Key resources table
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Mouse monoclonal anti-ß-Tubulin | Sigma-Aldrich | Cat#T4026; RRID:AB_477577 |
| Rabbit polyclonal anti-Munc13-1 | Synaptic Systems | Cat# 126 103; RRID:AB_887733 |
| Rabbit polyclonal anti-Munc13-1 (C-terminal #N395) | (Betz et al., 1997) | N/A |
| Rabbit polyclonal anti-Munc13-1 (N-terminal #40) | (Cooper et al., 2012; Varoqueaux et al., 2005) | N/A |
| Mouse monoclonal IgG1 anti-synaptophysin-1 (A100) | Synaptic Systems | Cat# 101 011; RRID:AB_887824 |
| Guinea pig polyclonal anti-Tyrosine hydroxylase (A111) | Synaptic Systems | Cat# 213 104, RRID:AB_2619897 |
| Rabbit polyclonal anti-Tyrosine hydroxylase (A66) | Millipore | Cat# AB152; RRID:AB_390204 |
| Rabbit monoclonal anti-GFP (A195) | Thermo Fisher Scientific | Cat# G10362; RRID:AB_2536526 |
| Mouse monoclonal IgG2a anti-Bassoon SAP7F407 (A85) | Enzo Life Sciences | Cat# ADI-VAM-PS003-F; RRID:AB_11181058 |
| Rabbit polyclonal antibody anti-RIM1 (A58) | Synaptic Systems | Cat# 140 003 RRID:AB_887774 |
| Rabbit polyclonal anti-ELKS2α (1029, A136) | (Held et al., 2016) | E3-1029 |
| Mouse monoclonal anti-HA-tag 6E2 (A250) | Cell Signaling Technology | Cat# 2367; RRID:AB_10691311 |
| Chemicals, Peptides, and Recombinant Proteins | ||
| Tetrodotoxin | Tocris | Cat. No. 1078 |
| Dihydro-β-erythroidine hydrobromide | Tocris | Cat. No. 2349 |
| Mammalian Protease Inhibitor Cocktail | Sigma | Cat# P8340 |
| Image-iT™ FX Signal Enhancer | Thermo Fisher Scientific | Cat# I36933 |
| Experimental Models: Cell Lines | ||
| Derivate of 129/Ola mouse embryonic stem cell line E14 | (Hooper et al., 1987) | RRID:CVCL_9108 |
| Mouse embryonic fibroblasts (MEF feeder cells) | Xenogen, Alameda, CA, USA | N/A |
| HEK293T | ATCC | Cat#: CRL-3216, RRID:CVCL_0063 |
| Experimental Models: Organisms/Strains | ||
| Mouse: Munc13-1fl/fl | This study | N/A |
| Mouse: Munc13-1 cKOgl | This study | N/A |
| Mouse: Munc13-1−/−; unc13atm1Bros | (Augustin et al., 1999) | MGI:2449468 |
| Mouse: Munc13-1mNeo/mNeo | (Rhee et al., 2002) | N/A; mutated Unc13a gene containing a neomycin cassette |
| Mouse: Munc13-2−/−; unc13btm1Rmnd | (Varoqueaux et al., 2002) | MGI:2449706 |
| Mouse: Munc13-3−/−; unc13ctm1Bros | (Augustin et al., 2001) | MGI:2449467 |
| Mouse: FLP deleter; Gt(ROSA)26Sortm1(FLP1)Dym | (Farley et al., 2000) | MGI:2429412 |
| Mouse: Ella-Cre; Tg(EIIa-cre)C5379Lmgd | (Lakso et al., 1996) | MGI:2137691 |
| Mouse: DATIRES-cre; B6.SJL-Slc6a3tm1.1(cre)Bkmm/J | (Backman et al., 2006) | JAX 006660, |
| Mouse: Munc13-1-EYFP; Unc13atm3Bros | (Kalla et al., 2006) | MGI:3695486 |
| Mouse: Liprin-α3−/− | (Wong et al., 2018) | N/A |
| Mouse: Liprin-α2fl/fl | (Emperador-Melero et al., 2021a) | MGI:2443834 |
| Mouse: RIM-BP1fl/fl; Tspoap1tm1Sud/J | (Acuna et al., 2015) | JAX: 023643 |
| Mouse: RIM-BP2fl/fl; Rimbp2tm1Sud/J | (Acuna et al., 2015) | JAX: 023644 |
| Mouse: RIM1fl/fl; Rims1tm3Sud/J | (Kaeser et al., 2008) | JAX:015832 |
| Mouse: RIM2fl/fl; Rims2tm1.1Sud/J | (Kaeser et al., 2011) | JAX:015833 |
| Recombinant DNA | ||
| BAC: BMQ-441L13 (pBACe3.6 backbone) | (Adams et al., 2005) | N/A |
| Targeting vector: PL253-M13-1loxP-e21-FRT-Neo-FRT-loxP-HSV-TK | This study | N/A |
| pAAV-hSyn-flex-RIM1-Zn-HA (p866) | This study | N/A |
| pAAV-hSyn-flex-RIM1-ΔZn-HA (p865) | This study | N/A |
| pAAV-hSyn-flex-RIM1-Zn-HA-C2B (p902) | This study | N/A |
| pAAV-hSyn-flex-RIM1-Zn-HA- Zn-C2BKE (p903) | This study | N/A |
| pAAV-hSyn-flex-RIM1-Zn-HA-PCP (p968) | This study | N/A |
| pAAV-hSyn-flex-oChIEF-citrine (p901) | Addgene, (Lin et al., 2009) | Plasmid# 50973, RRID:Addgene_50973 |
| pAAV-hSyn-flex-citrine (p906) | This study | N/A |
| Oligonucleotides | ||
| Munc13-1 genotyping: 5'-CTCTATGGCTTCTGAGGCGGAAA-3' |
This study | LabID:27121 |
| Munc13-1 genotyping: 5'-AGTTTTCATCTTGTAGCCCGAT-3' |
This study | LabID:27122 |
| Munc13-1 genotyping: 5'-CAACTGGCCAAGAACTAGAGGA-3' |
This study | LabID:27123 |
| Munc13-1 genotyping: 5'-GCACGGAATGTTGAATGGTCTT -3' |
This study | LabID:27124 |
| Munc13-1 splice variants, exon 20; sense: 5’-CCATCCGGCTTCACATCAGT-3’ | This study | LabID:24609 |
| Munc13-1 splice variants, exon 23; antisense: 5’-CGATCCGCAGAGAATTGTGTAGC-3’ | This study | LabID:34504 |
| Munc13-1 splice variants, exon 25; antisense: 5’-TACTCCCGGCCATAGAGCTCA-3’ | This study | LabID:34505 |
| Software and Algorithms | ||
| Custom machine learning code for TH morphology | This study | DOI: 10.5281/zenodo.5576622 |
| Custom MATLAB code | (Liu et al., 2018) | DOI: 10.5281/zenodo.5576635 |
| DNASTAR Lasergene 13 | DNASTAR | RRID:SCR_011854; https://www.dnastar.com/software/lasergene/ |
| GraphPad Prism | GraphPad | RRID:SCR_002798; http://www.graphpad.com/ |
| Fiji | Schindelin et al., 2012 | RRID:SCR_002285, https://imagej.net/Fiji/Downloads |
| DeltaVision OMX imaging system | GE Healthcare | RRID: SCR_019956; https://scicrunch.org/resources/Tools/record/nlx_144509-1/SCR_019956/resolver?q=SCR_019956&l=SCR_019956 |
| Olympus Fluoview FV10-ASW | Olympus Fluoview | RRID:SCR_014215, http://www.photonics.com/Product.aspx?PRID=47380 |
| pClamp | Molecular Devices | RRID:SCR_011323, http://www.moleculardevices.com/products/software/pclamp.html |
| Imaris 9.0.2 | Oxford Instruments | RRID:SCR_007370 http://www.bitplane.com/imaris/imaris |