Cerebellar lesion up-regulates P2X1 and P2X2 purinergic receptors in precerebellar nuclei
Section snippets
Animals and surgical procedures
Experiments were performed using 21 adult Wistar rats (body weight 200–250 g; Harlan, Italy); 18 animals were used for cerebellar lesions (three for immunohistochemistry at each time point: days 7, 14, 21, 28 and 35; three for western blot on day 14) and three as controls for immunohistochemistry. Experiments were performed according to the guidelines of the Italian law for care and use of experimental animals, approved by the Italian Ministry of Health. All efforts were made to minimize both
P2X1 and P2X2 receptor expressions in normal animals
In the precerebellar nuclei of control animals, we observed a widespread network of lightly labeled fibers with no neuronal staining in P2X1-immunoreacted material. In P2X2-immunoreacted material, we observed lightly stained neurons and some sparse positive fibers. In particular, although we did not observe P2X2-immunopositive cells in the ECu, we observed sparse P2X2-immunopositive neurons in the SpV and in the IO. In the LRn, the immunostaining pattern was characterized by a widespread and
Discussion
Our data show a transient and time-dependent up-regulation of P2X1 and P2X2 expression in the precerebellar nuclei of the experimental side after axotomy. Side differences in the number of immunoreactive neurons in the precerebellar nuclei became evident on day 7 and persisted until 35 days after the lesion. In the IO the number of P2X1-immunopositive neurons peaked at 28 days, whereas the number of P2X2-immunopositive neurons peaked at 14 days post-lesion (Fig. 5). In general in IO and Pn,
Acknowledgements
We are grateful to Prof. G. Bernardi for his continuous support and encouragement. This work was supported by Italian Ministry of Health Grant RA0085M-2000 to M.M. and by a grant from MURST Cofin 2001 on ‘Purinoceptors and Neuroprotection’ to C.V.
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