Pierucci-Alves Fernando, Akoyev Vladimir, Schultz Bruce D
Department of Anatomy & Physiology, Kansas State University, Manhattan, Kansas, USA.
Department of Anatomy & Physiology, Kansas State University, Manhattan, Kansas, USA
Physiol Rep. 2015 Apr;3(4). doi: 10.14814/phy2.12380.
The goal of this study was to test for expression of HCO3 (-) exchangers SLC26A3 and SLC26A6 in primary cultures of porcine vas deferens epithelial cells (1°PVD) and native porcine vas deferens. Quantitative RT-PCR revealed that mRNA coding for SLC26A6 was six times more abundant than mRNA coding for SLC26A3 in 1°PVD cells. Western blot analyses combined with surface biotinylation of 1°PVD demonstrated SLC26A3 and SLC26A6 immunoreactivities in whole-cell lysates and apical surfaces of monolayers. Laser scanning confocal microscopy (LSCM) of the 1°PVD cell monolayers demonstrated that SLC26A3 immunoreactivity was primarily in the apical region but present throughout the basal-apical cellular axis, whereas SLC26A6 immunoreactivity was present in the apical region and sometimes accumulated in the nuclear region. LSCM also demonstrated SLC26A3 and SLC26A6 immunoreactivities present along the entire apical lining of the native porcine vas deferens epithelium and in basal cells. The patterns and apparent abundance of SLC26A3 and SLC26A6 immunoreactivities in the proximal vas deferens were not different from the corresponding immunoreactivities in the distal region. There is no evidence of preferential expression of SLC26A3 or SLC26A6 in any portion of the vas deferens, as has been proposed for epithelia that secrete HCO3 (-) in other duct systems. Thus, vas deferens epithelia express transporters throughout the duct that can contribute to rapid alkalinization of the luminal contents as it has been demonstrated in vivo.
本研究的目的是检测猪输精管上皮细胞原代培养物（1°PVD）和天然猪输精管中HCO3(-)交换体SLC26A3和SLC26A6的表达。定量逆转录聚合酶链反应显示，在1°PVD细胞中，编码SLC26A6的mRNA丰度比编码SLC26A3的mRNA高6倍。蛋白质免疫印迹分析结合1°PVD的表面生物素化显示，在全细胞裂解物和单层细胞的顶端表面存在SLC26A3和SLC26A6免疫反应性。对1°PVD细胞单层进行激光扫描共聚焦显微镜（LSCM）检查显示，SLC26A3免疫反应性主要位于顶端区域，但在整个基底-顶端细胞轴上均有存在，而SLC26A6免疫反应性存在于顶端区域，有时在核区域积聚。LSCM还显示，SLC26A3和SLC26A6免疫反应性存在于天然猪输精管上皮的整个顶端内衬以及基底细胞中。输精管近端SLC26A3和SLC26A6免疫反应性的模式和明显丰度与远端区域相应的免疫反应性没有差异。没有证据表明输精管的任何部分优先表达SLC26A3或SLC26A6，正如在其他导管系统中分泌HCO3(-)的上皮细胞所提出的那样。因此，输精管上皮在整个管道中表达转运体，这有助于管腔内容物的快速碱化，正如在体内所证明的那样。
