College of Fisheries, Guangdong Ocean University, Zhanjiang 524088, China.
College of Fisheries, Guangdong Ocean University, Zhanjiang 524088, China.
Comp Biochem Physiol B Biochem Mol Biol. 2024 Apr-May;271:110950. doi: 10.1016/j.cbpb.2024.110950. Epub 2024 Feb 1.
Mitogen-activated protein kinases (MAPKs) are a class of protein kinases that regulate various physiological processes, and play a crucial role in maintaining the osmotic equilibrium of fish. The objective of this study was to identify and characterize the mapk family genes in cobia (Rachycentron canadum) and examine their expression profiles under different low salinity stress regimes (acute: from 30‰ to 10‰ in 1 h, sub-chronic: from 30‰ to 10‰ over 4 d). A total of 12 cobia mapk genes (Rcmapks) were identified and cloned, including six erk subfamily genes (Rcmapk1/3/4/6/7/15), three jnk subfamily genes (Rcmapk8/9/10) and three p38 mapk subfamily genes (Rcmapk 11/13/14). Domain analysis indicated that the RcMAPKs possessed the typical domains including S_TKc and PKc_like domain. Phylogenetic analysis revealed that the Rcmapks were most closely related to those of the turbot (Scophthalmus maximus). The tissue distribution of mapk genes in adult cobia and the expression patterns of Rcmapks under different low salinity stress regimes were investigated using quantitative real-time PCR (qRT-PCR). The results revealed that Rcmapk3/9/10/11/13/14 exhibited a relatively broad expression distribution across 14 different tissues. For all these genes the highest expression level was in the brain, except for Rcmapk14 (highly expressed in the stomach, gill, and skin). The genes Rcmapk1/6/15 showed significantly higher expression in the testis. Under acute low salinity stress, expression of Rcmapk1/3/6/7/9/11/13/14 was significantly altered in the gill, intestine, and trunk kidney, however, the aforementioned genes exhibited very different expression patterns among the three tissues. In the gill, most of the genes from the erk (Rcmapk3/6/7) and p38 mapk subfamily (Rcmapk11/13/14) were significantly up-regulated at almost all the time points (P < 0.05); Similarly, the expression of Rcmapk3/9/11/13/14 genes were significantly increased in the trunk kidney; while in the intestine, most of the altered genes (Rcmapk6/7/9/11/13/14) were significantly down-regulated at 1 h. Following the sub-chronic low salinity stress, expression of Rcmapk1/3/6/7/9/11/13/14 genes were significantly altered in all three tissues. These findings provide important reference data for elucidating the roles of cobia mapk family genes in response to low salinity stress.
丝裂原活化蛋白激酶(MAPKs)是一类调节各种生理过程的蛋白激酶,在维持鱼类渗透压平衡中起着关键作用。本研究的目的是鉴定和描述石斑鱼(Rachycentron canadum)中的 mapk 家族基因,并研究它们在不同低盐胁迫条件下的表达谱(急性:1 小时内从 30‰降至 10‰,亚慢性:4 天内从 30‰降至 10‰)。共鉴定和克隆了 12 种石斑鱼 mapk 基因(Rcmapks),包括 6 种 erk 亚家族基因(Rcmapk1/3/4/6/7/15)、3 种 jnk 亚家族基因(Rcmapk8/9/10)和 3 种 p38 mapk 亚家族基因(Rcmapk 11/13/14)。结构域分析表明,RcMAPKs 具有典型的结构域,包括 S_TKc 和 PKc_like 结构域。系统发育分析表明,Rcmapks 与大菱鲆(Scophthalmus maximus)的亲缘关系最为密切。采用实时定量 PCR(qRT-PCR)技术研究了 mapk 基因在成鱼中的组织分布以及 Rcmapks 在不同低盐胁迫条件下的表达模式。结果表明,Rcmapk3/9/10/11/13/14 在 14 种不同组织中具有广泛的表达分布。除了 Rcmapk14(在胃、鳃和皮肤中高表达)外,所有这些基因在大脑中的表达水平最高。Rcmapk1/6/15 在睾丸中的表达水平显著升高。在急性低盐胁迫下,Rcmapk1/3/6/7/9/11/13/14 在鳃、肠和肾中显著改变,但上述基因在三种组织中的表达模式差异很大。在鳃中,erk 亚家族(Rcmapk3/6/7)和 p38 mapk 亚家族(Rcmapk11/13/14)的大多数基因几乎在所有时间点都显著上调(P<0.05);同样,Rcmapk3/9/11/13/14 基因在肾中的表达显著增加;而在肠中,大多数改变的基因(Rcmapk6/7/9/11/13/14)在 1 小时时显著下调。亚慢性低盐胁迫后,Rcmapk1/3/6/7/9/11/13/14 基因在所有三种组织中的表达均显著改变。这些发现为阐明石斑鱼 mapk 家族基因在低盐胁迫反应中的作用提供了重要参考数据。