Choe Keith P, O'Brien Siobhan, Evans David H, Toop Tes, Edwards Susan L
Department of Zoology, University of Florida, Gainesville, Florida 32611, USA.
J Exp Zool A Comp Exp Biol. 2004 Aug 1;301(8):654-65. doi: 10.1002/jez.a.60.
As adults, anadromous lampreys migrate from seawater into freshwater rivers, where they require branchial ion (NaCl) absorption for osmoregulation. In teleosts and elasmobranchs, pharmological, immunohistochemical, and molecular data support roles for Na+/K+-ATPase (NPPase), carbonic anhydrase II (CAII), and vacuolar H+-ATPase (V-ATPase) in two different models of branchial ion absorption. To our knowledge, these transport-related proteins have not been studied in adult freshwater lampreys, and therefore it is not known if they are expressed, or have similar functions, in lampreys. The purpose of this study was to localize NPPase, CAII, and V-ATPase in the gills of adult freshwater lampreys and determine if any of these transport-related proteins are expressed in the same cells. Heterologous antibodies were used to localize the three proteins in gill tissue from pouched lamprey (Geotria australis). Immunoreactivity (IR) for all three proteins occurred between, and at the base of, lamellae in cells that match previous descriptions of mitochondrion-rich-cells (MRCs). NPPase-IR was always on the basolateral side of cells that did not stain for CAII or V-ATPase. In contrast, CAII-IR was always on the apical side of cells that also contained diffuse V-ATPase-IR. Therefore, we have identified two types of MRC in adult freshwater lamprey gills based on immunohistochemical staining for three transport proteins. A model of ion transport, based on our results, is proposed for adult freshwater lampreys.
成年溯河洄游七鳃鳗从海水洄游到淡水河流,在淡水中它们需要通过鳃吸收离子(氯化钠)来进行渗透压调节。在硬骨鱼和软骨鱼中,药理学、免疫组织化学和分子数据支持钠钾ATP酶(NPPase)、碳酸酐酶II(CAII)和液泡型H⁺-ATP酶(V-ATP酶)在两种不同的鳃离子吸收模型中发挥作用。据我们所知,这些与转运相关的蛋白质尚未在成年淡水七鳃鳗中进行研究,因此尚不清楚它们在七鳃鳗中是否表达或具有类似功能。本研究的目的是在成年淡水七鳃鳗的鳃中定位NPPase、CAII和V-ATP酶,并确定这些与转运相关的蛋白质是否在同一细胞中表达。使用异源抗体在袋七鳃鳗(Geotria australis)的鳃组织中定位这三种蛋白质。所有三种蛋白质的免疫反应性(IR)出现在薄片之间以及薄片基部的细胞中,这些细胞与先前对富含线粒体细胞(MRCs)的描述相符。NPPase-IR总是位于不被CAII或V-ATP酶染色的细胞的基底外侧。相比之下,CAII-IR总是位于也含有弥漫性V-ATP酶-IR的细胞的顶端一侧。因此,基于对三种转运蛋白的免疫组织化学染色,我们在成年淡水七鳃鳗的鳃中鉴定出了两种类型的MRC。基于我们的研究结果,提出了成年淡水七鳃鳗的离子转运模型。