| Annotated protein: | Sodium/hydrogen exchanger 6 (Na(+)/H(+) exchanger 6) (NHE-6) (Sodium/hydrogen exchanger) (Solute carrier family 9 member 6). Gene symbol: SLC9A6. Taxonomy: Rattus norvegicus (Rat). Uniprot ID: D3ZJ86 |
| antibody wiki: | |
| SynGO gene info: | SynGO data @ SLC9A6 |
| Ontology domain: | Biological Process |
| SynGO term: | regulation of proton loading (GO:1901546) |
| Annotated paper: | Lee U, et al. "SCAMP5 plays a critical role in axonal trafficking and synaptic localization of NHE6 to adjust quantal size at glutamatergic synapses" Proc Natl Acad Sci U S A. 2021 Jan 12;118(2):e2011371118 PMID:33372133 |
| Figure(s): | Figure 4 |
| Annotation description: | "Since NHE6 dissipates ΔpH and increases Δψ across the SV membrane, we hypothesized that the perturbed localization of NHE6 in SCAMP5 KD neurons would lead to hyperacidification of the SV lumen. ... We confirmed that pH-Lemon-TEV-Sty1 was expressed in SVs of cultured hippocampal neurons (SI Appendix, Fig. S7 A and B). We then removed the surface fraction of pH-Lemon-TEV-Syt1 by applying a nonpermeable TEV protease solution (SI Appendix, Fig. S7 C-H), which allowed us to observe the pH-Lemon signals emanated only from SVs. Using this probe, we found that the luminal pH of SVs in SCAMP5 KD neurons was significantly lower than that in control neurons (Fig. 4 F-I and N). " "We then cotransfected pH-Lemon-TEV-Syt1 along with shRNA SCAMP5 or scrambled RNA into NHE6 KO neurons at days in vitro (DIV) 8 to 10, and the luminal pH of SVs was measured at DIV 14 to 16 (SI Appendix, Figs. S8H and S9). The efficiency of SCAMP5 KD in NHE6 KO cells was confirmed by immunoblotting and ICC in mouse hippocampal cell line HT-22 and cultured rat hippocampal neurons, respectively (SI Appendix, Fig. S10 A-C). We found that the luminal pHs of SVs in NHE6 KO and SCAMP5 KD + NHE6 KO neurons were significantly lower than that in control neurons (Fig. 4 J-N). However, SCAMP5 KD + NHE6 KO did not reduce the SV pH any further than NHE6 KO or SCAMP5 KD. These results indicate that the loss of NHE6 occluded the effect of SCAMP5 KD. Together, our results show that the luminal pH of SVs is regulated through proper NHE6 localization mediated by SCAMP5, and ectopic mislocalization of NHE6 in SCAMP5 KD neurons is responsible for hyperacidification of the SV lumen. " |
| Evidence tracking, Biological System: | Cultured neurons |
| Evidence tracking, Protein Targeting: | Genetic transformation (eg; knockout, knockin, mutations) |
| Evidence tracking, Experiment Assay: | Whole-cell patch clamp pHluorin assay |
| Annotator(s): | Frank Koopmans (ORCID:0000-0002-4973-5732) 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 |
| SynGO annotation ID: | 5541 |
| Dataset release (version): | 20231201 |
| View annotation as GO-CAM model: |  |