Title: A practical method for the radioisotope labeling of rabbit astrocytes with 3H-thymidine
Abstract: A practical method for the radioisotope labeling aimed at the study of the proliferative behavior of astrocytes was described. It consisted in injecting 20 μCi of 3H-thymidine into the vitreous body and tracing by autoradiography labeled astrocytes located both inside and outside the retina, e.g. optic nerve and neighboring parts of the central nervous system. The paraffin sections were immunostained for glial fibrillary acidic protein (GFAP) previous to autoradiographic processing. The semiquantitative analysis of labeled astrocytes was carried out on autoradiographs of semithin sections of rabbits killed as early as 6 h and as late as 3 months after the single intravitreal injection of 3H-thymidine. Compared with the technique of labeling astrocytes by systemic administration (single injection or continuous infusion) of 3H-thymidine into small animals, the method described herein has the following outstanding features: (i) it is much more economical in terms of the amount of labeled precursor used per animal; (ii) the labeling of the astrocytes is obtained as early as 6 h and remains up to 3 months after injection; (iii) the immunolabeling of the astrocytes is compatible with autoradiography; (iv) it is less risky to the experimental animal and to the environment; (v) it can be used in animals much larger than rats or mice.
Publication Year: 2001
Publication Date: 2001-11-01
Language: en
Type: article
Indexed In: ['crossref', 'pubmed']
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