Oleskevich S, Descarries L, Lacaille J C
(Départment de physiologie), Faculté de médecine, Université de Montréal, Québec, Canada.
J Neurosci. 1989 Nov;9(11):3803-15. doi: 10.1523/JNEUROSCI.09-11-03803.1989.
A recently developed radioautographic technique, based on the uptake labeling of monoamine terminals (axonal varicosities) in vitro, was used to quantify the noradrenaline (NA) innervation in adult rat hippocampus. After incubation of brain slices with 1 microM 3H-NA, the NA varicosities were visualized as small aggregates of silver grains, in light microscope radioautographs prepared at 3 equidistant horizontal levels across the ventral 2/3 of the hippocampus. Using a computer-assisted image analyzer, counts were obtained from the subiculum (SUB), 3 sectors of Ammon's horn (CA1, CA3-a, CA3-b) and 3 sectors of the dentate gyrus (DG-medial blade, crest, and lateral blade), every lamina being sampled in each region. After a double correction for duration of radioautographic exposure and section thickness, and following measurement of varicosity diameter in electron microscope radioautographs, it was possible to express these results in number of terminals per volumetric unit of tissue. It was thus found that the overall density of hippocampal NA innervation averages 2.1 million varicosities/mm3 of tissue, a value almost twice as high as that in cerebral cortex. This innervation is 20% denser ventrally than dorsally and is heterogeneous both in terms of regional and laminar distribution. SUB and DG are more strongly innervated than Ammon's horn, wherein CA1 has the lowest overall density. In SUB and CA1, there is a clear predilection of NA varicosities for the stratum moleculare. In CA3, there is a narrow band of even stronger innervation in the stratum radiatum, near the apical border of the stratum pyramidale, contrasting with a 3 times lower density in this cell layer and the stratum oriens. In DG, the NA innervation is again the weakest in the cell body layer (granule) and exhibits an almost 3-fold greater density in the polymorph layer, the highest of all hippocampus. These figures allow for numerous correlations with other quantitative parameters--cytological, biochemical, and pharmacological--of NA function in the hippocampus. They also provide a strong basis for elucidating, at a cellular level, the action of NA in this part of the brain.
一种最近开发的放射自显影技术,基于体外单胺终末(轴突膨体)的摄取标记,被用于量化成年大鼠海马体中的去甲肾上腺素(NA)神经支配。在用1微摩尔3H-NA孵育脑片后,在横跨海马体腹侧2/3的3个等距水平制备的光学显微镜放射自显影片中,NA膨体被可视化为银颗粒的小聚集体。使用计算机辅助图像分析仪,从海马下托(SUB)、海马角的3个区域(CA1、CA3-a、CA3-b)和齿状回的3个区域(DG-内侧叶片、嵴和外侧叶片)进行计数,每个区域的每个层都进行采样。在对放射自显影曝光时间和切片厚度进行双重校正后,并在电子显微镜放射自显影片中测量膨体直径后,就有可能将这些结果表示为每单位组织体积中的终末数量。结果发现,海马体NA神经支配的总体密度平均为每立方毫米组织210万个膨体,这一数值几乎是大脑皮层的两倍。这种神经支配在腹侧比背侧密集20%,并且在区域和层状分布方面都是不均匀的。SUB和DG的神经支配比海马角更强,其中CA1的总体密度最低。在SUB和CA1中,NA膨体明显倾向于分子层。在CA3中,在放射层靠近锥体层顶端边界处有一条更强神经支配的窄带,与此细胞层和原层中低3倍的密度形成对比。在DG中,NA神经支配在细胞体层(颗粒层)中再次最弱,而在多形层中密度几乎高3倍,是整个海马体中最高的。这些数据允许与海马体中NA功能的其他定量参数——细胞学、生化和药理学参数——进行大量关联。它们也为在细胞水平阐明NA在大脑这一部分的作用提供了有力依据。