Lisovoski F, Cadusseau J, Akli S, Caillaud C, Vigne E, Poenaru L, Stratford-Perricaudet L, Pericaudet M, Kahn A, Peschanski M
INSERM CJF 91-02, Faculté de Médecine, Créteil, France.
Neuroreport. 1994 May 9;5(9):1069-72. doi: 10.1097/00001756-199405000-00013.
Adenovirus vectors containing a marker gene (lacZ from Escherichia coli) are potent for transferring the gene to neurones after intraparenchymal injections. Expression of the marker gene may lead to the synthesis of an enormous amount of beta-galactosidase which diffuses throughout the entire neurone, providing a 'Golgi-like' staining. This suggested that the technique may be used to study the morphology of specific neuronal populations. We have validated this hypothesis by analysing the postnatal development of motoneurones in the rat cervical cord. Injections of the viral suspension into one ventral horn were performed at different ages after birth. Histochemical staining using X-Gal revealed morphological changes occurring within the first 3 weeks with enlargement of the perikaryon and increased dendritic complexity. Immunoreactivity for CGRP was visualized in double-staining experiments. In vivo transfer of a marker gene therefore provides a new way to analyse neuronal morphology which allows selection of the cells to be studied and double-labelling with immunohistochemical markers.
含有标记基因（来自大肠杆菌的lacZ）的腺病毒载体在脑实质内注射后能有效地将基因转移到神经元。标记基因的表达可能导致大量β-半乳糖苷酶的合成，该酶会扩散到整个神经元，呈现出“高尔基样”染色。这表明该技术可用于研究特定神经元群体的形态。我们通过分析大鼠颈髓运动神经元的出生后发育来验证这一假设。在出生后的不同年龄，将病毒悬液注射到一侧腹角。使用X-Gal进行组织化学染色显示，在出生后的前三周内发生了形态变化，包括胞体增大和树突复杂性增加。在双重染色实验中观察到降钙素基因相关肽（CGRP）的免疫反应性。因此，标记基因的体内转移为分析神经元形态提供了一种新方法，该方法允许选择要研究的细胞并与免疫组织化学标记物进行双重标记。
