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To verify the possible role played by pig granulosa cells in the ovarian angiogenic process, we have developed a reliable in vitro system which allows the evaluation of endothelial sprouting and capillary growth in three-dimensional matrices. Granulosa cells collected from porcine follicles of different size were co-cultured with porcine aortic endothelial cells (PAEC) in a microcarrier-based fibrin gel system; after 2 and 5 days of co-culture, we determined the number and length of all endothelial sprouts; moreover, these parameters were quantified only in capillary-like structures, which were defined as continuous multicellular sprouts at least 200 µm long. In granulosa cells- PAEC co-cultures we observed an increase of angiogenic activity as compared to controls (PAEC alone). Granulosa cells from follicles of different size regulate angiogenesis differently: cells from the small follicle group significantly enhanced endothelial sprouting, while those from the large follicle group favoured mainly capillary elongation. Our observations seem therefore to suggest that the development and growth of thecal vascular bed is controlled by paracrine factors of granulosa cell origin that may induce the formation of a primitive capillary plexus during the early phases of antral follicle growth, which will be remodelled in more advanced phases of follicular development.
This study was designed to investigate levels of ETAR gene expression in the granulosa layer of broiler hens with different levels of plasma leptin and lipids (cholesterol and triacyglycerol). To induce different plasma leptin and lipid levels, the hens were fed high (20 and 40% more than recommended) and low (20% less than recommended) feed rations for 30 days. Variations of plasma leptin and lipids followed those found in the levels of feed intake and body weight in individual groups while the relative amount of ETAR mRNA increased in all groups. The effect, however, was significant (P<0.05) only for T+20% group. It is concluded that ETAR gene expression in follicular granulosa cells could be influenced by leptin in the broiler hens.
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