Kamatsuka Yosuke, Fukagawa Manami, Furuta Takahiro, Ohishi Akihiro, Nishida Kentaro, Nagasawa Kazuki
Department of Environmental Biochemistry, Kyoto Pharmaceutical University.
Biol Pharm Bull. 2014;37(12):1958-62. doi: 10.1248/bpb.b14-00000.
We previously demonstrated that the P2X7 receptor (P2X7R), a purinergic receptor, expressed by mouse cultured cortical astrocytes is constitutively activated without any exogenous stimulus, differing from the case of neurons. It is well known that astrocytic morphology differs between in vitro and in vivo situations, implying different functionalities. Brain acute slices are widely accepted as an in vitro experimental system that reflects in vivo cell conditions better than in vitro cell culture ones. We examined whether astrocytic P2X7Rs exhibited constitutive activation in mouse cortical slices. In acute cortical slices, P2X7R-immunoreactivity was detected in both glial fibrillary acidic protein-immunopositive astrocytes and microtubule-associated protein 2-immunopositive neurons. Astrocytic, but not neuronal, spontaneous uptake of propidium iodide, an indicator of P2X7R channel/pore activity, was inhibited by representative antagonists of P2X7R, but they had no effect on the uptake by astrocytes in membrane-permeabilized fixed slices. These findings indicate that astrocytes, but not neurons, in acute cortical slices exhibit constitutive activation of P2X7Rs under non-stimulated resting conditions as in the case of cell culture systems.
我们之前证明,嘌呤能受体P2X7受体(P2X7R)由小鼠培养的皮质星形胶质细胞表达,在没有任何外源刺激的情况下即组成性激活,这与神经元的情况不同。众所周知,星形胶质细胞的形态在体外和体内情况下有所不同,这意味着其功能也不同。脑急性切片作为一种体外实验系统被广泛接受,它比体外细胞培养能更好地反映体内细胞状况。我们研究了星形胶质细胞的P2X7R在小鼠皮质切片中是否表现出组成性激活。在急性皮质切片中,在胶质纤维酸性蛋白免疫阳性的星形胶质细胞和微管相关蛋白2免疫阳性的神经元中均检测到P2X7R免疫反应性。作为P2X7R通道/孔活性指标的碘化丙啶的星形胶质细胞而非神经元的自发摄取受到P2X7R代表性拮抗剂的抑制,但它们对膜通透固定切片中星形胶质细胞的摄取没有影响。这些发现表明,与细胞培养系统的情况一样,急性皮质切片中的星形胶质细胞而非神经元在未刺激的静息条件下表现出P2X7R的组成性激活。