| Annotated protein: | Early endosome antigen 1. Gene symbol: EEA1. Taxonomy: Mus musculus (Mouse). Uniprot ID: Q8BL66 |
| antibody wiki: | |
| SynGO gene info: | SynGO data @ EEA1 |
| Ontology domain: | Biological Process |
| SynGO term: | postsynaptic process involved in chemical synaptic transmission (GO:0099565) |
| Synapse type(s): | hippocampus, glutamatergic Schaffer collateral synapse (CA3->CA1) |
| Annotated paper: | Selak S, et al. "Human autoantibodies against early endosome antigen-1 enhance excitatory synaptic transmission" Neuroscience. 2006 Dec 28;143(4):953-64 PMID:17113235 |
| Figure(s): | Fig4 |
| Annotation description: | The reasoning behind this experiment was to use human-EEA1 antibodies to block the activity of EEA1in the neural synapse and conduct electrophysiological experiments to study the effect on synaptic transmission. Fig 4A: An immunolabelling experiment was carried out to verify the intracellular location of the introduced antibodies via the recording pipette. Visualization of recorded neurons was achieved by including biocytin into the recording pipette together with EEA1 or control human IgG antibodies. Internalized biocytin and human antibodies were detected by incubation with Alexa 488-conjugated streptavidin (yellow, inset) and Alexa 555-conjugated goat anti-human IgG, respectively (blue). Human EEA1 autoantibodies were detected in both soma and dendrites of the biocytin-labelled recorded cells. This suggests that the antibodies introduced during the recordings entered the cells, diffused into the dendrites and were able to reach and bind EEA1 at the post-synaptic nerve terminals (Fig2) within the time course of recording synaptic responses. Fig4B-C: Inclusion of human anti-EEA1 antibodies in the recording solution showed a more than twofold increase in the size of AMPAR-mediated EPSC than the values obtained in the presence of normal human IgG (green points) or in the absence of antibodies. Fig4D: The action of EEA1 antibodies appeared to be specific for excitatory receptors since no significant effect was detected when inhibitory post-synaptic currents (IPSCs) mediated by GABAA receptors were recorded. The specificity of anti-EEA1 antibody effects was demonstrated by the inclusion of denatured anti-EEA1 antibodies that had lost the EEA1 binding affinity. Inclusion of the denatured anti-EEA1 antibodies into the recording pipette in the same concentration as non-denatured antibodies (0.01 mg/ml) showed similar EPSC amplitude profiles as normal human IgG antibody. Fig4E: The PPF measure for AMPAR-mediated responses remained constant during the entire experiment. Fig5: To validate the proposed effects of EEA1 in excitatory neurotransmission, dominant-negative EEA1 deletion mutant was included in the recording pipette. Similar increase in the AMPAR-mediated EPSC was observed as compared to recordings with an absence in the polypeptide. The SynGO term was chosen on the basis of the available evidence the paper demonstrates. |
| Evidence tracking, Biological System: | Cultured neurons |
| Evidence tracking, Protein Targeting: | Antibody (detection) |
| Evidence tracking, Experiment Assay: | Confocal IP + WB/MSMS |
| Annotator(s): | Dnyanada Sahasrabudhe (ORCID:0000-0003-2916-7616) Guus Smit (ORCID:0000-0002-2286-1587) Matthijs Verhage (ORCID:0000-0002-2514-0216) |
| Lab: | Department of Functional Genomics, Department of Molecular and Cellular Neurobiology, Center for Neurogenomics and Cognitive Research, Vrije Universiteit Amsterdam, 1081 HV Amsterdam, The Netherlands |
| Additional literature: | The gene encoding early endosome antigen 1 (EEA1), is down-regulated in hypothalamic samples from Mecp2 KO mice (Chahrour et al. 2008). Fig 5: The levels of EEA1 mRNA and protein were lower in the hippocampus, cortex, thalamus and brainstem of symptomatic Mecp2 KO mice compared to age-matched WT controls. Fig 6B-C: Overexpressing EEA1in Mecp2 KO neurons significantly reduced mEPSC amplitudes. Fig 6B-E: Consistent with the recovery of only receptor endocytosis by EEA1 expression, scaling up mEPSC amplitudes in response to silencing with TTX was not recovered in EEA1-expressing Mecp2 KO neurons. Fig7: Schematic representation demonstrates the probable role of EEA1 in regulating AMPAR levels in the postsynapse. Taken altogether, the paper indicates that overexpression of EEA1 in Mecp2 KO neurons rescues impaired GluA1 endocytosis and as an after product effect chemical synaptic transmission. @ PMID:28621434 |
| SynGO annotation ID: | 4125 |
| Dataset release (version): | 20231201 |
| View annotation as GO-CAM model: |  |