Skip to main content
SpringerLink
Log in

Cloning and characterization of an apoptosis-associated gene in the human placenta

  • Published:
Apoptosis Aims and scope Submit manuscript

Abstract

Placenta is a transient feto-maternal association that develops during mammalian pregnancies. Human placental tissue during the first trimester of pregnancy is an actively dividing and differentiating tissue, while near term, it represents a fully differentiated unit performing many life-sustaining functions for the fetus. Previous studies have demonstrated that the percentage of placental cells that undergo apoptosis is greater at full term as compared to the first trimester of pregnancy. In this study, we undertook a study aimed at gaining an insight into the kind of genes expressed in the two developmentally distinct stages of gestation ie, the first trimester and term using Differential Display RT-PCR. Cloning and sequencing of one of the differentially expressed cDNAs from term placental tissue revealed that it is a novel gene, referred to as T-18 in the text. In this study, we also examined the regulation of this gene during apoptosis in the human placenta. A model for analysis of placental apoptosis was established by incubating placental villi in serum-free culture medium. It was observed that apoptosis occurred rapidly following incubation of placental villi without tropic support, and the proposed free-radical scavenger, superoxide dismutase (SOD) suppressed apoptosis in the placenta. Interestingly, the levels of T-18 mRNA increased significantly during spontaneous induction of apoptosis and decreased when apoptosis was blocked by SOD. These data clearly suggest that there is a strong correlation between the expression of T-18 and placental apoptosis and that T-18, may play a significant role in this process. Furthermore, the establishment of a defined in vitro explant culture model should facilitate elucidation of factors, which regulate apoptosis in human placenta.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Villee CA. Synthesis of proteins in the placenta. Gynecol Invest 1977; 8: 145–161.

    Google Scholar 

  2. Carter JE. Morphologic evidence of syncytial formation from cytotrophoblast cells. Obstet Gynecol 1964; 23: 647.

    Google Scholar 

  3. Petraglia F, Florio P, Nappi C, Genazzani AR. Peptide signaling in human placenta and membranes: Autocrine, paracrine, and endocrine mechanisms. Endocrine Reviews 1996; 17: 156–186.

    Google Scholar 

  4. Zeleznik AJ, Ihrig LL, Bassett SG. Developmental expression of Ca/Mg dependent endonuclease activity in rat granulosa and luteal cells. Endocrinol 1989; 125: 2218–2220.

    Google Scholar 

  5. Smith SC, Philip N, Baker DM, Malcolm ES. Placental apoptosis in normal human pregnancy. Am J Obstet & Gynecol 1997; 177: 57–65.

    Google Scholar 

  6. Uckan D, Steele A, Cherry B, Wang Y, Chamizo W, Koutsonikolis A, Gilbert-Barness E, Good RA. Trophoblast express Fas ligand: a proposed mechanism for immune privilege in placenta and maternal invasion. Molecular Human Reproduction 1997; 3: 655–662.

    Google Scholar 

  7. Payne SG, Smith SC, Davidge ST, Baker PN, Guilbert LJ. Death receptor Fas/Apo-1/CD95 expressed by human placental cytotrophoblasts does not mediate apoptosis. Biol Reprod 1999; 60(5): 1144–50.

    Google Scholar 

  8. Sakuragi N, Matsuo H, Coukos G, Furth EE, Bronner MP, VanArsdale CM, et al. Differentiation-dependent expression of the Bcl-2 proto-oncogene in the human trophoblast lineage. J Soc Gynecol Invest 1994; 1: 164–172.

    Google Scholar 

  9. Qiao S, Nagasaka T, Harada T, Nakashima N. p53, Bax and bcl-2 expression, and apoptosis in gestational trophoblast of complete hydatidiform mole. Placenta 1998; 19: 361–369.

    Google Scholar 

  10. Phillips TA, Ni J, Pan G, Ruben SM, Wei YF, Pace JL, Hunt JS. TRAIL (Apo-2L) and TRAIL receptors in human placentas: implications for immune privilege. J Immunol 1999; 162(10): 6053–9.

    Google Scholar 

  11. Smith SC, Philip N, Baker DM, Malcolm ES. Increased placental apoptosis in intrauterine growth restriction. Am J Obstet & Gynecol 1997; 177: 1395–1401.

    Google Scholar 

  12. Guo Ke, Wolf V, Dharmarajan AM, Feng Z, Bielke W, Surer, S, Friis, R. Apoptosis-associated gene expression in the corpus luteum of the rat. Biol Reprod 1998; 58: 739–746.

    Google Scholar 

  13. Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidium thiocyanate-phenol-chloroform. Anal Biochem 1987; 162: 156–159.

    Google Scholar 

  14. Gross-Bellard M, Oudet P, Chambon P. Isolation of highmolecular-weight DNA from mammalian cells. Eur J Biochem 1973; 36: 32–38.

    Google Scholar 

  15. Tilly JL, Hsueh AJ. Microsacle autoradiographic method for the qualitative and quantitative analysis of apoptotic DNA fragmentation. J Cell Physiol 1993; 154: 519–526.

    Google Scholar 

  16. Dharmarajan AM, Goodman SB, Tilly KI, Tilly JL. Apoptosis during functional corpus luteum regression: evidence of a role for chorionic gonadotropin in promoting luteal cell survival. Endocrine J (Endocrine) 1994; 2: 295–303.

    Google Scholar 

  17. Dharmarajan AM, Hisheh S, Singh B, Parkinson S, Tilly KI, Tilly JL. Anti-oxidants mimic the ability of chorionic gonadotropin to suppress apoptosis in the rabbit corpus luteum in vitro: A novel role for superoxide dismutase in regulating bax expression. Endocrinol 1999; 140(6): 2555–2561.

    Google Scholar 

  18. Tilly JL, Tilly KI, Kenton ML, Johnson AL. Expression of members of the bcl-2 gene family in the immature rat ovary:equine chorionic gonadotropin-mediated inhibition of granulosa cell apoptosis is associated with decreased bax and constitutive bcl-2 and bcl-x messenger ribonucleic acid levels. Endocrinol 1995; 136: 232–241.

    Google Scholar 

  19. Liang P, Pardee AB. Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction. Science 1992; 257: 967–971.

    Google Scholar 

  20. G. Huch, H.P Hohn, H.W. Denker. Identification of differentially expressed genes in the human trophoblast cells by differential display RT-PCR. Placenta 1998; 19: 557–67.

    Google Scholar 

  21. Cirelli N, Moens A, Lebrun P, et al. Apoptosis in human placenta is not increased during labor but can be massively induced in vitro. Biol Reprod 1999; 61(2): 458–63.

    Google Scholar 

  22. Watson AL, Skepper JN, Jauniaux E, Burton GJ. Susceptiblity of human placental syncytiotrophoblastic mitochondria to oxygen-mediated damage in relation to gestational age. J Clin Endocrinol Metab 1998; 83(5): 1697–705.

    Google Scholar 

  23. Tilly JL, Tilly KI. Inhibitors of oxidative stress mimic the ability of follicle-stimulating hormone to suppress apoptosis in cultured rat ovarian follicles. Endocrinol 1995; 136: 242–252.

    Google Scholar 

  24. Wang Y, Walsh SW. Placental mitochondria as a source of oxidative stress in pre-eclampsia. Placenta 1998; 19(8): 581–6

    Google Scholar 

  25. DiFederico E, Genbacev O, Fisher SJ. Preeclampsia is associated with widespread apoptosis of Placental cytotrophoblasts within the uterine wall. Am J Pathol 1999; 155(1): 293–301.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Biochemistry, Indian Institute of Science, Bangalore, India

    Rekha M. Rao & A. Jagannadha Rao

  2. Department of Anatomy and Human Biology, The University of Western Australia, Nedlands, Perth, Western Australia, Australia

    Arun M. Dharmarajan

Authors
  1. Rekha M. Rao
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Arun M. Dharmarajan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. A. Jagannadha Rao
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Rao, R.M., Dharmarajan, A.M. & Rao, A.J. Cloning and characterization of an apoptosis-associated gene in the human placenta. Apoptosis 5, 53–60 (2000). https://doi.org/10.1023/A:1009637726114

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1009637726114

Search

Navigation