Watson J T, Adkins-Regan E, Whiting P, Lindstrom J M, Podleski T R
Section of Neurobiology and Behavior, Cornell University, Ithaca, New York 14853.
J Comp Neurol. 1988 Aug 8;274(2):255-64. doi: 10.1002/cne.902740209.
We have localized nicotinic acetylcholine receptors in the zebra finch brain by using three 125I-labelled ligands: alpha bungarotoxin and two monoclonal antibodies to neuronal nicotinic receptors (MAb 35 of Tzardos et al., J. Biol. Chem., 250: 8635-8645, '81; and MAb 270 of Whiting and Lindstrom: J. Neurosci. 6: 3061-3069, '86). Unfixed brains from intact adult male and female zebra finches were prepared for in vitro autoradiography. Low-resolution film autoradiograms and high-resolution emulsion autoradiograms were prepared for each of the three ligands. The major brain structures that bind all three of the ligands are hippocampus; hyperstriatum dorsalis; hyperstriatum ventralis; nucleus lentiformis mesencephali; nucleus pretectalis, some layers of the optic tectum; nucleus mesencephalicus lateralis; pars dorsalis; locus ceruleus; and all cranial motor nuclei except nucleus nervi hypoglossi. The major structures labelled only by [125I]-alpha bungarotoxin binding included hyperstriatum accessorium and the nuclei: preopticus medialis, medialis hypothalami posterioris, semilunaris, olivarius inferior, and the periventricular organ. Of the song control nuclei, nucleus magnocellularis of the anterior neostriatum; hyperstriatum ventralis, pars caudalis; nucleus intercollicularis; and nucleus hypoglossus were labelled. The binding patterns of the two antibodies were similar to one another but not identical. Both labelled nucleus spiriformis lateralis and nucleus geniculatus lateralis, pars ventralis especially heavily and also labelled the nucleus habenula medialis; nucleus subpretectalis; nucleus isthmi, pars magnocellularis; nucleus reticularis gigantocellularis; nucleus reticularis lateralis; nucleus tractus solitarii; nucleus vestibularis dorsolateralis; nucleus vestibularis lateralis; nucleus descendens nervi trigemini; and the deep cerebellar nuclei. Lobus parolfactorius and nucleus vestibularis medialis were labelled by only MAb 270, whereas only MAb 35 labelled nucleus laminaris and the medial and lateral pontine nuclei. These data extend previous reports of cholinergic participation in the song system (Ryan and Arnold: J. Comp. Neurol. 202: 211-219, '81) to suggest that the zebra finch song system may contain several closely related nicotinic receptors. In several brain nuclei it appeared that certain anatomical portions of a nucleus or a certain class of neurons were specifically labelled. Furthermore, in certain cases, the labelling appeared to be clustered around Nissl-stained cell nuclei, thus suggesting that the receptors are concentrated on or in somata.
我们通过使用三种125I标记的配体,将烟碱型乙酰胆碱受体定位在斑胸草雀脑中:α-银环蛇毒素以及两种针对神经元烟碱型受体的单克隆抗体(Tzardos等人的单克隆抗体35,《生物化学杂志》,250: 8635 - 8645, '81;以及Whiting和Lindstrom的单克隆抗体270:《神经科学杂志》6: 3061 - 3069, '86)。对成年雄性和雌性完整斑胸草雀的未固定大脑进行体外放射自显影准备。针对这三种配体分别制备了低分辨率胶片放射自显影片和高分辨率乳胶放射自显影片。与所有三种配体结合的主要脑结构有海马体;背侧上纹状体;腹侧上纹状体;中脑豆状核;顶盖前核,视顶盖的一些层;中脑外侧核;背侧部;蓝斑;以及除舌下神经核外的所有颅运动核。仅由[125I]-α-银环蛇毒素结合标记的主要结构包括副上纹状体以及以下核团:内侧视前核、下丘脑后部内侧、半月核、下橄榄核以及脑室周器官。在发声控制核团中,新纹状体前部的大细胞核;腹侧上纹状体、尾侧部;间丘核;以及舌下神经核被标记。两种抗体的结合模式彼此相似但不完全相同。二者都特别强烈地标记了外侧螺旋核和外侧膝状体核、腹侧部,还标记了内侧缰核;顶盖前下核;峡核、大细胞部;巨细胞网状核;外侧网状核;孤束核;背外侧前庭核;外侧前庭核;三叉神经降核;以及小脑深部核团。嗅觉旁叶和内侧前庭核仅被单克隆抗体270标记,而仅单克隆抗体35标记了层状核以及脑桥内侧核和外侧核。这些数据扩展了先前关于胆碱能参与发声系统的报道(Ryan和Arnold:《比较神经学杂志》202: 211 - 219, '81),表明斑胸草雀发声系统可能包含几种密切相关的烟碱型受体。在几个脑核中,似乎一个核的某些解剖部分或某一类神经元被特异性标记。此外,在某些情况下,标记似乎聚集在尼氏染色的细胞核周围,因此表明受体集中在胞体上或胞体内。