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Ubiquitin is physiologically induced by interferons in luminal epithelium of porcine uterine endometrium in early pregnancy: global RT‐PCR cDNA in place of RNA for differential display screening 1
Author(s) -
Chwetzoff Serge,
d'Andrea Sabine
Publication year - 1997
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(97)00177-4
Subject(s) - northern blot , biology , microbiology and biotechnology , complementary dna , western blot , messenger rna , differential display , endometrium , rna , gene expression , blot , gene , biochemistry , endocrinology
Early in the course of pregnancy, at the preimplantation stage, the pig embryo is likely to exert a paracrine effect on the tissue intended to receive it, via the secretion of interferons. Our observations show that trophoblastic interferons induce an increase of some mRNAs in the epithelial cells of the gilt endometrium, which would illustrate this phenomenon. The increase of four mRNAs, whose corresponding cDNAs are dD1, dD2, dD3 and dD4, has been examined in this study. The method used is similar to Northern blot analysis except that mRNAs in the blot are replaced by cDNAs produced from total cellular poly(A) + mRNAs by global everse‐ ranscription olymerase hain eaction (RT‐PCR). Northern blot hybridization requires a considerable quantity of starting material – which we estimate in this study to be several million porcine endometrium cells – whereas the RT‐PCR‐based method gives comparable results starting with only a few cells – about 200. Using this method, the differential nature of dD1, dD2, dD3 and dD4 was shown. dD2 and dD3 correspond to genes already identified as interferon‐induced: the β 2 ‐microglobulin and inkel‐Biskis‐Reilly murine sarcoma virus‐ ssociated biquitously secreted protein (FAU). dD1 corresponds to a still unidentified gene. dD4 encodes for the porcine UbA 52 ubiquitin. Up to now, the increase in ubiquitin mRNA as a result of interferon effect has not been reported and is discussed in view of recent publications.