Objective: To investigate the expression of alpha7-nAChR on rat hippocampal astrocytes in vivo and in vitro.
Methods: Rat hippocampus was isolated and cut into 30-microm cryosections with anti-GFAP antibody staining followed by staining for alpha7-nAChR. The cultured astrocytes obtained from newborn rat (1-2 days old) hippocampus were identified with GFAP, and the expression of alpha7-nAChR was measured by fluorescein isothcyanate-tagged alpha-bungarotoxin staining analysis and double immunolabeling.
Results: The localization of alpha7-nAChR was visualized in green with fluorescein isothiocyanate labeled IgG, whereas that of GFAP in red with Texas red-labeled IgG in the hippocampal slices, and the yellow spots indicating colocalization of the two fluorescent probes was shown in a merging image. alpha-bungarotoxin-binding nicotinic receptors were clustered, and the colocalization of alpha7-nAChR and GFAP on the cultured hippocampal astrocytes was visualized with the two fluorescent probes.
Conclusion: The expression of alpha7-nAChR is identified on hippocampal astrocytes in vivo and in vitro.