Bernardini G, Peracchia C
Invest Ophthalmol Vis Sci. 1981 Aug;21(2):291-9.
Previous studies have shown that cell uncoupling is paralleled by an increase in tightness and crystallinity of gap junction particle arrays. Gap junction crystallinity is believed to be part of the uncoupling mechanism because it can be produced in gap junctions isolated from lens fibers on direct exposure to uncoupling agents such as divalent cations or hydrogen ions. Some doubts, however, have been raised on the capacity of lens fiber junctions to crystallize and uncouple in situ. The present study shows that the gap junctions of rat lens fibers indeed crystallize after a treatment that increases drastically the membrane permeability to ions. The treatment consists of a brief immersion of the lenses in liquid nitrogen, followed by incubation for several hours in Tyrode's solution at 37 degrees C. Immediately after liquid nitrogen treatment, the lenses start gaining sodium and calcium while losing potassium, and eventually become opaque. Addition of 10 mM EDTA to calcium and magnesium-free Tyrode's solutions inhibits particle crystallization and lens cataract, whereas low concentrations of EDTA (1 mM) are not effective. These findings, together with preliminary data on the capacity of lens fibers to heal over, indicate that the gap junctions of lens fibers are capable of crystallizing and uncoupling in situ.
先前的研究表明,细胞解偶联与间隙连接颗粒阵列的紧密性和结晶度增加同时发生。间隙连接结晶度被认为是解偶联机制的一部分,因为在直接暴露于二价阳离子或氢离子等解偶联剂的情况下,从晶状体纤维中分离出的间隙连接中可以产生结晶度。然而,对于晶状体纤维连接在原位结晶和解偶联的能力存在一些疑问。本研究表明,在大幅提高膜对离子通透性的处理后,大鼠晶状体纤维的间隙连接确实会结晶。该处理包括将晶状体短暂浸入液氮中,然后在37℃的台氏液中孵育数小时。液氮处理后,晶状体立即开始摄取钠和钙,同时丢失钾,最终变得不透明。向无钙镁的台氏液中添加10 mM EDTA可抑制颗粒结晶和晶状体白内障形成,而低浓度的EDTA(1 mM)则无效。这些发现,连同关于晶状体纤维愈合能力的初步数据,表明晶状体纤维的间隙连接能够在原位结晶和解偶联。