In vivo transfer of a marker gene to study motoneuronal development.
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.
Lisovoski
F
F
INSERM CJF 91-02, Faculté de Médecine, Créteil, France.
Cadusseau
J
J
Akli
S
S
Caillaud
C
C
Vigne
E
E
Poenaru
L
L
Stratford-Perricaudet
L
L
Pericaudet
M
M
Kahn
A
A
Peschanski
M
M
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D015740:Calcitonin Gene-Related Peptide / Q000276:immunology / Q000378:metabolism; D003712:Dendrites / Q000648:ultrastructure; D018014:Gene Transfer Techniques; D005819:Genetic Markers / Q000276:immunology; D005822:Genetic Vectors; D007150:Immunohistochemistry; D009046:Motor Neurons / Q000502:physiology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013116:Spinal Cord / Q000166:cytology / Q000201:enzymology / Q000254:growth & development*; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D005819:Genetic Markers; D001616:beta-Galactosidase; D015740:Calcitonin Gene-Related Peptide