Differential expression of cyclooxygenase 1 and cyclooxygenase 2 in the bovine oviduct.
The aim of the present study was to investigate the enzymes for the local prostaglandin (PG) biosynthesis present in the bovine oviduct during the estrous cycle to influence early reproductive events. Bovine oviducts were classified into four phases: pre-ovulatory, post-ovulatory, early-to-mid luteal, and late luteal phase, subdivided further into ipsi- or contralateral site and separated into ampulla or isthmus. Oviductal cells were gained by flushing the oviductal regions. Quantitative real-time reverse transcriptase-PCR was performed for the secretory and cytosolic phospholipases A(2) (sPLA(2)IB, cPLA(2)alpha, and cPLA(2)beta) and cyclooxygenases (COX-1 and COX-2) as the first step enzymes of PG synthesis. COX-1 and cPLA(2)beta showed significant highest mRNA expression around and before ovulation compared with the luteal phase respectively. sPLA(2)IB and cPLA(2)alpha mRNA expression was unregulated during the estrous cycle. Regional differences in mRNA content were found for sPLA(2)IB with higher mRNA expression in the ampulla than in the isthmus. Western blot analysis revealed the highest COX-1 protein content in the early-to-mid luteal phase. Immunohistochemistry demonstrated that COX-1 was localized in epithelial and smooth muscle cells, whereas COX-2 was only localized in epithelial cells. COX-2 showed a differential distribution within the epithelial cell layer suggesting a regulation on a cellular level, although the COX-2 mRNA and protein amounts did not vary throughout the estrous cycle. A COX activity assay of oviductal cells revealed that COX activity originated predominantly from COX-1 than from COX-2. Treatment of primary oviductal cells with 10 pg/ml 17beta-estradiol or 10 ng/ml progesterone resulted in a higher expression of COX-2 and cPLA(2)alpha, but not of the other enzymes. The expression pattern of these enzymes suggests that an estrous-cycle dependent and region-specific PG synthesis in the bovine oviduct may be required for a successful reproduction.
Odau
Simone
S
Institute of Veterinary-Biochemistry, Freie Universität Berlin, Oertzenweg 19b, 14163 Berlin, Germany.
Gabler
Christoph
C
Holder
Christoph
C
Einspanier
Ralf
R
D000818:Animals; D015153:Blotting, Western / Q000379:methods; D002417:Cattle / Q000378:metabolism*; D002478:Cells, Cultured; D051545:Cyclooxygenase 1 / Q000032:analysis / Q000235:genetics; D051546:Cyclooxygenase 2 / Q000032:analysis / Q000235:genetics; D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology; D030762:Estrous Cycle / Q000502:physiology*; D005260:Female; D015870:Gene Expression; D007150:Immunohistochemistry / Q000379:methods; D007527:Isoenzymes / Q000032:analysis / Q000235:genetics; D010057:Oviducts / Q000033:anatomy & histology / Q000201:enzymology*; D010060:Ovulation; D010741:Phospholipases A / Q000032:analysis / Q000235:genetics; D011451:Prostaglandin-Endoperoxide Synthases / Q000032:analysis* / Q000235:genetics; D011720:Pyrazoles / Q000494:pharmacology; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013876:Thiophenes / Q000494:pharmacology
D016861:Cyclooxygenase Inhibitors; D007527:Isoenzymes; D011720:Pyrazoles; D012333:RNA, Messenger; C115461:SC 560; D013876:Thiophenes; C064874:DuP 697; D051545:Cyclooxygenase 1; D051546:Cyclooxygenase 2; D011451:Prostaglandin-Endoperoxide Synthases; D010741:Phospholipases A