Holets V, Elde R
Neuroscience. 1982 May;7(5):1155-74. doi: 10.1016/0306-4522(82)91123-x.
The preganglionic sympathetic neurons in the intermediolateral cell column of the thoracic and upper lumbar segments of the spinal cord which innervate the chromaffin cells in the adrenal medulla, sympathoadrenal preganglionic neurons, were identified by the method of retrograde axonal transport of the fluorescent dyes Fast Blue and True Blue. In rats, Fast Blue or True Blue was injected into the medulla of the left adrenal gland. After a survival period of 5 days, the animals were perfusion fixed, the thoracic and lumbar spinal cord sectioned and processed for the immunofluorescent localization of met-enkephalin, neurophysin, oxytocin, serotonin, somatostatin and substance P immunoreactivity. Neuronal perikarya which were retrogradedly-labeled with Fast Blue or True Blue were observed in the intermediolateral cell column from the T1 to the L2 spinal cord segments. The distribution of the sympathoadrenal neurons was determined by counting the number of retrogradedly-labeled neurons per spinal cord segment. In the five animals used for quantifying the sympathoadrenal preganglionic neurons, the majority (72.3%) of the retrogradely-labeled neurons counted per spinal cord were located within the T7-T12 segments. The T9 segment contained the largest average number (20.1%) of retrogradely-labeled cells in a single segment. Met-enkephalin, serotonin and substance P immunoreactive fibers were prominent in the intermediolateral cell column, whereas oxytocin, neurophysin and somatostatin immunoreactive fibers were sparse. The met-enkephalin, serotonin and substance P fibers were seen surrounding both unlabeled and retrogradely-labeled neurons; somatostatin fibers appeared to preferentially contact retrogradely-labeled neurons; whereas, the neurophysin and oxytocin fibers were not found in proximity to retrogradely-labeled neurons. Met-enkephalin, neurophysin, oxytocin, somatostatin and substance P immunoreactivity were depleted in the intermediolateral cell column below the level of a spinal cord transection. Serotonin immunoreactivity was depleted in the intermediolateral cell column below the level of the transection for five to six segments, but sparse networks of immunoreactive fibers were observed in both the intermediolateral cell column and the ventral horn in more caudal segments. Met-enkephalin, serotonin, somatostatin and substance P immunoreactivity were decreased in both the contralateral and ipsilateral intermediolateral cell column below the level of a spinal cord hemisection, suggesting that both crossed and uncrossed descending pathways exist. Neurophysin and oxytocin immunoreactivity were depleted below the level of the hemisection in the ipsilateral intermediolateral cell column without noticeable decrease in the level of immunoreactivity in the contralateral intermediolateral cell column, suggesting that a decussation does not occur at the level of the spinal cord, but may exist above the level of the hemisection...
通过荧光染料快蓝和真蓝的逆行轴突运输方法,鉴定出脊髓胸段和上腰段中间外侧细胞柱中支配肾上腺髓质嗜铬细胞的节前交感神经元,即交感肾上腺节前神经元。在大鼠中,将快蓝或真蓝注入左肾上腺髓质。存活5天后,对动物进行灌注固定,将胸段和腰段脊髓切片,并进行甲硫氨酸脑啡肽、神经垂体素、催产素、5-羟色胺、生长抑素和P物质免疫反应性的免疫荧光定位。在T1至L2脊髓节段的中间外侧细胞柱中观察到被快蓝或真蓝逆行标记的神经元胞体。通过计算每个脊髓节段逆行标记神经元的数量来确定交感肾上腺神经元的分布。在用于量化交感肾上腺节前神经元的5只动物中,每个脊髓节段计数的逆行标记神经元大多数(72.3%)位于T7 - T12节段。T9节段在单个节段中包含的逆行标记细胞平均数量最多(20.1%)。甲硫氨酸脑啡肽、5-羟色胺和P物质免疫反应性纤维在中间外侧细胞柱中很突出,而催产素、神经垂体素和生长抑素免疫反应性纤维则很稀疏。甲硫氨酸脑啡肽、5-羟色胺和P物质纤维可见于未标记和逆行标记的神经元周围;生长抑素纤维似乎优先与逆行标记的神经元接触;而神经垂体素和催产素纤维在逆行标记的神经元附近未发现。在脊髓横断水平以下的中间外侧细胞柱中,甲硫氨酸脑啡肽、神经垂体素、催产素、生长抑素和P物质免疫反应性消失。在横断水平以下的中间外侧细胞柱中,5-羟色胺免疫反应性在五到六个节段中消失,但在更靠尾侧的节段中,在中间外侧细胞柱和腹角中均观察到稀疏的免疫反应性纤维网络。在脊髓半横断水平以下,对侧和同侧中间外侧细胞柱中甲硫氨酸脑啡肽、5-羟色胺、生长抑素和P物质免疫反应性均降低,这表明存在交叉和未交叉的下行通路。在同侧中间外侧细胞柱中,神经垂体素和催产素免疫反应性在半横断水平以下消失,而对侧中间外侧细胞柱中的免疫反应性水平没有明显降低,这表明在脊髓水平不存在交叉,但可能在半横断水平以上存在交叉……