Li Yan-feng, Zhuo Ye-hong, Bi Wei-na, Bai Yu-jing, Li Yan-na, Wang Zhi-jian
State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, Guangdong 510060, China.
Chin Med J (Engl). 2008 Nov 20;121(22):2272-7.
The cell layer of the ciliary epithelium is responsible for aqueous humor secretion and maintenance. Ion channels play an important role in these processes. The main aim of this study was to determine whether the well-characterized members of the Kv1 family (Kv1.3) contribute to the Kv currents in ciliary epithelium.
New Zealand White rabbits were maintained in a 12 hours light/dark cycle. Ciliary epithelium samples were isolated from the rabbits. We used Western blotting and immunocytochemistry to identify the expression and location of a voltage-gated potassium channel Kv1.3 in ciliary body epithelium. Membrane potential change after adding of Kv1.3 inhibitor margatoxin (MgTX) was observed with a fluorescence method.
Western blotting and immunocytochemical studies showed that the Kv1.3 protein expressed in pigment ciliary epithelium and nonpigment ciliary epithelium, however it seemed to express more in the apical membrane of the nonpigmented epithelial cells. One nmol/L margatoxin, a specific inhibitor of Kv1.3 channels caused depolarization of the cultured nonpigmented epithelium (NPE) membrane potential. The cytosolic calcium increased after NPE cell depolarization, this increase of cytosolic calcium was partially blocked by 12.5 micromol/L dantrolene and 10 micromol/L nifedipine. These observations suggest that Kv1.3 channels modulate ciliary epithelium potential and effect calcium dependent mechanisms.
Kv1.3 channels contribute to K+ efflux at the membrane of rabbit ciliary epithelium.
睫状体上皮细胞层负责房水的分泌与维持。离子通道在这些过程中发挥着重要作用。本研究的主要目的是确定Kv1家族中特征明确的成员(Kv1.3)是否对睫状体上皮中的Kv电流有贡献。
将新西兰白兔饲养在12小时光照/黑暗周期中。从兔子身上分离出睫状体上皮样本。我们使用蛋白质免疫印迹法和免疫细胞化学法来鉴定电压门控钾通道Kv1.3在睫状体上皮中的表达和定位。采用荧光法观察加入Kv1.3抑制剂玛格毒素(MgTX)后膜电位的变化。
蛋白质免疫印迹法和免疫细胞化学研究表明,Kv1.3蛋白在色素睫状体上皮和非色素睫状体上皮中均有表达,但似乎在非色素上皮细胞的顶端膜中表达更多。1 nmol/L的玛格毒素,一种Kv1.3通道的特异性抑制剂,可导致培养的非色素上皮(NPE)膜电位去极化。NPE细胞去极化后胞质钙增加,12.5 μmol/L的丹曲林和10 μmol/L的硝苯地平可部分阻断胞质钙的这种增加。这些观察结果表明,Kv1.3通道调节睫状体上皮电位并影响钙依赖机制。
Kv1.3通道有助于兔睫状体上皮细胞膜上的钾离子外流。