title: Divergent signaling mechanisms for venous versus arterial contraction as revealed by endothelin-1.
abstract: OBJECTIVE
  Venous function is underappreciated in its role in blood pressure determination, a physiologic parameter normally ascribed to changes in arterial function. Significant evidence points to the hormone endothelin-1 (ET-1) as being important to venous contributions to blood pressure. We hypothesized that the artery and vein should similarly depend on the signaling pathways stimulated by ET-1, specifically phospholipase C (PLC) activation. This produces two functional arms of signaling: diacylglycerol (DAG; protein kinase C [PKC] activation) and inositol trisphosphate (IP3) production (intracellular calcium release).
  
  
  METHODS
  The model was the male Sprague-Dawley rat. Isolated tissue baths were used to measure isometric contraction. Western blot and immunocytochemical analyses measured the magnitude of expression and site of expression, respectively, of IP3 receptors in smooth muscle/tissue. Pharmacologic methods were used to modify PLC activity and signaling elements downstream of PLC (IP3 receptors, PKC).
  
  
  RESULTS
  ET-1-induced contraction was PLC dependent in both tissues as the PLC inhibitor U-73122 significantly reduced contraction in aorta (86% ± 4% of control; P < .05) and vena cava (49% ± 11% of control; P < .05). However, ET-1-induced contraction was not significantly inhibited by the IP3 receptor inhibitor 2-aminoethoxydiphenylborane (100 μM) in vena cava (82% ± 8% of control; P = .23) but was in the aorta (55% ± 4% of control; P < .05). All three IP3 receptor isoforms were located in venous smooth muscle. IP3 receptors were functional in both tissues as the novel membrane-permeable IP3 analogue (Bt-IP3; 10 μM) contracted aorta and vena cava. Similarly, whereas the PKC inhibitor chelerythrine (10 μM) attenuated ET-1-induced contraction in vena cava and aorta (5% ± 2% and 50% ± 5% of control, respectively; P < .05), only the vena cava contracted to the DAG analogue 1-oleoyl-2-acetyl-sn-glycerol.
  
  
  CONCLUSIONS
  These findings suggest that ET-1 activates PLC in aorta and vena cava, but vena cava contraction to ET-1 may be largely IP3 independent. Rather, DAG—not IP3—may contribute to contraction to ET-1 in vena cava, in part by activation of PKC. These studies outline a fundamental difference between venous and arterial smooth muscle and further reinforce a heterogeneity of vascular smooth muscle function that could be taken advantage of for therapeutic development.
authors.affiliation: Department of Pharmacology and Toxicology, Michigan State University, East Lansing, Mich. Electronic address: tykockin@msu.edu.
authors.forename: Nathan R
authors.initials: NR
authors.lastname: Tykocki
authors.affiliation: Department of Pharmacology and Toxicology, Michigan State University, East Lansing, Mich.
authors.forename: BinXi
authors.initials: B
authors.lastname: Wu
authors.affiliation: Department of Pharmacology and Toxicology, Michigan State University, East Lansing, Mich.
authors.forename: William F
authors.initials: WF
authors.lastname: Jackson
authors.affiliation: Department of Pharmacology and Toxicology, Michigan State University, East Lansing, Mich.
authors.forename: Stephanie W
authors.initials: SW
authors.lastname: Watts
mesh_terms: D000818:Animals; D001011:Aorta / Q000187:drug effects* / Q000201:enzymology; D020013:Calcium Signaling / Q000187:drug effects*; D004075:Diglycerides / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D019332:Endothelin-1 / Q000494:pharmacology*; D004789:Enzyme Activation; D066298:In Vitro Techniques; D015544:Inositol 1,4,5-Trisphosphate / Q000378:metabolism; D053496:Inositol 1,4,5-Trisphosphate Receptors / Q000378:metabolism; D008297:Male; D011493:Protein Kinase C / Q000378:metabolism; D017207:Rats, Sprague-Dawley; D010738:Type C Phospholipases / Q000378:metabolism; D014661:Vasoconstriction / Q000187:drug effects*; D014662:Vasoconstrictor Agents / Q000494:pharmacology*; D014684:Venae Cavae / Q000187:drug effects* / Q000201:enzymology
chemical_list: D004075:Diglycerides; D019332:Endothelin-1; D053496:Inositol 1,4,5-Trisphosphate Receptors; D014662:Vasoconstrictor Agents; D015544:Inositol 1,4,5-Trisphosphate; D011493:Protein Kinase C; D010738:Type C Phospholipases
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