Isolation and characterization of ovine luteal pericytes and effects of nitric oxide on pericyte expression of angiogenic factors.
We have demonstrated that vascular endothelial growth factor (VEGF) is expressed in capillary pericytes of the developing corpus luteum (CL) and others have shown that basic fibroblast growth factor (FGF2) and angiopoietins (ANGPT) are present in the CL. VEGF and FGF2 target endothelial cells to initiate angiogenesis and stimulate nitric oxide (NO) production. Conversely, NO may increase VEGF expression by vascular smooth muscle cells and pericytes. To investigate the relationship between these angiogenic factors and NO in the CL, microvascular pericytes and endothelial cells were isolated from CL collected from superovulated ewes (n = 5) on d 9 of the estrous cycle. Pericytes were identified by their morphology in culture and by immunofluorescent staining for smooth muscle cell actin. Pericytes were incubated with or without varying doses of the NO-donor DETA-NO for 8 h. Then, total cellular RNA was extracted from the cells and evaluated for expression of mRNA for VEGF, FGF2, ANGPT1, ANGPT2, and NO receptor, guanylate cyclase 1, soluble beta3 (GUCY1B3), using real-time quantitative RT-PCR. NO caused a dose-dependent increase in VEGF (p < 0.001), FGF2 (p < 0.001), ANGPT2 (p < 0.06), and GUCY1B3 (p < 0.03) mRNA expression. Expression of mRNA for ANGPT1 in luteal pericytes was not affected by the NO treatment. These data provide further evidence of the role of the luteal pericyte and NO in angiogenic factor expression, and of the potential interactions of pericytes with endothelial cells via NO production.
Beckman
Joan D
JD
Department of Animal and Range Sciences, North Dakota State University, Fargo, ND 58105-5727, USA.
Grazul-Bilska
Anna T
AT
Johnson
Mary Lynn
ML
Reynolds
Lawrence P
LP
Redmer
Dale A
DA
D043925:Angiogenesis Inducing Agents / Q000378:metabolism*; D000809:Angiotensins / Q000378:metabolism; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D003338:Corpus Luteum / Q000166:cytology; D005260:Female; D051604:Guanylate Cyclase-Activating Proteins / Q000378:metabolism; D007150:Immunohistochemistry; D008077:Lipoproteins, LDL / Q000493:pharmacokinetics; D008184:Luteal Cells / Q000166:cytology* / Q000187:drug effects; D009569:Nitric Oxide / Q000096:biosynthesis / Q000494:pharmacology*; D020030:Nitric Oxide Donors / Q000494:pharmacology; D020286:Pericytes / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D012756:Sheep; D014226:Triazenes / Q000494:pharmacology
C105351:1-hydroxy-2-oxo-3,3-bis(2-aminoethyl)-1-triazene; D043925:Angiogenesis Inducing Agents; D000809:Angiotensins; D051604:Guanylate Cyclase-Activating Proteins; D008077:Lipoproteins, LDL; D020030:Nitric Oxide Donors; D012333:RNA, Messenger; D014226:Triazenes; C032629:acetyl-LDL; D009569:Nitric Oxide