Ravesloot J H, Van Houten R J, Ypey D L, Nijweide P J
Department of Cell Biology and Histology, Leiden University, The Netherlands.
J Bone Miner Res. 1991 Apr;6(4):355-63. doi: 10.1002/jbmr.5650060407.
Patch-clamp measurements done on excised membrane patches obtained from 1-5 day cultured embryonic chick osteoblasts, osteocytes, and periosteal fibroblasts revealed the existence of a high-conductance anion channel: 371 +/- 63 pS when measured under symmetrical 158 mM Cl- conditions. The channel frequently displayed subconductance levels. The ion selectivity of the channel expressed as the (an)ion to chloride permeability ratio was as follows: Cl- (1.0) greater than methylsulfate- (0.71) greater than gluconate- (0.25) greater than glutamate- (0.17) greater than Na+ = K+ (0.10). In addition, the channel had a significant permeability for inorganic phosphate ions. The channel was found in about 1% of the cell-attached patches, which indicates that the channel is under the control of as yet unknown intracellular factors. Once activated by patch excision, the channel was voltage dependent and active at potentials close to 0 mV. At potentials outside the range of +/- 10 mV channel activity decreased. This process proceeded faster at increasing membrane potentials of either polarity. Returning to potentials close to 0 mV caused reopening of the channels within seconds if the preceding voltage step led to complete closure of the channels. Channel activity did not depend noticeably on intracellular and extracellular CA2+ ions. The channel is not unique to (chick) osteogenic cells but has been demonstrated in excised patches obtained from excitable and other nonexcitable cells. Although its presence in a wide variety of cell types suggests that the channel plays a general role in as yet unknown cell physiologic processes, the channel may also have specific functions in osteogenic cells, for example providing a pathway for phosphate ions during mineralization.
对从培养1 - 5天的胚胎鸡成骨细胞、骨细胞和骨膜成纤维细胞获得的膜片进行膜片钳测量,结果显示存在一种高电导阴离子通道:在对称的158 mM Cl⁻条件下测量时,电导为371 ± 63 pS。该通道经常表现出亚电导水平。通道的离子选择性以(某)离子与氯离子的通透率之比表示如下:Cl⁻(1.0)>甲基硫酸根⁻(0.71)>葡萄糖酸根⁻(0.25)>谷氨酸根⁻(0.17)>Na⁺ = K⁺(0.10)。此外,该通道对无机磷酸根离子具有显著的通透性。在约1%的细胞贴附膜片中发现了该通道,这表明该通道受尚未知晓的细胞内因子调控。一旦通过膜片切除激活,该通道呈电压依赖性,在接近0 mV的电位下具有活性。在±10 mV范围之外的电位下,通道活性降低。在膜电位向任一极性增加时,这一过程进行得更快。如果先前的电压阶跃导致通道完全关闭,回到接近0 mV的电位会使通道在数秒内重新开放。通道活性对细胞内和细胞外Ca²⁺离子没有明显依赖性。该通道并非(鸡)成骨细胞所特有,在从可兴奋细胞和其他非可兴奋细胞获得的膜片中也得到了证实。尽管它在多种细胞类型中的存在表明该通道在尚未知晓的细胞生理过程中发挥着普遍作用,但该通道在成骨细胞中可能也具有特定功能,例如在矿化过程中为磷酸根离子提供一条通路。