神经型一氧化氮合酶在大菱鲆脑组织中的分布及定位
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摘要
【目的】探明神经型一氧化氮合酶(n NOS)在大菱鲆(Scophthalmus maximus)脑组织中的分布情况,为揭示大菱鲆脑组织中一氧化氮(NO)的生理功能提供形态学资料。【方法】分别采用NADPH-d组织化学染色法和免疫组织化学法对大菱鲆脑组织中的n NOS进行定位研究。【结果】NADPH-d组织化学染色结果显示,大菱鲆大脑皮质中有蓝色的神经元和神经纤维存在。神经元呈梭形、锥形等形状,神经纤维呈串珠状且无序交织;小脑中NOS阳性神经元在分子层分布稀疏,在颗粒层分布密集,浦肯野细胞胞核淡染。经免疫组织化学法染色后,可观察到大脑皮质中n NOS免疫组化反应阳性物质呈深棕色;小脑皮质的分子层、颗粒层及浦肯野细胞层均有n NOS阳性神经元分布,浦肯野细胞呈免疫组织化学阳性反应。【结论】大菱鲆脑组织中的NOS类型主要限于n NOS,且n NOS在神经活动中发挥重要作用,也进一步证实NO在不同物种中具有一些共同的作用,但不同物种或相同物种不同组织中NO分布及含量存在差异。
【Objective】The present experiment was conducted to investigate distribution of neuronal nitric oxide synthase(n NOS), in order to provide important morphological data for further revealing physiological function of nitric oxide(NO) in brain tissue of Scophthalmus maximus. 【Method 】NADPH-d and immunohistochemical staining methods were used to locate the n NOS in the brain tissue. 【 Result 】 NADPH- d staining results showed that numerous blue neurons and nerve fibers existed in the cerebra of S. maximus. The neurons were in various forms, such as fusiform, subuliform and so on, and the nerve fibers were like beads and interweaved disorderly. The positive neurons of n NOS were distributed sparsely in the molecular layer and distributed densely in the granular layer, and the nucleuses of Purkinje cells were slightly stained. In addition, the immunohistochemical positive reaction material of n NOS in the cerebral cortex was observed to be dark brown after immunohistochemical staining. The positive neurons of n NOS were found to be distributed in the molecular layer, granular layer and Purkinje cell layer. Moreover, Purkinje cells tested positive for immunohistochemistry. 【Conclusion 】The NOS in the brain tissue of S. maximus mainly belongs to type of n NOS, and n NOS can play an important role in the neural activity. Furthermore, it is further confirmed that NO has some common effects on different species, but there exist still some difference in distribution and content of NO between different species or between different tissues of same species.
【Objective】The present experiment was conducted to investigate distribution of neuronal nitric oxide synthase(n NOS), in order to provide important morphological data for further revealing physiological function of nitric oxide(NO) in brain tissue of Scophthalmus maximus. 【Method 】NADPH-d and immunohistochemical staining methods were used to locate the n NOS in the brain tissue. 【 Result 】 NADPH- d staining results showed that numerous blue neurons and nerve fibers existed in the cerebra of S. maximus. The neurons were in various forms, such as fusiform, subuliform and so on, and the nerve fibers were like beads and interweaved disorderly. The positive neurons of n NOS were distributed sparsely in the molecular layer and distributed densely in the granular layer, and the nucleuses of Purkinje cells were slightly stained. In addition, the immunohistochemical positive reaction material of n NOS in the cerebral cortex was observed to be dark brown after immunohistochemical staining. The positive neurons of n NOS were found to be distributed in the molecular layer, granular layer and Purkinje cell layer. Moreover, Purkinje cells tested positive for immunohistochemistry. 【Conclusion 】The NOS in the brain tissue of S. maximus mainly belongs to type of n NOS, and n NOS can play an important role in the neural activity. Furthermore, it is further confirmed that NO has some common effects on different species, but there exist still some difference in distribution and content of NO between different species or between different tissues of same species.
引文
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马倩倩,孙敬锋,孙萌,张胜根,邢克智.2013.半滑舌鳎脑组织中神经型一氧化氮合成酶免疫组织化学定位研究[J].水产科学,32(10):609-612.Ma Q Q,Sun J F,Sun M,Zhang S G,Xing K Z.2013.Immunohistochemical localization of neuronal nitric oxide synthase in brain tissue of semi-smooth sole Cynoglossus semilaevis Gunthe[rJ].Fisheries Science,32(10):609-612.
沈伟哉,郭国庆,邢旭光,余菁.2002.大鼠脑干神经元型一氧化氮合成酶免疫阳性神经元的分布[J].解剖学研究,24(2):138-140.Shen W Z,Guo G Q,Xing X G,Yu J.2002.The distribution of neuronal nitric oxide synthase immunopositive neurons in brain stem of rats[J].Anatomy Research,24(2):138-140.
孙虎山,王宜艳,王晓安,孙修勤,李光友.2005.栉孔扇贝神经节一氧化氮合酶的组织化学和免疫组化定位[J].中国水产科学,12(1):20-23.Sun H S,Wang Y Y,Wang X A,Sun X Q,Li G Y.2005.Histochemical and immunohistochemical localization of nitric oxide synthase in nervous ganglia of scallop Chlamys farreri[J].Journal of Fishery Sciences of China,12(1):20-23.
王晓安,蒋小满.2005.口虾蛄(Oratosquilla oratoria)神经系统一氧化氮合酶的NADPH-d组织化学定位研究[J].神经解剖学杂志,21(5):547-552.Wang X A,Jiang X M.2005.NADPH-d histochemicalloical localization of nitric oxide synthase in the nervous system of Oratosquilla oratoria[J].Chinese Journal of Neuroanatomy,21(5):547-552.
王晓安,陆宇燕,李丕鹏.2007.虎斑颈槽蛇脑一氧化氮合酶NADPH-d组织化学定位研究[J].四川动物,26(2):284-287.Wang X A,Lu Y Y,Li P P.2007.Histochemistry localization of nitric oxide synthase by using NADPH-d in brain of Rhabdophis tigrinus[J].Sichuan Journal of Zoology,26(2):284-287.
邬晓敏,金成.2005.一氧化氮在中枢神经系统发育中的作用[J].解剖科学进展,11(4):358-361.Wu X M,Jin C.2005.Roles of nitric oxide in the development of central nervous system[J].Progress of Anatomical Sciences,11(4):358-361.
张朝再,董雷,牟凤辉,杨晓虹,孙薇.2014.神经型一氧化氮合酶抑制剂的研究进展[J].药学学报,49(6):781-788.Zhang C Z,Dong L,Mu F H,Yang X H,Sun W.2014.Progress in the studies on neuronal nitric oxide synthase inhibitors[J].Acta Pharmaceutica Sinica,49(6):781-788.
Aley K O,Mccarter G,Levine J D.1998.Nitric oxide signaling in pain and nociceptor sensitization in the ra[tJ].The Journal of Neuroscience,18(17):7008-7014.
Biswas S P,Jadhao A G,Bhoyar R C,Palande N V,Sinh D P.2015.Neuroanatomical localization of nitric oxide synthase(n NOS)in the central nervous system of carp,Labeo rohita during post-embryonic developmen[tJ].International Journal of Developmental Neuroscience,46:14-26.
Gourdon I,Guérin M C,Torreilles J,Roch P.2001.Nitric oxide generation by hemocytes of the mussel Mytilus galloprovincialis[J].Nitric Oxide,5(1):1-6.
Hope B T,Michael G J,Knigge K M,Vincent S R.1991.Neuronal NADPH diaphorase is a nitric oxide synthase[J].Proceedings of the National Academy of Sciences of the United States of America,88(7):2811-2814.
Ling S,Zhou J,Rudd J A,Hu Z,Fang M.2012.The expression of neurona l nitric oxide synthase in the brain of the mouse during embryogenesis[J].Anatomical Record,295(3):504-514.
Mishra O P,Mishra R,Ashraf Q M,Delivoria-Papadopoulos M.2006.Nitric oxide-mediated mechanism of neuronal nitric oxide synthase and inducible nitric oxide synthase expression during hypoxia in the cerebral cortex of newborn piglets[J].Neuroscience,140(3):857-863.
Rao S,Lin Z,Drobyshevsky A,Chen L,Ji X,Ji H,Yang Y,Yu L,Derrick M,Silverman R B,Tan S.2011.Involvement of neuronal nitric oxide synthase in ongoing fetal brain injury following near-term rabbit hypoxia-ischemia[J].Developmental Neuroscience,33(3-4):288-298.
Salchner P,Lubec G,Engelmann M,Orlando G F,Wolf G,Sartori S B,Hoeger H,Singewald N.2004.Genetic functional inactivation of neuronal nitric oxide synthase affects stressrelated Fos expression in specific brain regions[J].Cellular and Molecular Life Sciences,61(12):1498-1506.
Singru P S,Sakharkar A J,Subhedar N.2003.Neuronal nitric oxide synthase in the olfactory system of an adult teleost fish Oreochromis mossambicu[sJ].Brain Research,977(2):157-168.
Singru P S,Sakharkar A J,Mazumdar M,Subhedar N.2007.Neuronal nitric oxide synthase immunoreactivity in forebrain,pineal,and pituitary of Oreochromis mossambicus(tilapia)[J].Fish Physiology and Biochemistry,33(4):297-309.
Yu S Y,Zhang M H,Luo J X,Zhang L,Shao Y,Li G.2013.Curcumin ameliorates memory deficits via neuronal nitric oxide synthase in aged mice[J].Progress in Neuro-Psychopharmacology&Biological Psychiatry,45:47-53.
Zou S,Somvanshi R K,Paik S,Kumar U.2015.Colocalization of cannabinoid receptor 1 with somatostatin and neuronal nitric oxide synthase in rat brain hypothalamus[J].Journal of Molecular Neuroscience,55(2):480-491.
梁润梅.2003.神经系统的NO及其生物学功能[J].生物学教学,28(4):3-5.Liang R M.2003.NO of nervous system and its biological function[J].Biology Teaching,28(4):3-5.
马倩倩,孙敬锋,孙萌,张胜根,邢克智.2013.半滑舌鳎脑组织中神经型一氧化氮合成酶免疫组织化学定位研究[J].水产科学,32(10):609-612.Ma Q Q,Sun J F,Sun M,Zhang S G,Xing K Z.2013.Immunohistochemical localization of neuronal nitric oxide synthase in brain tissue of semi-smooth sole Cynoglossus semilaevis Gunthe[rJ].Fisheries Science,32(10):609-612.
沈伟哉,郭国庆,邢旭光,余菁.2002.大鼠脑干神经元型一氧化氮合成酶免疫阳性神经元的分布[J].解剖学研究,24(2):138-140.Shen W Z,Guo G Q,Xing X G,Yu J.2002.The distribution of neuronal nitric oxide synthase immunopositive neurons in brain stem of rats[J].Anatomy Research,24(2):138-140.
孙虎山,王宜艳,王晓安,孙修勤,李光友.2005.栉孔扇贝神经节一氧化氮合酶的组织化学和免疫组化定位[J].中国水产科学,12(1):20-23.Sun H S,Wang Y Y,Wang X A,Sun X Q,Li G Y.2005.Histochemical and immunohistochemical localization of nitric oxide synthase in nervous ganglia of scallop Chlamys farreri[J].Journal of Fishery Sciences of China,12(1):20-23.
王晓安,蒋小满.2005.口虾蛄(Oratosquilla oratoria)神经系统一氧化氮合酶的NADPH-d组织化学定位研究[J].神经解剖学杂志,21(5):547-552.Wang X A,Jiang X M.2005.NADPH-d histochemicalloical localization of nitric oxide synthase in the nervous system of Oratosquilla oratoria[J].Chinese Journal of Neuroanatomy,21(5):547-552.
王晓安,陆宇燕,李丕鹏.2007.虎斑颈槽蛇脑一氧化氮合酶NADPH-d组织化学定位研究[J].四川动物,26(2):284-287.Wang X A,Lu Y Y,Li P P.2007.Histochemistry localization of nitric oxide synthase by using NADPH-d in brain of Rhabdophis tigrinus[J].Sichuan Journal of Zoology,26(2):284-287.
邬晓敏,金成.2005.一氧化氮在中枢神经系统发育中的作用[J].解剖科学进展,11(4):358-361.Wu X M,Jin C.2005.Roles of nitric oxide in the development of central nervous system[J].Progress of Anatomical Sciences,11(4):358-361.
张朝再,董雷,牟凤辉,杨晓虹,孙薇.2014.神经型一氧化氮合酶抑制剂的研究进展[J].药学学报,49(6):781-788.Zhang C Z,Dong L,Mu F H,Yang X H,Sun W.2014.Progress in the studies on neuronal nitric oxide synthase inhibitors[J].Acta Pharmaceutica Sinica,49(6):781-788.
Aley K O,Mccarter G,Levine J D.1998.Nitric oxide signaling in pain and nociceptor sensitization in the ra[tJ].The Journal of Neuroscience,18(17):7008-7014.
Biswas S P,Jadhao A G,Bhoyar R C,Palande N V,Sinh D P.2015.Neuroanatomical localization of nitric oxide synthase(n NOS)in the central nervous system of carp,Labeo rohita during post-embryonic developmen[tJ].International Journal of Developmental Neuroscience,46:14-26.
Gourdon I,Guérin M C,Torreilles J,Roch P.2001.Nitric oxide generation by hemocytes of the mussel Mytilus galloprovincialis[J].Nitric Oxide,5(1):1-6.
Hope B T,Michael G J,Knigge K M,Vincent S R.1991.Neuronal NADPH diaphorase is a nitric oxide synthase[J].Proceedings of the National Academy of Sciences of the United States of America,88(7):2811-2814.
Ling S,Zhou J,Rudd J A,Hu Z,Fang M.2012.The expression of neurona l nitric oxide synthase in the brain of the mouse during embryogenesis[J].Anatomical Record,295(3):504-514.
Mishra O P,Mishra R,Ashraf Q M,Delivoria-Papadopoulos M.2006.Nitric oxide-mediated mechanism of neuronal nitric oxide synthase and inducible nitric oxide synthase expression during hypoxia in the cerebral cortex of newborn piglets[J].Neuroscience,140(3):857-863.
Rao S,Lin Z,Drobyshevsky A,Chen L,Ji X,Ji H,Yang Y,Yu L,Derrick M,Silverman R B,Tan S.2011.Involvement of neuronal nitric oxide synthase in ongoing fetal brain injury following near-term rabbit hypoxia-ischemia[J].Developmental Neuroscience,33(3-4):288-298.
Salchner P,Lubec G,Engelmann M,Orlando G F,Wolf G,Sartori S B,Hoeger H,Singewald N.2004.Genetic functional inactivation of neuronal nitric oxide synthase affects stressrelated Fos expression in specific brain regions[J].Cellular and Molecular Life Sciences,61(12):1498-1506.
Singru P S,Sakharkar A J,Subhedar N.2003.Neuronal nitric oxide synthase in the olfactory system of an adult teleost fish Oreochromis mossambicu[sJ].Brain Research,977(2):157-168.
Singru P S,Sakharkar A J,Mazumdar M,Subhedar N.2007.Neuronal nitric oxide synthase immunoreactivity in forebrain,pineal,and pituitary of Oreochromis mossambicus(tilapia)[J].Fish Physiology and Biochemistry,33(4):297-309.
Yu S Y,Zhang M H,Luo J X,Zhang L,Shao Y,Li G.2013.Curcumin ameliorates memory deficits via neuronal nitric oxide synthase in aged mice[J].Progress in Neuro-Psychopharmacology&Biological Psychiatry,45:47-53.
Zou S,Somvanshi R K,Paik S,Kumar U.2015.Colocalization of cannabinoid receptor 1 with somatostatin and neuronal nitric oxide synthase in rat brain hypothalamus[J].Journal of Molecular Neuroscience,55(2):480-491.
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