| Annotated protein: | H(+)/Cl(-) exchange transporter 3 (Chloride channel protein 3) (ClC-3) (Chloride transporter ClC-3). Gene symbol: CLCN3. Taxonomy: Rattus norvegicus (Rat). Uniprot ID: P51792 |
| antibody wiki: | |
| SynGO gene info: | SynGO data @ CLCN3 |
| Ontology domain: | Cellular Component |
| SynGO term: | integral component of postsynaptic membrane (GO:0099055) |
| Synapse type(s): | hippocampus, glutamatergic |
| Annotated paper: | Wang XQ, et al. "CLC-3 channels modulate excitatory synaptic transmission in hippocampal neurons" Neuron. 2006 Oct 19;52(2):321-33 PMID:17046694 |
| Figure(s): | Fig. 5, Fig. 6 |
| Annotation description: | Wang et al performed biotinylation experiments in order to verify plasma membrane expression of ClC-3. Shortly, the authors use membrane impermeable NHS-biotin derivate in order to biotinylate plasma membrane proteins exposed to the extracellular surface of cultured rat hippocampal neurons (Fig. 5A). Furthermore, ClC-3 expression on the synaptic plasma membrane surface was examined by preparation of synaptic vesicle and membrane fractions from rat hippocampi followed by immunodetection of ClC-3 and plasma membrane and SV marker proteins (Fig. 5B). In addition, in a double immunogold labeling EM experiment on isolated plasma membrane fraction, PSD95 and ClC-3 surface expression was confirmed. In order to validate the postsynaptic expression of ClC-3, as determined biochemically and by EM, Wang et al perfomed immunofluorescene experiments on cultured rat hippocampal neurons and showed colocalization of the chloride channel with PSD95 and NMDAR (Fig. 6 A and B). Association of ClC-3 with postsynaptic membranes was confirmed biochemically by immunoprecipitation of PSD95 and ClC-3 and Western blot detection of postsynaptic and plasma membrane proteins (Fig. 6C). 6/2/2018 Pim - Based on the known transmembrane topology of the protein, the "integral component of ... membrane" term was chosen. |
| Evidence tracking, Biological System: | Intact tissue Cultured neurons |
| Evidence tracking, Protein Targeting: | Antibody (detection) |
| Evidence tracking, Experiment Assay: | Electron Microscopy Confocal Western blot Biochemical fractionation (generic) IP + WB/MSMS |
| Annotator(s): | Momchil Ninov (ORCID:0000-0002-0808-7003) Mahdokht Kohansalnodehi (ORCID:0000-0002-3898-5197) Reinhard Jahn (ORCID:0000-0003-1542-3498) |
| Lab: | Department of Neurobiology, Max Planck Institute for Biophysical Chemistry, 37077 Göttingen, Germany |
| Additional literature: | The lab of Deborah J Nelson, which examined the postsynaptic expression of ClC-3 in the primary reference, showed in this paper (Fig. 1) by means of immunofluorescence labelling in hippocampus cryosections of WT and CLC-3 KO mice that ClC-3 colocalizes with the detection of PSD95 and NMDAR. @ PMID:23165767 |
| SynGO annotation ID: | 381 |
| Dataset release (version): | 20231201 |
| View annotation as GO-CAM model: |  |