仿刺参肠道神经定位初步研究
|
摘要
本文以仿刺参(Apostichopus japonicus)肠道组织为研究材料,利用HE染色、米利根染色和免疫组化技术对其组织学结构及神经定位进行了观察和分析,初步探讨了仿刺参肠道的神经分布。HE染色结果表明,仿刺参肠壁由外向内分别为:体腔上皮层、肌肉层、内部结缔组织层和肠腔上皮层;和后肠相比,其中肠内壁常形成高而窄,且方向单一的褶皱,在结构上扩大了吸收面积,与其主要起消化吸收作用这一功能一致。米利根染色表明肠壁的纵向肌肉、肠腔上皮层和体腔上皮层均有神经成分存在,棘皮动物神经特异性抗体1E11的定位研究结果同样显示其免疫活性广泛而强烈,阳性位点集中于体腔上皮层、肌肉层和内部结缔组织层,且在肌肉层中表达最强,推测其可能参与肠道的再生过程及长期夏眠萎缩后的肠道快速修复过程。本研究为仿刺参神经生物学的研究提供了理论支持,同时也为仿刺参特殊生理机制的研究提供神经学参考资料。
In this study,the intestines of sea cucumber(Apostichopus japonicus)were used as experimental materials,and HE staining,Milligan staining and immunohistochemistry were applied to studying the histological structures and neural localization of sea cucumberintestine.The HE staining results showed that the intestine of sea cucumber is composed of 4 layers,coelomic epithelium,muscle,inner connective tissue and epithelium from outside to inside.Compared with the posterior intestine,the inner wall of mid intestine often forms many high and narrow folds in the same direction,expanding the absorption area.Such structure is consistent with the major function of mid intestine,digestion and absorption.The Milligan staining showed that the nerves exist in the longitudinal muscle,the coelomic epithelium layer and the epithelium of the intestine.The neuronal localization results of 1 E11,the neural specific antibody in echinoderm,showed that the 1 E11 immunoreactivity is wide and strong,which concentrates on the coelomic epithelium,muscle,and inner connective tissue layers,and the strongest in muscle layer.It was speculated that such immunoreactivity may be involved in the regeneration of the intestinal tract and in the rapid repair process of the intestinal tract after long-term aestivation atrophy.The present work provided a theoretical foundation for the future research on neurobiology and laid a foundation for the study onthe special physiological mechanism of A.japonicus.
In this study,the intestines of sea cucumber(Apostichopus japonicus)were used as experimental materials,and HE staining,Milligan staining and immunohistochemistry were applied to studying the histological structures and neural localization of sea cucumberintestine.The HE staining results showed that the intestine of sea cucumber is composed of 4 layers,coelomic epithelium,muscle,inner connective tissue and epithelium from outside to inside.Compared with the posterior intestine,the inner wall of mid intestine often forms many high and narrow folds in the same direction,expanding the absorption area.Such structure is consistent with the major function of mid intestine,digestion and absorption.The Milligan staining showed that the nerves exist in the longitudinal muscle,the coelomic epithelium layer and the epithelium of the intestine.The neuronal localization results of 1 E11,the neural specific antibody in echinoderm,showed that the 1 E11 immunoreactivity is wide and strong,which concentrates on the coelomic epithelium,muscle,and inner connective tissue layers,and the strongest in muscle layer.It was speculated that such immunoreactivity may be involved in the regeneration of the intestinal tract and in the rapid repair process of the intestinal tract after long-term aestivation atrophy.The present work provided a theoretical foundation for the future research on neurobiology and laid a foundation for the study onthe special physiological mechanism of A.japonicus.
引文
[1] Inoue M,Birenheide R,Koizumi O,et al.Localization of the neuropeptide NGIWYamide in the holothurian nervous system and its effects on muscular contraction[J].Proceedings of the Royal Society of London B:Biological Sciences,1999,266(1423):993-1000.
[2]朱峰.仿刺参Apostichopus japonicus胚胎发育和主要系统的组织学研究[D].青岛:中国海洋大学,2009.Zhu F.The Study on Embryonic Development and Major Organs’Histology of Sea Cucumber Apostichopus japonicus[D].Qingdao:Ocean University of China,2009.
[3]严芳,胡炜,李成林,等.饲料中添加活性酵母制剂对刺参生长、免疫力和抗病力的影响[J].动物营养学报,2017,29(2):583-589.Yan F,Hu W,Li C L,et al.Effects of dietary active yeast products on growth,immunity and disease resistance of sea cucumber(Apostichopus japonicas Selenka)[J].Chinese Journal of Animal Nutrition,2017,29(2):583-589.
[4]梁晓旺.刺参自溶过程中体壁胶原组织变化的研究[D].大连:大连工业大学,2015.Liang X W.Changes of Collagenous Tissue in Body Wall of Autolytic Sea Cucumber Stichopus japonicus[D].Dalian:Dalian Polytechnic University,2015.
[5]赵业.刺参Apostichopus japonicus(Selenka)夏眠期间基因表达模式及DNA甲基化基础特征研究[D].北京:中国科学院研究生院,2015.Zhao Y.Histological Study on the Digestive Tract of the Apostichopus japonicus in Aestivation[J].Beijing:Graduate University of Chinese Academy of Sciences,2015.
[6]徐冬雪.仿刺参热胁迫响应的分子调控特征研究与SNP标记筛选[D].北京:中国科学院大学,2016.Xu D X.Research of Molecular Mechanisms of Heat Shock Response and Heat-resistance SNP in Sea Cucumbers Apostichopus japonicus[D].Beijing:University of Chinese Academy of Sciences,2016.
[7] Yang H,Hamel J F,Mercier A,et al.The Sea Cucumber Apostichopus japonicus:History,Biology and Aquaculture[M].USA:Academic Press,2015.
[8] Iwakoshi E.Comparative aspects of structure and action of bioactive peptides isolated from the seacucumber Stichopus japonicus[J].Peptide Chemistry,1995,48(5):261-264.
[9] Tamori M,Saha A K,Matsuno A,et al.Stichopin-containing nerves and secretory cells specific to connective tissues of the sea cucumber[J].Proceedings of the Royal Society of London B:Biological Sciences,2007,274(1623):2279-2285.
[10] Nakajima Y,Kaneko H,Murray G,et al.Divergent patterns of neural development in larval echinoids and asteroids[J].Evolution&Development,2004,6(2):95-104.
[11]杨红生.刺参生物学:理论与实践[M].北京:科学出版社,2014.Yang H S.The Biology of Sea Cucumber Apostichopus japonicus:Theory and Practice[M].Beijing:Science Press,2014.
[12]苏琳,陈慕雁,王天明,等.刺参夏眠不同时期划分以及前肠各组织层厚度定量分析[J].海洋科学,2012,36(12):1-5.Su L,Chen M Y,Wang T M,et al.The division of different stages of aestivation and quantitative analysis of tissue layers thickness of anterior intestine in sea cucumber Apostichopus japonicus[J].Marine Science,2012,36(12):1-5.
[13] Burke R D,Osborne L,Wang D,et al.Neuron-specific expression of a synaptotagmin gene in the sea urchin Strongylocentrotus purpuratus[J].Journal of Comparative Neurology,2006,496(2):244-251.
[14]崔龙波,董志宁,陆瑶华.仿刺参消化系统的组织学和组织化学研究[J].动物学杂志,2000,35(6):2-4.Cui L B,Dong Z N,Lu Y H.Histological and histochemical studies on the digestive system of Apostichopus japonicus[J].Chinese Journal of Zoology,2000,35(6):2-4.
[15] García-Arrarás J E,Díaz-Miranda L,Torres I I,et al.Regeneration of the enteric nervous system in the sea cucumber Holothuria glaberrima[J].Journal of Comparative Neurology,1999,406(4):461-475.
[16]赵鹏,杨红生,孙丽娜.仿刺参(Apostichopusjaponicus)摄食和运动器官的结构与功能[J].海洋通报,2013,32(2):178-183.Zhao P,Yang H S,Sun L N.Study on the structures and functions of feeding and locomotor organs of sea cucumber Apostichopus japonicus[J].Marine Science Bulletin,2013,32(2):178-183.
[17] Inoue M,Tamori M,Motokawa T.Innervation of holothurian body wall muscle:Inhibitory effects and localization of 5-HT[J].Zoological Science,2002,19(11):1217-1222.
[18] Saha A K,Tamori M,Inoue M,et al.NGIWYamide-induced contraction of tube feet and distribution of NGIWYamide-like immunoreactivity in nerves of the starfish Asterina pectinifera[J].Zoological Science,2006,23(7):627-632.
[19] Lin M,Mita M,EgertováM,et al.Cellular localization of relaxin-like gonad-stimulating peptide expression in Asterias rubens:New insights into neurohormonal control of spawning in starfish[J].Journal of Comparative Neurology,2017,525(7):1599-1617.
[20]郑法新.刺参器官再生的组织学与相关基因研究[D].青岛:中国海洋大学,2006.Zheng F X.The Study of HistoIogy of Organ Regeneration and Regeneration-related Gene of Apostichopus japonicus(SeIenka).[D].Qingdao:Ocean University of China,2009.
[21] Anderson J M.Studies on visceral regeneration in sea-stars.II.Regeneration of pyloric caeca in Asteriidae,with notes on the source of cells in regenerating organs[J].The Biological Bulletin,1965,128(1):1-23.
[22] Díaz-Miranda L,Price D A,Greenberg M J,et al.Characterization of two novel neuropeptides from the sea cucumber Holothuria glaberrima[J].The Biological Bulletin,1992,182(2):241-247.
[23] Díaz-Miranda L,García-Arrarás J E.Pharmacological action of the heptapeptide GFSKLYFamide in the muscle of the sea cucumber Holothuria glaberrima(Echinodermata)[J].Comparative Biochemistry and Physiology Part C:Pharmacology,Toxicology and Endocrinology,1995,110(2):171-176.
[24]李润玲,丁君,张玉勇,等.刺参(Apostichopus japonicus)夏眠期间消化道的组织学研究[J].海洋环境科学,2006,25(4):15-19.Li R L,Ding J,Zhang Y Y,et al.Histological study on the digestive tract of the Apostichopus japonicus in aestivation[J].Environmental Science of Sea,2006,25(4):15-19.
[2]朱峰.仿刺参Apostichopus japonicus胚胎发育和主要系统的组织学研究[D].青岛:中国海洋大学,2009.Zhu F.The Study on Embryonic Development and Major Organs’Histology of Sea Cucumber Apostichopus japonicus[D].Qingdao:Ocean University of China,2009.
[3]严芳,胡炜,李成林,等.饲料中添加活性酵母制剂对刺参生长、免疫力和抗病力的影响[J].动物营养学报,2017,29(2):583-589.Yan F,Hu W,Li C L,et al.Effects of dietary active yeast products on growth,immunity and disease resistance of sea cucumber(Apostichopus japonicas Selenka)[J].Chinese Journal of Animal Nutrition,2017,29(2):583-589.
[4]梁晓旺.刺参自溶过程中体壁胶原组织变化的研究[D].大连:大连工业大学,2015.Liang X W.Changes of Collagenous Tissue in Body Wall of Autolytic Sea Cucumber Stichopus japonicus[D].Dalian:Dalian Polytechnic University,2015.
[5]赵业.刺参Apostichopus japonicus(Selenka)夏眠期间基因表达模式及DNA甲基化基础特征研究[D].北京:中国科学院研究生院,2015.Zhao Y.Histological Study on the Digestive Tract of the Apostichopus japonicus in Aestivation[J].Beijing:Graduate University of Chinese Academy of Sciences,2015.
[6]徐冬雪.仿刺参热胁迫响应的分子调控特征研究与SNP标记筛选[D].北京:中国科学院大学,2016.Xu D X.Research of Molecular Mechanisms of Heat Shock Response and Heat-resistance SNP in Sea Cucumbers Apostichopus japonicus[D].Beijing:University of Chinese Academy of Sciences,2016.
[7] Yang H,Hamel J F,Mercier A,et al.The Sea Cucumber Apostichopus japonicus:History,Biology and Aquaculture[M].USA:Academic Press,2015.
[8] Iwakoshi E.Comparative aspects of structure and action of bioactive peptides isolated from the seacucumber Stichopus japonicus[J].Peptide Chemistry,1995,48(5):261-264.
[9] Tamori M,Saha A K,Matsuno A,et al.Stichopin-containing nerves and secretory cells specific to connective tissues of the sea cucumber[J].Proceedings of the Royal Society of London B:Biological Sciences,2007,274(1623):2279-2285.
[10] Nakajima Y,Kaneko H,Murray G,et al.Divergent patterns of neural development in larval echinoids and asteroids[J].Evolution&Development,2004,6(2):95-104.
[11]杨红生.刺参生物学:理论与实践[M].北京:科学出版社,2014.Yang H S.The Biology of Sea Cucumber Apostichopus japonicus:Theory and Practice[M].Beijing:Science Press,2014.
[12]苏琳,陈慕雁,王天明,等.刺参夏眠不同时期划分以及前肠各组织层厚度定量分析[J].海洋科学,2012,36(12):1-5.Su L,Chen M Y,Wang T M,et al.The division of different stages of aestivation and quantitative analysis of tissue layers thickness of anterior intestine in sea cucumber Apostichopus japonicus[J].Marine Science,2012,36(12):1-5.
[13] Burke R D,Osborne L,Wang D,et al.Neuron-specific expression of a synaptotagmin gene in the sea urchin Strongylocentrotus purpuratus[J].Journal of Comparative Neurology,2006,496(2):244-251.
[14]崔龙波,董志宁,陆瑶华.仿刺参消化系统的组织学和组织化学研究[J].动物学杂志,2000,35(6):2-4.Cui L B,Dong Z N,Lu Y H.Histological and histochemical studies on the digestive system of Apostichopus japonicus[J].Chinese Journal of Zoology,2000,35(6):2-4.
[15] García-Arrarás J E,Díaz-Miranda L,Torres I I,et al.Regeneration of the enteric nervous system in the sea cucumber Holothuria glaberrima[J].Journal of Comparative Neurology,1999,406(4):461-475.
[16]赵鹏,杨红生,孙丽娜.仿刺参(Apostichopusjaponicus)摄食和运动器官的结构与功能[J].海洋通报,2013,32(2):178-183.Zhao P,Yang H S,Sun L N.Study on the structures and functions of feeding and locomotor organs of sea cucumber Apostichopus japonicus[J].Marine Science Bulletin,2013,32(2):178-183.
[17] Inoue M,Tamori M,Motokawa T.Innervation of holothurian body wall muscle:Inhibitory effects and localization of 5-HT[J].Zoological Science,2002,19(11):1217-1222.
[18] Saha A K,Tamori M,Inoue M,et al.NGIWYamide-induced contraction of tube feet and distribution of NGIWYamide-like immunoreactivity in nerves of the starfish Asterina pectinifera[J].Zoological Science,2006,23(7):627-632.
[19] Lin M,Mita M,EgertováM,et al.Cellular localization of relaxin-like gonad-stimulating peptide expression in Asterias rubens:New insights into neurohormonal control of spawning in starfish[J].Journal of Comparative Neurology,2017,525(7):1599-1617.
[20]郑法新.刺参器官再生的组织学与相关基因研究[D].青岛:中国海洋大学,2006.Zheng F X.The Study of HistoIogy of Organ Regeneration and Regeneration-related Gene of Apostichopus japonicus(SeIenka).[D].Qingdao:Ocean University of China,2009.
[21] Anderson J M.Studies on visceral regeneration in sea-stars.II.Regeneration of pyloric caeca in Asteriidae,with notes on the source of cells in regenerating organs[J].The Biological Bulletin,1965,128(1):1-23.
[22] Díaz-Miranda L,Price D A,Greenberg M J,et al.Characterization of two novel neuropeptides from the sea cucumber Holothuria glaberrima[J].The Biological Bulletin,1992,182(2):241-247.
[23] Díaz-Miranda L,García-Arrarás J E.Pharmacological action of the heptapeptide GFSKLYFamide in the muscle of the sea cucumber Holothuria glaberrima(Echinodermata)[J].Comparative Biochemistry and Physiology Part C:Pharmacology,Toxicology and Endocrinology,1995,110(2):171-176.
[24]李润玲,丁君,张玉勇,等.刺参(Apostichopus japonicus)夏眠期间消化道的组织学研究[J].海洋环境科学,2006,25(4):15-19.Li R L,Ding J,Zhang Y Y,et al.Histological study on the digestive tract of the Apostichopus japonicus in aestivation[J].Environmental Science of Sea,2006,25(4):15-19.