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.
| Original language | English |
|---|---|
| Pages (from-to) | 77-83 |
| Number of pages | 7 |
| Journal | Journal of Neuroscience Methods |
| Volume | 198 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - May 15 2011 |
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Keywords
- Adult mouse cortex
- Astrocytes
- Gliosomes
- Synaptosomes
- Two-electrode voltage clamp
- Xenopus oocytes
ASJC Scopus subject areas
- Neuroscience(all)
Cite this
Characterization of GABA A receptors expressed in glial cell membranes of adult mouse neocortex using a Xenopus oocyte microtransplantation expression system. / Bernareggi, Annalisa; Grilli, Massimo; Marchi, Mario; Limatola, Cristina; Ruzzier, Fabio; Eusebi, Fabrizio.
In: Journal of Neuroscience Methods, Vol. 198, No. 1, 15.05.2011, p. 77-83.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Characterization of GABA A receptors expressed in glial cell membranes of adult mouse neocortex using a Xenopus oocyte microtransplantation expression system
AU - Bernareggi, Annalisa
AU - Grilli, Massimo
AU - Marchi, Mario
AU - Limatola, Cristina
AU - Ruzzier, Fabio
AU - Eusebi, Fabrizio
PY - 2011/5/15
Y1 - 2011/5/15
N2 - 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.
AB - 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.
KW - Adult mouse cortex
KW - Astrocytes
KW - Gliosomes
KW - Synaptosomes
KW - Two-electrode voltage clamp
KW - Xenopus oocytes
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UR - http://www.scopus.com/inward/citedby.url?scp=79955521739&partnerID=8YFLogxK
U2 - 10.1016/j.jneumeth.2011.03.011
DO - 10.1016/j.jneumeth.2011.03.011
M3 - Article
C2 - 21439322
AN - SCOPUS:79955521739
VL - 198
SP - 77
EP - 83
JO - Journal of Neuroscience Methods
JF - Journal of Neuroscience Methods
SN - 0165-0270
IS - 1
ER -