TY - JOUR
T1 - The expression of Argonaute2 and related microRNA biogenesis proteins in normal and hypoxic trophoblasts
AU - Donker, Rogier B.
AU - Mouillet, Jean François
AU - Nelson, D. Michael
AU - Sadovsky, Yoel
N1 - Funding Information:
We are grateful to Elena Sadovsky, Lynne Collins and Aaron Barton for their technical assistance during these studies, to Baosheng Chen and Timothy Schaiff for discussions and to Lori Rideout for assistance during manuscript preparation. This work was supported by the Jan Kornelis de Cock Foundation, Foundation De Drie Lichten and the Groningen University Institute for Drug Exploration (to R.B.D.) and NIH R21-HD053878 and R01-HD045675 (to Y.S.). R.B.D. was also supported by a Scholarship for Talented Students from the Dutch Ministry of Education.
PY - 2007/4
Y1 - 2007/4
N2 - Endogenous microRNAs (miRNAs) post-transcriptionally regulate mRNA and protein expression during tissue development and function. Whereas adaptation to environmental insults are tightly regulated in human tissues, the role of miRNAs and miRNA biogenesis proteins in this context is inadequately explored. We sought to analyse the expression of the key RNAi enzyme Argonaute2 (Ago2) and other miRNA biogenesis proteins in human trophoblasts during differentiation and in hypoxic environment. Using an in vitro analysis of primary term human trophoblasts, we identified the expression of the core miRNA biogenesis proteins in human villous trophoblasts, with expression levels unaffected by cellular differentiation. We found that the miRNA biosynthetic pathway was functional and produced miRNAs, with miR-93 up-regulated and miR-424 down-regulated in hypoxic environment. In contrast, hypoxia did not alter the expression of key miRNA machinery proteins. The pivotal miRNA processing enzyme Ago2, along with its interacting protein DP103, were expressed in normal placentas as well as in placentas from pregnancies complicated by placental hypoperfusion that resulted in fetal growth restriction. Ago2 and DP103 co-immunoprecipitated, and did not limit trophoblast response to hypoxic stress. We concluded that the core miRNA machinery proteins are expressed and functional in human trophoblasts. The influence of hypoxia on the expression of a subset of placental miRNA species is unlikely to reflect altered expression of key miRNA biogenesis proteins.
AB - Endogenous microRNAs (miRNAs) post-transcriptionally regulate mRNA and protein expression during tissue development and function. Whereas adaptation to environmental insults are tightly regulated in human tissues, the role of miRNAs and miRNA biogenesis proteins in this context is inadequately explored. We sought to analyse the expression of the key RNAi enzyme Argonaute2 (Ago2) and other miRNA biogenesis proteins in human trophoblasts during differentiation and in hypoxic environment. Using an in vitro analysis of primary term human trophoblasts, we identified the expression of the core miRNA biogenesis proteins in human villous trophoblasts, with expression levels unaffected by cellular differentiation. We found that the miRNA biosynthetic pathway was functional and produced miRNAs, with miR-93 up-regulated and miR-424 down-regulated in hypoxic environment. In contrast, hypoxia did not alter the expression of key miRNA machinery proteins. The pivotal miRNA processing enzyme Ago2, along with its interacting protein DP103, were expressed in normal placentas as well as in placentas from pregnancies complicated by placental hypoperfusion that resulted in fetal growth restriction. Ago2 and DP103 co-immunoprecipitated, and did not limit trophoblast response to hypoxic stress. We concluded that the core miRNA machinery proteins are expressed and functional in human trophoblasts. The influence of hypoxia on the expression of a subset of placental miRNA species is unlikely to reflect altered expression of key miRNA biogenesis proteins.
KW - Ago2
KW - Hypoxia
KW - MicroRNA biogenesis
KW - Placenta
KW - Trophoblast
UR - http://www.scopus.com/inward/record.url?scp=34147116217&partnerID=8YFLogxK
U2 - 10.1093/molehr/gam006
DO - 10.1093/molehr/gam006
M3 - Article
C2 - 17327266
AN - SCOPUS:34147116217
SN - 1360-9947
VL - 13
SP - 273
EP - 279
JO - Molecular human reproduction
JF - Molecular human reproduction
IS - 4
ER -