Expression of prolactin gene in mouse placenta during late pregnancy: detection of mRNA and its translation product.
To examine the existence of PRL messenger ribonucleic acid (mRNA) in the mouse placenta during late pregnancy, reverse transcriptase-polymerase chain reaction (RT-PCR) and Southern blot analysis were carried out followed by nucleotide sequence analysis of cDNA. Total RNA extracted from each tissue was reverse-transcribed, followed by PCR with two oligonucleotide primers specific or a part of mouse PRL (mPRL) cDNA. An amplified RT-PCR product of predicted size was detected in all samples from the placenta of days 16 and 18 pregnant mice. This product was specifically hybridized with a probe overlapping an entire sequence of mPRL cDNA in Southern blot analysis. Nucleotide sequence analysis also provided evidence that the amplified cDNA had a nucleotide sequence completely identical to the mPRL cDNA sequence reported previously. Furthermore, mPRL with a slightly bigger molecular weight than that of pituitary PRL was detected in the placenta of days 12, 14, 16 and 18 pregnancy by immunoblot analysis. These results suggest that PRL mRNA and its translation product are synthesized in mouse placenta during late pregnancy.
Harigaya
T
T
Laboratory of Functional Anatomy, Faculty of Agriculture, Meiji University, Kanagawa, Japan.
Komori
M
M
Kawakami
T
T
Watanabe
H
H
Abe
M
M
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D017931:DNA Primers / Q000737:chemistry; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000235:genetics*; D015151:Immunoblotting; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D010920:Placenta / Q000737:chemistry / Q000378:metabolism*; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D011388:Prolactin / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis* / Q000235:genetics
D017931:DNA Primers; D012333:RNA, Messenger; D011388:Prolactin