| Annotated protein: | Ras-related protein Rab-5C. Gene symbol: RAB-5. Taxonomy: Caenorhabditis elegans (Worm). Uniprot ID: P91857 |
| antibody wiki: | |
| SynGO gene info: | SynGO data @ RAB5A SynGO data @ RAB5B SynGO data @ RAB5C |
| Ontology domain: | Biological Process |
| SynGO term: | presynaptic dense core vesicle exocytosis (GO:0099525) |
| Synapse type(s): | spinal cord, acetylcholinergic |
| Annotated paper: | Sasidharan N, et al. "RAB-5 and RAB-10 cooperate to regulate neuropeptide release in Caenorhabditis elegans" Proc Natl Acad Sci U S A. 2012 Nov 13;109(46):18944-9 PMID:23100538 |
| Figure(s): | Figure 1, S1, S2, S3, S4 and S5 |
| Annotation description: | Figure 1A: DCV secretion from cholinergic motoneurons was assayed using a transgenic strain (nuIs183) expressing the proneuropeptide NLP-21 tagged to YFP exclusively in dorsally projecting cholinergic motoneurons. This strain has been successfully used to assay DCV biogenesis and release. At the trans-Golgi, the proneuropeptide NLP-21-YFP fusion is packaged into DCVs, where it is subsequently processed into active neuropeptides and soluble YFP. YFP-labeled DCVs are then transported to axonal release sites. After fusion, the soluble YFP is released into the body cavity. Here, it is constitutively endocytosed by macrophage-like cells, called coelomocytes. The YFP fluorescence in coelomocytes can, therefore, be used as an indirect readout for DCV release. Figure 1B and S1: To identify Rabs that are regulating DCV release, all viable rab mutants were screened for defects in NLP-21-YFP secretion resulting in low coelomocytes fluorescence. As a positive control, the paper uses unc-31/CAPS mutants, which are known to have defects in DCV release. Figure 1C: Expression of constitutively active RAB-5 Q78L mutant also led to strongly impaired DCV secretion. Figure S2: DCV secretion defect of rab-5 mutants were also confirmed using another DCV marker, the insulin-like neuropeptide INS-22-YFP. Figure S3: High-pressure freeze electron microscopic (HPF-EM) analysis showed normal DCV numbers at axonal release sites with a similar distribution and morphology compared with wild type. This indicates that Rab5 mutants do not affect the number of DCV but rather the secretion process. Figure S4: To demonstrate that the results derived from Figure 1 aren't due to disruption in coelomocyte function, rab-5 mutants were analyzed for the coelomocyte uptake of soluble Texas Red-conjugated BSA (TR-BSA) injected into the body cavity. Neither uptake nor the kinetics of endocytosis was affected in rab-5 mutants. Figure S5 D-F: In order to check if the defect in DCV exocytosis was cell-autonomous, tissue-specific knockdown of rab-5 by RNAi phenocopied the defect in NLP-21-YFP secretion with decreased YFP levels in coelomocytes. 20/01/2021 Dnyanada: Information on the species of the Rab5 cDNA used for generating genetic lines could not be retrieved. Hence, P91857 was chosen. |
| Evidence tracking, Biological System: | Intact tissue |
| Evidence tracking, Protein Targeting: | Genetic transformation (eg; knockout, knockin, mutations) RNAi / shRNA Antibody (detection) |
| Evidence tracking, Experiment Assay: | Wide-field fluorescence |
| 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 |
| SynGO annotation ID: | 4068 |
| Dataset release (version): | 20231201 |
| View annotation as GO-CAM model: |  |