Immunohistochemical Funicular Orientation of Peripheral Nerve

1.周围神经的神经索定位免疫组织化学方法应用于周围神经的神经索定位。用市售抗体和染色试剂盒对大鼠和猪的脊神经根进行染色。以抗胆碱乙酰转移酶抗体为第一抗体，用该方法可区分大鼠前、后脊神经根。然而，在前根和后根，组织化学方法比免疫组织化学方法更具优势。结扎大鼠坐骨神经和臀肌支，观察组织学和组织化学的逆行变化，生化方法观察到中央残端乙酰胆碱酯酶活性降低，表现为乙酰胆碱酯酶阳性轴突横截面积缩小。计算机图像分析结果显示，臀肌支中心残端轴突横截面积明显减少，有髓纤维数量明显增多。但对乙酰胆碱酯酶活性的电子显微镜观察显示，仅有10%~20%的有髓小轴突显示出酶活性，这种逆行变化随着时间的推移而发展，在大鼠坐骨神经和臀肌支结扎部位近端>15 mm和>5 mm处分别观察到。

1. Immunohistochemical funicular orientation of peripheral nerveImmunohistochemical method was introduced into the funicular orientation of peripheral nerve. We stained the spinal roots of rat and pig with commercialy obtainable antibodies and staining kit. An anti- choline acetyltransferase antibody was used as the first antibody.We could differentiate rat anterior and posterior spinal roots with this method. The histochemical method was, however, more advantageous than the immunohistochemical method in theanterior and posterior roots.2. Histological and histochemical study on the retrograde changes of injured peripheral nerveRat sciatic nerve and gluteal muscle branch were ligated and retrograde changes were observed histologically and histochemically.The decrease of acetylcholinesterase activity in the central stump, which has been observed by biochemical method, was observed as the decrease of teh cross-sectional area of the acetylcholinesterase positive axons. The decrease of the cross-sectional area of the axons in the central stump was confirmed statistically using micro-computer assisted image analyser.Small myelinated fibers have remarkably increased in number in the central stump of the ligated gluteal muscle branch. But electron microscopic observation of acetylcholinesterase activity revealed that only 10 % to 20 % of the small myelinated axons in the central stump showed the enzymatic activity.These retrograde changes progressed with the lapse of time after the ligationand were observed at the region more than 15 mm and more than 5 mm proximal to the ligated site in the ligated rat sciatic nerve and gluteal muscle branch respectively.