| Project/Area Number |
06454145
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
General physiology
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| Research Institution | Shinshu University School of Medicine |
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Principal Investigator |
OHHASHI Toshio The 1st Department of Physiology, Shinshu University School of Medicine, Professor, 医学部, 教授 (80020832)
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| Project Period (FY) |
1994 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥5,800,000 (Direct Cost: ¥5,800,000)
Fiscal Year 1995: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1994: ¥4,800,000 (Direct Cost: ¥4,800,000)
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| Keywords | lymph vessel / lymphatic endothelial cell / lymphatic smooth muscle / lymph node / nitric oxide / macrophage / Ca-transient / pH-transient / Ca^<2+>-transient / pH-transient / 誘導型一酸化窒素 / L‐nitro arginine methyl ester / dexamethasone / nifedipine |
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| Research Abstract |
The distal end of isolated dog thoracic duct was cannulated by a polyethylene catheter and then perfused with Rinaldi solution containing 250 U/ml collagenase and 0.5 mM EGTA.After a 10-min incubation with the solution, the thoracic duct was flushed with minimum essential medium containing 20% fetal bovine serum and then perfusate was centrifuged to obtain the denuded endothelial cells. The pellets were resuspended in MEM supplemented with fetal bovine serum and placed onto tissue culture dishes coated with type I collagen. The dog lymphatic endothelial cells grew with doubling time of about 58 hrs. The ploygonal cells had a classic "cobblestone " morphology at confluence. Most culyured cells expressed Factor VIII-related antigen and were shown to take up acetylated LDL via the "scavenger cell pathway " by use of fluorescence microscopy. The cultured lymphatic endothelial cells were loaded with fura-2 AM to measure Ca^<2+>-transient. An administration of 10^<-5>M histamine caused a significant increase of intracellular concentrations of Ca^<2+> in the cultured cells. A tenfold increase of [Ca^<2+>]i was observed 1 min after the administration. The [Ca^<2+>]i was gradually decreased and then kept at constant level by 4-5 min after the administration.
5-HT_1-like receptors exist in the monkey popliteal lymph nodes. Stimulation of these receptors produces an endogenous nitric oxide-dependent relaxation in lymph node smooth muscle through an activation of cytosolic guanylate cyclase in the cells. Unstimulated rat peritoneal macrophages cause a corelease of nitric oxide and vasodilative prostaglandins resulting in a marked relaxation of arterial smooth muscles
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