Perroteau I, Danger J M, Biffo S, Pelletier G, Vaudry H, Fasolo A
Dipartimento di Biologia Animale, Lab. Anatomia Comparata, Torino, Italy.
J Comp Neurol. 1988 Sep 15;275(3):309-25. doi: 10.1002/cne.902750302.
The newt brain represents a simplified model for the increasingly complex vertebrate neuronal organization. The localization of neuropeptide Y-like (NPY-like) containing neurons in the brain of Triturus cristatus was studied by means of indirect immunofluorescence, peroxidase-antiperoxidase, and avidin-biotin techniques using a highly specific antiserum. NPY-like positive cell bodies were observed in several areas, most notably in the telencephalon (primordium hippocampi and amygdaloid complex), the preoptic and suprachiasmatic areas, the hypothalamus, the dorsal thalamus, the tegmentum, and the rhombencephalon (laterolateral grey column and raphe area). Nerve fibres were particularly abundant in the pallium, striatum, septum, amygdaloid, preoptic neuropils, and pars intercalaris diencephali. Bundles of NPY-immunoreactive fibres also were visualized in the dorsal thalamus and in the posterior hypothalamus. The pars intermedia lacked any NPY-like positive fibres. Neuronal processes also were found in the tectum mesencephali and in the body of the cerebellum. A prominent NPY-like fibre network was observed in the octavolateralis. Concentrations of NPY measured by means of a specific radioimmunoassay were threefold higher in the hypothalamus (15.2 +/- 1.3 ng/mg proteins) than in the rhombencephalon (4.9 +/- 0.3) and the mesencephalon (4.3 +/- 0.2). The concentration found in the telencephalon was 2.1 +/- 0.3 ng/mg proteins. Sephadex G-50 gel chromatography of whole brain extracts indicated the presence of high molecular weight forms of NPY-like material in addition to the authentic peptide. Both amphibian and mammalian NPY peptides had an apparent molecular weight of 4,000 daltons, as evidenced by immunoblotting analysis. High-performance liquid chromatography demonstrated, however, that the newt peptide was slightly less hydrophobic than porcine NPY. The present findings indicate that NPY-immunoreactive neurons are widely distributed in the brain of urodeles. Our data indicate that the NPY molecule has been relatively well preserved during evolution.
蝾螈脑代表了日益复杂的脊椎动物神经元组织的一个简化模型。利用高度特异性抗血清，通过间接免疫荧光、过氧化物酶-抗过氧化物酶和抗生物素蛋白-生物素技术，研究了有冠欧螈脑中含神经肽Y样（NPY样）神经元的定位。在几个区域观察到NPY样阳性细胞体，最显著的是在端脑（海马原基和杏仁复合体）、视前区和视交叉上区、下丘脑、背侧丘脑、被盖以及后脑（外侧灰质柱和中缝区）。神经纤维在大脑皮质、纹状体、隔膜、杏仁核、视前神经毡和间脑中间部特别丰富。在背侧丘脑和下丘脑后部也可见到NPY免疫反应性纤维束。中间部没有任何NPY样阳性纤维。在中脑顶盖和小脑体中也发现了神经元突起。在八侧器中观察到一个显著的NPY样纤维网络。通过特异性放射免疫测定法测得的下丘脑NPY浓度（15.2±1.3 ng/mg蛋白质）比后脑（4.9±0.3）和中脑（4.3±0.2）高三倍。端脑中发现的浓度为2.1±0.3 ng/mg蛋白质。全脑提取物的Sephadex G - 50凝胶色谱表明，除了真实肽外，还存在高分子量形式的NPY样物质。免疫印迹分析表明，两栖类和哺乳类的NPY肽的表观分子量均为4000道尔顿。然而，高效液相色谱显示，蝾螈肽的疏水性略低于猪NPY。目前的研究结果表明，NPY免疫反应性神经元广泛分布于有尾两栖类动物的脑中。我们的数据表明，NPY分子在进化过程中得到了较好的保留。
