Departamento de Biologia, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, Brazil.
J Exp Biol. 2010 Nov 15;213(Pt 22):3894-905. doi: 10.1242/jeb.046870.
We evaluate osmotic and chloride (Cl(-)) regulatory capability in the diadromous shrimp Macrobrachium amazonicum, and the accompanying alterations in hemolymph osmolality and [Cl(-)], gill Na(+)/K(+)-ATPase activity, and expression of gill Na(+)/K(+)-ATPase α-subunit and V-ATPase B subunit mRNA during salinity (S) acclimation. We also characterize V-ATPase kinetics and the organization of transport-related membrane systems in the gill epithelium. Macrobrachium amazonicum strongly hyper-regulates hemolymph osmolality and [Cl(-)] in freshwater and in salinities up to 25‰ S. During a 10-day acclimation period to 25‰ S, hemolymph became isosmotic and hypo-chloremic after 5 days, [Cl(-)] alone remaining hyporegulated thereafter. Gill Na(+)/K(+)-ATPase α-subunit mRNA expression increased 6.5 times initial values after 1 h, then decreased to 3 to 4 times initial values by 24 h and to 1.5 times initial values after 10 days at 25‰ S. This increased expression was accompanied by a sharp decrease at 5 h then recovery of initial Na(+)/K(+)-ATPase activity within 24 h, declining again after 5 days, which suggests transient Cl(-) secretion. V-ATPase B-subunit mRNA expression increased 1.5-fold within 1 h, then reduced sharply to 0.3 times initial values by 5 h, and remained unchanged for the remainder of the 10-day period. V-ATPase activity dropped sharply and was negligible after a 10-day acclimation period to 21‰ S, revealing a marked downregulation of ion uptake mechanisms. The gill epithelium consists of thick, apical pillar cell flanges, the perikarya of which are coupled to an intralamellar septum. These two cell types respectively exhibit extensive apical evaginations and deep membrane invaginations, both of which are associated with numerous mitochondria, characterizing an ion transporting epithelium. These changes in Na(+)/K(+)- and V-ATPase activities and in mRNA expression during salinity acclimation appear to underpin ion uptake and Cl(-) secretion by the palaemonid shrimp gill.
我们评估了洄游性虾类亚马逊沼虾的渗透和氯离子(Cl(-))调节能力,以及伴随而来的血淋巴渗透压和[Cl(-)]、鳃 Na(+)/K(+)-ATP 酶活性、以及鳃 Na(+)/K(+)-ATP 酶 α-亚基和 V-ATPase B 亚基 mRNA 表达的变化,这些变化发生在盐度(S)驯化过程中。我们还描述了 V-ATPase 的动力学和鳃上皮细胞中运输相关膜系统的组织。亚马逊沼虾在淡水中和高达 25‰S 的盐度下强烈地过度调节血淋巴渗透压和[Cl(-)]。在适应 25‰S 的 10 天期间,血淋巴在第 5 天变得等渗和低氯,此后仅 Cl(-)仍未被调节。鳃 Na(+)/K(+)-ATP 酶 α-亚基 mRNA 表达在 1 小时后增加了初始值的 6.5 倍,然后在 24 小时内降至初始值的 3 到 4 倍,10 天后在 25‰S 时降至初始值的 1.5 倍。这种表达增加伴随着 5 小时时 Na(+)/K(+)-ATP 酶活性的急剧下降,然后在 24 小时内恢复到初始水平,5 天后再次下降,这表明存在短暂的 Cl(-)分泌。V-ATPase B-亚基 mRNA 表达在 1 小时内增加了 1.5 倍,然后在 5 小时内急剧降至初始值的 0.3 倍,在 10 天的剩余时间内保持不变。V-ATPase 活性在适应 21‰S 10 天后急剧下降,几乎检测不到,表明离子摄取机制明显下调。鳃上皮由厚的、顶端柱细胞翼缘组成,其细胞体与板层间隔相连。这两种细胞类型分别具有广泛的顶端外突和深膜内陷,都与许多线粒体相关,具有离子转运上皮的特征。在盐度驯化过程中 Na(+)/K(+)-和 V-ATPase 活性以及 mRNA 表达的这些变化似乎为 Palaemonid 虾类的鳃部离子摄取和 Cl(-)分泌提供了基础。