Department of Biological Sciences, National University of Singapore, Kent Ridge, Singapore, 117543, Republic of Singapore.
Natural Sciences and Science Education, National Institute of Education, Nanyang Technological University, 1 Nanyang Walk, Singapore, 637616, Republic of Singapore.
J Comp Physiol B. 2018 Sep;188(5):765-777. doi: 10.1007/s00360-018-1161-6. Epub 2018 Apr 24.
Ammonium transporters (AMTs) can participate in ammonia uptake or excretion across the plasma membrane of prokaryotic, plant and invertebrate cells. The giant clam, Tridacna squamosa, harbors nitrogen-deficient symbiotic zooxanthellae, and normally conducts light-enhanced ammonia absorption to benefit the symbionts. Nonetheless, it can excrete ammonia when there is a supply of exogenous nitrogen or exposed to continuous darkness. This study aimed to elucidate the role of AMT1 in the ctenidium of T. squamosa by cloning and characterizing the AMT1/AMT1, determining its subcellular localization, and examining changes in its transcript and protein expression levels in response to light exposure. The cDNA coding sequence of AMT1 from T. squamosa consisted of 1527 bp and encoded 508 amino acids of 54.6 kDa. AMT1-immunofluorescence was detected mainly at the apical epithelium of ctenidial filaments, and it decreased significantly after 12 h of exposure to light. By contrast, the epithelial cells surrounding the tertiary water channels in the ctentidium, which are known to exhibit light-enhanced glutamine synthetase expression and take part in the assimilation of exogenous ammonia in light, did not display any AMT1-immunolabelling. Furthermore, the transcript level and protein abundance of ctenidial AMT1/AMT1 decreased significantly at the 6th and 12th h of light exposure. Taken together, these results indicate that AMT1 might participate in ammonia excretion instead of ammonia absorption and assimilation in T. squamosa. It is probable that the expression levels of AMT1/AMT1 need to be down-regulated during light exposure to achieve light-enhanced ammonia uptake.
铵转运蛋白(AMTs)可以参与原核生物、植物和无脊椎动物细胞的跨质膜氨吸收或排泄。巨蛤 Tridacna squamosa 拥有缺乏氮的共生甲藻,并正常进行光增强氨吸收以利于共生体。尽管如此,当有外源氮供应或暴露于持续黑暗中时,它可以排泄氨。本研究旨在通过克隆和表征 AMT1/AMT1,确定其亚细胞定位,并检查其转录物和蛋白质表达水平在暴露于光下时的变化,来阐明 AMT1 在 T. squamosa 中鳃丝中的作用。T. squamosa 的 AMT1 的 cDNA 编码序列由 1527bp 组成,编码 54.6kDa 的 508 个氨基酸。AMT1-免疫荧光主要在鳃丝纤毛的顶上皮细胞中检测到,在暴露于光 12 小时后显著减少。相比之下,在鳃丝中,三级水通道周围的上皮细胞没有显示任何 AMT1-免疫标记,这些细胞已知表现出光增强谷氨酰胺合成酶表达,并参与光下外源氨的同化。此外,鳃丝 AMT1/AMT1 的转录水平和蛋白丰度在光暴露的第 6 小时和第 12 小时显著降低。综上所述,这些结果表明 AMT1 可能参与氨排泄而不是氨吸收和同化在 T. squamosa 中。在光暴露期间,AMT1/AMT1 的表达水平可能需要下调,以实现光增强的氨吸收。