Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
Microtransplantation of Synaptic Membranes to Reactivate Human Synaptic Receptors for Functional Studies.
Miller B
,
Powell A
,
Gutierrez BA
,
Limon A
.
???displayArticle.abstract???
Excitatory and inhibitory ionotropic receptors are the major gates of ion fluxes that determine the activity of synapses during physiological neuronal communication. Therefore, alterations in their abundance, function, and relationships with other synaptic elements have been observed as a major correlate of alterations in brain function and cognitive impairment in neurodegenerative diseases and mental disorders. Understanding how the function of excitatory and inhibitory synaptic receptors is altered by disease is of critical importance for the development of effective therapies. To gain disease-relevant information, it is important to record the electrical activity of neurotransmitter receptors that remain functional in the diseased human brain. So far this is the closest approach to assess pathological alterations in receptors' function. In this work, a methodology is presented to perform microtransplantation of synaptic membranes, which consists of reactivating synaptic membranes from snap frozen human brain tissue containing human receptors, by its injection and posterior fusion into the membrane of Xenopus laevis oocytes. The protocol also provides the methodological strategy to obtain consistent and reliable responses of α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) and γ-aminobutyric acid (GABA) receptors, as well as novel detailed methods that are used for normalization and rigorous data analysis.
???displayArticle.pubmedLink???
35938847
???displayArticle.pmcLink???PMC10729793 ???displayArticle.link???J Vis Exp ???displayArticle.grants???[+]
Bae,
Synapses in neurodegenerative diseases.
2017, Pubmed
Bae,
Synapses in neurodegenerative diseases.
2017,
Pubmed
Bond,
The quantification of gene expression in an animal model of brain ischaemia using TaqMan real-time RT-PCR.
2002,
Pubmed
Calvo,
Cationic modulation of rho 1-type gamma-aminobutyrate receptors expressed in Xenopus oocytes.
1994,
Pubmed
,
Xenbase
Crespin,
Injection of insect membrane in Xenopus oocyte: An original method for the pharmacological characterization of neonicotinoid insecticides.
2016,
Pubmed
,
Xenbase
Cutting,
Cloning of the gamma-aminobutyric acid (GABA) rho 1 cDNA: a GABA receptor subunit highly expressed in the retina.
1991,
Pubmed
,
Xenbase
Eusebi,
Microtransplantation of ligand-gated receptor-channels from fresh or frozen nervous tissue into Xenopus oocytes: a potent tool for expanding functional information.
2009,
Pubmed
,
Xenbase
Furcila,
A Study of Amyloid-β and Phosphotau in Plaques and Neurons in the Hippocampus of Alzheimer's Disease Patients.
2018,
Pubmed
Hodge,
Conserved cell types with divergent features in human versus mouse cortex.
2019,
Pubmed
Lauterborn,
Increased excitatory to inhibitory synaptic ratio in parietal cortex samples from individuals with Alzheimer's disease.
2021,
Pubmed
Le Mauff,
Nicotinic acetylcholine receptors in the synganglion of the tick Ixodes ricinus: Functional characterization using membrane microtransplantation.
2020,
Pubmed
,
Xenbase
Limon,
Microtransplantation of neurotransmitter receptors from postmortem autistic brains to Xenopus oocytes.
2008,
Pubmed
,
Xenbase
Limon,
Properties of GluR3 receptors tagged with GFP at the amino or carboxyl terminus.
2007,
Pubmed
,
Xenbase
Lin-Moshier,
The Xenopus oocyte: a single-cell model for studying Ca2+ signaling.
2013,
Pubmed
,
Xenbase
Marsal,
Incorporation of acetylcholine receptors and Cl- channels in Xenopus oocytes injected with Torpedo electroplaque membranes.
1995,
Pubmed
,
Xenbase
Martinez-Torres,
Expression of gamma-aminobutyric acid rho 1 and rho 1 Delta 450 as gene fusions with the green fluorescent protein.
2001,
Pubmed
,
Xenbase
Mazzo,
Reconstitution of synaptic Ion channels from rodent and human brain in Xenopus oocytes: a biochemical and electrophysiological characterization.
2016,
Pubmed
,
Xenbase
Miledi,
Expression of functional neurotransmitter receptors in Xenopus oocytes after injection of human brain membranes.
2002,
Pubmed
,
Xenbase
Newman,
Isolation of Xenopus Oocytes.
2018,
Pubmed
,
Xenbase
Ochoa-de la Paz,
Dopamine and serotonin modulate human GABAρ1 receptors expressed in Xenopus laevis oocytes.
2012,
Pubmed
,
Xenbase
Palma,
GABA(A)-current rundown of temporal lobe epilepsy is associated with repetitive activation of GABA(A) "phasic" receptors.
2007,
Pubmed
,
Xenbase
Palma,
Microtransplantation of membranes from cultured cells to Xenopus oocytes: a method to study neurotransmitter receptors embedded in native lipids.
2003,
Pubmed
,
Xenbase
Perrais,
Antagonism of recombinant and native GluK3-containing kainate receptors.
2009,
Pubmed
Preece,
Quantifying mRNA in postmortem human brain: influence of gender, age at death, postmortem interval, brain pH, agonal state and inter-lobe mRNA variance.
2003,
Pubmed
Preece,
An optimistic view for quantifying mRNA in post-mortem human brain.
2003,
Pubmed
Sandoval,
Antagonistic effects of TrkB and p75(NTR) on NMDA receptor currents in post-synaptic densities transplanted into Xenopus oocytes.
2007,
Pubmed
,
Xenbase
Sanna,
Expression of native GABAA receptors in Xenopus oocytes injected with rat brain synaptosomes.
1996,
Pubmed
,
Xenbase
Sanna,
Functional changes in rat nigral GABA(A) receptors induced by degeneration of the striatonigral GABAergic pathway: an electrophysiological study of receptors incorporated into Xenopus oocytes.
1998,
Pubmed
,
Xenbase
Scaduto,
Preservation of global synaptic excitatory to inhibitory ratio during long postmortem intervals.
2020,
Pubmed
Sive,
Microinjection of Xenopus oocytes.
2010,
Pubmed
,
Xenbase
Soualah,
GABAA Receptor Subunit Composition Drives Its Sensitivity to the Insecticide Fipronil.
2021,
Pubmed
,
Xenbase
Spitzer,
A Rosetta stone for analysis of human membrane protein function.
2008,
Pubmed
Stan,
Human postmortem tissue: what quality markers matter?
2006,
Pubmed
Taoufik,
Synaptic dysfunction in neurodegenerative and neurodevelopmental diseases: an overview of induced pluripotent stem-cell-based disease models.
2018,
Pubmed
Varol,
HYDRA: Revealing heterogeneity of imaging and genetic patterns through a multiple max-margin discriminative analysis framework.
2017,
Pubmed
Wu,
GABA(A) receptor-mediated excitation in dissociated neurons from human hypothalamic hamartomas.
2008,
Pubmed
Zeppillo,
Functional impairment of cortical AMPA receptors in schizophrenia.
2022,
Pubmed
Zhao,
Architecture and subunit arrangement of native AMPA receptors elucidated by cryo-EM.
2019,
Pubmed
Zwart,
Microtransplantation of human brain receptors into oocytes to tackle key questions in drug discovery.
2019,
Pubmed
,
Xenbase
