Bicarbonate exchangers SLC26A3 and SLC26A6 are localized at the apical membrane of porcine vas deferens epithelium.
Bicarbonate exchangers SLC26A3 and SLC26A6 are localized at the apical membrane of porcine vas deferens epithelium.
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DOI：
10.14814/phy2.12380
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发表时间：
2015-04
影响因子：
2.5
通讯作者：
Schultz BD
中科院分区：
文献类型：
--
作者：
Pierucci-Alves F;Akoyev V;Schultz BD
本研究的目的是检测−交换子SLC26A3和SLC26A6在原代培养的猪输精管上皮细胞(1°PVD)和天然猪输精管上皮细胞中的表达。定量RT-PCR结果显示，在1°PVD细胞中，SLC26A6编码基因的mRNA丰度是SLC26A3基因编码基因的6倍。免疫印迹分析结合1°PVD的表面生物素标记，发现SLC26A3和SLC26A6在单层的全细胞裂解产物和根尖表面有免疫反应。1°PVD细胞单层的激光扫描共聚焦显微镜(LSCM)显示SLC26A3免疫反应主要位于顶端，但存在于整个基底-顶端细胞轴，而SLC26A6免疫反应主要位于顶端，有时聚集在核区。LSCM还显示SLC26A3和SLC26A6免疫阳性反应存在于猪输精管上皮的整个顶层和基底细胞中。SLC26A3和SLC26A6在输精管近端和远端的免疫反应模式和表达丰度与输精管远端无明显差异。没有证据表明SLC26A3或SLC26A6在输精管的任何部分优先表达，就像在其他导管系统中分泌HCO3−的上皮细胞所建议的那样。因此，输精管上皮细胞在整个输卵管中表达转运蛋白，这可以促进管腔内容物的快速碱化，这一点已经在体内得到证实。
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.