KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Rabbit anti-Phox2a (1:10,000 from frozen stock; Figures 1, 2, 3, 4, 5, and 6) | Jean-François Brunet (École normale supérieure, Paris, France) | (Tiveron et al., 1996) RRID: AB_2315159 |
| Rabbit anti-Phox2a (Figure 7; and 1:1000 for mouse, 1:500 for human embryo) | Abcam | Cat#: Ab155084 Lot#: GR117345–3 RRID: N/A |
| Rabbit anti-Phox2b (1:10,000 from frozen stock) | Jean-François Brunet (École normale supérieure, Paris, France) | (Pattyn et al., 1997) RRID: AB_2315160 |
| Rabbit anti-RFP (red fluorescent protein) (1:1000) | Rockland | Cat#: 600-401-379 RRID: AB_2209751 |
| Mouse anti-NeuN (1:1000) | Millipore | Cat#: MAB377 RRID: AB_2298772 |
| Guinea Pig anti-vGlut2 (1:1000) | Synaptic Systems | Cat#: 135–404 RRID: AB_887884 |
| Rat anti-Bromodeoxyuridine (BrdU) (1:10,000) | Abcam | Cat# ab6326: RRID: AB_305426 |
| Goat anti-rTrkA (1:1000 for mouse, 1:500 for human embryo) | R&D Systems | Cat#: Af1056 RRID: AB_2283049 |
| Mouse anti-Islet1 (1:100) | Developmental Studies Hybridoma Bank (DSHB) | Cat#: 39.3F7 RRID: AB_1157901 |
| Mouse anti-Pax7 (1:100) | DSHB | Cat#: pax7 RRID: AB_528428 |
| Mouse anti-Lhx2 (1:100) | DSHB | Cat#: Lhx2–1C11 RRID: AB_2618817 |
| Mouse anti-Nkx6.1 (1:100) | DSHB | Cat#: F55A10 RRID: AB_532378 |
| Mouse anti-Neurofilament-L (NF-L) | DSHB | Cat#: 2H3 RRID: AB_531793 |
| Goat anti-hPax2 (1:1000 for mouse, 1:500 for human embryo) | R&D Systems | Cat#: AF3364 RRID: AB_10889828 |
| Guinea Pig anti-Lbx1 (1:10000 for mouse, 1:5000 for human embryo) | Carmen Birchmeier (Max Delbruck Center, Berlin, Germany) | RRID: AB_2532144 |
| Guinea Pig anti-Lmx1b (1:10000 for mouse, 1:5000 for human embryo) | Carmen Birchmeier (Max Delbruck Center, Berlin, Germany) | RRID: AB_2314752 |
| Guinea Pig anti-Tlx3 (1:10000 in mouse, 1:10000 for chick, 1:5000 for human embryo) | Carmen Birchmeier (Max Delbruck Center, Berlin, Germany) | RRID:AB_2532145 |
| Guinea pig anti-PRDM13 (1:1000) | Takahisa Furukawa (Osaka University, Osaka, Japan) | (Watanabe et al., 2015) RRID: N/A |
| Guinea pig anti-PTF1A (1:10000) | Jane Johnson (University of Texas Southwestern, Dallas, United States) | TX507 RRID: N/A |
| Mouse anti-MYC (1:1000) | Abcam | Cat# ab32 RRID: AB_303599 |
| Goat anti-Brn3b (Pou4F2), (1:1000 for mouse, 1:500 for human embryo) | Santa Cruz Biotechnology | Cat#: sc-6026 RRID: AB_673441 |
| Mouse anti-Ascl1 (1:100) | Santa Cruz Biotechnology | Cat#: sc-390794 RRID: N/A |
| Mouse anti-Ascl1 (1:100) | Santa Cruz Biotechnology | Cat#: sc-374550 RRID: AB_10985986 |
| Mouse anti-Ascl1 (1:100) | Santa Cruz Biotechnology | Cat# sc-374104 RRID: AB_10918561 |
| Goat anti-hRobo3 | R&D Systems | Cat# AF3076 RRID: AB_2181865 |
| Goat anti-mDcc | R&D Systems | Cat# AF844 RRID: AB_2089765 |
| Rat anti-Pou6F2 (1:2000) | Jay Bikoff (Thomas Jessell Laboratory, HHMI Columbia University, New York, United States) | Cat# CU1796 RRID: AB_2665427 |
| Rabbit anti-Shox2 (1:200) | Laskaro Zagoraiou (Thomas Jessell Laboratory, HHMI Columbia University, New York, United States) | (Dougherty et al., 2013) RRID: N/A |
| Sheep anti-FoxP2 (1:2000) | R&D Systems | Cat#: AF5647 RRID: AB_2107133 |
| Alexa 488 Donkey anti-Rabbit (1:500) | Jackson Immunoresearch Laboratories | Cat#: 711-545-152 Lot#: 141848 RRID: AB_2313584 |
| Alexa 488 Donkey anti-Guinea Pig (1:500) | Jackson Immunoresearch Laboratories | Cat#: 706-545-148 Lot#: 138058 RRID: AB_2340472 |
| Alexa 488 Donkey anti-Mouse (1:500) | Jackson Immunoresearch Laboratories | Cat#: 715-545-150 Lot#: 136831 RRID: AB_2340846 |
| Alexa 488 Donkey anti-Goat (1:500) | Jackson Immunoresearch Laboratories | Cat#: 705-545-147 Lot#: 136089 RRID: AB_2336933 |
| Alexa 488 Donkey anti-Rat (1:500) | Jackson Immunoresearch Laboratories | Cat#: 712-545-153 Lot#: 138117 RRID: AB_2340684 |
| Alexa 488 Donkey anti-Sheep (1:500) | Jackson Immunoresearch Laboratories | Cat#: 713-545-003 Lot#: N/A RRID: AB_2340744 |
| Cy3 Donkey anti-Rat (1:500) | Jackson Immunoresearch Laboratories | Cat#: 712-165-153 Lot#: 139289 RRID: AB_2340667 |
| Cy3 Donkey anti-Rabbit (1:500) | Jackson Immunoresearch Laboratories | Cat#: 711-165-152 Lot#: 138270 RRID: AB_2307443 |
| Cy3 Donkey anti-Mouse (1:500) | Jackson Immunoresearch Laboratories | Cat#: 715-165-150 Lot#: N/A RRID: AB_2340813 |
| Cy3 Donkey anti-Goat (1:500) | Jackson Immunoresearch Laboratories | Cat#: 705-165-147 Lot#: 134527 RRID: AB_2307351 |
| Cy5 Donkey anti-Rabbit (1:500) | Jackson Immunoresearch Laboratories | Cat#: 711-175-152 Lot#: 138336 RRID: AB_2340607 |
| Cy5 Donkey anti-Mouse (1:500) | Jackson Immunoresearch Laboratories | Cat#: 715-175-150 Lot#: 135323 RRID: AB_2340819 |
| Cy5 Donkey anti-Goat (1:500) | Jackson Immunoresearch Laboratories | Cat#: 705-175-147 Lot#: 134531 RRID: AB_2340415 |
| Cy5 Donkey anti-Guinea Pig (1:500) | Jackson Immunoresearch Laboratories | Cat#: 706-175-148 Lot#: 136607 RRID: AB_2340462 |
| Chemicals, Peptides, and Recombinant Proteins | ||
| NeuroTrace 435/455 Blue Fluorescent Nissl Stain | Thermofisher Scientific | Cat#: N21479 RRID: N/A |
| NeuroTrace 500/525 Green Fluorescent Nissl Stain | Thermofisher Scientific | Cat#: N21480 RRID: N/A |
| 2-Hydroxystilbene-4,4′-dicarboxamidine (fluorogold) | Thermofisher Scientific | Cat#: H22845 RRID: N/A |
| Alexa 488-conjugated Choleratoxin B | Thermofisher Scientific | Cat#: C22841 Lot#: 2038245 RRID: N/A |
| 5-bromo-2′-deoxyuridine (BrdU) | Thermofisher Scientific | Cat#: B23151 Lot#: 1916418 RRID: N/A |
| Paraformaldehyde | Millipore Sigma | Cat#: P6148 RRID: N/A |
| (E)-Capsaicin | Tocris | Cat#: 0462 Lot#: 7A/218361 RRID: N/A |
| 4′,6-diamidino-2-phenylindole (DAPI) | Thermo Fisher Scientific | Cat# D1306 |
| Mowiol (Polyvinyl alcohol) | Millipore Sigma | Cat#: 81381 RRID: N/A |
| Experimental Models: Organisms/Strains | ||
| Mice - Phox2aCre | This manuscript | RRID: N/A |
| Mice - HoxB8Cre (Tg(Hoxb8-cre)1403Uze) | Hanns Ulrich Zeilhofer, ETH Zürich, Zurich, Switzerland) | Cat#: MGI: 4881836 RRID: N/A |
| Mice - Cdx2FlpO (Tg(CDX2-flpo)#Gld) | Martyn Goulding (Salk Institute, San Diego, United States) | Cat#: MGI: 5911680 RRID: N/A |
| Mice - Ai14 (B6;129S6-Gt(ROSA)26Sortm14(CAG-tdTomato)Hze/J) | The Jackson Laboratory | Cat#: JAX:007908 RRID: IMSR_JAX:007908 |
| Mice - Ai65 (B6;129S-Gt(ROSA)26Sortm65.1(CAG-tdTomato)Hze/J) | The Jackson Laboratory | Cat#: JAX:021875 RRID: IMSR_JAX:021875 |
| Mice - Phox2alox (B6D2.129S2-Phox2atm2Jbr/Orl) | European Mutant Mouse Archive (EMMA) | Cat#: EM:04758 RRID:IMSR_EM:04758 |
| Mice – TrkA−/− (Ntrk1tm1Par) | Lino Tessarollo (National Cancer Institute, Frederick, MD, United States) | (Liebl et al., 2000) Cat#: MGI: 1933963 RRID: N/A |
| Mice – Ptf1atm1(cre)Wri | Christopher Wright (Vanderbilt University, Nashville, United States) | (Kawaguchi et al., 2002) Cat#: MGI: 2387812 RRID: N/A |
| Mice - Ascl1tm1Reed/J | The Jackson Laboratory | Cat# JAX:012881 RRID: IMSR_JAX:012881 |
| Mice - C57BL/6J | The Jackson Laboratory | Cat# JAX:000664 RRID: IMSR_JAX:000664 |
| Mice - 129S1/SvImJ | The Jackson Laboratory | Cat# JAX:002448 RRID: IMSR_JAX:002448 |
| Mice - B6C3F1/J | The Jackson Laboratory | Cat# JAX: 100010 RRID: IMSR_JAX: 100010 |
| Oligonucleotides | ||
| RNA Scope Probe - Mm-Tac1 C1 | Advanced Cell Diagnostics | Cat#: 410351 Lot#: 18354A RRID: N/A |
| RNA Scope Probe - Mm-Phox2a C2 | Advanced Cell Diagnostics | Cat#: 520371-C2 Lot#: N/A RRID: N/A |
| RNA Scope Probe - Mm-Lmx1b C3 | Advanced Cell Diagnostics | Cat#: 412931-C3 Lot#: N/A RRID: N/A |
| RNA Scope Probe - Mm-Nms C1 | Advanced Cell Diagnostics | Cat#: 472331 Lot#: TBD RRID: N/A |
| RNA Scope Probe - Mm-Tm4sf4 C1 | Advanced Cell Diagnostics | Cat#: 819831 Lot#: N/A RRID: N/A |
| RNA Scope Probe - Mm-Zim1 C1 | Advanced Cell Diagnostics | Cat#: 819821 Lot#: N/A RRID: N/A |
| RNA Scope Probe - Mm-Scn9a C1 | Advanced Cell Diagnostics | Cat#: 313341 Lot#: N/A RRID: N/A |
| RNA Scope Probe - Mm-Pdzrn3 C1 | Advanced Cell Diagnostics | Cat#: 517061 Lot#: 17269A RRID: N/A |
| RNA Scope Probe - Mm-Syt4 C1 | Advanced Cell Diagnostics | Cat#: 574731 Lot#: N/A RRID: N/A |
| RNA Scope Probe - Mm-VIP C1 (Vasoactive Intestinal Polypeptide) | Advanced Cell Diagnostics | Cat#: 415961 Lot#: 19045A RRID: N/A |
| RNA Scope Probe - Mm-Sst C1 | Advanced Cell Diagnostics | Cat#: 404631 Lot#: N/A RRID: N/A |
| RNA Scope Probe - Mm-TacR3 C1 | Advanced Cell Diagnostics | Cat#: 481671 Lot#: 18254A RRID: N/A |
| RNA Scope Probe - Mm-Crh C1 | Advanced Cell Diagnostics | Cat#: 316091 Lot#: N/A RRID: N/A |
| RNA Scope Probe - Mm-Slc17A6 C1 (vGlut2) | Advanced Cell Diagnostics | Cat#: 319171 Lot#: 19052B RRID: N/A |
| RNA Scope Probe - Mm-Slc32A1 C1 (vGAT) | Advanced Cell Diagnostics | Cat#: 319191 Lot#: 19057A RRID: N/A |
| RNA Scope Probe - Mm-TacR1 C1 | Advanced Cell Diagnostics | Cat#: 428781 Lot#: 19057A RRID: N/A |
| RNA Scope Probe - Mm-Cck C1 (Cholecystokinin) | Advanced Cell Diagnostics | Cat#: 402271 Lot#: 19057A RRID: N/A |
| RNA Scope Probe - Mm-Lypd1 C1 | Advanced Cell Diagnostics | Cat#: 318361 Lot#: 18353B RRID: N/A |
| RNA Scope Probe - Mm-Gal C1 (Galanin) | Advanced Cell Diagnostics | Cat#: 400961 Lot#: 18277C RRID: N/A |
| RNA Scope Probe - Mm-pDyn C1 (preproDynorphin) | Advanced Cell Diagnostics | Cat#: 318771 Lot#: 18303A RRID: N/A |
| RNA Scope Probe - Mm-pNoc C1 (prepronociceptin) | Advanced Cell Diagnostics | Cat#: 437881 Lot#: 19016B RRID: N/A |
| Genotyping primers | ||
| Cre-1: 5′-AGG TGT AGA GAA GGC ACT TAG C −3′ Expected band size: 412 bp (only one band) | This manuscript | N/A |
| Cre-2: 5′-CTA ATC GCC ATC TTC CAG CAG G-3′ | This manuscript | N/A |
| FLPo-1: 5′-TGA GCT TCG ACA TCG TGA AC-3′ Expected band size: 350 bp (only one band) | Martyn Goulding (Salk Institute, San Diego, United States) | N/A |
| FLPo-2: 5′-ACA GGG TCT TGG TCT TGG TG −3′ | Martyn Goulding (Salk Institute, San Diego, United States) | N/A |
| Ai14–1: 5′-TCA ATG GGC GGG GGT CGT T-3′ Expected band sizes: WT: 350 bp, Mutant: 250 bp | This manuscript | N/A |
| Ai14–2: 5′-CTC TGC TGC CTC CTG GCT TCT-3′ | This manuscript | N/A |
| Ai14–3: 5′-CGA GGC GGA TCA CAA GCA ATA-3′ | This manuscript | N/A |
| Ai65 WT1: “oIMR9020” 5′-AAG GGA GCT GCA GTG GAG TA-3′ Expected band size: 315 bp | The Jackson Laboratory | N/A |
| Ai65 WT2: “oIMR9021” 5′-CCG AAA ATC TGT GGG AAG TC-3′ | The Jackson Laboratory | N/A |
| Ai65 Mutant1: “oIMR9103” 5′-GGC ATT AAA GCA GCG TAT CC-3′ Expected band size: 297 bp | The Jackson Laboratory | N/A |
| Ai65 Mutant2: “oIMR9105” 5′-CTG TTC CTG TAC GGC ATG G-3′ | The Jackson Laboratory | N/A |
| Phox2afl-1: 5′-GCC TCC AAC TCC ATA TTC C-3′ Expected band sizes: WT: 150bp, Flox: 200bp | Jean-François Brunet (École normale supérieure, Paris, France) | N/A |
| Phox2afl-2: 5′-ATC AGG AGT CAG TCG TCT G −3′ | Jean-François Brunet (École normale supérieure, Paris, France) | N/A |
| TrkA-WT-5′: 5′-TGT ACG GCC ATA GAT AAG CAT-3′ Expected WT band size: 160 bp | Lino Tessarollo (National Cancer Institute, Frederick, MD, United States) | N/A |
| TrkA-WT-3′: 5′-TTG CAT AAC TGT GTA TTT CAC-3′ | Lino Tessarollo (National Cancer Institute, Frederick, MD, United States) | N/A |
| TrkA-mutant (pGKneopolyA) forward primer: 5′-CGC CTT CTT GAC GAG TTC TTC TG-3′ Expected mutant band size: 550 bp | Lino Tessarollo (National Cancer Institute, Frederick, MD, United States) | N/A |
| Recombinant DNA | ||
| Plasmid: pMiWIII-Myc- ASCL1 | Jane Johnson (University of Texas Southwestern, Dallas, United States) | N/A |
| Plasmid: pMiWIII-Myc- PTF1A | Jane Johnson (University of Texas Southwestern, Dallas, United States) | N/A |
| Plasmid: pMiWIII-Prdm13 | Jane Johnson (University of Texas Southwestern, Dallas, United States) | N/A |
| Plasmid: pMiWIII-Myc-tag | Jane Johnson (University of Texas Southwestern, Dallas, United States) | N/A |
| Plasmid: pMCSIII-ePhox2a | This manuscript | N/A |
| Software and Algorithms | ||
| Graphpad Prism 8 (macOS) - Version 8.3.0 | Graphpad Software | https://www.graphpad.com/scientific-software/prism RRID: SCR_002798 |
| Bioseb T2CT - Version 2.2.4 | Bioseb | https://www.bioseb.com/en/pain-thermal-allodynia-hyperalgesia/897-thermal-place-preference-2-temperatures-choice-nociception-test.html RRID: N/A |
| ImageJ - Version 2.0.0 | National Institutes of Health | https://imagej.nih.gov/ij/ RRID: SCR_003070 |
| Photoshop and Illustrator | Adobe | N/A |
| Excel for Mac 2011 v14.7.7 | Microsoft | N/A |
| Aegisub v3.2.2 | Aegisub | http://www.aegisub.org/ |
| Other | ||
| Stereotaxic frame and digital display | David Kopf Instruments | Cat#s: 940, 960, 1770, 900C, 922, 933-B RRID: N/A |
| Stereotaxic syringe pump with Micro4 controller | David Kopf Instruments | Cat#s: UMP3-1, 1770-C RRID: N/A |
| Touch Test Sensory Evaluators (0.008, 0.02, 0.04, 0.07, 0.16, 0.4, 0.6, 1, 1.4, 2g) | North Coast Medical | Cat#: NC12775–01 - NC12775–10 RRID: N/A |
| IITC Hot Cold-Plate Analgesia Meter for Mice and Rats | IITC | Cat#: PE34 RRID: N/A |
| Thermal Place Preference, 2 Temperatures Choice Nociception Test | Bioseb | Cat#: BIO-T2CT RRID: N/A |
| iPhone SE-Version 12.3.1 | Apple | Cat#: iPhone SE. RRID: N/A |
| Micro Toothless Alligator Test Clip (Copper Plated) | https://www.amazon.com/Toothless-Alligator-Copper-Plated-Microscopic/dp/B0187MIUU4 | Cat#: CECOMINOD005515 RRID: N/A |
| Confocal Microscope | Zeiss | Cat#: LSM-710 |
| Confocal Microscope | Leica | Cat#: SP8 |
| Epifluorescence Upright Microscope | Leica | Cat#: DM6000 |
| Epifluorescence Upright Microscope | Leica | Cat#: DM6 |
| Epifluorescence Dissecting Stereomicroscope | Leica | Cat#: MZ16FA |