ADENYLATE-CYCLASE RESPONSES AND BIOCHEMICAL-CHARACTERIZATION OF PRIMARY RABBIT PROXIMAL TUBULAR CELL-CULTURES AND LLC-PK1 CELLS

被引：10

作者：

TOUTAIN, H ^{[1
]}

COURJAULT, F ^{[1
]}

VAUCLINJACQUES, N ^{[1
]}

MORIN, JP ^{[1
]}

机构：

[1] INSERM, U-295, UER MED PHARM ROUEN, BP 97, ST ETIENNE DU ROUVRAY, FRANCE

来源：

RENAL FAILURE | 1990年 / 12卷 / 03期

关键词：

D O I：

10.3109/08860229009065561

中图分类号：

R5 [内科学]; R69 [泌尿科学（泌尿生殖系疾病）];

学科分类号：

1002 ; 100201 ;

摘要：

Freshly isolated rabbit proximal tubules (PT), confluent primary rabbit proximal tubule cultures (PTC) and LLC-PK1 cells were characterised. Brushborder enzyme activities were lower in PTC than in LLC-PK1: ratios were 0.026 for alkaline phosphatase (AP), 0.458 for alanine aminopeptidase (AAP) and 0.514 for gamma-glutamyl transpeptidase (GGT). PT/PTC ratios were 79.7 for AP, 7.96 for AAP and 3.45 for GGT. Specific activities of hexokinase (HK) and lactate dehydrogenase (LDH) were high in cultured cells as compared to PT: PT/PTC ratios were 0.063 and 0.033, while PTC/LLC-PK1 ratios were 0.406 and 1.19 for HK and LDH respectively. PTC/LLC-PK1 ratios were 2.21 forNa/K ATPase, 2.07 for succinate dehydrogenase, 1.12 for cathepsin B, 0.607 for N-acetyl-βD-glucosaminidase and 8.98 for gluta-thione-S-transferase. Adenylate cyclase response to parathormone (PTH), was similar in PTC and PT, but stimulated/basal ratios were higher in PT than in PTC. LLC-PK1 cells were stimulated by thyrocalcitonin(SCT),arginin-vasopressin (AVP) and PTH; stimulated/basal ratios ranked AVP > PTH > SCT. Differences between both types of cultures affect the choice of in vitro model for nephrotoxicity studies. © 1990 Informa UK Ltd All rights reserved: reproduction in whole or part not permitted.

引用

页码：177 / 182

页数：6

共 28 条

[1] Husted R.F., Welsch M.J., Stokes J.B., Variability of functional characteristics of MDCK cells, Am J Physiol, 250, pp. C214-C221, (1986)
[2] Wohlwend A., Vassa U.I.D., Belin D., Orci L., LLC-PK1 cells: cloning of phenotypically stable subpopulations, Am J Physiol, 250, pp. C682-C687, (1986)
[3] Haggerty J.G., Agarwal N., Cragoe E.J., Adelberg E.A., Slayman C.W., LLC-PK1 mutant with increased Na+-H+ exchange and decreased sensitivity to amiloride, Am J Physiol, 255, pp. C495-C501, (1988)
[4] Toutain H., Fillastre J.P., Morin J.P., Preparative free flow electrophoresis for the isolation of two populations of proximal cells from the rabbit kidney, Eur J Cell Biol, 49, pp. 274-280, (1989)
[5] Gstraunthaler G., Pfaller W., Kotanko P., Biochemical characterization of renal epithelial cell cultures (LLC-PK1 and MDCK), Am J Physiol, 248, pp. F536-F544, (1985)
[6] Brendel K., Meezan E., Isolation and properties of a pure preparation of proximal kidney tubules obtained without collagenase treatment, Fed Proc, 34, (1975)
[7] Bradford M.M., A rapid and sensitive method for quantification of microgram quantities of protein utilizing the principle of protein dye binding, Anal Biochem, 72, pp. 248-254, (1976)
[8] Hjelle J.T., Morin J.P., Trouet A., Analytical cell fractionation of isolated rabbit renal proximal tubules, Kidney Int, 20, pp. 71-77, (1981)
[9] Bergmeyer H.U., Hexokinase assay, Methods of enzymatic analysis, pp. 473-475, (1974)
[10] Bergmeyer H.U., Bemt E., Lactate dehydrogenase UV-assay with pyruvate and NADH, Methods of enzymatic analysis, pp. 574-579, (1974)

← 1 2 3 →