The fluxes of Na were measured on isolated coprodeal mucosa at 1--220 mM-Na from hens on low (L) and high (H) Na diets with the purpose of finding the location and characteristics of Na sites activated in the cellular pathway by L. 2. The influx across the brush border, JNamc, and the transmural fluxes, JNasm and JNams, were determined. Effects on these fluxes of ouabain, 10(-3) M in the serosal solution, and amiloride, 10(-4) M in the mucosal solution, were studied for both dietary states. 3. JNamc was 5--22 (L) and 0--0.8 (H) muequiv/cm2.hr at 130 mM-Na corrected for the paracellular flux of Na. The JNamc (H) is tenfold smaller than found by Choshniak, Munck & Skadhauge (1977). This discrepancy is at present inexplicable. Amiloride completely inhibited JNamc (L). Preincubation in 0 or 130 mM-Na had no effect on JNamc. Ouabain reduced JNamc (L) by only about 37% after preincubation at 130 mM-Na. The Kt of JNamc was 5.1 (L) and 50.6 (H) mM-Na. 4. JNasm was 50 (H) and 61 (L) n-equiv/cm2.hr at 6.5 mM-Na. Ouabain increased JNasm by 360% in the low Na state. The increased JNasm was inhibited 74--100% by amiloride. This is interpreted as a ouabain induced Na-Na exchange at the basolateral Na-K-ATPase and an almost complete block of JNacm by amiloride. A similar exchange of Na at the basolateral membrane in the high-Na state was revealed by 'opening' the brush border for Na with monensin added to the mucosal solution. Amiloride in itself prevented a 50% recirculation of Na via the paracellular route and back across the cells in the low Na state. 5. JNams was 5.6 (L) muequiv/cm2.hr and 187 (L) microA/cm2 at 6.5 mM-Na. Amiloride reduced these values to 0.4 muequiv/cm2.hr and 5.8 microA/cm2. On addition of amiloride the transmural resistance in (L) coprodea at 130 mM-Na increased from 140 to 190 and it remained unchanged at 260 omega cm2 in (H) coprodea. The resistance of (L) birds, 163, was not affected by ouabain, 166 (L) omega cm2. 6. 20:1 NaCl dilution potentials at the mucosal side of 17--18 mV (L) and nearly zero (H) had half-times around 1 sec. Amiloride eliminated completely these diffusion potentials. The short half-time indicates a location in the brush border of sodium specific sites induced by the low-Na diet. This conclusion is oppsite to that described by Choshniak et al. (1977). 7. Ion selectivity, voltage--current and conductance--concentration relations in the presence of amiloride indicated a weakly cation selective and highly hydrated pathway, which was also thick and with neutral sites. This fits a paracellular route with the limiting barrier for ions at the tight junction.
摘要
以低钠(L)和高钠(H)日粮喂养的母鸡为研究对象,在1 - 220 mM - Na浓度下测定分离的泄殖腔黏膜上的钠通量,目的是确定低钠日粮在细胞途径中激活的钠位点的位置和特性。2. 测定了跨刷状缘的通量JNamc以及跨膜通量JNasm和JNams。研究了在两种日粮状态下,浆膜溶液中10⁻³ M哇巴因和黏膜溶液中10⁻⁴ M氨氯吡脒对这些通量的影响。3. 在130 mM - Na时,经钠的细胞旁通量校正后,JNamc在低钠组为5 - 22,高钠组为0 - 0.8 μeq/cm²·hr。高钠组的JNamc比乔什尼亚克、蒙克和斯卡德豪格(1977年)所发现的值小10倍。目前这种差异无法解释。氨氯吡脒完全抑制了低钠组的JNamc。在0或130 mM - Na中预孵育对JNamc没有影响。在130 mM - Na预孵育后,哇巴因仅使低钠组的JNamc降低约37%。JNamc的Kt值在低钠组为5.1,高钠组为50.6 mM - Na。4. 在6.5 mM - Na时,JNasm在高钠组为50,低钠组为61 n - eq/cm²·hr。在低钠状态下,哇巴因使JNasm增加了360%。氨氯吡脒使增加的JNasm受到74% - 100%的抑制。这被解释为哇巴因诱导基底外侧钠钾ATP酶处的钠 - 钠交换,以及氨氯吡脒几乎完全阻断JNacm。通过在黏膜溶液中添加莫能菌素“打开”刷状缘对钠的通透性,揭示了高钠状态下基底外侧膜上类似的钠交换。在低钠状态下,氨氯吡脒本身阻止了约50%的钠通过细胞旁途径再循环并穿过细胞。5. 在6.5 mM - Na时,JNams在低钠组为5.6 μeq/cm²·hr,电流为187 μA/cm²。氨氯吡脒将这些值分别降低至0.4 μeq/cm²·hr和5.8 μA/cm²。添加氨氯吡脒后,130 mM - Na时低钠组泄殖腔的跨膜电阻从140增加到190,高钠组泄殖腔的跨膜电阻在260 Ω·cm²保持不变。低钠组鸡的电阻为163,不受哇巴因影响,仍为166(低钠组)Ω·cm²。6. 黏膜侧20:1 NaCl稀释电位在低钠组为17 - 18 mV,高钠组几乎为零,半衰期约为1秒。氨氯吡脒完全消除了这些扩散电位。短半衰期表明低钠日粮诱导的钠特异性位点位于刷状缘。这一结论与乔什尼亚克等人(1977年)所描述的相反。7. 在氨氯吡脒存在下的离子选择性、电压 - 电流和电导 - 浓度关系表明存在一条弱阳离子选择性且高度水合的途径,该途径也较厚且具有中性位点。这符合细胞旁途径,其在紧密连接处对离子具有限制屏障。