Mathias R T, Riquelme G, Rae J L
Department of Physiology and Biophysics, State University of New York, Stony Brook 11794-8661.
J Gen Physiol. 1991 Dec;98(6):1085-103. doi: 10.1085/jgp.98.6.1085.
Fiber cells of the lens are electrically and diffusionally interconnected through extensive gap junctions. These junctions allow fluxes of small solutes to move between inner cells and peripheral cells, where the majority of transmembrane transport takes place. We describe here a method utilizing two intracellular microelectrodes to measure the cell to cell resistance between fiber cells at any given distance into the intact lens. We also use ion-sensitive microelectrodes to record intracellular pH at various depths in the intact lens. We find that gap junctions connecting inner fiber cells differ in pH sensitivity as well as normal coupling resistance from those connecting peripheral cells. The transition occurs in a zone between 500 and 650 microns into the lens. Fiber cells peripheral to this zone have a specific coupling resistance of 1.1 omega cm2, whereas those inside have a specific coupling resistance of 2.7 omega cm2. However, when the cytoplasm of fiber cells is acidified by bubbling with CO2, peripheral cells uncouple and the cell to cell resistance goes up more than 40-fold, whereas junctions inside this zone are essentially unaffected by changes in intracellular pH. In a normal frog lens, the intracellular pH in fiber cells near the lens surface is 7.02, a value significantly alkaline to electrochemical equilibrium. Our data suggest that Na/H exchange and perhaps other Na gradient-dependent mechanisms in the peripheral cells maintain this transmembrane gradient. Deep in the lens, the fiber cell cytoplasm is significantly more acidic (pHi 6.81) due to influx of hydrogen across the inner fiber cell membranes and production of H+ by the inner fiber cells. Because of the normally acid cytoplasm of interior fiber cells, their loss of gap junctional sensitivity to pH may be essential to lens survival.
晶状体的纤维细胞通过广泛的缝隙连接在电学和扩散方面相互连接。这些连接允许小溶质通量在内层细胞和外周细胞之间移动,而大多数跨膜运输发生在外周细胞。我们在此描述一种利用两个细胞内微电极来测量完整晶状体中任意给定距离处纤维细胞之间的细胞间电阻的方法。我们还使用离子敏感微电极来记录完整晶状体不同深度处的细胞内pH值。我们发现连接内层纤维细胞的缝隙连接在pH敏感性以及与连接外周细胞的缝隙连接的正常偶联电阻方面存在差异。这种转变发生在晶状体内部500至650微米之间的区域。该区域外周的纤维细胞具有1.1Ω·cm²的特定偶联电阻,而内部的纤维细胞具有2.7Ω·cm²的特定偶联电阻。然而,当通过用CO₂鼓泡使纤维细胞的细胞质酸化时,外周细胞解偶联,细胞间电阻增加超过40倍,而该区域内部的连接基本上不受细胞内pH变化的影响。在正常青蛙晶状体中,晶状体表面附近纤维细胞的细胞内pH值为7.02,该值相对于电化学平衡呈显著碱性。我们的数据表明,外周细胞中的Na/H交换以及可能其他依赖Na梯度的机制维持了这种跨膜梯度。在晶状体深处,由于氢跨内层纤维细胞膜的流入以及内层纤维细胞产生H⁺,纤维细胞的细胞质酸性明显更强(细胞内pH值为6.81)。由于内部纤维细胞的细胞质通常呈酸性,它们对pH的缝隙连接敏感性丧失可能对晶状体的存活至关重要。