Posts Tagged ‘Cyclooxygenase

The fetal membranes, chorioallantois, and amnion from cows at term expressed COX-2 mRNA, but the signals were weaker than in cotyledons (not shown). Figure 2 shows a representative gel for the protected fragments of COX-2 mRNA and GAPDH in intercarun-cular endometrium, myometrium, and cervical mucosa. COX-2 mRNA expression in endometrium was variable and rather weak during […]

COX-2 Expression in Tissues of Pregnant Cows Total RNA was prepared from myometrium, endometrium, caruncles, fetal cotyledons, and cervical mucosa (n = 3). Amnion and chorioallantoic membranes were examined only from term pregnant animals. It was necessary to use 20 ^g of total RNA for each hybridization reaction to obtain detectable signals from tissues other […]

COX-1 Expression in Tissues of Pregnant Cows No signals for COX-1 mRNA were detected by RPA in 20 ^g of total RNA from tissues of pregnant cows after exposure on XOMAT-AR (Eastman Kodak, Rochester, NY) films for 1 wk. After exposure for 3-7 days on BioMax (Eastman Kodak, Rochester, NY) film, which is three times […]

Ribonuclease protection assay (RPA). The RPAs were performed using a commercial kit (Ambion, Austin, TX) according to the instructions for the kit. Ten or twenty micrograms of total RNA of each sample was hybridized with the COX-2-specific or COX-1-specific cRNA probes and, as control for equal loading on the gel, bovine GAPDH-specific cRNA in the […]

For bovine COX-1, the antisense probe was prepared by in vitro transcription from the Sp6 RNA polymerase promoter out of the PCR fragment. The probe for COX-1 had a length of 301 bases, which included the multiple cloning site of 82 bases and protected an RNA sequence of 219 bases. COX-2 fragment was inserted into […]

Detection of Specific Gene Transcripts by RNase Protection Assay (RPA) Preparation of polymerase chain reaction-amplified cDNA fragments for bovine COX-1 and -2 and the corresponding cRNA probes. The sequence of bovine COX-1 mRNA and COX-2 mRNA had not been described at the time this study was begun. Oligonucleotides suitable for use in PCR assay were […]

OT Challenges The six cows used for this experiment were paired according to gestation length; one cow in each pair was injected i.v. with 200 IU OT and the other with saline i.v. Two hours after OT injection the cows were exsanguinated, and a sample of intercaruncular endometrium was removed, frozen in liquid nitrogen, and […]