Frindt G, Palmer L G
Department of Physiology, Cornell University Medical College, New York, New York 10021.
Am J Physiol. 1989 Jan;256(1 Pt 2):F143-51. doi: 10.1152/ajprenal.1989.256.1.F143.
Low-conductance, K-selective channels were identified in the apical membrane of the rat cortical collecting tubule (CCT) by use of the patch-clamp technique. Isolated, split tubules were bathed in K gluconate medium to depolarize the cell while keeping the intracellular K concentration high. With the patch-clamp pipette containing predominantly either Na+ or Li+ but no K, outward currents were observed through channels that had a single-channel conductance (g) of 9 pS and a probability of being open (Po) of greater than 0.9, independent of the voltage (+/- 40 mV) applied to the pipette (Vp). Similarly, only outward currents were observed when the patch was excised into high-K solution, implying a high selectivity of the channel for K+. When 1 mM BaCl2 was added to the pipette, Po decreased to 0.36 at Vp = 0; however, g was not changed but the channels flickered rapidly between open and blocked states; Po decreased as Vp was made positive, and increased as Vp was made negative. With the pipette filled with KCl + 1 mM Ba, the channels conducted K+ in both directions. The inward currents (at positive Vp were larger than the outward currents (at negative Vp) and g near Vp = 0 increased to 25 pS. When the pipette was filled with RbCl + 1 mM Ba the inward and outward currents were similar in magnitude, suggesting that the channels can conduct Rb, although not as well as K. With the tubules bathed in NaCl Ringer solution and the pipette containing KCl, inward currents were observed that could be attributed to the same pathway for K.(ABSTRACT TRUNCATED AT 250 WORDS)
运用膜片钳技术,在大鼠皮质集合管(CCT)的顶端膜中鉴定出了低电导、钾选择性通道。分离的、劈开的小管浸泡在葡萄糖酸钾培养基中,以使细胞去极化,同时保持细胞内高钾浓度。膜片钳微管主要含有Na⁺或Li⁺但不含K⁺,通过单通道电导(g)为9 pS且开放概率(Po)大于0.9的通道观察到外向电流,该电流与施加于微管的电压(Vp)(±40 mV)无关。同样,当膜片切除后置于高钾溶液中时,仅观察到外向电流,这意味着该通道对K⁺具有高选择性。当向微管中加入1 mM BaCl₂时,在Vp = 0时Po降至0.36;然而,g未改变,但通道在开放和阻断状态之间快速闪烁;Po随着Vp变为正值而降低,随着Vp变为负值而增加。当微管中充满KCl + 1 mM Ba时,通道在两个方向上都传导K⁺。内向电流(在正Vp时)大于外向电流(在负Vp时),且Vp = 0附近的g增加到25 pS。当微管中充满RbCl + 1 mM Ba时,内向和外向电流大小相似,这表明通道可以传导Rb,尽管不如传导K⁺那样好。当小管浸泡在NaCl林格溶液中且微管中含有KCl时,观察到内向电流,该电流可归因于K⁺的相同传导途径。(摘要截断于250字)