Hostname: page-component-cd9895bd7-dzt6s Total loading time: 0 Render date: 2024-12-23T18:53:07.916Z Has data issue: false hasContentIssue false
  • English
  • Français

Dose- and time-dependent effects of TNFα and actinomycin D on cell death incidence and embryo growth in mouse blastocysts

Published online by Cambridge University Press:  01 August 2007

D. Fabian ,
S. Juhás ,
G. Il'ková  and
J. Koppel
D. Fabian*
Affiliation:
Institute of Animal Physiology, Slovak Academy of Sciences, Košice, Slovakia
S. Juhás
Affiliation:
Institute of Animal Physiology, Slovak Academy of Sciences, Košice, Slovakia
G. Il'ková
Affiliation:
Institute of Animal Physiology, Slovak Academy of Sciences, Košice, Slovakia
J. Koppel
Affiliation:
Institute of Animal Physiology, Slovak Academy of Sciences, Košice, Slovakia
*
All correspondence to: Dušan Fabian, Institute of Animal Physiology, Slovak Academy of Sciences, Šoltésovej 4/6, 04001 Košice, Slovakia. Tel: +421 55 728 78 41. Fax: +421 55 728 78 42. e-mail: [email protected]
Get access Rights & Permissions [Opens in a new window]

Summary

This study was undertaken to obtain information about characteristics of different types of induced apoptosis in preimplantation embryos. Freshly isolated mouse blastocysts were cultured in vitro with the addition of two apoptotic inductors – TNFα and actinomycin D – at various doses and times. The average number of nuclei and the percentage of dead cells were evaluated in treated embryos. Classification of dead cells was based on morphological assessment of their nuclei evaluated by fluorescence microscopy, the detection of specific DNA degradation (TUNEL assay), the detection of active caspase-3 and cell viability assessed by propidium iodide staining. The addition of both apoptotic inductors into culture media significantly increased cell death incidence in blastocysts. Their effects were dose and time dependent. Lower concentrations of inductors increased cell death incidence, usually without affecting embryo growth after 24 h culture. Higher concentrations of inductors caused wider cell damage and also retarded embryo development. In all experiments, the negative effect of actinomycin D on blastomere survival and blastocyst growth was greater than the effect of TNFα. Furthermore, the addition of actinomycin D into culture media increased cell death incidence even after 6 h culture. Differences resulted probably from diverse specificity of apoptotic inductors. The majority of dead cells in treated blastocysts were of apoptotic origin. Morphological and biochemical features of apoptotic cell death induced by both TNFα and actinomycin D were similar and had homologous profile. In blastomeres, similarly to somatic cells, the biochemical pathways of induced apoptosis included activation of caspase-3 and internucleosomal DNA fragmentation.

Keywords

Actinomycin DCaspase-3Induced apoptosisMouse blastocystsTNFα
Type
Research Article
Information
Zygote , Volume 15 , Issue 3 , August 2007 , pp. 241 - 249
Copyright
Copyright © Cambridge University Press 2007

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Abdelhaleem, M. (2003). The actinomycin D-induced apoptosis in BCR-ABL-positive K562 cells is associated with cytoplasmic translocation and cleavage of RNA helicase A. Anticancer Res. 23, 485–90.Google ScholarPubMed
Argiles, J.M., Carbo, N. & Lopez-Soriano, F.J. (1997). TNF and pregnancy: the paradigm of a complex interaction. Cytokine Growth Factor Rev. 8, 181–8.CrossRefGoogle ScholarPubMed
Baran, V., Fabian, D., Rehak, P. & Koppel, J. (2003). Nucleolus in apoptosis-induced mouse preimplantation embryos. Zygote 11, 271–83.CrossRefGoogle ScholarPubMed
Bedaiwy, M.A., Falcone, T., Goldberg, J.M., Attaran, M., Sharma, R., Miller, K., Nelson, D.R. & Agarwal, A. (2005). Relationship between cytokines and the embryotoxicity of hydrosalpingeal fluid. J. Assist. Reprod. Genet. 22, 161–5.CrossRefGoogle ScholarPubMed
Bellier, S., Chastant, S., Adenot, P., Vincent, M., Renard, J.P. & Bensaude, O. (1997). Nuclear translocation and carboxyl-terminal domain phosphorylation of RNA polymerase II delineate the two phases of zygotic gene activation in mammalian embryos. EMBO J. 16, 6250–62.CrossRefGoogle ScholarPubMed
Ben-Yair, E., Less, A., Lev, S., Ben-Yehoshua, L. & Tartakovsky, B. (1997). Tumour necrosis factor alpha binding to human and mouse trophoblast. Cytokine 9, 830–6.CrossRefGoogle ScholarPubMed
Byrne, A.T., Southgate, J., Brison, D.R. & Leese, H.J. (2002). Effects of insulin-like growth factors I and II on tumour-necrosis-factor-alpha-induced apoptosis in early murine embryos. Reprod. Fertil. Dev. 14, 79–83.CrossRefGoogle ScholarPubMed
Fabian, D., Rehak, P., Czikkova, S., Il'kova, G., Baran, V. & Koppel, J. (2003). Induced cell death of preimplantation mouse embryos cultured in vitro evaluated by comet assay. Theriogenology 60, 691–706.CrossRefGoogle ScholarPubMed
Fabian, D., Il'kova, G., Rehak, P., Czikkova, S., Baran, V. & Koppel, J. (2004). Inhibitory effect of IGF-I on induced apoptosis in mouse preimplantation embryos cultured in vitro. Theriogenology 61, 745–55.CrossRefGoogle ScholarPubMed
Fabian, D., Gjorret, J.O., Berthelot, F., Martinat-Botte, F. & Maddox-Hyttel, P. (2005). Ultrastructure and cell death of in vivo derived and vitrified porcine blastocysts. Mol. Reprod. Dev. 70, 155–65.CrossRefGoogle ScholarPubMed
Fabian, D., Koppel, J. & Maddox-Hyttel, P. (2005). Apoptotic processes during mammalian preimplantation development. Theriogenology 64, 221–31.CrossRefGoogle ScholarPubMed
Gjorret, J.O., Knijn, H.M., Dieleman, S.J., Avery, B., Larsson, L.I. & Maddox-Hyttel, P. (2003). Chronology of apoptosis in bovine embryos produced in vivo and in vitro. Biol. Reprod. 69, 1193–200.CrossRefGoogle ScholarPubMed
Hunt, J.S. (1993). Expression and regulation of the tumour necrosis factor-alpha gene in the female reproductive tract. Reprod. Fertil. Dev. 5, 141–53.CrossRefGoogle ScholarPubMed
Ju, E.J., Kwak, D.H., Lee, D.H., Kim, S.M., Kim, J.S., Kim, S.M., Choi, H.G., Jung, K.Y., Lee, S.U., Do, S.I., Park, Y.I. & Choo, Y.K. (2005). Pathophysiological implication of ganglioside GM3 in early mouse embryonic development through apoptosis. Arch. Pharm. Res. 28, 1057–64.CrossRefGoogle ScholarPubMed
Kleeff, J., Kornmann, M., Sawhney, H. & Korc, M. (2000). Actinomycin D induces apoptosis and inhibits growth of pancreatic cancer cells. Int. J. Cancer. 86, 399–407.3.0.CO;2-G>CrossRefGoogle ScholarPubMed
Kurzawa, R., Glabowski, W. & Wenda-Rozewicka, L. (2001). Evaluation of mouse preimplantation embryos cultured in media enriched with insulin-like growth factors I and II, epidermal growth factor and tumor necrosis factor alpha. Folia. Histochem. Cytobiol. 39, 245–51.Google ScholarPubMed
Lachapelle, M.H., Miron, P., Hemmings, R., Falcone, T., Granger, L., Bourque, J. & Langlais, J. (1993). Embryonic resistance to tumour necrosis factor-alpha mediated cytotoxicity: novel mechanism underlying maternal immunological tolerance to the fetal allograft. Hum. Reprod. 8, 1032–8.CrossRefGoogle Scholar
Lasek, W., Giermasz, A., Kuc, K., Wankowicz, A., Feleszko, W., Golab, J., Zagozdzon, R., Stoklosa, T. & Jakobisiak, M. (1996). Potentiation of the anti-tumor effect of actinomycin D by tumor necrosis factor alpha in mice: correlation between in vitro and in vivo results. Int. J. Cancer. 66, 374–9.3.0.CO;2-B>CrossRefGoogle ScholarPubMed
Lawits, J.A. & Biggers, J.D. (1993). Culture of preimplantation embryos. In: Guide to Techniques in Mouse Development, Wassarman PM, DePhamphilis ML (Ed.), San Diego, Academic Press, pp. 153–64.CrossRefGoogle Scholar
Pampfer, S., Wuu, Y.D., Vanderheyden, I. & De Hertogh, R. (1994). Expression of tumor necrosis factor-alpha (TNF alpha) receptors and selective effect of TNF alpha on the inner cell mass in mouse blastocysts. Endocrinology 134, 206–12.CrossRefGoogle ScholarPubMed
Pampfer, S., Vanderheyden, I., McCracken, J.E., Vesela, J. & De Hertogh, R. (1997). Increased cell death in rat blastocysts exposed to maternal diabetes in utero and to high glucose or tumor necrosis factor-alpha in vitro. Development 124, 4827–36.CrossRefGoogle ScholarPubMed
Perry, R.P. & Kelley, D.E. (1970). Inhibition of RNA synthesis by actinomycin D: characteristic dose-response of different RNA species. J. Cell. Physiol. 76, 127–39.CrossRefGoogle ScholarPubMed
Pivko, J., Grafanau, P. & Kubovicova, E. (2002). Bovine abnormal preimplantation embryos: analysis of segregated cells occurring in the subzonal space and/or blastocoele cavity for their nuclear morphology and persistence of RNA synthesis. Zygote 10, 141–7.CrossRefGoogle ScholarPubMed
Rivera-Leon, R. & Gerbi, S.A. (1997). Delocalization of some small nucleolar RNPs after actinomycin D treatment to deplete early pre-rRNAs. Chromosoma 105, 506–14.CrossRefGoogle ScholarPubMed
Sanford, T.R., De, M. & Wood, G.W. (1992). Expression of colony-stimulating factors and inflammatory cytokines in the uterus of CD1 mice during days 1 to 3 of pregnancy. J. Reprod. Fertil. 94, 213–20.CrossRefGoogle ScholarPubMed
Soto, P., Natzke, R.P. & Hansen, P.J. (2003). Actions of tumor necrosis factor-alpha on oocyte maturation and embryonic development in cattle. Am. J. Reprod. Immunol. 50, 380–8.CrossRefGoogle ScholarPubMed
Takeuchi, R., Hoshijima, H., Onuki, N., Nagasaka, H., Chowdhury, S.A., Kawase, M. & Sakagami, H. (2005). Effect of anticancer agents on codeinone-induced apoptosis in human cancer cell lines. Anticancer Res. 25, 4037–41.Google ScholarPubMed
Tomanek, M., Kopecny, V. & Kanka, J. (1986). Studies on RNA synthesis in early pig embryos. Histochem. J. 18, 140. Abstract.Google Scholar
Torchinsky, A., Markert, U.R. & Toder, V. (2005). TNF-alpha-mediated stress-induced early pregnancy loss: a possible role of leukemia inhibitory factor. Chem. Immunol. Allergy 89, 62–71.CrossRefGoogle ScholarPubMed
Van Der Velden, A.W., Destree, O.H.J., Voorma, H.O. & Thomas, A.A. (2000). Controlled translation initiation on insulin-like growth factor 2-leader 1 during Xenopus laevis embryogenesis. Int. J. Dev. Biol. 44, 843–50.Google ScholarPubMed
Whiteside, E.J., Boucaut, K.J., Teh, A., Garcia-Aragon, J., Harvey, M.B. & Herington, A.C. (2003). Elevated concentration of TNF-alpha induces trophoblast differentiation in mouse blastocyst outgrowths. Cell. Tissue. Res. 314, 275–80.CrossRefGoogle ScholarPubMed
Williams, S.T., Smith, A.N., Cianci, C.D., Morrow, J.S. & Brown, T.L. (2003). Identification of the primary caspase 3 cleavage site in alpha II-spectrin during apoptosis. Apoptosis 8, 353–61.CrossRefGoogle ScholarPubMed
Wuu, Y.D., Pampfer, S., Vanderheyden, I., Lee, K.H. & De Hertogh, R. (1998). Impact of tumor necrosis factor alpha on mouse embryonic stem cells. Biol. Reprod. 58, 1416–24.CrossRefGoogle ScholarPubMed
Wuu, Y.D., Pampfer, S., Becquet, P., Vanderheyden, I., Lee, K.H. & De Hertogh, R. (1999). Tumor necrosis factor alpha decreases the viability of mouse blastocysts in vitro and in vivo. Biol. Reprod. 60, 479–83.CrossRefGoogle ScholarPubMed
Yamazaki, Y., Tsuruga, M., Zhou, D., Fujita, Y., Shang, X., Dang, Y., Kawasaki, K. & Oka, S. (2000). Cytoskeletal disruption accelerates caspase-3 activation and alters the intracellular membrane reorganization in DNA damage-induced apoptosis. Exp. Cell. Res. 259, 64–78.CrossRefGoogle ScholarPubMed