Muñoz M, Muñoz A, Marín O, Alonso J R, Arévalo R, Porteros A, González A
Department of Cell Biology, University Complutense of Madrid, Spain.
J Comp Neurol. 1996 Mar 25;367(1):54-69. doi: 10.1002/(SICI)1096-9861(19960325)367:1<54::AID-CNE5>3.0.CO;2-I.
The distribution of NADPH-diaphorase (ND) activity was histochemically investigated in the brain of the frog Rana perezi. This technique provides a highly selective labeling of neurons and tracts. In the telencephalon, labeled cells are present in the olfactory bulb, pallial regions, septal area, nucleus of the diagonal band, striatum, and amygdala. Positive neurons surround the preoptic and infundibular recesses of the third ventricle. The magnocellular and suprachiasmatic hypothalamic nuclei contain stained cells. Numerous neurons are present in the anterior, lateral anterior, central, and lateral posteroventral thalamic nuclei. Positive terminal fields are organized in the same thalamic areas but most conspicuously in the visual recipient plexus of Bellonci, corpus geniculatum of the thalamus, and the superficial ventral thalamic nucleus. Labeled fibers and cell groups are observed in the pretectal area, the mesencephalic optic tectum, and the torus semicircularis. The nuclei of the mesencephalic tegmentum contain abundant labeled cells and a conspicuous cell population is localized medial and caudal to the isthmic nucleus. Numerous cells in the rhombencephalon are distributed in the octaval area, raphe nucleus, reticular nuclei, sensory trigeminal nuclei, nucleus of the solitary tract, and, at the obex levels, the dorsal column nucleus. Positive fibers are abundant in the superior olivary nucleus, the descending trigeminal, and the solitary tracts. In the spinal cord, a large population of intensely labeled neurons is present in all fields of the gray matter throughout its rostrocaudal extent. Several sensory pathways were heavily stained including part of the olfactory, visual, auditory, and somatosensory pathways. The distribution of ND-positive cells did not correspond to any single known neurotransmitter or neuroactive molecule system. In particular, abundant codistribution of ND and catecholamines is found in the anuran brain. Double labeling techniques have revealed restricted colocalization in the same neurons and only in the posterior tubercle and locus coeruleus. If ND is in amphibians a selective marker for neurons containing nitric oxide synthase, as generally proposed, with this method the neurons that may synthesize nitric oxide would be identified. This study provides evidence that nitric oxide may be involved in novel tasks, primarily related to forebrain functions, that are already present in amphibians.
采用组织化学方法研究了泽陆蛙脑内还原型辅酶Ⅱ黄递酶（ND）的活性分布。该技术能对神经元和神经束进行高度选择性标记。在端脑，标记细胞见于嗅球、大脑皮质区、隔区、斜角带核、纹状体和杏仁核。阳性神经元环绕第三脑室的视前隐窝和漏斗隐窝。下丘脑的大细胞和视交叉上核含有染色细胞。丘脑前核、丘脑前外侧核、丘脑中央核和丘脑后外侧腹核有大量神经元。阳性终末野分布于相同的丘脑区域，但在贝龙奇视觉接受丛、丘脑外侧膝状体和丘脑腹外侧浅核最为明显。在顶盖前区、中脑视顶盖和半规管隆起可见标记纤维和细胞群。中脑被盖核含有丰富的标记细胞，在峡核的内侧和尾侧有一群明显的细胞。延髓内的许多细胞分布于听神经核区、中缝核、网状核、三叉神经感觉核、孤束核，在闩平面还有薄束核。上橄榄核、三叉神经降支和孤束中有丰富的阳性纤维。在脊髓中，灰质各节段均有大量强标记神经元。包括部分嗅觉、视觉、听觉和躯体感觉通路在内的几条感觉通路被重度染色。ND阳性细胞的分布与任何已知的单一神经递质或神经活性分子系统均不对应。特别是在无尾两栖类动物脑中发现ND与儿茶酚胺大量共分布。双重标记技术显示二者仅在同一神经元的后结节和蓝斑有限共定位。如果如普遍认为的那样，在两栖动物中ND是含一氧化氮合酶神经元的选择性标记，那么用这种方法就可以识别可能合成一氧化氮的神经元。本研究提供了证据，表明一氧化氮可能参与了主要与前脑功能相关的新任务，而这些任务在两栖动物中已经存在。
