拟穴青蟹视神经节孕酮受体的免疫识别

研究表明,在甲壳类中可能存在类固醇的调控系统,然而核受体在进化的过程中是否依赖配体作用一直存在着争论.应用免疫印迹和免疫组织化学方法对拟穴青蟹(Scylla paramamosain)视神经节孕酮受体(PR)进行免疫识别.结果发现,拟穴青蟹视神经节中存在PR免疫阳性反应;其分子量约为70 ku,该物质广泛分布于视神经节的神经细胞核内.PR在拟穴青蟹视神经节的发现暗示着其类似于脊椎动物调节机制的存在;PR在神经细胞核中的定位表明其为核受体,很有可能在基因水平上参与拟穴青蟹视神经节的生理活动调节.     

拟穴青蟹GnRH-R的免疫识别

促性腺激素释放激素(GnRH)及其受体(GnRH-R)对生殖具有重要的调控作用.迄今无脊椎动物GnRH及GnRH-R的研究尚少.本研究采用兔抗人GnRH-R、兔抗果蝇GnRH-R和兔抗海鞘GnRH-R的抗体,应用免疫印迹和免疫共沉淀技术对拟穴青蟹(Scyllaparamamosain)GnRH-R进行了免疫识别,所得结果如下:1)经免疫印迹检测,拟穴青蟹脑、胸神经团、视神经节和精巢中共有的免疫阳性条带分子质量为45~55 ku.脑和胸神经团中另有一条36 ku条带;在精巢中,兔抗人GnRH-R和兔抗果蝇GnRH-R抗体只有一条28 ku条带,而兔抗海鞘GnRH-R抗体显示有28,36和30 ku的条带.2)利用免疫共沉淀技术,分离出与GnRH-R抗体相结合的两种物质,分子质量分别为38.1和54.0 ku,分别与哺乳动物和非哺乳动物GnRH-R的分子质量接近.该发现提示拟穴青蟹体内存在GnRH-R,为深入了解甲壳动物生殖调控机理提供了理论依据.     

雌激素受体在锯缘青蟹视神经节的免疫识别

利用免疫细胞化学SABC法对锯缘青蟹(Scylla serrata)视神经节雌激素受体(ER)进行定位研究.结果表明:视神经节的视外髓、视内髓和视端髓存在ER免疫阳性细胞.视神经节存在的ER免疫阳性细胞可以为雌激素提供结合位点.本研究为雌激素参与视神经节神经内分泌调节作用提供了形态学依据.     

中国鲎脑神经节5-羟色胺免疫组化的定位观察

中国鲎(Tachypleus tridentatus)是一种具有特殊研究意义的海洋动物.本研究应用免疫组织化学方法对中国鲎脑神经节中5-羟色胺(5-HT)的分布进行了定位观察,发现5-HT免疫阳性主要分布在视神经层和视神经髓的神经纤维,中间体以及蕈形体周边的细胞群中,而蕈形体中未发现免疫阳性.经比较,中国鲎5-HT的分布与美洲鲎(Limulus polyphemus)相似,而与昆虫、甲壳类存在一些区别.本研究可为中国鲎的神经生物学提供基础资料.     

多环芳烃—屈对三疣梭子蟹免疫影响的初步研究

研究了多环芳烃类物质屈(chrysene)对三疣梭子蟹的毒性作用.测定了该有机污染物对三疣梭子蟹血细胞密度(THC)、溶菌酶活力(LZM)、酸性磷酸酶活力(ACP)、碱性磷酸酶活力(AKP)、超氧化物歧化酶活力(SOD)、酚氧化酶活力(PO)这几种免疫因子的影响;研究了其对三疣梭子蟹免疫影响的特点.屈对三疣梭子蟹各酶类的影响有随屈浓度的增加和作用时间的加长而增强的趋势.SOD对屈最为敏感,其次是溶菌酶、血细胞密度和PO,ACP和AKP受其影响最小.     

锯缘青蟹视神经节FSH和LH的免疫识别

甲壳动物的视神经节含有性腺抑制激素(Gonad inhibiting hormone, GIH),长期以来视神经节被理解为生殖抑制效应.本研究应用脊椎动物卵泡刺激素(Fllicle stimulating hormone,FSH)抗体和黄体生成素(Luteinizing hormone,LH)抗体,从锯缘青蟹(Scylla serrata)视神经节的视外髓、视内髓和视端髓检测到FSH和LH免疫细胞化学阳性反应.锯缘青蟹视神经节中的FSH和LH免疫阳性物质,可能参与了生殖神经内分泌调节.     

壳寡糖对三疣梭子蟹免疫力的影响

为研究壳寡糖对三疣梭子蟹免疫功能的影响,设置了含壳寡糖0.0、0.5、1.0、1.5和2.0g/kg(分别为T0、T1、T2、T3和T4组)的5组试验饲料饲喂三疣梭子蟹,分别在第10 d、20 d和30 d取血清进行酚氧化酶(PO)、酸性磷酸酶(ACP)、碱性磷酸酶(AKP)、总超氧化物歧化酶(T-SOD)、溶菌酶(LSZ)和过氧化物酶(POD)活性的测定,试验结束时测定血细胞的密度.结果显示,在基础饲料中添加适量的壳寡糖可以显著性提高ACP、T-SOD、LSZ、POD活力(P＜0.05),对PO和AKP活力促进效果不显著(P＞0.05).壳寡糖可以显著提高三疣梭子蟹血细胞密度(P＜0.05),对3种血细胞所占比例有一定的影响(P＞0.05).综合考虑,T2、T3组对三疣梭子蟹免疫促进效果较好,建议基础饲料中壳寡糖添加量为1.0～1.5g/kg.     

P物质在正常与致聋白腰文鸟前脑X区表达水平的差异研究

通过免疫组织化学方法对正常和致聋不同时程的成年雄性白腰文鸟X区P物质的表达进行了定量分析,结果发现:致聋后鸟的X区P物质表达量显著减少.X区是鸣禽鸣唱学习的重要核团,P物质也是一种重要的神经调质,提示X区P物质可能参与了鸣禽鸣唱可塑性的调节或鸣唱模式的维持.     

舟山海域池塘养殖和野生三疣梭子蟹抱卵量的比较研究

系统研究和比较池塘养殖和野生三疣梭子蟹人工繁殖亲本的体重、甲壳长、甲壳宽、抱卵量、生殖力和单个卵重等指标,结果表明:1.人工育苗用野生三疣梭子蟹的体重、甲壳长、甲壳宽、抱卵量显著高于池塘养殖蟹(P<0.05);2.野生三疣梭子蟹的平均生殖力(Fecundity,粒/g)和单个卵重也大于池塘养殖梭子蟹,但是由于变异系数较大,并无统计学的差异(P>0.05);3.池塘养殖三疣梭子蟹的抱卵量与体重、甲壳长和甲壳宽相关系数高于野生梭子蟹,无论野生还是池塘养殖梭子蟹的抱卵量与体重、甲壳长和甲壳宽呈正相关,且抱卵量与甲壳长的相关系数最高。从而,总结出了利用三疣梭子蟹的甲壳长和甲壳宽来预测其抱卵量的方法。     

2个海域野生三疣梭子蟹肌肉中营养成分比较

通过分析比较2个不同海域野生三疣梭子蟹(Portunus trituberculatus)肌肉的营养成分,对其营养价值进行综合评价。采用国家标准方法测定肌肉中的水分、灰分、粗蛋白、粗脂肪、脂肪酸、氨基酸、矿物质元素、核苷酸、甜菜碱和胆固醇,根据联合国粮农组织(FAO)标准进行营养品质评价,并针对营养组成、风味物质含量同产地之间的关联进行对比分析。结果表明:2个不同海域的三疣梭子蟹在蛋白质、氨基酸、核苷酸、甜菜碱等营养组成上差异显著。浙江台州海域野生雄蟹肌肉蛋白质含量、必需氨基酸含量和总氨基酸含量均高于山东日照海域野生雄蟹,差异显著(P＜0.05);浙江台州海域野生三疣梭子蟹肌肉中肌苷酸含量、甜菜碱含量均显著高于山东日照海域(P＜0.05)。研究发现,2个海域野生三疣梭子蟹肌肉中营养成分含量存在一定差异,并对其风味及营养价值产生影响,可能与环境因素、遗传基因、摄食饵料等因素有关。     

Immunocytological evidence for the presence of vertebrate FSH and LH-like substances in the brain and thoracic ganglion of the swimming crab, Portunus trituberculatus

Investigating the distribution of follicle stimulating hormone (FSH) and luteinizing hormone (LH) in the swimming crab, Portunus trituberculatus, helps in the understanding of its reproductive regulation mechanism. This study has detected the location of vertebrate FSH- and LH-like substances in the brain and the thoracic ganglion mass with immunocytochemical techniques. Their immunoreaction is very weak or negative during the immature period and the post-spawning period, while it is stronger in the mature period. Localization of FSH-like immunoreactive cells resembles that of LH-tike cells. These immunoreactive cells were observed chiefly in the anterior median cell-cluster of the protocerebrum with stronger immunostaining. There were only a few in the inboard cell-cluster of the olfactory lobe of the deutocerebrum and the posterior cell-cluster of the tritocerebrum. Immunoreactive cells were located in all the three ganglia in the thoracic ganglion mass. These results indicated that substances resembling the vertebrate FSH and LH exist in P. trituberculatus, and their expression has some correlation with the developmental stage of ovaries, thus they might engage in the ovary development and ovulation.     

Immunocytological evidence for the presence of vertebrate FSH- and LH-like substances in the brain and thoracic ganglion of the swimming crab，Portunus trituberculatus

Investigating the distribution of follicle stimulating hormone （FSH） and luteinizing hormone （LH） in the swimming crab, Portunus trituberculatus, helps in the understanding of its reproductive regulation mechanism. This study has detected the location of vertebrate FSH- and LH-like substances in the brain and the thoracic ganglion mass with immunocytochemical techniques. Their immunoreaction is very weak or negative during the immature period and the post-spawning period, while it is stronger in the mature period. Localization of FSH-like immunoreactive cells resembles that of LH-like cells. These immunoreactive cells were observed chiefly in the anterior median cell-duster of the protocerebrum with stronger immunostaining. There were only a few in the inboard cell-cluster of the olfactory lobe of the deutocerebrum and the posterior cell-cluster of the tritocerebrum. Immunoreactive cells were located in all the three ganglia in the thoracic ganglion mass. These results indicated that substances resembling the vertebrate FSH and LH exist in P. trituberculatus, and their expression has some correlation with the developmental stage of ovaries, thus they might engage in the ovary development and ovulation.     

Immunorecognition of estrogen and androgen receptors in the brain and thoracic ganglion mass of mud crab, Scylla paramamosain

The brain and the thoracic ganglion of a crustacean can synthesize and secrete gonad-stimulating hormone (GSH) which stimulates the maturation of gonad. In the previous experiments, sex steroid hormones (estradiol, testosterone, progesterone, etc.) have been detected from the crustacean. However, the feedback regulation of sex steroid hormones on the brain and the thoracic ganglion of the crustacean has not been reported so far. In the present experiment, monoclonal antibodies were applied to investigate the immunorecognition of estrogen receptor (ER) and androgen receptor (AR) in the brain and the thoracic ganglion mass of Scylla paramamosain. The results showed that the distribution of the immunopositive substances of ER and AR was extremely similar. They distributed in the protocerebrum, deutocerebrum and tritocerebrum of the brain, and mainly in protocerebrum. In the thoracic ganglion mass, immunopositive substances distributed in the subesophageal ganglion, thoracic ganglion and abdominal ganglion, and mostly in subesophageal ganglion. Immunopositive substances of ER and AR mostly existed in the cytoplasm of neurons. The present study will provide morphological evidence for the origin and the evolution of ER and AR. In addition, the immunoreactivities of ER and AR suggested that the estrogen and androgen may be involved in the feedback regulation of crustacean neuroendocrine.     

Immunorecognition of estrogen and androgen receptors in the brain and thoracic ganglion mass of mud crab, Scylla paramamosain

The brain and the thoracic ganglion of crustacean can synthesize and secrete gonad-stimulating hormone (GSH) which stimulates the maturation of gonad. In the previous experiments, sex steroid hormones (estradiol, testosterone, progesterone, etc.) have been detected from crustacean. However, the feedback regulation of sex steroid hormones on the brain and thoracic ganglion of crustacean has not been reported so far. In the present experiment, monoclonal antibodies were applied to investigate the immunorecognition of estrogen receptor (ER) and androgen receptor (AR) in the brain and thoracic ganglion mass of Scylla paramamosain. The results showed that the distribution of the immunopositive substances of ER and AR was extremely similar. They distributed in the protocerebrum, deutocerebrum and tritocerebrum of the brain, and mainly in protocerebrum. In the thoracic ganglion mass, immunopositive substances distributed in the subesophageal ganglion, thoracic ganglion and abdomen ganglion, and mostly in subesophageal ganglion. Immunopositive substances of ER and AR mostly existed in the cytoplasm of neurons. The present study will provide morphological evidence for the origin and evolution of ER and AR. In addition, the immunoreactivities of ER and AR suggested that the estrogen and androgen may be involved in the feedback regulation of crustacean neuroendocrine.     

Immunorecognition of estrogen and androgen receptors in the brain and thoracic ganglion mass of mud crab, Scylla paramamosain

The brain and the thoracic ganglion of crustacean can synthesize and secrete gonad-stimulating hormone (GSH) which stimulates the maturation of gonad. In the previous experiments, sex steroid hormones (estradiol, testosterone, progesterone, etc.) have been detected from crustacean. However, the feedback regulation of sex steroid hormones on the brain and thoracic ganglion of crustacean has not been reported so far. In the present experiment, monoclonal antibodies were applied to investigate the immunorecognition of estrogen receptor (ER) and androgen receptor (AR) in the brain and thoracic ganglion mass of Scylla paramamosain. The results showed that the distribution of the immunopositive substances of ER and AR was extremely similar. They distributed in the protocerebrum, deutocerebrum and tritocerebrum of the brain, and mainly in protocerebrum. In the thoracic ganglion mass, immunopositive substances distributed in the subesophageal ganglion, thoracic ganglion and abdomen ganglion, and mostly in subesophageal ganglion. Immunopositive substances of ER and AR mostly existed in the cytoplasm of neurons. The present study will provide morphological evidence for the origin and evolution of ER and AR. In addition, the immunoreactivities of ER and AR suggested that the estrogen and androgen may be involved in the feedback regulation of crustacean neuroendocrine.     

Substance P-immunoreactive retinal ganglion cells and their central axon terminals in the rabbit

Retinal ganglion cells are the projection neurons that link the retina to the brain. Peptide immunoreactive cells in the ganglion cell layer (GCL) of the mammalian retina have been noted but their identity has not been determined. We now report that, in the rabbit, 25-35% of all retinal ganglion cells contain substance P-like (SP) immunoreactivity. They were identified by either retrograde transport of fluorescent tracers injected into the superior colliculus, or by retrograde degeneration after optic nerve section. SP immunoreactive cells are present in all parts of the retina and have medium to large cell bodies with dendrites that ramify extensively in the proximal inner plexiform layer. Their axons terminate in the dorsal lateral geniculate nucleus, superior colliculus and accessory optic nuclei, and these terminals disappear completely after contralateral optic nerve section and/or eye enucleation. In the dorsal lateral geniculate nucleus large, beaded, immunoreactive axons and varicosities make up a narrow plexus just below the optic tract, where they define a new geniculate lamina. The varicosities make multiple synaptic contacts with dendrites of dorsal lateral geniculate nucleus projection neurons and presumptive interneurons in complex glomerular neuropil. This is direct evidence that some mammalian retinal ganglion cells contain substance P-like peptides and strongly suggests that, in the rabbit, substance P (or related tachykinins) may be a transmitter or modulator in a specific population or populations of retinal ganglion cells.     

榆紫叶甲 Ambrostoma quadriim-pressum脑及咽下神经节GABA阳性神经元分布模式的构建

采用树脂石蜡(CP)组织包埋切片技术,结合免疫组化链霉菌抗生物素蛋白-过氧化物酶(SP)双染法,初步构建了榆紫叶甲脑及咽下神经节GABA阳性神经元的分布模式.结果显示,榆紫叶甲脑结构比较特殊:视叶不发达,但具内髓板;嗅叶不发达;蕈形体冠高度退化,而根叶相对发达;中央复合体结构明显;后脑经由围咽神经束与咽下神经结相连,咽下神经结非常发达.榆紫叶甲的所有髓区几乎都有GABA免疫阳性纤维分布.一侧脑半球及咽下神经节共存在约2 800个GABA阳性神经元,分布广泛,且趋向于集群分布.该研究表明榆紫叶甲的脑结构及GABA阳性神经元的分布模式与榆紫叶甲的生活习性、进化地位等密切相关.