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Submitted on March 14, 2005
Accepted on May 23, 2005
Department of Animal Sciences, Faculty of Agricultural, Food and Environmental Quality Sciences, The Hebrew University of Jerusalem, Rehovot 76100, Israel. Institute of Anatomy, University of Leipzig, Liebigstrasse 13, D-04103, Leipzig, Germany. Department of Pharmacology, University of California, Irvine, CA 92697, USA
* To whom correspondence should be addressed. E-mail: rina.meidan{at}huji.ac.il.
A highly vascular endocrine gland, the corpus luteum (CL) is an excellent model for the study of angiogenic factors. Prokineticins (PK-1 and 2), also termed endocrine-gland-derived VEGF and BV8 are newly identified proteins described as selective angiogenic mitogens. We previously identified PK binding sites - two closely homologous G protein-coupled receptors (PK-R1 and PK-R2) in human and bovine ovarian cells, but their function remained unknown. In this study we examined the presence and effects of PKsin CL-derived endothelial and steroidogenic cell types (LEC and LSC, respectively). PK-1 mRNA were identified in CL and follicles by real-time PCR, using primers specific for the bovine PK-1 sequence (retrieved from Bos taurus whole genome shotgun database). PKs were potent angiogenic mitogens for LEC: they enhanced cell proliferation, elevated [3H]-thymidine incorporation, MAPK activation and c-jun/fos mRNA expression. The effects of PK proteins on cell survival were examined by nuclear morphology (DAPI staining), measurement of DNA fragmentation (TUNEL assay) and caspase-3 cleavage. Results obtained by these techniques demonstrated that PKs protected LEC from serum starvation-induced apoptosis. Stress conditions such as serum withdrawal, TNF
and hypoxia markedly increased PK-R2 expression, whereas mRNA levels of PK-R1 remained unchanged. These suggest that the anti-apoptotic effect of PK-1 on LEC may be mediated via PK-R2. PK-1 increased VEGF mRNA expression by LSC implying that it could also indirectly, via VEGF, affect luteal angiogenesis. Together, these findings suggest an important role for PK-1 in luteal function by acting as a mitogen and survival factor in LEC.
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