| 摘要: |
| 热休克蛋白(heat shock proteins, HSPs)在鱼类的应激与免疫反应中发挥重要的生理调控作用，HSP70是该家族的重要成员。为探讨热休克蛋白在大洋性经济鱼类黄条鰤 (Seriola aureovittata)生长发育中的生理作用，本研究克隆获得了黄条鰤hsp70基因的全长cDNA序列，并采用定量PCR技术测定了其组织分布及在早期生长发育过程中的表达特征。结果显示，黄条鰤 hsp70基因的cDNA序列全长为2 332 bp，其中，5′-UTR长度为187 bp，ORF长度为1 920 bp，3′-UTR长度为225 bp，编码639个氨基酸，蛋白质分子量为70.1 kDa，等电点为5.16。黄条鰤 hsp70 mRNA的组织表达具有性别二态性差异，其中，在雌性鳃、心、脾脏和卵巢组织中显著高表达(P<0.05)，且以卵巢中表达量最高；雄性垂体、鳃、头肾和精巢组织显著高表达(P<0.05)，且以鳃中表达量最高。胚胎发育过程的表达检测显示，在卵裂前的受精卵中可检测到hsp70的表达，表明其具有亲本遗传的特性。同时，在胚胎发育过程的各个时期都可检测到hsp70 mRNA的表达，且在低囊胚期之前的各发育阶段一直保持较低表达水平，在原肠前期开始显著上调表达(P<0.05)，其后保持相对较高表达水平，至胚胎孵化出膜期达峰值。在仔稚幼鱼中，hsp70 mRNA在初孵仔鱼和1 d仔鱼中高表达，其后在4 d仔鱼中显著降低(P<0.05)，其后显著上调表达，至15 d仔鱼达峰值，其后在20 d仔鱼显著下降，并在25 d后稚鱼和幼鱼中保持相对较低表达水平。研究结果可为深入认识黄条鰤hsp70基因的结构特征、发生发育及其早期生长发育阶段的表达调控功能提供依据。 |
| 关键词: 黄条鰤 hsp70 基因克隆 表达调控 早期生活史 |
| DOI：10.19663/j.issn2095-9869.20220311003 |
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| Molecular cloning and temporal expression pattern of hsp70 gene during the early life stages of Seriola aureovittata |
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FANG Lu1,2,3, XU Yongjiang2,3, CUI Aijun2,4, JIANG Yan2,5, WANG Bin2,6, ZHOU Heting1,2,3, LIU Xuezhou2,3
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1.School of Fisheries and Life Sciences, Shanghai Ocean University, Shanghai 201306, China;2.Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences;3.Joint Laboratory for Deep Blue Fishery Engineering of Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao 266071, China;4.Joint Laboratory for Deep Blue Fishery Engineering of Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao 266072, China;5.Joint Laboratory for Deep Blue Fishery Engineering of Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao 266073, China;6.Joint Laboratory for Deep Blue Fishery Engineering of Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao 266074, China
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| Abstract: |
| Seriola aureovittata is a long-distance migratory oceanic species belonging to the Carangidae family; it is also known as yellowtail kingfish. It inhabits temperate and subtropical marine waters worldwide. We reached a significant breakthrough in S. aureovittata seedlings production in 2017, enabling current massively juvenile rearing in China, which became a promising candidate for the farming industry, especially for the rapidly growing open ocean aquaculture. However, sudden larvae death caused by environmental changes-related stresses during the early life stages of S. aureovittata leads to great losses. Thus, exploring the physiological mechanism of larvae responses to environmental stresses during seedlings' production under artificial breeding conditions is urgently needed. Heat shock proteins (HSPs) play an important function in the physiological regulation of stress and immune responses in vertebrates, including fish. HSP70 is a member of the HSPs family studied in dozens of fish species, with physiological roles in protein homeostasis, DNA protection, and stress tolerance enhancement, among others. To investigate the possible physiological effects of HSPs on early growth and development of S. aureovittata, we cloned and obtained the full-length cDNA sequence of the hsp70 gene. Furthermore, the structure and spatial and temporal expression patterns of hsp70 during embryonic development, larval and juvenile growth and development of S. aureovittata were determined. The results showed that the full-length cDNA sequence of the hsp70 gene contains 2 332 bp, wherein the 5'-UTR is 187 bp, the ORF is 1 920 bp, and the 3'-UTR is 225 bp in length. A 639 amino acids protein with a molecular weight of 70.1 kDa is encoded by hsp70. The hsp70 spatial expression exhibited a sex dimorphism pattern, with significantly high expression levels in the gill, heart, spleen, and ovary in females, whereas in males, the significantly high expression levels were found in the pituitary, gill, head kidney, and testis. Notably, the highest hsp70 expression levels were observed in the ovary of females and the testis of males. During the S. aureovittata embryonic development, the hsp70 mRNA was detected in fertilized eggs before cleavage, indicating that hsp70 is parentally inherited. Additionally, the hsp70 mRNA could be detected at all stages of embryonic development, wherein the lowest expression levels were observed before the low blastula stage, with a significant increase at the early gastrula stage maintained until the hatching stage. The high levels of hsp70 mRNA were detected in one-day-old larva, which decreased in four-day-old larva, followed by an increase in 15-day-old larva. Remarkably, the hsp70 mRNA level once again decreased in 20-day-old larva, maintaining an average level until 60-day-old juveniles. The results from the present study may provide insights into the origin and physiological function of hsp70 during the early life stages of S. aureovittata. |
| Key words: Seriola aureovittata hsp70 Gene cloning Expression and regulation Early life stages |
