| Project/Area Number |
03454246
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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 |
Circulatory organs internal medicine
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| Research Institution | TOHOKU UNIVERSITY |
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Principal Investigator |
ABE Keishi Tohoku Univ. The 2nd Dep. Int. Med. Professor, 医学部, 教授 (60004777)
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| Co-Investigator(Kenkyū-buntansha) |
KOZUKI Masahiro Tohoku Univ. The 2nd Dep. Int. Med. Assist. Prof., 医学部・附属病院, 助手 (70234698)
OMATA Ken Tohoku Univ. The 2nd Dep. Int. Med. Assist. Prof., 医学部・附属病院, 助手 (50194634)
根東 義明 東北大学医学部, 附属病院, 助手 (00221250)
毛代 昇 (主代 昇) 東北大学, 医学部, 助手 (80237792)
角田 一男 東北大学, 医学部, 助手 (50217361)
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| Project Period (FY) |
1991 – 1993
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| Project Status |
Completed (Fiscal Year 1993)
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| Budget Amount *help |
¥6,200,000 (Direct Cost: ¥6,200,000)
Fiscal Year 1993: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1992: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1991: ¥3,400,000 (Direct Cost: ¥3,400,000)
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| Keywords | Cyochrome P450 / HETE / EET / insulin / RT-PCR / prosraglandin E_2 receptor / chloride channel / チトクロームP450 / Epoxygenase / ω / ω-1 Hydroxylase / Dahlラット / microperfusion / 顕微蛍光測定 / 単離尿細管 / 微小潅流法 / amiloride / 細胞内pH / NaーH対輸送体 / 経上皮電位(Vte) / NaーKー2Cl共輸送体 |
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| Research Abstract |
1. Cytochrome P450 represents the third metabolic pathway of arachidonic acid giving rise to several biologically active compounds, such as 19 HETE, 20 HETE and EETs and their corresponding DHETs. The kidney is the rich source of these metabolites and these metabolites have a wide and contrasting spectrum of biological and renal effects, from vasodilation to vasoconstriction and from inhibition to stimulation of Na-K-ATPase, . Their relative production rates may influence not only renal hemodynamics but also pro- and anti-hypertensive mechanisms of hypertension.
2. The effect of insulin on intracellular pH reguration in microperfused rabbit S2 proximal straight tubules were examined. According to our results, insulin stimulates luminal Na-H exchange.
3. The cDNA encodes 365 amino acids with 97% sequence identify to mouse EP3a receptor (EP3alpha) were isolated. Specific binding of [^3H]PGE_2 was found in COS-7 cells transfected with the cDNA (Kd=3.2nM) and was displaced with unlabeled pro
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staglandins in the order of PGE_2=PGE_1>>PGF_<2alpha>>PGD_2. In situ hybridization revealed expression of rEP_3 receptor principally in the medulla. Moreover, the reverse transcription and polymerase chain reaction (RT-PCR) using dissected nephron segments showed the receptor expression specifically in medullary and cortical thick ascending limbs of Henle's loop, cortical collecting ducts and inner medullary collecting ducts. Moreover, another isoform of EP_3 subtype (EP_<3beta>) with different carboxyl-terminal domains have been characterized.
4. A cDNA encoding MDCK-type chloride channel (RKCL) was isolated. Electrophysiological analysis using Xenopus oocyte expression system showed the functional character of chloride channel. RKCL mRNA is widely expressed in various tissues. In situ hybridization revealed that RKCL mRNA is more expressed in the renal medulla and papilla than in the cortex. RT-PCR showed that main sites of the RKCL mRNA expression are ascending thin limbs of Henle's loop and inner medullary collecting ducts. Under dehydration, renal RKCL mRNA expression level was not statistically changed up to five days. Less
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