星点东方鲀胚胎发育的观察
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摘要
为探讨星点东方鲀(Takifgu niphobles)作为海水鱼遗传生物学和鱼类行为学研究实验动物的可行性提供科学依据,选取北戴河海域自然成熟的星点东方鲀亲鱼,经人工催产授精,受精卵在水温20. 0～21. 0℃条件下,于300 L玻璃钢水槽循环水中孵化。参考暗纹东方鲀(Takifugu obscure)、青鳉(Oryzias layioes)和斑马鱼(Barchydanio rerio var)胚胎发育分期形态指标,按分裂球数目、大小,胚盘形状、延展、下包,神经系统发育,体节数目,视觉和听觉器官的发育以及血液循环系统的不同发育阶段,记录了星点东方鲀完整的胚胎发育过程。研究结果表明,星点东方鲀受精卵卵径约为756～863μm,胚胎发育可分为33期,主要分期与对应时间点为:2细胞期:受精后1 h 40 min;囊胚期:受精后8 h 6 min;原肠期:受精后13 h 48 min;心脏原基期:受精后58h 45 min;孵化期:受精后99 h 31 min;初孵仔鱼期:受精后108 h 31 min。值得注意的是,在32细胞期,观察到了与青鳉相同的边缘细胞和部分水平分裂。本实验中的星点东方鲀卵径在东方鲀属鱼中是最小的,也明显小于日本的星点东方鲀。本研究星点东方鲀胚胎发育不仅快于暗纹东方鲀和红鳍东方鲀(Takifgu rubripes),而且大大快于日本种群。
The naturally mature grass puffer Takifugu niphobles used in this study were collected from Beidaihe area of the Bohai Sea. Artificially inseminated eggs of grass puffer were incubated in the 300 L fiberglass tank with overflow water at a rate about 1 ～ 2 L·min-1,and the water temperature was controlled at 20. 0 ～ 21. 0℃ by electric heater; the embryonic development was carefully observed by light microscopy.Based on the principal morphological characters of medaka Oryzias latipes and Takifugu Obscuru,features like: the number and size of blastomeres,form of the blastoderm,extent of epiboly,development of the central nervous system,number and form of somites,optic and otic development,heart development were observed,and the embryonic development process was divided into 32 stages. The diameter of fertilized eggs was 756 ～ 863 μm. The main stages and corresponding time point were as followings: 2-cell: 1 h 40 min after fertilization( AF); blastula stage: 8 h 6 min AF; gastrula period: 13 h 48 min AF; somite stage: 28 h 43 min AF; heart anlage stage: 58 h 45 min AF; hatching stage: 99 h 31 min AF; 50% hatching stage: 108 h31 min AF. It was close to medaka that the horizontal division and the marginal cell were observed at 32-cell stage. The size of the fertilized egg was the smallest among the congeneric species of Takifugu,and was even much smaller than the individuals of the same species in Japan. The embryonic development of grass puffer was faster than that of T. Obscuru and Takifugu rubripes and even faster than the Japanese population of the same species.
The naturally mature grass puffer Takifugu niphobles used in this study were collected from Beidaihe area of the Bohai Sea. Artificially inseminated eggs of grass puffer were incubated in the 300 L fiberglass tank with overflow water at a rate about 1 ～ 2 L·min-1,and the water temperature was controlled at 20. 0 ～ 21. 0℃ by electric heater; the embryonic development was carefully observed by light microscopy.Based on the principal morphological characters of medaka Oryzias latipes and Takifugu Obscuru,features like: the number and size of blastomeres,form of the blastoderm,extent of epiboly,development of the central nervous system,number and form of somites,optic and otic development,heart development were observed,and the embryonic development process was divided into 32 stages. The diameter of fertilized eggs was 756 ～ 863 μm. The main stages and corresponding time point were as followings: 2-cell: 1 h 40 min after fertilization( AF); blastula stage: 8 h 6 min AF; gastrula period: 13 h 48 min AF; somite stage: 28 h 43 min AF; heart anlage stage: 58 h 45 min AF; hatching stage: 99 h 31 min AF; 50% hatching stage: 108 h31 min AF. It was close to medaka that the horizontal division and the marginal cell were observed at 32-cell stage. The size of the fertilized egg was the smallest among the congeneric species of Takifugu,and was even much smaller than the individuals of the same species in Japan. The embryonic development of grass puffer was faster than that of T. Obscuru and Takifugu rubripes and even faster than the Japanese population of the same species.
引文
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[19]MIYAKI K,TYUDU H,WATANABE T.Treatment of tiger puffer,Takifugu yubripes,eggs with tannic acid to eliminate their adhesiveness for seed propagation[J].Aquaculture Science,1996,46(1):97-100
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[23]CHEN Y.Induced ovulation and embryonic development of ocellated puffer,Takifugu ocellatus[J].Journal of Applied Ichthyology,2005(21):136-140.
[24]SUZUKI Y,TAKEMURA A.Comparison of hormonal treatments for induction of oocyte maturation and ovulation in the tiger puffer,Takifugu rubripes[J].Aquaculture Science,1996,44(1):85-90
[25]SHI Y,ZHANG G,LIU J,et al.Induced ovulation in cultured tawny puffer Takifugu flavidus by injections of LHRH-a.[J].Journal of Applied Ichthyology,2012(28):22-25.
[26]DOI H,SONOYAMA T,IMAI C,et al.Early development of five tiger puffer species of the genus Takifugu[J].Journal of National Fisheries University,2014,62(3):109-114
[27]YU C F,YU P H F.The annual toxicological profile of two common puffer fish,Takifugu niphobles(Jordan and Snyder)and Takifugu alboplumbeus(Richardson),collected along Hong Kong coastal waters[J].Toxicon,2002(40):313-316
[28]LIN W F,HWANG D F.Analysis of poisoning cases,monitoring and risk warning for marine toxins(TTX,PSP and CTXs)in Taiwan[J].Journal of Food and Drag Analysis,2012(4):764-771.
[29]UJI S,KUROKAWA T,HASHIMOTO H.Embryogenic staging of fugu,Takifugu rubripes,and expression profiles of aldh1a2,aldh1a3 and cyp26a1[J].Development Growth&Differentiation,2011(53):715-725.
[2]王奎旗,陈梅,高天翔.东方鲀属鱼类的分类与区系分布研究[J].青岛海洋大学学报,2001(6):855-860.WANG K Q,CHEN M,GAO T X.Study on taxonomy and fauna of Takifugu[J].Journal of Ocean University of Qingdao,2001(6):855-860.
[3]叶宁,吴晓东,张苇.湛江高桥红树林区鱼类调查[J].广东海洋大学学报.2007,27(6):55-61.YE N,WU X D,ZHANG W.Survey of fishes in Gaoqiao mangrove zone in Zhanjiang[J].Journal of Guangdong Ocean University.2007,27(6):55-61.
[4]MIYAKI K.Biological study of hybrid pufferfishes of the genus Takifugu,Tetraodontidae[D].Nagasaki:Nagasaki University,1992.
[5]EIJI M,TAKESHI Y,HIROYUKI D,et al.Aggregating behavior of the grass puffer,Takifugu niphobles,observed in aquarium during the spawning period[J].Zoological Science,2002(27):559-564.
[6]MATSUMURA K.Tetrodotoxin as a pheromone[J].Nature,1995(375):563-564.
[7]ANDO H,SHAHJAHAN M,HATTORI A.Molecular neuroendocrine basis of lunar-related spawning in grass puffer[J].General and Comparative Endocrinology,2013(181):211-214.
[8]ITOI S,ISHIZAKA K,MITSUOKA R,et al.Seasonal changes in the tetrodotoxin content of the pufferfish Takifugu niphobles[J].Toxicon,2016(114):53-58.
[9]ITOI S,YOSHIKAWA S,ASAHINA K,et al.Larval pufferfish protected by maternal tetrodotoxin[J].Toxicon,2014(78):35-40.
[10]HAMASAKI M,TAKEUCHI Y,MIYAKI K,et al.Gonadal development and fertility of triploid grass puffer Takifugu niphobles induced by cold shock treatment[J].Marine Biotechnology,2013(15):133-144.
[11]APARICIO S,CHAPMAN J,STUPKA E,et al.Whole-genome shotgun assembly and analysis of the genome of Fugu rubripes.[J].Science,2002,297(5585):1301-1310.
[12]周巍,杨焕明,刘国仰.河豚鱼与人类基因组计划[J].遗传,1997,19(6):37-40.ZHOU W,YANG H M,LIU G Y.Pufferfish and human genome projects[J].Hereditas,1997 19(6):37-40.
[13]GILLIGAN P,BRENNER S,VENKATESH B.Fugu and human sequence comparison identifies novel human genes and conserved non-coding sequences[J].Gene,2002(294):35-44.
[14]VENKATACHALAM K.The molecular paradigm of human complexity[J].Cloning Transgenes,2013(3):1-2.
[15]YAMANOUE Y,MIYA M,MATSUURA K,et al.Explosive speciation of Takifugu:Another use of fugu as a model system for evolutionary biology[J].Molecular Biology and Evolution,2009(26):623-629.
[16]WITTBRODT J,SHIMA A,SCHARTL M.Medaka-a model organism from the far East[J].Nature Reviews Genetics,2002(3):53-64.
[17]吴端生,王宗保.鱼类实验动物开发与应用研究的现状及展望[J].中国实验动物学杂志,2000,10(2):103-108.WU D S,WANG Z B.Current state and prospects of application and exploitation of fish as laboratory animal[J].Chinese Journal of Laboratory Animal Science,2000,10(2):103-108.
[18]MATSUI H.The use of fish models to study human neurological disorders[J].Neuroscience Research,2017(120):1-7.
[19]MIYAKI K,TYUDU H,WATANABE T.Treatment of tiger puffer,Takifugu yubripes,eggs with tannic acid to eliminate their adhesiveness for seed propagation[J].Aquaculture Science,1996,46(1):97-100
[20]莫根永,胡庚东,周彦锋.暗纹东方鲀胚胎发育的观察[J].淡水渔业,2009,39(6):22-27.MO G Y,HU G D,ZHOU Y F.Observation of the embryonic development of Takifugu obscurus[J].Fresh Water Fisheries,2009,39(6):22-27.
[21]IWAMATSU T.Stages of normal development in the medaka Oryzias latipes[J].Mechanisms of Development,2004(121):605-18.
[22]KIMMEL C B,BALLARD W,KIMMEL S R.Stages of embryonic development of the zebrafish[J].Developmental Dynamics,1995(203):253-310.
[23]CHEN Y.Induced ovulation and embryonic development of ocellated puffer,Takifugu ocellatus[J].Journal of Applied Ichthyology,2005(21):136-140.
[24]SUZUKI Y,TAKEMURA A.Comparison of hormonal treatments for induction of oocyte maturation and ovulation in the tiger puffer,Takifugu rubripes[J].Aquaculture Science,1996,44(1):85-90
[25]SHI Y,ZHANG G,LIU J,et al.Induced ovulation in cultured tawny puffer Takifugu flavidus by injections of LHRH-a.[J].Journal of Applied Ichthyology,2012(28):22-25.
[26]DOI H,SONOYAMA T,IMAI C,et al.Early development of five tiger puffer species of the genus Takifugu[J].Journal of National Fisheries University,2014,62(3):109-114
[27]YU C F,YU P H F.The annual toxicological profile of two common puffer fish,Takifugu niphobles(Jordan and Snyder)and Takifugu alboplumbeus(Richardson),collected along Hong Kong coastal waters[J].Toxicon,2002(40):313-316
[28]LIN W F,HWANG D F.Analysis of poisoning cases,monitoring and risk warning for marine toxins(TTX,PSP and CTXs)in Taiwan[J].Journal of Food and Drag Analysis,2012(4):764-771.
[29]UJI S,KUROKAWA T,HASHIMOTO H.Embryogenic staging of fugu,Takifugu rubripes,and expression profiles of aldh1a2,aldh1a3 and cyp26a1[J].Development Growth&Differentiation,2011(53):715-725.