Broadhead C L, O'Sullivan U T, Deacon C F, Henderson I W
Department of Animal and Plant Sciences, University of Sheffield, U.K.
J Mol Endocrinol. 1992 Oct;9(2):103-14. doi: 10.1677/jme.0.0090103.
The presence of atrial natriuretic peptide (ANP) and the nature of its binding sites were studied in fresh-water (FW)- and seawater (SW)-adapted eels using a heterologous analogue, that of the rat (rANP). Rat ANP-like immunoreactivity was demonstrated in the cardiac atria and ventricles of both FW and SW eels, and electron-dense ANP-like granules were observed. The atria and ventricles of FW eels contained significantly more granules than those of SW animals and, in both types, the atria were more granular than the ventricles. Specific binding sites for rANP were demonstrated by displacement and uptake experiments using labelled rANP in dispersed eel branchial cell preparations, enriched in chloride cells. The concentration of rANP required to produce a 50% inhibition of binding in FW cells was significantly lower than that in SW cells. Scatchard analyses revealed the presence of two classes of binding site in SW eel branchial cells but only a single class of receptor in FW cells. The affinity of the FW receptor was not significantly different from that of the SW high affinity site. Rat ANP stimulated the production of cyclic GMP (cGMP) in a dose-dependent manner, and both basal and stimulated levels of cGMP were significantly greater in SW branchial cells. These studies suggest that ANP is involved in the adaptation of the euryhaline eel to differing environmental salinities; the levels of the peptide in the heart alter with changing salinity, and the nature of the receptors in the sodium chloride-transporting epithelium of the gill changes in response to the need either to eliminate or to absorb sodium chloride.
利用大鼠心房利钠肽(rANP)这种异源类似物,对适应淡水(FW)和海水(SW)的鳗鱼体内心房利钠肽(ANP)的存在情况及其结合位点的性质进行了研究。在适应淡水和海水的鳗鱼的心脏心房和心室中均检测到了大鼠ANP样免疫反应性,并且观察到了电子致密的ANP样颗粒。适应淡水的鳗鱼的心房和心室中的颗粒明显多于适应海水的鳗鱼,并且在这两种类型中,心房的颗粒都比心室更多。通过在富含氯化物细胞的分散鳗鱼鳃细胞制剂中使用标记的rANP进行置换和摄取实验,证明了rANP的特异性结合位点。在淡水细胞中产生50%结合抑制所需的rANP浓度明显低于海水细胞。Scatchard分析显示,海水鳗鱼鳃细胞中存在两类结合位点,而淡水细胞中只有一类受体。淡水受体的亲和力与海水高亲和力位点的亲和力没有显著差异。大鼠ANP以剂量依赖性方式刺激环磷酸鸟苷(cGMP)的产生,并且海水鳃细胞中cGMP的基础水平和刺激水平均明显更高。这些研究表明,ANP参与了广盐性鳗鱼对不同环境盐度的适应;心脏中该肽的水平随盐度变化而改变,并且鳃中氯化钠转运上皮细胞中受体的性质会根据消除或吸收氯化钠的需求而发生变化。