Cell membranes isolated from nervous tissue can be easily injected into Xenopus oocytes, thereby effectively “microtransplanting” functional neurotransmitter receptors. This technique therefore allows a direct functional characterization of the original membrane receptor/ion channel proteins and the associated molecules while still embedded in their natural lipid environment. Cell membranes will contain components from different types of cells, i.e. neurons and glial cells, expressing their own receptors, with possibly different properties. To study the receptor properties of a single cell type, we injected oocytes with membranes isolated only from glia (gliosomes) of adult mouse neocortex and we focused our work on GABAA receptors incorporated in the oocyte cell membrane. We found that GABAA-activated currents allowed a good biophysical and pharmacological characterization of glial GABAA receptors. Therefore, the microtransplantation of gliosomes into oocytes can represent a good model to study the electrical and pharmacological properties of adult glial cells under different physiological and pathological conditions. Moreover, since gliosomes can be isolated from frozen tissues, this approach can be extended to post-mortem human tissues

Characterization of GABA(A) receptors expressed in glial cell membranes of adult mouse neocortex using a Xenopus oocyte microtransplantation expression system.

BERNAREGGI, Annalisa;RUZZIER, FABIO;
2011-01-01

Abstract

Cell membranes isolated from nervous tissue can be easily injected into Xenopus oocytes, thereby effectively “microtransplanting” functional neurotransmitter receptors. This technique therefore allows a direct functional characterization of the original membrane receptor/ion channel proteins and the associated molecules while still embedded in their natural lipid environment. Cell membranes will contain components from different types of cells, i.e. neurons and glial cells, expressing their own receptors, with possibly different properties. To study the receptor properties of a single cell type, we injected oocytes with membranes isolated only from glia (gliosomes) of adult mouse neocortex and we focused our work on GABAA receptors incorporated in the oocyte cell membrane. We found that GABAA-activated currents allowed a good biophysical and pharmacological characterization of glial GABAA receptors. Therefore, the microtransplantation of gliosomes into oocytes can represent a good model to study the electrical and pharmacological properties of adult glial cells under different physiological and pathological conditions. Moreover, since gliosomes can be isolated from frozen tissues, this approach can be extended to post-mortem human tissues
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11368/2327619
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