title: Maturation enhances fluid shear-induced activation of eNOS in perfused ovine carotid arteries.
abstract: The present study tests the hypothesis that age-dependent increases in endothelial vasodilator capacity are due to maturational increases in endothelial nitric oxide (NO) synthesis and release. Intact 4-cm carotid artery segments taken from term fetal lambs and nonpregnant adult sheep were perfused by using a closed system that enabled independent control of flow and inflow pressure and facilitated complete recovery of all NO released. Fluid shear stress induced a graded release of NO (in nmol NO x min x cm(-2) of luminal surface area) that was significantly greater in adult (890 +/- 140) than in fetal (300 +/- 40) carotid arteries at corresponding values of shear stress (5.9 +/- 0.3 dyn/cm2) but was independent of inflow pressure in both age groups. These age-related differences in NO release were not attributable to corresponding differences in endothelial NO synthase (eNOS) abundance, as eNOS protein levels (in ng of eNOS/cm2 of luminal surface area) were similar in adult (14 +/- 2) and fetal (12 +/- 1) arteries. Adult (80 +/- 15) and fetal (89 +/- 32) levels of eNOS mRNA (in 10(6) copies/cm2 of luminal surface area) were also similar. However, when NO release was normalized relative to the associated mass of eNOS protein to estimate eNOS-specific activity in situ, this value (in nmol NO x microg of eNOS(-1) x min(-1)) was significantly greater in adult (177 +/- 44) than in fetal (97 +/- 36) arteries when the endothelium was maximally activated by A-23187. Similarly, the slope of the relation between fluid shear stress and estimated eNOS-specific activity (in nmol NO x microg of eNOS(-1) x min(-1) per dyn/cm2) was also significantly greater in adult (6.8 +/- 0.1) than in fetal (2.9 +/- 0.1) arteries, which suggests that eNOS may be more sensitive to or more efficiently coupled to activating stimuli in adult compared with fetal arteries. We conclude that maturational increases in endothelial vasodilator capacity are attributable to age-dependent increases in NO release secondary to elevated eNOS-specific activity and involve more efficient coupling between endothelial activation and enhancement of eNOS activity in adult compared with fetal arteries.
authors.affiliation: Center for Perinatal Biology, Loma Linda Univ., Loma Linda, CA 92350, USA.
authors.forename: Charles Ray
authors.initials: CR
authors.lastname: White
authors.forename: Mohammad Wael
authors.initials: MW
authors.lastname: Hamade
authors.forename: Koushan
authors.initials: K
authors.lastname: Siami
authors.forename: Melody M
authors.initials: MM
authors.lastname: Chang
authors.forename: Anandit
authors.initials: A
authors.lastname: Mangalwadi
authors.forename: John A
authors.initials: JA
authors.lastname: Frangos
authors.forename: William J
authors.initials: WJ
authors.lastname: Pearce
mesh_terms: D000244:Adenosine Diphosphate / Q000494:pharmacology; D000375:Aging / Q000502:physiology*; D000818:Animals; D015153:Blotting, Western; D000001:Calcimycin / Q000494:pharmacology; D002339:Carotid Arteries / Q000254:growth & development* / Q000502:physiology*; D004789:Enzyme Activation / Q000502:physiology; D005260:Female; D005333:Fetus / Q000502:physiology; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D008297:Male; D009569:Nitric Oxide / Q000378:metabolism; D011247:Pregnancy; D012333:RNA, Messenger / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D033081:Shear Strength; D012756:Sheep
chemical_list: D012333:RNA, Messenger; D009569:Nitric Oxide; D000001:Calcimycin; D000244:Adenosine Diphosphate