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 GABA(A) receptors incorporated in the oocyte cell membrane. We found that GABA(A)-activated currents allowed a good biophysical and pharmacological characterization of glial GABA(A) 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. (C) 2011 Elsevier B.V. All rights reserved.
Characterization of GABA(A) receptors expressed in glial cell membranes of adult mouse neocortex using a Xenopus oocyte microtransplantation expression system / Annalisa, Bernareggi; Massimo, Grilli; Mario, Marchi; Limatola, Cristina; Fabio, Ruzzier; Eusebi, Fabrizio. - In: JOURNAL OF NEUROSCIENCE METHODS. - ISSN 0165-0270. - STAMPA. - 198:1(2011), pp. 77-83. [10.1016/j.jneumeth.2011.03.011]
Characterization of GABA(A) receptors expressed in glial cell membranes of adult mouse neocortex using a Xenopus oocyte microtransplantation expression system
LIMATOLA, Cristina;EUSEBI, Fabrizio
2011
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 GABA(A) receptors incorporated in the oocyte cell membrane. We found that GABA(A)-activated currents allowed a good biophysical and pharmacological characterization of glial GABA(A) 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. (C) 2011 Elsevier B.V. All rights reserved.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.