刺参体腔细胞类型及其死亡率与温度的关系
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摘要
应用光镜和透射电镜技术研究了刺参体腔细胞的类型,结合流式细胞术分析了刺参体腔细胞的亚群。光学显微镜下,根据细胞形态将体腔细胞分为球型细胞、变形细胞、淋巴样细胞和纺锤细胞;透射电镜下,根据细胞结构将体腔细胞分为颗粒细胞和无颗粒细胞。利用流式细胞仪通过测试前向角散射光(FSC)和侧向角散射光(SSC)两个参数,分别代表细胞的大小和细胞的颗粒度,绘制双参数(SSC×FSC)散点图和等高图,得出刺参体腔细胞可以分为颗粒细胞和无颗粒细胞两大群,其中无颗粒细胞是主要细胞类群,占体腔细胞总数的65%,而颗粒细胞占31%。采用流式细胞仪研究了刺参体腔细胞死亡率与温度的关系。结果表明,在0~20℃下海参体腔细胞死亡率变化不大,在高温条件下(20~30℃),细胞破裂解体速度加快,细胞存活时间较短。
The types of coelomocytes in Apostichopus japonicas were determined by light microscopy and transmission electron microscopy,and the subsets of coelomocytes were analyzed by flow cytometry.Coelomocytes could be divided into spherical cell,amoebocytes,lymphoid cell and spindle cell according to the morphology under light microscopy;besides,the coelomocytes could be identified as granulosa cell and agranulocytes based on the cellular structure under the transmission electron microscopy.The population of coelomocytes in sea cucumber Apostichopus japonicas was further analyzed by flow cytometry.Two parameters of light-scatter were set up such as light-scatter(FSC)and side light-scatter(SSC).The analysis results were expressed as the dot and the contour plots of two parameters(FSC & SSC),the single parameter(FSC & SSC)histogram of coelomocytes for Apostichopus japonicas.Apostichopus japonicas coelomocytes were divided into two populations of granulocytes and agranulocytes by flow cytometry,and agranulocytes were major population accounting for 65% whereas granulocytes accounted for 31%.The relationship between mortality of coelomocytes and temperature was invested based on flow cytometry analysis.Results showed that the mortality of sea cucumber coelomocyte did not change significantly at low temperature range(0-20℃),while higher temperature(20-30℃)accelerated cell rupture sharply,leading to the shorter cell survival duration.
The types of coelomocytes in Apostichopus japonicas were determined by light microscopy and transmission electron microscopy,and the subsets of coelomocytes were analyzed by flow cytometry.Coelomocytes could be divided into spherical cell,amoebocytes,lymphoid cell and spindle cell according to the morphology under light microscopy;besides,the coelomocytes could be identified as granulosa cell and agranulocytes based on the cellular structure under the transmission electron microscopy.The population of coelomocytes in sea cucumber Apostichopus japonicas was further analyzed by flow cytometry.Two parameters of light-scatter were set up such as light-scatter(FSC)and side light-scatter(SSC).The analysis results were expressed as the dot and the contour plots of two parameters(FSC & SSC),the single parameter(FSC & SSC)histogram of coelomocytes for Apostichopus japonicas.Apostichopus japonicas coelomocytes were divided into two populations of granulocytes and agranulocytes by flow cytometry,and agranulocytes were major population accounting for 65% whereas granulocytes accounted for 31%.The relationship between mortality of coelomocytes and temperature was invested based on flow cytometry analysis.Results showed that the mortality of sea cucumber coelomocyte did not change significantly at low temperature range(0-20℃),while higher temperature(20-30℃)accelerated cell rupture sharply,leading to the shorter cell survival duration.
引文
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[13]WANG F Y,YANG H S,GAO F,et al.Effects of acute temperature or salinity stress on the immune response in sea cucumber Apostichopus japonicus[J].Comparative Biochemistry and Physiology,2008,151(4):491-498.
[2]王淑娴,叶海斌,于晓清,等.海参的免疫机制研究[J].安徽农业科学,2012,40(25):12553-12555.
[3]CANICCATTI C,MIGLIETTA A.Arylsulphatase in echinoderm immunocompetent cells[J].Histochemical Journal,1989,21(7):419-424.
[4]廖玉麟.中国动物志(一卷)[M].北京:科学出版社,1997.
[5]ELISEIKINA M G,MAGARLAMOY T Y.Coelomocyte morphology in the Holothurians Apostichopus japonicus(Aspidochirota:Stichopodidae)and Cucumaria japonica(Dendrochirota:Cucumariidae)[J].Russian Journal of Marine Biology,2002,28(3):197-202.
[6]刘晓云,谭金山,包振民,等.刺参体腔细胞的超微结构观察[J].电子显微学报,2005,24(6):613-615.
[7]DONAGHY L,KIM B K,HONG H K,et al.Flow cytometry studies on the populations and immune parameters of the hemocytes of the Suminoe oyster,Crassostrea ariakensis[J].Fish and Shellfish Immunology,2009,27(2):296-301.
[8]徐海圣,徐步进.甲壳动物细胞及体液免疫机理的研究进展[J].大连海洋大学学报,2001,16(1):49-56.
[9]周永灿,潘金培.贝类细胞和体液的防御机制研究进展[J].水产学报,1997,12(4):449-454.
[10]孙敬锋,冼健安,张铁军,等.流式细胞术比较研究三种对虾血细胞的分群[J].淡水渔业,2008,38(3):18-22.
[11]许秀芹,王宜艳,孙虎山.流式细胞术比较研究4种双壳贝类血细胞的分群[J].海洋湖沼通报,2006(1):46-50.
[12]陈勇,高峰,刘国山,等.温度、盐度和光照周期对刺身生长及行为的影响[J].水产学报,2007(5):687-691.
[13]WANG F Y,YANG H S,GAO F,et al.Effects of acute temperature or salinity stress on the immune response in sea cucumber Apostichopus japonicus[J].Comparative Biochemistry and Physiology,2008,151(4):491-498.