2000 Fiscal Year Final Research Report Summary
Molecularbiological analysis of the function and localization of membrane proteins.
| Project/Area Number |
09671151
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Kidney internal medicine
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| Research Institution | University of Tokyo |
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Principal Investigator |
TAKAICHI Kenmei University of Tokyo Branch Hospital, Lecturer, 医学部・附属病院・分院, 講師 (00175423)
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| Project Period (FY) |
1997 – 2000
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| Keywords | sodium-proton exchanger / NHE1 / LLC-PK1 cells / OK cells / MDCK cells / collagen gel / DNA tip / fibronectin / MDCK細胞 |
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| Research Abstract |
The appropriate localization of membrane proteins in the apical or basolateral is mandatory for the physiologic transport of the solutes in the epithelia. Derangement of the localization of membrane proteins has been observed in the pathological situations such as renal cysts. We examined the localization of sodium-proton exchanger (NHE1), one of membrane proteins, in the renal tubular epithelial cells (LLC-PK1 cells, OK cells) cultured on the filter membranes coated with collagen. Antisense-Oligo of NHE1 decreased intrinsic NHE1 activity only in the basolateral membranes in LLC-PK1 cells. NHE1 activity was detected only in the basolateral membranes in OK cells transfected with human NHE1 cDNA.These results show that NHE1 is localized in the basolateral membranes in the renal epitelia in the physiologic situations. MDCK cells cultured in the collagen gel form cysts but not tubules without hepatic growth factor (HGF). We isolated the MDCK cells (C4) that form tubules without HGF and cells (C1) that form cysts. The differential expression of mRNA in these two kinds of cells was examined using DNA tip, in which more than 9,000 different cDNAs were arrayed. The expression of fibronectin was decreased in C1, which indicated the significance of fibronectin for tubular formation of MDCK cells.
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