Isolation and characterization of ovine luteal pericytes and effects of nitric oxide on pericyte expression of angiogenic factors

Joan D. Beckman, Anna T. Grazul-Bilska, Mary Lynn Johnson, Lawrence P. Reynolds, Dale A. Redmer

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

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 NOdonor 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 β3 (GUCY1B3), using real-time quantitative RT-PCR. NO caused a dosedependent 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.

Original languageEnglish (US)
Pages (from-to)467-476
Number of pages10
JournalEndocrine
Volume29
Issue number3
DOIs
StatePublished - Jun 1 2006

Fingerprint

Pericytes
Angiogenesis Inducing Agents
Corpus Luteum
Sheep
Nitric Oxide
Fibroblast Growth Factor 2
Vascular Endothelial Growth Factor A
Endothelial Cells
Messenger RNA
Smooth Muscle Myocytes
DEET
Angiopoietins
Estrous Cycle
Vascular Smooth Muscle
Actins
Real-Time Polymerase Chain Reaction
RNA
Staining and Labeling

Keywords

  • Angiogenesis
  • Corpus luteum
  • Nitric oxide
  • Pericytes

Cite this

Isolation and characterization of ovine luteal pericytes and effects of nitric oxide on pericyte expression of angiogenic factors. / Beckman, Joan D.; Grazul-Bilska, Anna T.; Johnson, Mary Lynn; Reynolds, Lawrence P.; Redmer, Dale A.

In: Endocrine, Vol. 29, No. 3, 01.06.2006, p. 467-476.

Research output: Contribution to journalArticle

Beckman, Joan D. ; Grazul-Bilska, Anna T. ; Johnson, Mary Lynn ; Reynolds, Lawrence P. ; Redmer, Dale A. / Isolation and characterization of ovine luteal pericytes and effects of nitric oxide on pericyte expression of angiogenic factors. In: Endocrine. 2006 ; Vol. 29, No. 3. pp. 467-476.
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AB - 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 NOdonor 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 β3 (GUCY1B3), using real-time quantitative RT-PCR. NO caused a dosedependent 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.

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