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Messenger RNA and Protein Expression in the Uteroplacental Unit of Mice with Pregnancy Loss1
Department of Embryology and Teratology, Sackler School of Medicine, Tel Aviv University, Ramat Aviv, Tel Aviv, Israel
An elevated expression of TNF-
in embryonic microenvironment was
found to be associated with postimplantation loss. In this work, we
examined the pattern of TNF- expression at both the mRNA and the
protein level as well as the distribution of TNF- receptor mRNA in
the uteroplacental unit of mice with induced (cyclophosphamide-treated)
or spontaneous (CBA/J x DBA/2J mouse combination) pregnancy loss.
RNase protection analysis demonstrated an increase in TNF- mRNA
expression in the placentae of mice with pregnancy loss compared with
that in control mice. TNF- messages were localized to the uterine
epithelium and stroma as well as the giant and spongiotrophoblast cells
of the placenta. The intensity of the hybridization signal in placentae
of mice with pregnancy loss was substantially higher than that in
control mice. The up-regulation of TNF- mRNA was accompanied by an
increase in the expression of TNF- receptor I mRNA in the same cell
populations. The elevation of TNF- production was also demonstrated
at the protein level. Western blot analysis showed an increased level
of the 18- and 26-kDa TNF- protein species in the uteroplacental
unit of mice with pregnancy loss. Immunostaining revealed
TNF--positive leukocytes located in the uterus and placenta.
Finally, we found that immunization of mice with
cyclophosphamide-induced pregnancy loss while decreasing the resorption
rate in these females resulted in a decline in TNF- expression at
the fetomaternal interface. These data clearly suggest an involvement
of TNF- in pathways leading to both spontaneous and induced
placental death.
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