自噬抑制、增效对曼氏无针乌贼性腺自噬、凋亡及雌雄标记基因的影响  

Effects of Autophagy Inhibition(CQ)and Autophagy Potentiation(RAPA)on Autophagy,Apoptosis,and Sex-Marker Genes in Testis and Ovary of Sepiella japonica

作　　者：罗俊 陈龙 张涛[3] 陈爽 赵一凤 杨昺玥 杜庆乐 宋伟华 柳意樊 LUO Jun;CHEN Long;ZHANG Tao(National Engineering Research Center for Marine Facilities Aquaculture of Zhejiang Ocean University,Zhoushan 316022;School of Marine Science and Technology of Zhejiang Ocean University,Zhoushan 316022;Zhejiang Marine Fisheries Research Institute,Key Lab of Mariculture and Enhancement of Zhejiang Province,Zhoushan 316021,China)

机构地区：[1]浙江海洋大学国家海洋设施养殖工程技术研究中心,浙江舟山316022 [2]浙江海洋大学海洋科学与技术学院,浙江舟山316022 [3]浙江省海洋水产研究所,浙江省海水增养殖重点实验室,浙江舟山316021

出　　处：《浙江海洋大学学报（自然科学版）》2024年第5期373-381,共9页Journal of Zhejiang Ocean University：Natural Science

基　　金：浙江省自然科学基金项目(LY20C19000);浙江省重点研发计划项目(2020C02004);国家自然科学基金项目(31702321)。

摘　　要：氯喹(chloroquine,CQ)和雷帕霉素(rapamycin,RAPA)是实验室常用的自噬抑制剂和增效剂。小鼠、斑马鱼等模式生物中的研究发现改变自噬水平会对精子和卵子发生产生影响。以曼氏无针乌贼为对象,通过建立精巢和卵巢的体外自噬抑制(CQ)和自噬增效(RAPA)组织培养模型,系统比较不同浓度CQ和RAPA处理后精巢、卵巢中自噬(LC3、BECN1)、凋亡(p53、Cas3)、无脊椎动物缝隙连接蛋白基因(Inx4)、雄性标记(Sox3、Sox30)和雌性标记(Nanos-1、FTZ-F1)基因表达的影响。结果表明CQ处理引起曼氏无针乌贼的精巢和卵巢的自噬水平显著抑制,自噬标记基因LC3显著升高,BECN1显著下降;Inx4的变化趋势与自噬标记基因基本一致,表明Inx4与自噬之间存在密切关系;凋亡标记基因p53和Cas3显著升高,表明自噬抑制引起凋亡通路被激活;雄性标记基因Sox4和Sox30的表达变化相对滞后于自噬和凋亡相关基因,表明自噬抑制对配子发生也产生了显著影响。自噬增效(RAPA)同样引起了自噬标记基因(LC3、BECN1)、无脊椎动物缝隙连接蛋白基因(Inx4)、凋亡基因(p53、Cas3)、雄性(Sox3、Sox30)和雌性标记基因(Nanos-1、FTZ-F1)均发生了显著变化,表明自噬对曼氏无针乌贼精子发生和卵子发生有重要作用。研究结果首次在海洋无脊椎动物中揭示了Inx4与自噬之间存在密切关系,且二者对配子发生有直接影响,为进一步研究自噬和Inx4对包括头足类在内的海洋无脊椎动物配子发生的调控作用提供了重要的基础数据。Chloroquine(CQ)and rapamycin(RAPA)are commonly used autophagy inhibitors and potentiators in the laboratory.Studies in model organisms such as mice and zebrafish have found that altering the level of autophagy can have an effect on spermatogenesis and oogenesis.In this study,we used Sepiella japonica to systematically compare the effects of autophagy(LC3,BECN1),apoptosis(p53,Cas3),invertebrate gap function protein gene(Inx4),male marker(Sox3,Sox30)and female marker(Nanos-1,FTZ-F1)gene expression after treatment with different concentrations of CQ and RAPA in testis and ovaries by establishing in vitro autophagy-inhibiting(CQ)and autophagy-potentiating(RAPA)tissue-culture models in testis and ovary.The results showed that CQ treatment caused a significant inhibition of autophagy level in testis and ovary of S.japonica,a significant increase of autophagy marker gene LC3 and a significant decrease of BECN1;the trend of change of Inx4 was basically consistent with that of autophagy marker genes,suggesting that there was a close relationship between Inx4 and autophagy;apoptosis marker genes p53 and Cas3 were significantly elevated,indicating that the The expression changes of male marker genes Sox4 and Sox30 lagged behind those of autophagy and apoptosis-related genes,indicating that autophagy inhibition also had a significant effect on gametogenesis.Autophagy potentiation(RAPA)likewise caused significant changes in autophagy marker genes(LC3,BECN1),invertebrate gap function protein gene(Inx4),apoptosis genes(p53,Cas3),male marker genes(Sox3,Sox30)and female marker genes(Nanos-1,FTZ-F1),suggesting that autophagy exerted a significant effect on S.japonica spermatogenesis and oogenesis.The results of this study reveal for the first time in marine invertebrates the close relationship between Inx4 and autophagy,and the direct influence of both on gametogenesis,which provides important basic data for further research on the regulation of autophagy and Inx4 on gametogenesis in marine invertebrates including cephalopods.

关 键 词：氯喹 雷帕霉素 自噬 曼氏无针乌贼 

分 类 号：S927.4[农业科学—渔业资源]