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a Department of Animal Science, Obihiro University of Agriculture and Veterinary Medicine, Obihiro 080-8555, Japan
b Institute of Physiology, Technical University of Munich, D-80350 Freising-Weihenstephan, Germany
c Laboratory of Reproductive Endocrinology, Faculty of Agriculture, Okayama University, Okayama 700-8530, Japan
When Ang I was added to the MVE at a concentration of 10-9 M, it was converted to Ang II (21%). The production of Ang II from Ang I time-dependently rose for 24 h. Addition of captopril (an inhibitor of Ang-converting enzyme [ACE]) to the MVE cultures significantly inhibited Ang II production from 6 h to 24 h (P < 0.05). Addition of estradiol-17ß (E2) + vascular endothelial growth factor and E2 + basic fibroblast growth factor to MVE cultures increased Ang II production, whereas E2 or growth factors alone had no effect. Specific transcription for AT1R and AT2R was detected in bovine CL and MVE. There were no significant changes in Ang II tissue concentration or AT1R mRNA expression using reverse transcription-polymerase chain reaction during the estrous cycle. In contrast, AT2R mRNA expression decreased during the midluteal phase (P < 0.05) and increased to the highest level during the late luteal phase (P < 0.05).
Results demonstrated that Ang II is generated from Ang I in MVE isolated from the developing bovine CL, indicating that MVE have ACE activity. In addition, mRNA expression for Ang II receptors was detected in the bovine CL and the luteal MVE. These results suggest that Ang II is produced by actions of the local renin-angiotensin system, at least in part, on MVE in the bovine CL, and that this peptide may be involved in the regulation of luteal function during early development and luteolysis.
Recent findings suggest that the ovarian renin-angiotensin system regulates ovarian function through the paracrine/autocrine actions of angiotensin (Ang) II. The aims of this study were to investigate 1) the endothelial cell capacity to convert Ang I to Ang II, 2) the effects of endocrine and paracrine/autocrine factors on Ang II production in microvascular endothelial cells (MVE) derived from the developing corpora lutea (CL), and 3) the relationship between Ang II peptide concentration and expression of mRNA for angiotensin type 1 and 2 receptors (ATR1 and AT2R) in the bovine CL at different stages of the estrous cycle.
1 This study was supported by a Grant-in-Aid for Scientific Research (C) and the Japan-Germany joint research project of the Japan Society for the Promotion of Science, the Novartis Foundation (Japan) for the Promotion of Science (A.M.), and the German Research Foundation (Scha 257/14-1). K.H. was supported by H. Wilhelm Schaumann-Stiftung, and A.M. was supported by Alexander von Humbolt Stiftung.
2 Correspondence. FAX: 49 8161 714204; physio{at}pollux.weihenstephan.de
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