Nitric oxide–cyclic guanosine monophosphate signaling in the local circuit of the cricket abdominal nervous system
Nitric oxide–cyclic guanosine monophosphate signaling in the local circuit of the cricket abdominal nervous system
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DOI：
10.1016/j.neuroscience.2008.09.032
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发表时间：
2008-12
期刊：
影响因子：
3.3
通讯作者：
H. Aonuma;Y. Kitamura;K. Niwa;H. Ogawa;K. Oka
中科院分区：
文献类型：
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作者：
H. Aonuma;Y. Kitamura;K. Niwa;H. Ogawa;K. Oka
观察了蟋蟀腹末神经节（TAG）内一氧化氮（NO）电位供体神经元和一氧化氮反应靶神经元的分布。免疫印迹法检测神经系统一氧化氮合酶（NOS）的表达，采用通用型一氧化氮合酶（uNOS）抗体，提供约130 kDa的蛋白带。免疫组织化学使用uNOS抗体检测胞体位于TAG外侧区域的神经元。这些神经元将其神经元分支扩展到背正中区或背外侧区。采用NADPH-diaphorase组织化学方法证实nos神经元的分布。细胞体和染色神经元分支的分布与uNOS免疫组化相似。采用no诱导的cGMP免疫组化方法揭示no应答的靶神经元。免疫组化染色的大部分细胞体出现在TAG的背侧。背正中区部分未配对的神经元细胞体染色强烈。部分轴突支配各神经根的传出神经元染色强烈。采用NO电极检测TAG中NO的生成情况。我们发现一氧化氮的产生通常维持在70 nM的基础浓度。清除的血红蛋白从神经节释放一氧化氮。用生理盐水替代血红蛋白后，一氧化氮浓度部分恢复。应用作为NOS底物的10 μM l-精氨酸可使NO释放量增加约10 nM。此外，兴奋性神经递质乙酰胆碱（ACh）在基础水平70 nM的基础上，也增加了约40-50 nM的NO生成。荧光NO-指示剂光学成像显示，乙酰胆碱诱导的一氧化氮释放在TAG外缘区短暂增强，这是nos免疫反应神经元的细胞体所在区域。这些结果表明，ACh通过ACh在TAG中激活的神经元事件加速NO的产生。
The distribution of potential nitric oxide (NO) donor neurons and NO-responsive target neurons was revealed in the terminal abdominal ganglion (TAG) of the cricket. The expression of nitric oxide synthase (NOS) in the nervous system was examined by Western blotting using universal nitric oxide synthase (uNOS) antibody that gave about a 130 kDa protein band. Immunohistochemistry using the uNOS antibody detected neurons whose cell bodies are located at the lateral region of the TAG. These neurons expanded their neuronal branches into the dorsal–median region or the dorsal–lateral region of the TAG. NADPH-diaphorase histochemistry was performed to confirm the distribution of NOS-containing neurons. The distributions of cell bodies and stained neuronal branches were similar to those revealed by uNOS immunohistochemistry. NO-induced cGMP immunohistochemistry was performed to reveal NO-responsive target neurons. Most of the cell bodies stained by immunohistochemistry appeared at the dorsal side of the TAG. At the dorsal–median region, some unpaired neuronal cell bodies were strongly stained. Some efferent neurons whose axon innervate into each nerve root were strongly stained. The generation of NO in the TAG was detected by NO electrode. We found that NO is generally produced to maintain a basal concentration of 70 nM. Hemoglobin scavenged released NO from the ganglion. The concentration of NO was partly recovered when hemoglobin was replaced by normal saline. Application of 10 μM l-arginine that is a substrate of NOS increased NO release by approximately 10 nM. Furthermore, an excitatory neurotransmitter acetylcholine (ACh) also increased NO generation by approximately 40–50 nM in concentration in addition to the basal level of 70 nM. Optical imaging with fluorescent NO-indicator demonstrated that ACh-induced enhancement of NO release was transiently observed in the outer-edge region of TAG, where cell bodies of NOS-immunoreactive neurons were located. These results suggest that ACh accelerates NO production via neuronal events activated by ACh in the TAG.