Influence of Cell Culture Conditions and Passage Number on the Response of Membrane Voltage to ATP and Angiotensin II in Rat Mesangial Cells
作者:
Joachim Gloy,
Rainer Greger,
Peter Schollmeyer,
Markus Huber,
Hermann Pavenstädt,
期刊:
Kidney and Blood Pressure Research
(Karger Available online 1994)
卷期:
Volume 17,
issue 2
页码: 62-72
ISSN:1420-4096
年代: 1994
DOI:10.1159/000173789
出版商: S. Karger AG
关键词: Mesangial cells;Cell culture conditions;Extracellular ATP;Angiotensin II;Fetal calf serum
数据来源: Karger
摘要:
The influence of passage number and different culture conditions on the effect of ATP and angiotensin II (A II) on membrane voltage (Vm) of rat mesangial cells (MC) was examined with the patch clamp technique in slow and fast whole cell recordings. MC were characterized immunologically and grown in standard medium in primary culture (PC) and long-term culture up to passage 21 in the presence of 90 g/l fetal calf serum (LTC/ +FCS) or without or with 5 g/l FCS for 1-3 days (LTC/-FCS). In all three series the studies were performed in a FCS-free Ringer-like solution. Vm of MC did not differ in the series (PC: -49 ± lmV, n = 151; LTC/+FCS: -52 ± 1 mV, n = 49; LTC/-FCS: -51 ± 1 mV, n = 44). In primary culture and long-term cultured MC up to passage 8, FCS (ED50≈5 g/l), ATP (ED50 ≈ 2 × 10-6 mol/l) and AII (ED50 ≈ 5 × 10-10 mol/l) induced a depolarization of Vm. Reduction of extracellular Cl- concentration (from 145 to 32 mmol/l) had no effect on Vm but led to an increased depolarization of Vm by FCS, ATP and A II. In long-term cultured MC above passage 8 grown with 90 g/l FCS both ATP and AII induced a concentration-dependent hyperpolarization of Vm, which was attenuated in increased extracellular K+ concentration (from 3.6 to 33.6 mmol/l). In long-term cultured MC beyond passage 8, grown without or with a reduced FCS concentration of 5 g/l, ATP and A II led to a transient depolarization of Vm, which was increased in the presence of 32 mmol/l extracellular Cl-. The depolarization was followed by a hyperpolarization, which was attenuated in the presence of increased extracellular K+. The data indicate that vasoactive agents depolarize Vm of MC in primary culture by activating a Cl- conductance whereas they hyperpolarize Vm by activation of a K+ conductance in long-term cultured MC grown with FCS. The latter effect was partially reversed when FCS w
点击下载:
PDF
(1730KB)
返 回