Nguyen Ha-Van, Stuart-Tilley Alan, Alper Seth L, Melvin James E
Center for Oral Biology, University of Rochester Medical Center, Rochester, New York 14642, USA.
Am J Physiol Gastrointest Liver Physiol. 2004 Feb;286(2):G312-20. doi: 10.1152/ajpgi.00158.2003. Epub 2003 Sep 4.
Large volumes of saliva are generated by transepithelial Cl(-) movement during parasympathetic muscarinic receptor stimulation. To gain further insight into a major Cl(-) uptake mechanism involved in this process, we have characterized the anion exchanger (AE) activity in mouse serous parotid and mucous sublingual salivary gland acinar cells. The AE activity in acinar cells was Na(+) independent, electroneutral, and sensitive to the anion exchange inhibitor DIDS, properties consistent with the AE members of the SLC4A gene family. Localization studies using a specific antibody to the ubiquitously expressed AE2 isoform labeled acini in both parotid and sublingual glands. Western blot analysis detected an approximately 170-kDa protein that was more highly expressed in the plasma membranes of sublingual than in parotid glands. Correspondingly, the DIDS-sensitive Cl(-)/HCO(3)(-) exchanger activity was significantly greater in sublingual acinar cells. The carbonic anhydrase antagonist acetazolamide markedly inhibited, whereas muscarinic receptor stimulation enhanced, the Cl(-)/HCO(3)(-) exchanger activity in acinar cells from both glands. Intracellular Ca(2+) chelation prevented muscarinic receptor-induced upregulation of the AE, whereas raising the intracellular Ca(2+) concentration with the Ca(2+)-ATPase inhibitor thapsigargin mimicked the effects of muscarinic receptor stimulation. In summary, carbonic anhydrase activity was essential for regulating Cl(-)/HCO(3)(-) exchange in salivary gland acinar cells. Moreover, muscarinic receptor stimulation enhanced AE activity through a Ca(2+)-dependent mechanism. Such forms of regulation may play important roles in modulating fluid and electrolyte secretion by salivary gland acinar cells.
在副交感神经毒蕈碱受体刺激过程中,大量唾液通过跨上皮Cl(-)移动产生。为了进一步深入了解参与此过程的主要Cl(-)摄取机制,我们对小鼠浆液性腺泡状腮腺和黏液性腺泡状舌下唾液腺细胞中的阴离子交换器(AE)活性进行了表征。腺泡细胞中的AE活性不依赖于Na(+),呈电中性,且对阴离子交换抑制剂DIDS敏感,这些特性与SLC4A基因家族的AE成员一致。使用针对普遍表达的AE2亚型的特异性抗体进行的定位研究标记了腮腺和舌下腺中的腺泡。蛋白质印迹分析检测到一种约170 kDa的蛋白质,其在舌下腺的质膜中比在腮腺中表达更高。相应地,DIDS敏感的Cl(-)/HCO(3)(-)交换器活性在舌下腺泡细胞中明显更高。碳酸酐酶拮抗剂乙酰唑胺显著抑制,而毒蕈碱受体刺激增强了来自两个腺体的腺泡细胞中的Cl(-)/HCO(3)(-)交换器活性。细胞内Ca(2+)螯合可防止毒蕈碱受体诱导的AE上调,而用Ca(2+)-ATP酶抑制剂毒胡萝卜素提高细胞内Ca(2+)浓度则模拟了毒蕈碱受体刺激的效果。总之,碳酸酐酶活性对于调节唾液腺泡细胞中的Cl(-)/HCO(3)(-)交换至关重要。此外,毒蕈碱受体刺激通过Ca(2+)依赖性机制增强AE活性。这种调节形式可能在调节唾液腺泡细胞的液体和电解质分泌中起重要作用。