Boo Mel V, Hiong Kum C, Choo Celine Y L, Cao-Pham Anh H, Wong Wai P, Chew Shit F, Ip Yuen K
Department of Biological Sciences, National University of Singapore, Kent Ridge, Singapore, Republic of Singapore.
Natural Sciences and Science Education, National Institute of Education, Nanyang Technological University, Singapore, Republic of Singapore.
PLoS One. 2017 Oct 19;12(10):e0186865. doi: 10.1371/journal.pone.0186865. eCollection 2017.
Na+/K+-ATPase (NKA) is essential for maintaining the Na+ and K+ gradients, and supporting the secondary active transport of certain ions/molecules, across the plasma membrane of animal cells. This study aimed to clone the NKA α-subunit (NKAα) from the inner mantle adjacent to the extrapallial fluid of Tridacna squamosa, to determine its subcellular localization, and to examine the effects of light exposure on its transcript level and protein abundance. The cDNA coding sequence of NKAα from T. squamosa comprised 3105 bp, encoding 1034 amino acids with an estimated molecular mass of 114 kDa. NKAα had a basolateral localization along the shell-facing epithelium of the inner mantle. Exposure to 12 h of light led to a significantly stronger basolateral NKAα-immunofluorescence at the shell-facing epithelium, indicating that NKA might play a role in light-enhanced calcification in T. squamosa. After 3 h of light exposure, the transcript level of NKAα decreased transiently in the inner mantle, but returned to the control level thereafter. In comparison, the protein abundance of NKAα remained unchanged at hour 3, but became significantly higher than the control after 12 h of light exposure. Hence, the expression of NKAα in the inner mantle of T. squamosa was light-dependent. It is probable that a higher expression level of NKA was needed in the shell-facing epithelial cells of the inner mantle to cope with a rise in Na+ influx, possibly caused by increases in activities of some Na+-dependent ion transporters/channels involved in light-enhanced calcification.
钠钾ATP酶(NKA)对于维持动物细胞质膜两侧的钠钾离子梯度以及支持某些离子/分子的继发性主动运输至关重要。本研究旨在从鳞砗磲外套膜靠近外套腔液的内层克隆NKAα亚基(NKAα),确定其亚细胞定位,并研究光照对其转录水平和蛋白质丰度的影响。鳞砗磲NKAα的cDNA编码序列长3105 bp,编码1034个氨基酸,估计分子量为114 kDa。NKAα沿外套膜内层面向贝壳的上皮细胞呈基底外侧定位。光照12小时后,面向贝壳的上皮细胞基底外侧NKAα免疫荧光显著增强,表明NKA可能在鳞砗磲光照增强钙化过程中发挥作用。光照3小时后,外套膜内层NKAα的转录水平短暂下降,但随后恢复到对照水平。相比之下,NKAα的蛋白质丰度在3小时时保持不变,但光照12小时后显著高于对照。因此,鳞砗磲外套膜内层NKAα的表达依赖于光照。可能需要在内套膜面向贝壳的上皮细胞中更高水平地表达NKA,以应对可能由参与光照增强钙化的一些钠依赖性离子转运体/通道活性增加导致的钠离子内流增加。