Kunzelmann K, Allert N, Kubitz R, Breuer W V, Cabantchik Z I, Normann C, Schumann S, Leipziger J, Greger R
Physiologisches Institut der Albert-Ludwigs-Universität Freiburg, Germany.
Pflugers Arch. 1994 Aug;428(1):76-83. doi: 10.1007/BF00374754.
HT29 cells were preincubated with forskolin (10(-5) mol/l, FORHT) or phorbol 12-myristate 13-acetate (PMA) (10(-7) mol/l, PMAHT) for 20 h, which has been shown previously and is also shown here, to upregulate and downregulate, respectively, the expression of the cystic fibrosis transmembrane conductance regulator (CFTR). CFPAC-1 cells underwent the same protocols. HT29 cells were examined by slow (SWC) and fast (FWC) whole-cell patch-clamp techniques. The results of SWC and FWC were indistinguishable and were pooled. CFPAC-1 cells were examined with FWC. The membrane voltage (V) of FORHT was -41.8 +/- 1.4 mV (n = 77) and that of PMAHT was -43.6 +/- 2.4 mV (n = 76). The conductance (G) of FORHT (9.4 +/- 0.9 nS, n = 77) was significantly larger than that of PMAHT (3.7 +/- 0.4 nS, n = 76). Acute application of forskolin (10(-5) mol/l, FOR) plus 0.5 mmol/l 8-(4-chlorophenylthio)-cAMP (cAMP) depolarized V by 12 (FORHT) and 8 (PMAHT) mV, respectively. The acute increase of G by FOR plus cAMP was by 7.6 +/- 1.9 nS for FORHT (n = 22) and only 2.2 +/- 1 nS for PMAHT (n = 13). ATP (10(-4) mol/l) depolarized V in both types of cells. It enhanced G by 16.7 +/- 4.1 nS in FORHT (n = 14) and significantly less (by 5.5 +/- 1.2 nS, n = 14) in PMAHT. Also the G increase lasted longer in FORHT. Neurotensin (NT, 10(-8) mol/l) also had a stronger and longer lasting effect in FORHT.(ABSTRACT TRUNCATED AT 250 WORDS)
将HT29细胞与毛喉素（10⁻⁵ mol/l，FORHT）或佛波酯12 -肉豆蔻酸酯13 -乙酸酯（PMA）（10⁻⁷ mol/l，PMAHT）预孵育20小时，此前研究及本研究均表明，这分别上调和下调了囊性纤维化跨膜传导调节因子（CFTR）的表达。CFPAC - 1细胞采用相同方案处理。通过慢（SWC）和快（FWC）全细胞膜片钳技术检测HT29细胞。SWC和FWC的结果无差异，合并分析。用FWC检测CFPAC - 1细胞。FORHT的膜电压（V）为 - 41.8±1.4 mV（n = 77），PMAHT的膜电压为 - 43.6±2.4 mV（n = 76）。FORHT的电导（G）为9.4±0.9 nS（n = 77），显著大于PMAHT的电导（3.7±0.4 nS，n = 76）。急性施加毛喉素（10⁻⁵ mol/l，FOR）加0.5 mmol/l 8 -（4 -氯苯基硫代）-环磷酸腺苷（cAMP）分别使FORHT的V去极化12 mV、PMAHT的V去极化8 mV。FOR加cAMP使FORHT的G急性增加7.6±1.9 nS（n = 22），而PMAHT仅增加2.2±1 nS（n = 13）。ATP（10⁻⁴ mol/l）使两种细胞的V去极化。它使FORHT的G增加16.7±4.1 nS（n = 14），在PMAHT中增加显著较少（5.5±1.2 nS，n = 14）。而且FORHT中G的增加持续时间更长。神经降压素（NT，10⁻⁸ mol/l）在FORHT中也有更强且更持久的作用。（摘要截断于250字）
