Institute of Cellular and Organismic Biology, Academia Sinica, Nankang, Taipei, Taiwan 11529, Republic of China.
Am J Physiol Cell Physiol. 2011 Feb;300(2):C295-307. doi: 10.1152/ajpcell.00263.2010. Epub 2010 Dec 1.
Similar to mammalian proximal tubular cells, H(+)-ATPase rich (HR) cells in zebrafish skin and gills are also responsible for Na(+) uptake and acid secretion functions. However, the basolateral transport pathways in HR cells are still unclear. In the present study, we tested the hypothesis if there are specific slc4 members involved in basolateral ion transport pathways in HR cells. Fourteen isoforms were identified in the zebrafish(z) slc4 family, and the full-length cDNAs of two novel isoforms, zslc4a1b (anion exchanger, zAE1b) and zslc4a4b (Na(+)/HCO(3)(-) cotransporter, zNBCe1b), were sequenced. mRNA signals of zslc4a1b and zslc4a4b were mainly detected in certain groups of ionocytes in zebrafish skin/gills. Further double immunocytochemistry or in situ hybridization demonstrated that zAE1b, but not zNBCe1b, was localized to basolateral membranes of HR cells. Acclimation to low-Na(+) or acidic environments stimulated the mRNA expression of zslc4a1b in zebrafish gills, and loss-of-function of zslc4a1b with specific morpholinos caused significant decreases in both the whole body Na(+) content and the skin H(+) activity in the morphants. On the basis of these results, it was concluded that zAE1b, but not zNBCe1b, is involved in the basolateral transport pathways in Na(+) uptake/acid secretion mechanisms in zebrafish HR cells.
类似于哺乳动物的近端肾小管细胞,斑马鱼皮肤和鳃中的 H(+)-ATPase 丰富 (HR) 细胞也负责 Na(+)摄取和酸分泌功能。然而,HR 细胞的基底外侧转运途径仍不清楚。在本研究中,我们测试了一个假设,即是否有特定的 slc4 成员参与 HR 细胞的基底外侧离子转运途径。在斑马鱼(z) slc4 家族中鉴定出了 14 种同工型,并且两种新型同工型的全长 cDNA,zslc4a1b(阴离子交换体,zAE1b)和 zslc4a4b(Na(+)/HCO(3)(-)共转运体,zNBCe1b)被测序。zslc4a1b 和 zslc4a4b 的 mRNA 信号主要在斑马鱼皮肤/鳃中的某些离子细胞群中检测到。进一步的双重免疫细胞化学或原位杂交表明,zAE1b 而不是 zNBCe1b 定位于 HR 细胞的基底外侧膜。适应低 Na(+)或酸性环境刺激斑马鱼鳃中 zslc4a1b 的 mRNA 表达,并且特异性 morpholinos 引起的 zslc4a1b 功能丧失导致形态发生体中全身 Na(+)含量和皮肤 H(+)活性显著降低。基于这些结果,可以得出结论,zAE1b 而不是 zNBCe1b 参与了斑马鱼 HR 细胞中 Na(+)摄取/酸分泌机制的基底外侧转运途径。