In our previous study, we reported that the levels of m2 muscarinic acetylcholine receptor and gammaaminobutyric acid receptor α1 in motoneurons were downregulated in the axotomized adult facial nucleus. These results led us to speculate that the phenomenon was attributable to an interruption of the retrograde supply of glial cell line-derived neurotrophic factor (GDNF). To investigate this possibility, in this study we determined the levels of GDNF in the adult rat axotomized facial nucleus. Western blotting revealed that the amounts of GDNF in the transected facial nucleus significantly decreased from 6 h to 7 d post-insult and returned to control levels at 3-5 weeks post-insult. Immunohistochemical study of the facial nucleus at 3 d post-insult indicated that many cells were positively stained by anti-GDNF antibody in the control facial nucleus, but the degree of staining was lower in the axotomized facial nucleus. The anti-GDNF antibody-stained cells in the facial nucleus were almost coincident with the anti-NMDA receptor 3B subunit (NR3B) antibody-positive cells, suggesting that the former were motoneurons. Together, these results demonstrated that GDNF levels in injured facial motoneurons transiently decreased from 6 h to 7 d post-insult but subsequently returned to the control levels.