Immunofluorescence and biochemical techniques to detect nuclear localization of ciliary neurotrophic factor in glial cells.
Ciliary neurotrophic factor (CNTF) promotes the survival of several populations of neurons, including sensory and motor neurons. It is mainly produced by Schwann cells and astrocytes and exerts its biological function via a specific membrane receptor. We recently determined the nuclear localization of CNTF in producing cells, after transfection and in the heterologous system of Xenopus oocytes. In the present paper, we describe in detail the techniques for the detection of CNTF in the nucleus of rat astrocytes, transfected cells, isolated nuclei and injected Xenopus oocytes.
Bajetto
A
A
Advanced Biotechnology Center, Istituto Nazionale per la Ricerca sul Cancro, Largo Rosanna Benzi 10, 16132, Genova, Italy. bajetto@sirio.cba.unige.it
Barbero
S
S
Bonavia
R
R
Chimini
G
G
Schettini
G
G
D000818:Animals; D000911:Antibodies, Monoclonal; D001253:Astrocytes / Q000737:chemistry*; D019556:COS Cells; D002467:Cell Nucleus / Q000737:chemistry*; D020934:Ciliary Neurotrophic Factor / Q000032:analysis* / Q000276:immunology; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D005910:Glioma; D008845:Microinjections; D018613:Microscopy, Confocal / Q000379:methods; D009865:Oocytes / Q000502:physiology; D011233:Precipitin Tests / Q000379:methods; D012333:RNA, Messenger; D051381:Rats; D017207:Rats, Sprague-Dawley; D014162:Transfection / Q000379:methods*; D014407:Tumor Cells, Cultured; D014982:Xenopus laevis
D000911:Antibodies, Monoclonal; D020934:Ciliary Neurotrophic Factor; D012333:RNA, Messenger