Key resources table.
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Gene (Mus musculus) | Stxbp5 | NCBI | 78808 | syntaxin binding protein 5 (tomosyn) |
| Gene (Mus musculus) | Stxbp5l | NCBI | 207227 | syntaxin binding protein 5-like (tomosyn-2) |
| Genetic reagent (Mus musculus) | C57BL/6J-Stxbp5lox/Stxbp5llox | This paper | - | Generation of the mouse model is described in Materials and Methods (Mice) |
| Genetic reagent (Mus musculus) | C57BL/6 J | Charles River | Strain code 631 | |
| Genetic reagent (Rattus norvegicus) | Wistar (Crl:WI) | Charles River | Strain code: 003 | Used for preparation of glia feeder layer |
| Antibody | anti-syntaxin (mouse monoclonal) | Sigma | S0664 | 1:2000 |
| Antibody | anti-SNAP25 (mouse monoclonal) | Covance | SMI-81R | 1:1000 |
| Antibody | anti-VAMP2 (mouse monoclonal) | Synaptic Systems | 104211 | 1:2000 |
| Antibody | anti-tomosyn (rabbit polyclonal) | Synaptic Systems | 183103 | 1:1000 (WB) 1:500 (ICC) |
| Antibody | anti-tomosyn-2 (rabbit polyclonal) | Synaptic Systems | 183203 | 1:1000 |
| Antibody | anti-alpha-tubulin (mouse monoclonal) | Synaptic Systems | 302211 | 1:1000 |
| Antibody | anti-actin (mouse monoclonal) | Merck | MAB1501 | 1:1000 |
| Antibody | anti-synaptophysin (guinea pig polyclonal) | Synaptic Systems | 101004 | 1:1000 |
| Antibody | anti-MAP2 (chicken polyclonal) | Abcam | ab5392 | 1:5000 |
| Antibody | anti-IA-2 (PTPRN) (mouse monoclonal) | Merck | MABS469 | 1:100 |
| Antibody | anti-CHGA (rabbit polyclonal) | Synaptic Systems | 259003 | 1:500 |
| Antibody | anti-CHGB (rabbit polyclonal) | Synaptic Systems | 259103 | 1:500 |
| Antibody | anti-BDNF (mouse monoclonal) | DSHB hybridoma product | BDNF #9 (supernatant)1 | 1:4 |
| Antibody | anti-BDNF (mouse monoclonal) | Biosensis | BSENM-1736–50 (clone 3C11) | 1:500 |
| Antibody | anti-LAMP1 (rat monoclonal) | DSHB hybridoma product | 1D4B (supernatant)2 | 1:20 (ICC) 1:200 (WB) |
| Antibody | anti-LIMP2 (rabbit polyclonal) | Novus Biologicals | NB400-129 | 1:1000 |
| Antibody | anti-GAPDH (rabbit polyclonal) | Elabscience | E-AB-40337 | 1:2000 |
| Antibody | anti-TMEM87A (rabbit polyclonal) | Novus Biologicals | NBP1-90532 | 1:100 |
| Antibody | anti-TGN38 (sheep polyclonal) | Bio-Rad | AHP499G | 1:100 |
| Recombinant DNA reagent | pLenti-Syn(pr)-Cre-mCherry | Wolzak et al., 2022 | - | |
| Recombinant DNA reagent | pLenti-Syn(pr)-ΔCre-mCherry | Wolzak et al., 2022 | - | Cre lacking catalytic domain |
| Recombinant DNA reagent | pLenti-Syn(pr)-pre-NPY-pHluorin | Nassal et al., 2022 | - | |
| Recombinant DNA reagent | pLenti-mScarlet-T2A-tomosyn | This paper | - | mouse isoform m (NM_001081344). Generation of this reagent is described in Materials and methods (Expression constructs) |
| Recombinant DNA reagent | pLenti-mScarlet-T2A-tomosyn(aa1-1047) | This paper | - | Generation of this reagent is described in Materials and methods (Expression constructs) |
| Recombinant DNA reagent | pLenti-Syn(pr)-mCherry | This paper | - | Generation of this reagent is described in Materials and methods (Expression constructs) |
| Recombinant DNA reagent | pCMV-Streptavidin-KDEL-IRES-EGFP-NPY | Emperador-Melero et al., 2018 | - | Construct used for RUSH assay |
| Sequence-based reagent | qPCR primers are listed in Table 1 | |||
| Peptide, recombinant protein | 2.5% trypsin | gibco | 15090046 | |
| Peptide, recombinant protein | papain | Worthington Biochemical Corporation | LS003127 | |
| Peptide, recombinant protein | poly-L-ornithine | Sigma | P4957 | |
| Peptide, recombinant protein | laminin | Sigma | L2020 | |
| Peptide, recombinant protein | rat tail collagen | BD Biosciences | 354236 | |
| Peptide, recombinant protein | poly-D-lysine | Sigma | P6407 | |
| Commercial assay or kit | SensiFast cDNA Synthesis Kit | Meridian Bioscience | BIO-65054 | |
| Commercial assay or kit | SensiFast SYBR Lo-ROX Kit | Meridian Bioscience | BIO-94020 | |
| Chemical compound, drug | leupeptin | Hello Bio | HB3958 | |
| Chemical compound, drug | biotin | Sigma | B4501 | |
| Software, algorithm | MATLAB | MathWorks | - | |
| Software, algorithm | Prism | GraphPad | - | |
| Software, algorithm | Fiji/ImageJ | NIH | - |