Peracchia C, Dulhunty A F
J Cell Biol. 1976 Aug;70(2 pt 1):419-39. doi: 10.1083/jcb.70.2.419.
Electrical uncoupling of crayfish septate lateral giant axons is paralleled by structural changes in the gap junctions. The changes are characterized by a tighter aggregation of the intramembrane particles and a decrease in the overall width of the junction and the thickness of the gap. Preliminary measurements indicate also a decrease in particle diameter. The uncoupling is produced by in vitro treatment of crayfish abdominal cords either with a Ca++, Mg++-free solution containing EDTA, followed by return to normal saline (Van Harreveld's solution), or with VAn Harreveld's solution containing dinitrophenol (DNP). The uncoupling is monitored by the intracellular recording of the electrical resistance at a septum between lateral giant axons. The junctions of the same septum are examined in thin sections; those of other ganglia of the same chain used for the electrical measurements are studied by freeze-fracture. In controls, most junctions contain a more or less regular array of particles repeating at a center to center distance of approximately 200 A. The overall width of the junctions is approximately 200 A and the gap thickness is 40-50 A. Vesicles (400-700 A in diameter) are closely apposed to the junctional membranes. In uncoupled axons, most junctions contain a hexagonal array of particles repeating at a center to center distance of 150-155 A. The overall width of the junctions is approximately 180 A and the gap thickness is 20-30 A. These junctions are usually curved and are rarely associated with vesicles. Isolated, PTA-stained junctions, also believed to be uncoupled, display similar structural features. There are reasons to believe that the changes in structure and permeability are triggered by an increase in the intracellular free Ca++ concentration. Most likely, the changes in permeability are caused by conformational changes in some components of the intramembrane particles at the gap junctions.
小龙虾分隔的外侧巨轴突的电去耦联与间隙连接的结构变化同时发生。这些变化的特征是膜内颗粒聚集更紧密,连接的总宽度和间隙厚度减小。初步测量还表明颗粒直径减小。去耦联是通过用含乙二胺四乙酸(EDTA)的无钙、无镁溶液对小龙虾腹神经索进行体外处理,然后再回到生理盐水(范哈雷维尔德溶液),或用含二硝基苯酚(DNP)的范哈雷维尔德溶液来产生的。通过细胞内记录外侧巨轴突之间隔膜处的电阻来监测去耦联。在薄片中检查同一隔膜的连接;用于电测量的同一神经链其他神经节的连接则通过冷冻断裂进行研究。在对照中,大多数连接含有或多或少规则排列的颗粒,颗粒中心间距约为200埃。连接的总宽度约为200埃,间隙厚度为40 - 50埃。囊泡(直径400 - 700埃)紧密贴附于连接膜。在去耦联的轴突中,大多数连接含有六边形排列的颗粒,颗粒中心间距为150 - 155埃。连接的总宽度约为180埃,间隙厚度为20 - 30埃。这些连接通常是弯曲的,很少与囊泡相关。分离的、经磷钨酸染色的连接,也被认为是去耦联的,显示出类似的结构特征。有理由相信结构和通透性的变化是由细胞内游离钙离子浓度的增加引发的。很可能,通透性的变化是由间隙连接处膜内颗粒某些成分的构象变化引起的。