Kishimoto J, Keverne E B, Hardwick J, Emson P C
Department of Neurobiology, AFRC, Babraham Institute, Cambridge, UK.
Eur J Neurosci. 1993 Dec 1;5(12):1684-94. doi: 10.1111/j.1460-9568.1993.tb00236.x.
The distribution of nitric oxide synthase (NOS) in the mouse olfactory bulb and olfactory epithelium, including the vomeronasal organ, was studied using an anti-NOS antibody, NADPH diaphorase histochemistry and in situ hybridization with NOS specific antisense oligonucleotide probes. Interneurons containing NOS protein and mRNA, and exhibiting NADPH diaphorase activity were detected in the plexiform layer of the main olfactory bulb and the granule cell layer of main and accessory olfactory bulbs. Periglomerular cells and granule cells in the main olfactory bulb were also NOS positive with diaphorase and immunostaining for NOS. In contrast, no evidence for NOS expression was found either in the main olfactory epithelium or in the vomeronasal organ, in spite of the strong diaphorase staining of the surface of the main olfactory epithelium. Polymerase chain reaction amplification experiments for detection of NOS gene expression further indicated that NOS is expressed in the olfactory bulb but not in either the main olfactory epithelium or vomeronasal organ. Use of an antibody raised against another enzyme, NADPH-P450 oxidoreductase, showed that this protein was strongly expressed in the olfactory epithelium. Activity of this enzyme may account for the diaphorase histochemical staining of the epithelia. An involvement of neuronal nitric oxide synthase in signalling in olfactory receptor neurons is therefore doubtful, although NOS is clearly expressed in neurons in both main and accessory olfactory bulbs.
利用抗一氧化氮合酶（NOS）抗体、NADPH 黄递酶组织化学以及与 NOS 特异性反义寡核苷酸探针进行原位杂交，研究了一氧化氮合酶（NOS）在小鼠嗅球和嗅上皮（包括犁鼻器）中的分布。在主嗅球的丛状层以及主、副嗅球的颗粒细胞层中，检测到了含有 NOS 蛋白和 mRNA 并表现出 NADPH 黄递酶活性的中间神经元。主嗅球中的球周细胞和颗粒细胞经黄递酶和 NOS 免疫染色也呈 NOS 阳性。相比之下，尽管主嗅上皮表面有强烈的黄递酶染色，但在主嗅上皮或犁鼻器中均未发现 NOS 表达的证据。用于检测 NOS 基因表达的聚合酶链反应扩增实验进一步表明，NOS 在嗅球中表达，但在主嗅上皮或犁鼻器中均不表达。使用针对另一种酶 NADPH - P450 氧化还原酶产生的抗体表明，该蛋白在嗅上皮中强烈表达。这种酶的活性可能是上皮细胞黄递酶组织化学染色的原因。因此，尽管 NOS 在主、副嗅球的神经元中均有明显表达，但神经元型一氧化氮合酶在嗅觉受体神经元信号传导中的作用仍值得怀疑。
