Rapid reportLocalization of the pannexin1 protein at postsynaptic sites in the cerebral cortex and hippocampus
Section snippets
Animals
C57/BL6 mice, Wistar rats and Chinchilla rabbits were bred and housed under standard conditions and maintained in a 12-h light/dark cycle in the animal core facility of the Medical Faculty of the Ruhr-University Bochum. Animal handling was performed in accordance with the German animal protection law and with permission of local authorities. The number of animals used was kept to the minimum necessary for evaluating the experimental data. All efforts were made to minimize animal suffering.
Light and electron microscopical immunohistochemistry
Results
To determine the localization of Panx1 in mouse brain tissues we produced an antibody directed to a carboxy-terminal GST-fusion protein (Panx1-CT), and used a further Panx1 chicken antibody (4515). Specificity of the latter antibody has been described in a recent paper (Locovei et al., 2006a). Western blot analysis using the Panx1-CT antibody revealed a prominent band at approximately 50 kDa in extracts from different brain areas (Fig. 1A). Immunolabeling of brain sections displayed a strong
Discussion
At first glance our observation of a postsynaptic localization of Panx1 seems to be incompatible with the suggested gap junction forming ability as ascribed for Panx1 from paired oocytes experiments (Bruzzone et al., 2003). As indicated above our data do not exclude a localization of Panx1 at gap junctional sites since neuronal gap junctions are hard to detect and can be unambiguously recognized only at optimal ultrastructural preservation (Fukuda and Kosaka, 2003). This predicament is
Acknowledgments
This work was supported by a grant from the Deutsche Forschungsgemeinschaft to R.D. and G.Z (De 292/11–4) and a NIH grant to G.D. (GM48610). S.B. has a stipend of the Graduate School (GRK 736) Ruhr University Bochum (Germany). We like to express our gratitude to Dr. D. Krause-Finkeldey, K. Ladage, M. Loebbecke-Schumacher, and L. Augustinowski for technical assistance. Special thanks to Dr. H. Moneyer and Dr. J. von Englehardt for providing the Panx1 knockout tissue.
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