Identification of gestationally regulated genes in rat myometrium by use of messenger ribonucleic acid differential display


      Objective We hypothesized that the proteins contributing to myometrial changes during gestation could be identified indirectly by analyzing the changing pattern of messenger ribonucleic acid expression in the myometrium during pregnancy. Study design Ribonucleic acid was extracted from myometrium of timed pregnant Sprague-Dawley rats on days 12, 16, 20, 21, and 22 of pregnancy and on day 1 post partum. The technique of messenger ribonucleic acid differential display, a simple and sensitive polymerase chain reaction–based method for rapidly identifying messenger ribonucleic acids whose levels increase or decrease, was performed with the nine different anchoring primers (oligodeoxythymidine11 VN: V = G, A, or C; N = G, A, or C) in combination with 24 different 10-base oligonucleotides of random sequence. The polymerase chain reaction products were separated by electrophoresis on a 5% polyacrylamide sequencing gel, and those whose levels changed were then cloned, sequenced, and compared with those in the GenBank database to determine whether they corresponded to a known sequence in the database or were novel. Semiquantitative reverse transcriptase–polymerase chain reaction was used to confirm differential expression of selected products. Results: Messenger ribonucleic acid differential display revealed >500 polymerase chain reaction products that were differentially expressed during gestation, 179 of which were cloned and sequenced. Of these, 157 were from messenger ribonucleic acids whose levels increased during gestation, and 22 were from transcripts that decreased. Eighty-seven (49%) were related to sequences in the GenBank database, of which 62 (35%) were from messenger ribonucleic acids encoding known proteins and 25 (14%) corresponded to known expressed sequence tags. The technique of semiquantitative reverse transcriptase–polymerase chain reaction confirmed the increased expression of messenger ribonucleic acids encoding β-tropomyosin, type II phosphatidyl inositol-4-phosphate 5-kinase, and a novel myometrial messenger ribonucleic acid named RPU0901AC. Conclusion: Messenger ribonucleic acid differential display is a simple and sensitive method for rapidly identifying myometrial messenger ribonucleic acids that are differentially regulated during pregnancy. The identification of these differentially expressed messenger ribonucleic acids may lead to a better understanding of the molecular basis of normal and abnormal parturition.


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        • Huszar G
        • Roberts M
        Biochemistry and pharmacology of myometrium and labor.
        Am J Obstet Gynecol. 1982; 142: 225-237
        • Garfield RE
        • Puri CP
        • Csapo AI
        Endocrine, structural, and functional changes in the uterus during premature labor.
        Am J Obstet Gynecol. 1982; 142: 21-27
        • Mackenzie LW
        • Garfield RE
        Effects of 17β-estradiol on myometrial gap junction and pregnancy in the rat.
        Can J Physiol Pharmacol. 1985; 64: 462-466
        • Fuchs A
        • Periyasamy S
        • Alexandrova M
        • Soloff M
        Correlation between oxytocin receptor concentration and responsiveness to oxytocin in pregnant rat myometrium: effects of ovarian steroids.
        Endocrinology. 1983; 113: 742-749
        • Larcher A
        • Neculcea J
        • Breton C
        • Arslan A
        • Rozen F
        • Russo C
        • et al.
        Oxytocin receptor gene expression in the rat uterus during pregnancy and the estrous cycle and in response to gonadal steroid treatment.
        Endocrinology. 1995; 136: 5350-5356
        • Liang P
        • Pardee AB
        Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction.
        Science. 1992; 257: 967-971
        • Tokuyama Y
        • Takeda J
        Use of 33P-labeled primer increases the sensitivity and specificity of mRNA differential display.
        BioTechniques. 1995; 18: 424-425
        • Altschul SF
        • Boguski MS
        • Gish W
        • Wootton JC
        Issues in searching molecular sequence databases.
        Nature Genet. 1994; 6: 119-129
        • Alberts B
        • Bray D
        • Lewis J
        • Raff M
        • Roberts K
        • Watson JD
        The cell nucleus.
        in: Molecular biology of the cell. Garland, New York1989: 481-549
        • Skalli O
        • Vandekerckhove J
        • Gabbiani G
        Actin-isoform patterns as a marker of normal or pathological smooth-muscle and fibroblastic tissues.
        Differentiation. 1987; 33: 232-238
        • Gohari P
        • Berkowitz RL
        • Hobbins JC
        Prediction of intrauterine growth retardation by determination of total intra-uterine volume.
        Am J Obstet Gynecol. 1977; 127: 255-260
        • Uldbjerg N
        • Ekman G
        • Malmstrom A
        • Olsson K
        • Ulmsten U
        Ripening of the human uterine cervix related to changes in collagen, glycosaminoglycans, and collagenolytic activity.
        Am J Obstet Gynecol. 1983; 147: 662-666
        • Imada K
        • Ito A
        • Itoh Y
        • Nagase H
        • Mofi Y
        Progesterone increases the production of tissue inhibitor of metalloproteinases-2 in rabbit uterine cervical fibroblasts.
        FEBS Lett. 1994; 341: 109-112
        • Phillippe M
        The relationship between oxytocin, phophoinositide-specific phospholipase C, and phasic myometrial contractions.
        J Soc Gynecol Invest. 1994; 1: 49-54