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Published on: May 18, 2009
Characteristics of P2X7 receptors from human B lymphocytes expressed in Xenopus oocytes
M Klapperstück1, C Büttner, T Böhm
1Julius-Bernstein-Institut für Physiologie, Martin-Luther-Universität Halle-Wittenberg, Halle/Saale, Germany. manuela.klapperstueck@medizin.uni-halle.de
Abstract:
Human B lymphocytes express an ATP-gated ion channel (P2Z receptor), which shares similarities with the recently identified P2X7 receptor. Using gene specific primers, we have now isolated P2X7 cDNA from the total RNA of human B lymphocytes. This hP2X7 receptor subtype was expressed in Xenopus oocytes and electrophysiologically characterized. The hP2X7 receptor is similar to, but does not completely match, P2Z of human B cells. The hP2X7 receptors resemble the P2Z receptors with regard to the ATP concentration of half maximal activation, reproducibility, permeation characteristics and lack of desensitization of the ATP-evoked currents. However, in contrast to the native lymphocytic P2Z receptor, the time course of activation of hP2X7 displayed an additional linearly increasing current component. Furthermore, a second, small and slowly deactivating current component exists only in hP2X7 expressed in oocytes. The activation and deactivation kinetics as well as permeation characteristics of hP2X7 are different from rat P2X7 recently expressed in oocytes. Unlike in mammalian cells, hP2X7 expressed in Xenopus oocytes is not sufficient to induce large non-selective pores.
Insights
Researchers identified and characterized the human P2X7 receptor (hP2X7) in B lymphocytes. While similar to the native P2Z receptor, hP2X7 exhibits distinct activation and current properties when expressed in Xenopus oocytes.
Area of Science:
- Immunology
- Molecular Biology
- Neuroscience
Background:
- Human B lymphocytes possess an ATP-gated ion channel, the P2Z receptor.
- This P2Z receptor shares characteristics with the recently identified P2X7 receptor.
Purpose of the Study:
- To isolate and characterize the P2X7 receptor cDNA from human B lymphocytes.
- To electrophysiologically analyze the properties of the human P2X7 (hP2X7) receptor expressed in Xenopus oocytes.
Main Methods:
- Isolation of P2X7 cDNA from human B lymphocyte RNA using gene-specific primers.
- Expression of hP2X7 in Xenopus oocytes.
- Electrophysiological characterization of expressed hP2X7 receptors.
Main Results:
- hP2X7 cDNA was successfully isolated from human B lymphocytes.
- Expressed hP2X7 receptors showed similarities to native P2Z receptors in ATP sensitivity and lack of desensitization.
- Distinct differences were observed, including a linearly increasing current component and a slowly deactivating current in hP2X7 expressed in oocytes.
- hP2X7 expressed in oocytes did not induce large non-selective pores, unlike in mammalian cells.
Conclusions:
- The hP2X7 receptor subtype has been identified and characterized.
- While sharing similarities, hP2X7 exhibits unique electrophysiological properties compared to the native P2Z receptor and rat P2X7.
- The functional differences highlight the complexity of P2X7 receptor subtypes and their expression systems.
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