| 摘要: |
| 采用组织化学、免疫组化和生物化学方法对栉孔扇贝外套膜中的一氧化氮合酶(EC1.14.13.39，NOS)活性进行了研究。组织化学显示结果表明，帆状部上皮细胞呈NOS强阳性，血细胞和神经纤维呈阳性；外套触手内有较多近圆形细胞和大量波浪形神经纤维呈NOS强阳性；边缘膜内有大量神经细胞和神经纤维分别呈NOS强阳性和阳性，外套环走肌束附近和血管内皮周围均有大量近圆形的强阳性细胞分布；中央膜有大量近圆形阳性细胞聚集成团块或分散分布。免疫组化显示表明，血细胞、帆状部和边缘膜上皮细胞呈神经型NOS(nNOS)和内皮型NOS(eNOS)弱阳性，而呈诱导型NOS(iNOS)阳性。生化测定结果表明，总NOS(tNOS)活力和一氧化氮(NO)含量均为中央膜最高，边缘膜次之，外套触手较低，帆状部最低，其中结构型NOS(cNOS)活力也是帆状部的最低，而iNOS活力则是外套触手的最低。栉孔扇贝外套膜可能是其神经系统中NOS的发生中心，NO-NOS体系可在其免疫防御和调节等方面发挥重要的作用。 |
| 关键词: 栉孔扇贝 外套膜 一氧化氮合酶 一氧化氮 酶活性 |
| DOI： |
| 分类号: |
| 基金项目:国家973计划项目资助,G1999012005号 |
附件 |
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| ACTIVITIES OF NITRIC OXIDE SYNTHASE IN THE MANTLE OF SCALLOP CHLAMYS FARRERI |
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SUN Hu-Shan,WANG Yi-Yan,WANG Xiao-An,SUN Xiu-Qin,LI Guang-You
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1.College of Life Science,Ludong University,Yantai,264025;2.First Institute of Oceanography,State Oceanic Administration,Qingdao,266061
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| Abstract: |
| The nitric oxide synthase (ECl.l4.l3.39; NOS) in the mantle of scallop Chlamys farreri (obtained from Zhifu Bay, of the north of Yellow Sea; shell length 45-50 mm) were studied with methods of histochemical localization, immunohistochemical localization and spectrophotometric assay. The aim of this study is to determine the role of NOS in the immune response of molluscs.
The histochemical analysis results were as follow. In the inner mantle fold, the epithelial cells were strongly NOS-positive, the haemocytes and nerve fibers in the connective tissue were NO S-positive, the muscle fibers were weakly NOS-positive. In the mantle tentacle, many round cells and wavelike nerve fibers presented strongly NOS-positivity. In the margin of mantle, there were many strongly NOS positive nerve cells and positive nerve fibers; abundant strongly NOS-positive round cells were detected near the pallial retractor muscle and inner epithelia of blood vessel. In the center of mantle, the distribution of NOS-positive cells were very different in different parts, abundant round NOS-positive cells distributed in gathering or scattering in the epithelial tissue, there were many round NOS-positive cells near the inner epithelia of blood vessel too.
The immunohistochemical analysis showed that the haemocytes and epithelial cells of the inner mantle fold and margin of mantle presented weakly positive reaction of neuronal nitric oxide synthase (nNOS) and endothelial nitric oxide synthase (eNOS), and presented positive reaction of inducible nitric oxide synthase (iNO S). The nerve cells, nerve fibers and inner epithelial cells of blood vessel presented nNO S, eNOS and iNOS negative reaction.
The biochemical test showed that both NOS and NO were detected in each parts of mantle. The enzyme activity level of total nitric oxide synthase (?\TOS) and contents of NO in center of mantle were the highest in all parts of the mantle, the ?\TOS activity and contents of NO in mantle margin were higher than the mantle tentacle or the inner mantle fold, and the lowest part was the inner mantle fold. In both the central mantle and inner mantle fold, the iNOS activity was higher than cNOS activity; in the margin of mantle, the iNOS activity level was close to the cNOS activity level; in the mantle tentacle, the iNOS activity level was much lower than the cNOS activity level.
The results suggested that the NO-NOS system in mantle of C. farreri may play an important role in its immunological defense and immunoregulation, and the mantle may be the produce center of NOS. |
| Key words: Scallop Chlamys farreri, Mantle, Nitric oxide synthase, Nitric Oxide, Enzyme activity |
