Spike R C, Todd A J, Johnston H M
Department of Anatomy, University of Glasgow, United Kingdom.
J Comp Neurol. 1993 Sep 15;335(3):320-33. doi: 10.1002/cne.903350303.
The enzyme NADPH diaphorase is present in many spinal neurons, and is thought to correspond to nitric oxide synthase. In order to determine which types of neuron in the spinal cord contain this enzyme, we have carried out a combined enzyme histochemical and immunocytochemical study with antibodies to GABA, glycine, and choline acetyltransferase. Two hundred rats were tested for GABA- and glycine-like immunoreactivity. The majority of these neurons (207/224) were GABA-immunoreactive and 139 were also glycine-immunoreactive. NADPH diaphorase-positive neurons in laminae I and II generally showed both types of immunoreactivity, while those in deeper laminae of the dorsal horn and around the central canal either showed both types or else were only GABA-immunoreactive. Since GABA and acetylcholine are thought to coexist in spinal neurons, NADPH diaphorase staining was combined with immunostaining for choline acetyltransferase. Immunoreactive neurons in laminae III and IV were all NADPH diaphorase-positive, while only some of those around the central canal and in the deeper laminae of the dorsal horn were positive. Choline acetyltransferase-immunoreactive neurons in the intermediolateral cell column (presumed sympathetic preganglionic neurons) were often NADPH diaphorase-positive, whereas those in the ventral horn (presumed motoneurons) were not. NADPH diaphorase-positive cells in the intermediolateral cell column were not immunoreactive with GABA or glycine antibodies.
NADPH 黄递酶存在于许多脊髓神经元中，被认为与一氧化氮合酶相对应。为了确定脊髓中哪些类型的神经元含有这种酶，我们使用针对γ-氨基丁酸（GABA）、甘氨酸和胆碱乙酰转移酶的抗体进行了酶组织化学和免疫细胞化学联合研究。对 200 只大鼠进行了 GABA 和甘氨酸样免疫反应性检测。这些神经元中的大多数（207/224）呈 GABA 免疫反应性，其中 139 个也呈甘氨酸免疫反应性。I 层和 II 层中的 NADPH 黄递酶阳性神经元通常表现出两种免疫反应性，而背角深层和中央管周围的那些神经元要么表现出两种类型，要么仅呈 GABA 免疫反应性。由于 GABA 和乙酰胆碱被认为在脊髓神经元中共存，因此将 NADPH 黄递酶染色与胆碱乙酰转移酶免疫染色相结合。III 层和 IV 层中的免疫反应性神经元均为 NADPH 黄递酶阳性，而中央管周围和背角深层中的只有一些呈阳性。中间外侧细胞柱（推测为交感神经节前神经元）中的胆碱乙酰转移酶免疫反应性神经元通常为 NADPH 黄递酶阳性，而腹角中的（推测为运动神经元）则不是。中间外侧细胞柱中的 NADPH 黄递酶阳性细胞与 GABA 或甘氨酸抗体无免疫反应性。
