T Viergutz1, B Löhrke. 1. Department of Reproductive Biology, Research Institute of the Biology for Farm Animals, Dummerstorf, Germany. viergutz@fbn-dummerstorf.de
Abstract
UNLABELLED: Proliferation of granulosa cells and their withdrawal from the cell cycle may regulate follicular ovulation. Antagonists of platelet-activating factor (PAF) and its receptor (PAFr) inhibit follicle rupture. OBJECTIVES: Thus, PAF and PAFr may be involved in proliferative regulation of granulosa cells; however, expression of PAFr in these cells is unknown. MATERIALS AND METHODS: The aim of this study was to investigate the presence of PAFr and the effect of PAF on proliferation of cultured bovine granulosa cells using real-time polymerase chain reaction to assay steady-state level of mRNA, immunocytochemistry to quantify PAFr protein and proliferating cell nuclear antigen protein by flow cytometry. RESULTS: We found that granulosa cells express PAFr transcripts and protein. PAF presence did not change the concentration of PAFr mRNA or PAFr protein. Granulosa cells responded to PAF doses of 10 and 50 nm with increasing proportions of cells entering G0/G1 phase, as well as a significant expansion of total cell numbers. Rise in G0/G1-phase cells was accompanied by a decline in proliferating cell nuclear antigen protein expression, and these effects could be suspended by simultaneous PAFr blockage. The results provide clear evidence for expression of PAFr in bovine granulosa cells and its functional involvement in PAF/PAFr-mediated stimulation of cell recruitment. CONCLUSIONS: PAF antagonists are suggested to disturb this regulative activity of PAF and to contribute in this way to blockage of ovulation.
UNLABELLED: Proliferation of granulosa cells and their withdrawal from the cell cycle may regulate follicular ovulation. Antagonists of platelet-activating factor (PAF) and its receptor (PAFr) inhibit follicle rupture. OBJECTIVES: Thus, PAF and PAFr may be involved in proliferative regulation of granulosa cells; however, expression of PAFr in these cells is unknown. MATERIALS AND METHODS: The aim of this study was to investigate the presence of PAFr and the effect of PAF on proliferation of cultured bovine granulosa cells using real-time polymerase chain reaction to assay steady-state level of mRNA, immunocytochemistry to quantify PAFr protein and proliferating cell nuclear antigen protein by flow cytometry. RESULTS: We found that granulosa cells express PAFr transcripts and protein. PAF presence did not change the concentration of PAFr mRNA or PAFr protein. Granulosa cells responded to PAF doses of 10 and 50 nm with increasing proportions of cells entering G0/G1 phase, as well as a significant expansion of total cell numbers. Rise in G0/G1-phase cells was accompanied by a decline in proliferating cell nuclear antigen protein expression, and these effects could be suspended by simultaneous PAFr blockage. The results provide clear evidence for expression of PAFr in bovine granulosa cells and its functional involvement in PAF/PAFr-mediated stimulation of cell recruitment. CONCLUSIONS:PAF antagonists are suggested to disturb this regulative activity of PAF and to contribute in this way to blockage of ovulation.
Authors: E Baldi; L Bonaccorsi; G Finetti; M Luconi; M Muratori; T Susini; G Forti; M Serio; M Maggi Journal: J Steroid Biochem Mol Biol Date: 1994-06 Impact factor: 4.292