D.A., Liu, W., Galinovic-Schwartz, V. and Van De Water, T.R. (1996) Retinoic acid-induced embryopathy of the mouse inner ear. Teratology, 53, 292–303.[CrossRef][Web of Science][Medline]
Gardner, R.L. and Davies, T.J. (1993) Lack of coupling between onsets of giant transformation and genome endoreduplication in the mural trophectoderm of the mouse blastocysts. J. Dev. Zool., 265, 54–60.
Glozak, M.A. and Rogers, M.B. (2001) Retinoic acid- and bone morphogenetic protein 4- induced apoptosis in P19 embryonal carcinoma cells requires p27. Exp. Cell Res., 268, 128–138.[CrossRef][Web of Science][Medline]
Gudas, L.J. (1992) Retinoids and vertebrate development. J. Biol. Chem., 269, 15399–15402.
Guzey, M., Demirpence, E., Criss, W. and DeLuca, H.F. (1998) Effects of retinoic acid (all-trans and 9-cis) on tumor progression in small cell lung carcinoma. Biochem. Biophys. Res. Commun., 242, 369–375.[CrossRef][Web of Science][Medline]
Hardy, K., Handyside, A.H. and Winston, R.M.L. (1989) The human blastocyst: cell number, death and allocation during late preimplantation development in vitro. Development, 107, 597–604.[Abstract]
Hofmann, C. and Eichele, G. (1994) Retinoids in development. In Sporn, M.B., Roberts, A.B. and Goodman, D.S. (eds), The Retinoids. Biology, Chemistry, and Medicine, 2nd edn. Raven Press, New York, pp. 387–441.
Huang, F.J. and Lin, Y.L. (2001) Effects of retinoic acid on pre-implantation embryo development in mice. Chang Gung Med. J., 24, 681–687.[Medline]
Huang, F.J., Wu, T.C. and Tsai, M.Y. (2001) Effect of retinoic acid on implantation and post-implantation development of mouse embryos in vitro. Hum. Reprod., 16, 2171–2176.
Kim, D.G., Jo, B.H., You, K.R. and Ahn, D.S. (1996) Apoptosis induced by retinoic acid in Hep 3B cells in vitro. Cancer Lett., 107, 149–159.[CrossRef][Web of Science][Medline]
Kochhar, D.M. (1973) Limb development in mouse embryos. I. Analysis of teratogenic effects of retinoic acid. Teratology, 7, 289–295.[CrossRef][Web of Science][Medline]
Kochhar, D.M., Penner, J. and Tellone, C. (1984) Comparative teratogeneic activities of two retinoids: Effects on palate and limb development. Teratogen Carcin. Mut., 4, 377–387.
Kochhar, D.M., Jiang, H., Harnish, D.C. and Soprano, D.R. (1993) Evidence that retinoic acid-induced apoptosis in the mouse limb bud core mesenchymal cells is gene-mediated. Prog. Clin. Biol. Res., 383, 815–825.
Kraft, J.C., Löfberg, B., Chahoud, I., Bochert, G. and Nau, H. (1989) Teratogenicity and placental transfer of all-trans-, 13-cis-, 4-oxo-all-trans-, and 4-oxo-13-cis-retinoic acid after administration of a low oral dose during organogenesis in mice. Toxicol. Appl. Pharm., 100, 162–176.[CrossRef][Web of Science][Medline]
Lee, K.K., Li, F.C., Yung, W.T., Kung, J.L., Ng, J.L. and Cheah, K.S. (1994) Influence of digits, ectoderm, and retinoic acid on chondrogenesis by mouse interdigital mesoderm in culture. Dev. Dyn., 201, 297–309.[Web of Science][Medline]
Lu, H., Jin, Y. and Dong, S. (1999) Ectomesenchymal stem cells and all-trans-retinoic acid induced apoptosis. Chin. J. Dent. Res. 2, 36–42.[Medline]
Mangiarotti, R., Danova, M., Alberici, R. and Pellicciari, C. (1998) All-trans retinoic acid (ATRA)-induced apoptosis is preceded by G1 arrest in human MCF-7 breast cancer cells. Br. J. Cancer, 77, 186–191.[Web of Science][Medline]
Mohan, M., Malayer, J.R., Geisert, R.D. and Morgan, G.L. (2001) Expression of retinal-binding protein messenger RNA and retinoic acid receptors in preattachment bovine embryos. Mol. Reprod. Dev., 60, 289–296.[CrossRef][Web of Science][Medline]
Morriss-Kay, G.M., Murphy, P., Hill, R.E. and Davidson, D.R. (1991) Effects of retinoic acid excess on expression of Hox-2.9 and Krox-20 and on morphological segmentation in the hindbrain of mouse embryos. EMBO J. 10, 2985–2995.[Web of Science][Medline]
Okazawa, H., Shimizu, J., Kamei, M., Imafuku, I., Hamada, H. and Kanazawa, I. (1996) Bcl-2 inhibits retinoic acid-induced apoptosis during the neural differentiation of embryonal stem cells. J. Cell Biol., 132, 955–968.
Pamfer, S., De Hertogh, R., Vanderheyden, I., Michiels, B. and Vercheval, M. (1990) Decreased inner cell mass proportion in blastocysts from diabetic rats. Diabetes, 39, 471–476.[Abstract]
Shum, A.S., Poon, L.L., Tang, W.W., Koide, T., Chan, B.W., Leung, Y.C., Shiroishi, T. and Copp, A.J. (1999) Retinoic acid induces down-regulation of Wnt-3a, apoptosis and diversion of tail bud cells to a neural fate in the mouse embryo. Mech. Dev. 84, 17–30.[CrossRef][Web of Science][Medline]
Solary, E., Bertrand, R. and Pommier, Y. (1994) Apoptosis of human leukemic HL-60 cells induced to differentiate by phorbol ester treatment. Leukemia, 8, 792–797.[Web of Science][Medline]
Sporn, M.B. and Robert, A.B. (1991) Interactions of retinoids and transforming growth factor-beta in regulation of cell differentiation and proliferation. Mol. Endocrinol., 5, 3–7.
Srivastava, R.K., Srivastava, A.R., Cho-Chung, Y.S. and Longo, D.L. (1999) Synergistic effects of retinoic acid and 8-C1-cAMP on apoptosis require caspase-3 activation in human ovarian cancer cells. Oncogene, 18, 1755–1763.[CrossRef][Web of Science][Medline]
Stafford, D.L., Lussier, M.R., Sank, A.C. and Shuler, C.F. (1995) In vitro model of syndactyly replicates the morphologic features observed in vivo. Plast. Reconstr. Surg., 96, 1169–1176.[Web of Science][Medline]
Stewart, S., Yi, S., Kassabian, G., Mayo, M., Sank, A. and Shuler, C. (2000) Changes in expression of the lysosomal membrane glycoprotein, LAMP-1 in interdigital regions during embryonic mouse limb development, in vivo and in vitro. Anat. Embryol. (Berl.), 201, 483–490.[CrossRef][Medline]
Suwa, F., Jin, Y., Lu, H., Li, X., Tipoe, G.L., Lau, T.Y., Tamada, Y., Kuroki, K. and Fang, Y.R. (2001) Alteration of apoptosis in cleft palate formation and ectomesenchymal stem cells influenced by retinoic acid. Okajimas Folia Anat. Jpn., 78, 179–186.[Medline]
Tamagawa, M., Mortia, J. and Naruse, I. (1995) Effects of all-trans-retinoic acid on limb development in the genetic polydactyly mouse. J. Toxicol. Sci., 20, 383–393.[Medline]
Tarrade, A., Schoonjans, K., Guibourdenche, J., Bidart, J.M., Vidaud, M., Auwerx, J., Rochette-Egly, C. and Evain-Brion, D. (2001) PPARã/RXRá heterodimers are involved in human CGâ synthesis and human trophoblast differentiation. Development, 142, 4504–4514.
Witschi, E. (1972) Characterization of developmental stages. Part II. Rat. In Biology Data Book, 2nd edn. Federation of American Societies of Experimental Biologies, Washington, DC, vol. 1, pp. 178–180.
Wu, T.C., Wang, L. and Wan, Y.J. (1992) Differential expression of retinoic acid receptor mRNA during mouse embryogenesis. Dev. Growth Differ., 34, 685–691.[CrossRef]
Yan, J., Tanaka, S., Oda, M., Makino, T., Ohgane, J. and Shiota, K. (2001) Retinoic acid promotes differentiation of trophoblast stem cells to a giant cell fate. Dev. Biol., 235, 422–432.[CrossRef][Web of Science][Medline]
Submitted on April 2, 2002; resubmitted on July 29, 2002. accepted on September 6, 2002
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