Key resources table.
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Genetic reagent (Mus musculus) | Ntrk1Flag | (Sharma et al., 2010) PMID: 20696380 |
Mice were handled and housed in accordance with Harvard Medical School and Johns Hopkins University IACUC guidelines | |
| Genetic reagent (M. musculus) | Th2a-CreER | (Abraira et al., 2017) PMID: 28041852 |
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| Genetic reagent (M. musculus) | Rab7flox | (Roy et al., 2013) PMID: 23615463 |
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| Antibody | NeuN | Abcam (Cambridge, MA) | 1:1000 | |
| Antibody | Rab5 | 1:500 | ||
| Antibody | Rab7 | 1:200 for ICC; 1:1000 for immunoblot | ||
| Antibody | Lamp1 | 1:1000 | ||
| Antibody | P-Trk Y490 | Cell signaling(Danvers, MA) | 1:2000; 1:100 for immunoEM | |
| Antibody | P-Trk Y785 | |||
| Antibody | P-PLCγ | 1:500; 1:50 for immunoEM | ||
| Antibody | VAChT | Enzo Life Sciences (Farmingdale, NY) | 1:1000 | |
| Antibody | Homer1 | Synaptic Systems (Germany) | 1:500 | |
| Antibody | Flag | Sigma (St. Louis, MO) | 1 ug/ml | |
| Antibody | Alexa conjugated secondary antibodies (488, 555, 647) | Thermo Fisher Scientific (Waltham, MA) | 1:1000 | |
| Antibody | IgG F(ab’)2–6 nm/10 nm gold secondary antibodies | Aurion (Netherlands) | 1:50 | |
| Antibody | Protein A-5nm/10 nm gold | Made by The Harvard Medical School EM Facility | 1:50 | |
| Recombinant DNA reagent | FUW | (Lois et al., 2002) PMID: 11786607 |
Addgene 14882 | |
| Recombinant DNA reagent | APEX2 | (Lam et al., 2015) PMID: 25419960 |
Addgene 49385 | |
| Recombinant DNA reagent | FUW-EGFP-Rab5 | This paper | ||
| Recombinant DNA reagent | FUW-EGFP-Rab7 | This paper | ||
| Recombinant DNA reagent | FUW-CD63-EGFP | This paper | ||
| Recombinant DNA reagent | FUW-CD63-mCherry | This paper | ||
| Recombinant DNA reagent | FUW-RILP-EGFP | This paper | ||
| Recombinant DNA reagent | FUW-APEX2-Rab5 | This paper | ||
| Recombinant DNA reagent | FUW-APEX2-Vps35 | This paper | ||
| Recombinant DNA reagent | FUW-Flag-TrkB/A-WT | This paper | The TrkB/A chimeric receptor comprises the extracellular domain of TrkB (nucleotide 1–1242) and the transmembrane and intracellular domains of TrkA (nucleotide 1156–2400) |
|
| Recombinant DNA reagent | FUW-Flag-TrkB/A-F592A | This paper | ||
| Sequence-based reagent | Primers for genotypingTh2a-CreER | CATGCCCATATCCAATCTCC and CTGGAGCGCATGCAGTAGTA |
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| Sequence-based reagent | Primers for genotyping Rab7flox | CTCACTCACTCCTAAATGG and TTAGGCTGTATGTATGTGC |
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| Sequence-based reagent | shRNAs for Rab7 | GAAGTTCAGTAACCAGTACAA; GCGGCAGTATTCTGTACAGTA; GCCCTTAAACAGGAAACAGAA; TGAACCCATCAAACTGGACAA; TGCTGTGTTCTGGTGTTTGAT | ||
| Sequence-based reagent | shRNAs for RILP | CAGCTATGCAGGAGGCTTAAC; AGATCAAGGCCAAGATGTTAG; CCAGAATTTCTTTGGCTTATG; TTCAGCAGGGAAGAGCTTAAG; AGGAGCGGAATGAGCTCAAAG | ||
| Sequence-based reagent | Scrambled shRNA | CCTAAGGTTAAGTCGCCCTCG | ||
| Chemical compound, drug | K252a | EMD Millipore (Billerica, MA) | ||
| Chemical compound, drug | 1NMPP1 | |||
| Chemical compound, drug | Nocodazole | |||
| Chemical compound, drug | Saponin | MP Biomedicals (Santa Ana, CA) | ||
| Chemical compound, drug | 3.3’-Diaminobenzidine (DAB) | |||
| Chemical compound, drug | 6 nm gold-conjugated transferrin, CTB, BSA | Electron Microscopy Sciences (Hatfield, PA) |