大鼠发育中皮质脊髓束内顺行运输的辣根过氧化物酶的超微结构观察

Ultrastructural visualization of anterogradely transported horseradish peroxidase in developing corticospinal tract of rat.

作者信息

Joosten E A, Gribnau A A, Dederen P J

出版信息

J Histochem Cytochem. 1987 May;35(5):623-6. doi: 10.1177/35.5.2435786.

DOI:10.1177/35.5.2435786

Abstract

Until now a satisfactory method for electron microscopic (EM) detection of anterogradely transported horseradish peroxidase (HRP) in developing neural tissue, using sensitive chromogen tetramethylbenzidine (TMB), has not been described. Use of the stabilizing agent ammoniumheptamolybdate (AHM), in combination with a modified prolonged osmication [4 hr at pH 5.0 in 0.1 M phosphate buffer (PB)] made possible visualization of HRP-TMB-(AHM) reaction product at the ultrastructural level in outgrowing corticospinal tract (CST) fibers of young postnatal rat. This reaction product appeared to be very distinctive and clearly detectable, making ultrastructural identification of HRP-labeled outgrowing CST fibers in rat spinal cord rather easy. In addition, the procedure described in this report preserves the ultrastructural details of the developing neural tissue.

摘要

迄今为止，尚未有关于在发育中的神经组织中使用灵敏显色剂四甲基联苯胺（TMB）进行电子显微镜（EM）检测顺行运输辣根过氧化物酶（HRP）的满意方法的描述。使用稳定剂七钼酸铵（AHM），结合改良的延长锇酸固定法[在0.1 M磷酸盐缓冲液（PB）中pH 5.0下固定4小时]，使得在新生大鼠生长中的皮质脊髓束（CST）纤维的超微结构水平上能够观察到HRP-TMB-（AHM）反应产物。该反应产物显得非常独特且易于检测，使得在大鼠脊髓中对HRP标记的生长中的CST纤维进行超微结构鉴定变得相当容易。此外，本报告中描述的方法保留了发育中神经组织的超微结构细节。