Mouat M F, Cantrell A C, Manchester K L
Department of Biochemistry, University of the Witwatersrand, Johannesburg, South Africa.
Biosci Rep. 1995 Aug;15(4):173-84. doi: 10.1007/BF01540451.
The effect has been studied of various media, hormones and of amino acids on the membrane potential of rat hepatoma cells in culture measured by microelectrode impalement. Cells in Eagle's minimal essential medium plus 5% serum had a value which varied daily from about 5-8 mV, inside negative. The membrane potential of rat hepatocytes was measured to be 8.7 +/- 0.2 mV, inside negative. The membrane potential of the hepatoma cells was decreased by insulin and increased by glucagon. Membrane potential was unaffected by change of medium to Hanks' or Earle's balanced salt solutions or deprivation of serum. It was, however, reduced in cells in phosphate-buffered saline and by reduction of pH. The former effect was shown to be due to the higher [Na+] of phosphate-buffered saline as opposed to the other media. Addition of alanine, glycine, serine, proline and methylaminoisobutyrate all reduced membrane potential by 2-3 mV. Smaller decreases were seen with methionine, leucine and phenylalanine, but none with glutamine, threonine, BCH (2-aminonorborane-2-carboxylic acid) and D-alanine. The results are compared with the effects of similar conditions on aminoisobutyrate uptake. Whilst there was a correlation under some conditions there was not under others. It is concluded that for the hepatoma cells factors additional to the membrane potential must exert some influence on the capacity for amino acid transport.
通过微电极刺入法对培养的大鼠肝癌细胞的膜电位进行了研究,观察了各种培养基、激素和氨基酸的影响。在含有5%血清的伊格尔氏最低限度基本培养基中的细胞,其膜电位值每天有所不同,约为5 - 8mV,膜内为负。测得大鼠肝细胞的膜电位为8.7±0.2mV,膜内为负。肝癌细胞的膜电位受胰岛素降低、胰高血糖素升高的影响。将培养基换成汉克斯氏或厄尔氏平衡盐溶液或去除血清,膜电位不受影响。然而,在磷酸盐缓冲盐溶液中的细胞以及pH值降低时,膜电位会降低。前者的影响被证明是由于磷酸盐缓冲盐溶液中较高的[Na⁺],与其他培养基不同。添加丙氨酸、甘氨酸、丝氨酸、脯氨酸和甲基氨基异丁酸均使膜电位降低2 - 3mV。甲硫氨酸、亮氨酸和苯丙氨酸使膜电位降低幅度较小,而谷氨酰胺、苏氨酸、2 - 氨基降冰片烷 - 2 - 羧酸(BCH)和D - 丙氨酸则没有影响。将这些结果与类似条件对氨基异丁酸摄取的影响进行了比较。虽然在某些条件下存在相关性,但在其他条件下则不存在。得出的结论是,对于肝癌细胞,除膜电位外的其他因素必定对氨基酸转运能力产生一些影响。