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Cellular distribution of NKCC2 in the gastric mucosa and its response to short-term osmotic shock

机译：NKCC2在胃粘膜中的细胞分布及其对短期渗透压的反应

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摘要

The Na+-K+-2Cl- cotransporter-2 (NKCC2) has long been recognized as a “kidney-specific” transporter and is important in salt reabsorption. NKCC2 has been found in the gastric mucosa; however, its cellular distribution and function remain obscure. The present study characterized the distribution pattern of NKCC2 in mammalian gastric mucosa and investigated its response to osmotic challenge. Reverse transcription with the polymerase chain reaction, Western blot and immunofluorescence were used to determine NKCC2 expression and localization. The effect of osmotic shock on NKCC2 expression was studied in isolated gastric mucosa with variable osmolarity treatment. Results from all of the above studies were compared with those of NKCC1. Our data indicated that NKCC1 and NKCC2 were expressed in the gastric mucosa of rat, mouse and human. The mRNA transcripts and proteins for NKCC1 and NKCC2 were broadly expressed in the rat gastric mucosa. In rat and mouse, NKCC1 was largely confined to the lower part of the oxyntic and pyloric gland areas, whereas NKCC2 extended throughout the gastric glands. NKCC1 immunoreactivity was strongly expressed in the parietal and chief cells but was weaker in the mucous cells. NKCC2 was abundantly located in the parietal and mucous cells but faintly distributed in the chief cells. Hypertonic treatment increased the protein level of NKCC1 and caused evident membrane translocation. In contrast, NKCC2 was significantly downregulated and no obvious membrane translocation was observed. Thus, NKCC2 displayed a more ubiquitous distribution in the gastric mucosa and might work coordinately with NKCC1 to maintain cell volume homeostasis under hypertonic conditions.
机译：Na + -K + -2Cl -共转运蛋白2（NKCC2）长期以来被认为是“肾脏特异性”转运蛋白，在盐的重吸收中很重要。在胃粘膜中发现了NKCC2。但是，其细胞分布和功能仍然不清楚。本研究表征了NKCC2在哺乳动物胃粘膜中的分布模式，并研究了其对渗透挑战的反应。用聚合酶链反应，蛋白质印迹和免疫荧光的逆转录来确定NKCC2的表达和定位。研究了渗透压休克对可变渗透压处理的离体胃黏膜中NKCC2表达的影响。将以上所有研究的结果与NKCC1的结果进行了比较。我们的数据表明NKCC1和NKCC2在大鼠，小鼠和人的胃粘膜中表达。 NKCC1和NKCC2的mRNA转录物和蛋白在大鼠胃粘膜中广泛表达。在大鼠和小鼠中，NKCC1很大程度上局限于氧化性和幽门腺的下部，而NKCC2则遍及整个胃腺。 NKCC1免疫反应性在顶细胞和原代细胞中强烈表达，但在粘液细胞中较弱。 NKCC2大量位于顶细胞和粘液细胞中，但在主要细胞中微弱分布。高渗治疗增加了NKCC1的蛋白质水平，并引起明显的膜移位。相反，NKCC2被显着下调并且没有观察到明显的膜移位。因此，NKCC2在胃粘膜中显示出更普遍的分布，并且可能与NKCC1协同作用以在高渗条件下维持细胞体积的稳态。

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