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比较大西洋鲑(Salmo salar)幼鱼和成鱼鳃组织中 Na(+)/K (+) ATP 酶α亚基基因的表达。

Comparison between parr and smolt Atlantic salmon (Salmo salar) α subunit gene expression of Na(+)/K (+) ATPase in gill tissue.

机构信息

Laboratoire de Physiologie des Poissons, Institut National de la Recherche Agronomique, Campus de Beaulieu, 35042, Rennes Cedex, France.

出版信息

Fish Physiol Biochem. 1996 Feb;15(1):29-39. doi: 10.1007/BF01874835.

Abstract

Increases in branchial Na(+)/K(+) ATPase activity during seawater adaptation of euryhaline fish species, have been well documented. During the parr-smolt transformation of salmonids this activity increases two to five fold and is used as an indicator of the transformation. In order to improve the understanding of differences in enzyme activity found between Atlantic salmonSalmo salar parr and smolt fish, we investigated the gene expression of the Na(+)/K(+) ATPase α-subunit(s) in gill tissue. Gill mRNAs were analyzed and quantified at distinct time points using Northern and Dot blot techniques. We amplified by PCR, a conserved region of the cDNA encoding the Na(+)/K(+) ATPase α-subunit of the rainbow troutOncorhynchus mykiss. The PCR products (670 bp) were cloned and all independent clones showed a sequence corresponding to the α subunit of the Na(+)/K(+) ATPase. The fragments obtained appeared as a heterogenous population of three sequences showing, when compared between each other, 86 to 93% identity. This suggests that different allelic forms of the α-subunit are expressed in gill tissue. Hybridization studies performed with these PCR probes revealed two mRNA species, a major 3.7 kb transcript and a minor transcript of 1.8 kb. Enhanced 3.7 kb transcript levels are concurrent with elevated enzyme activity in smolts during the March and April parrsmolt transformation of Atlantic salmon. Interestingly, our study disclosed that smolt fish only displayed a two-fold increase in transcript levels when compared to parr whereas enzyme activity showed a 4 to 5 fold increase. This suggests that the increase in the 3.7 kb mRNA content of gill tissue is probably not the only mediator leading to the rise in enzyme activity during parr-smolt transformation.

摘要

海水鱼类在适应海水的过程中鳃 Na(+)/K(+)ATP 酶活性增加的现象已得到充分证实。在鲑鱼的幼鱼变熟鱼过程中,这种活性增加了两到五倍,并被用作转化的指标。为了更好地理解大西洋鲑鱼幼鱼和熟鱼之间发现的酶活性差异,我们研究了鳃组织中 Na(+)/K(+)ATP 酶 α-亚基的基因表达。使用 Northern 和斑点印迹技术在不同时间点分析和定量鳃组织的 mRNA。我们通过 PCR 扩增了虹鳟 Oncorhynchus mykiss cDNA 编码 Na(+)/K(+)ATP 酶 α-亚基的保守区域。PCR 产物(670bp)被克隆,所有独立的克隆均显示与 Na(+)/K(+)ATP 酶的 α 亚基相对应的序列。获得的片段显示出三种序列的异质群体,彼此之间的相似度为 86%至 93%。这表明不同的 α-亚基等位基因形式在鳃组织中表达。用这些 PCR 探针进行的杂交研究显示出两种 mRNA 物种,主要的 3.7kb 转录物和次要的 1.8kb 转录物。在大西洋鲑鱼 3 月至 4 月幼鱼变熟鱼的转化过程中,增强的 3.7kb 转录物水平与酶活性的升高同时发生。有趣的是,我们的研究表明,与幼鱼相比,熟鱼的转录物水平仅增加了两倍,而酶活性则增加了 4 到 5 倍。这表明鳃组织中 3.7kb mRNA 含量的增加可能不是导致幼鱼变熟鱼转化过程中酶活性升高的唯一因素。

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