Department of Molecular and Environmental Bioscience, Graduate School, Hanyang University, Seoul 133-791, South Korea.
Comp Biochem Physiol A Mol Integr Physiol. 2012 Apr;161(4):436-42. doi: 10.1016/j.cbpa.2012.01.004. Epub 2012 Jan 9.
Salinity is an important parameter that affects survival and metabolism in fish. In fish, pituitary growth hormone (GH) regulates physiological functions including adaptation to different salinity as well as somatic growth. GH is stimulated by growth hormone-releasing hormone (GHRH) and exerts its function via binding to growth hormone receptor (GHR). As Kryptolebias marmoratus is a euryhaline fish, this species would be a useful model species for studying the adaptation to osmotic stress conditions. Here, we cloned GH, -GHR, somatolactin (SL), and somatolactin receptor (SLR) genes, and analyzed their expression patterns in different tissues and during early developmental stages by using real-time RT-PCR. We also further examined expression of them after acclimation to different salinity. Tissue distribution studies revealed that Km-GH and -SL mRNAs were remarkably expressed in brain and pituitary, whereas Km-GHR and -SLR mRNAs were predominantly expressed in liver, followed by gonad, muscle, pituitary, and brain. During embryonic developmental stages, the expression of their mRNA was increased at stage 3 (9 dpf). The Km-GH and -SL mRNA transcripts were constantly elevated until stage 5 (5h post hatch), whereas Km-GHR and -SLR mRNA levels decreased at this stage. After we transferred K. marmoratus from control (12 psu) to hyper-osmotic condition (hyperseawater, HSW; 33 psu), Km-GH, -SL, and GHR mRNA levels were enhanced. In hypo-osmotic conditions like freshwater (FW), Km-GH and -SL expressions were modulated 24 h after exposure, and Km-SLR transcripts were significantly upregulated. This finding suggests that Km-GH and -SL may be involved in the osmoregulatory mechanism under hyper-osmotic as well as hypo-osmotic stress. This is the first report on transcriptional modulation and relationship of GH, GHR, SL, and SLR during early development and after salinity stress. This study will be helpful to a better understanding on molecular mechanisms of adaptation response to salt stress in euryhaline fish.
盐度是影响鱼类生存和代谢的一个重要参数。在鱼类中,垂体生长激素(GH)调节包括适应不同盐度以及躯体生长在内的生理功能。GH 受生长激素释放激素(GHRH)的刺激,并通过与生长激素受体(GHR)结合发挥其功能。由于 Kryptolebias marmoratus 是一种广盐性鱼类,因此该物种将是研究适应渗透胁迫条件的有用模式物种。在这里,我们克隆了 GH、-GHR、生长激素抑制素(SL)和生长激素抑制素受体(SLR)基因,并通过实时 RT-PCR 分析了它们在不同组织和早期发育阶段的表达模式。我们还进一步研究了它们在适应不同盐度后的表达情况。组织分布研究表明,Km-GH 和 -SL mRNAs 在脑垂体中表达显著,而 Km-GHR 和 -SLR mRNAs 在肝脏中表达最多,其次是性腺、肌肉、垂体和脑。在胚胎发育阶段,其 mRNA 的表达在第 3 期(9 dpf)增加。Km-GH 和 -SL mRNA 转录物一直升高到第 5 期(孵化后 5 小时),而 Km-GHR 和 -SLR mRNA 水平在该阶段下降。当我们将 K. marmoratus 从对照(12 psu)转移到高渗条件(高渗海水,HSW;33 psu)时,Km-GH、-SL 和 GHR mRNA 水平升高。在低渗条件下,如淡水(FW),暴露后 24 小时 Km-GH 和 -SL 的表达被调节,Km-SLR 转录物显著上调。这表明 Km-GH 和 -SL 可能参与高渗和低渗胁迫下的渗透压调节机制。这是 GH、GHR、SL 和 SLR 在早期发育和盐度胁迫后转录调节及其关系的首次报道。本研究将有助于更好地理解广盐鱼类适应盐胁迫的分子机制。