Nighorn A, Gibson N J, Rivers D M, Hildebrand J G, Morton D B
Arizona Research Laboratories, Division of Neurobiology, University of Arizona, Tucson, Arizona 85721, USA.
J Neurosci. 1998 Sep 15;18(18):7244-55. doi: 10.1523/JNEUROSCI.18-18-07244.1998.
The nitric oxide (NO)-cGMP signaling system is thought to play important roles in the function of the olfactory system in both vertebrates and invertebrates. One way of studying the role of NO in the nervous system is to study the distribution and properties of NO synthase (NOS), as well as the soluble guanylyl cyclases (sGCs), which are the best characterized targets of NO. We study NOS and sGC in the relatively simple and well characterized insect olfactory system of the hawkmoth, Manduca sexta. We have cloned Manduca sexta nitric oxide synthase (MsNOS) and two sGCs (MsGCalpha1 and MsGCbeta1), characterized their basic biochemical properties, and studied their expression in the olfactory system. The sequences of the Manduca genes are highly similar to their mammalian homologs and show similar biochemical properties when expressed in COS-7 cells. In particular, we find that MsGC functions as an obligate heterodimer that is stimulated significantly by NO. We also find that MsNOS has a Ca2+-sensitive NO-producing activity similar to that of mammalian neuronal NOS. Northern and in situ hybridization analyses show that MsNOS and the MsGCs are expressed in a complementary pattern, with MsNOS expressed at high levels in the antennae and the MsGCs expressed at high levels in a subset of antennal lobe neurons. The expression patterns of these genes suggest that the NO-sGC signaling system may play a role in mediating communication between olfactory receptor neurons and projection neurons in the glomeruli of the antennal lobe.
一氧化氮(NO)-环磷酸鸟苷(cGMP)信号系统被认为在脊椎动物和无脊椎动物的嗅觉系统功能中发挥着重要作用。研究NO在神经系统中作用的一种方法是研究一氧化氮合酶(NOS)以及可溶性鸟苷酸环化酶(sGCs)的分布和特性,sGCs是NO最具特征的作用靶点。我们在相对简单且特征明确的烟草天蛾(Manduca sexta)昆虫嗅觉系统中研究NOS和sGC。我们克隆了烟草天蛾一氧化氮合酶(MsNOS)和两种sGC(MsGCalpha1和MsGCbeta1),表征了它们的基本生化特性,并研究了它们在嗅觉系统中的表达。烟草天蛾基因的序列与它们的哺乳动物同源物高度相似,并且在COS-7细胞中表达时表现出相似的生化特性。特别地,我们发现MsGC作为一种专一性异二聚体发挥作用,受到NO的显著刺激。我们还发现MsNOS具有类似于哺乳动物神经元NOS的对Ca2+敏感的产NO活性。Northern杂交和原位杂交分析表明,MsNOS和MsGCs以互补模式表达,MsNOS在触角中高水平表达,而MsGCs在触角叶神经元的一个子集中高水平表达。这些基因的表达模式表明,NO-sGC信号系统可能在介导嗅觉受体神经元与触角叶小球中的投射神经元之间的通讯中发挥作用。