Plasmid Preparation:Article Title: Whole-brain afferent input mapping to functionally distinct brainstem noradrenaline cell types.
Article Snippet: .. Plasmid for adeno-associated virus-CAG promoter-flip-excisionflippase (FLP) recognition site-TVA receptor fused to mCherry (pAAVCAG-FLEX(FRT)-TC, titer estimated to be 1.893 and 9.56 × 1012 GC/ml, Schwarz et al., 2015) was provided by Dr. Liqun Luo (Addgene plasmid #67827, RRID:Addgene_67827). .. Plasmid for AAV-FRT site-human histone 2B-human influenza hemaggluttin (HA)-2A self-cleaving peptide-glicoprotein N2c strain (pAAV-CBA-FRT-H2B-3xHA-P2A-N2cG, titer estimated to be 5.28 ×1012 GC/ml and 1.09 ×1013 GC/ml) was produced and package in our lab. Plasmid for the envelope glycoprotein of subgroup A-pseudotyped CVS-N2cΔG fused with EGFP (EnvA-CVS-N2cΔG-EGFP, titer estimated to be 2 ×107 GC/ml, Reardon et al., 2016) was provided by Dr. Thomas Jessell (Addgene plasmid #73461, RRID:Addgene_73461).
Virus:Article Title: Whole-brain afferent input mapping to functionally distinct brainstem noradrenaline cell types.
Article Snippet: .. Plasmid for adeno-associated virus-CAG promoter-flip-excisionflippase (FLP) recognition site-TVA receptor fused to mCherry (pAAVCAG-FLEX(FRT)-TC, titer estimated to be 1.893 and 9.56 × 1012 GC/ml, Schwarz et al., 2015) was provided by Dr. Liqun Luo (Addgene plasmid #67827, RRID:Addgene_67827). .. Plasmid for AAV-FRT site-human histone 2B-human influenza hemaggluttin (HA)-2A self-cleaving peptide-glicoprotein N2c strain (pAAV-CBA-FRT-H2B-3xHA-P2A-N2cG, titer estimated to be 5.28 ×1012 GC/ml and 1.09 ×1013 GC/ml) was produced and package in our lab. Plasmid for the envelope glycoprotein of subgroup A-pseudotyped CVS-N2cΔG fused with EGFP (EnvA-CVS-N2cΔG-EGFP, titer estimated to be 2 ×107 GC/ml, Reardon et al., 2016) was provided by Dr. Thomas Jessell (Addgene plasmid #73461, RRID:Addgene_73461).
Article Title: Encoding the glucose identity by discrete hypothalamic neurons via the gut-brain axis.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Goat polyclonal anti-Fos Santa Cruz Cat# sc-52-g; RRID: AB_2629503 Rabbit anti-CRF Forver Vale Lab, Salk Institute Cat# CRF; RRID: AB_2314231 Rabbit anti-alpha-CGRP Bachem Cat# T-4032; RRID: AB_518147 Donkey anti-Goat IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 647 Thermo Fisher Scientific Cat# A-21447; RRID: AB_2535864 Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 594 Thermo Fisher Scientific Cat# A-21207; RRID: AB_141637 Donkey anti-rabbit IgG (H+L) highly cross-adsorbed secondary antibody, Alexa Fluor 647 Thermo Fisher Scientific Cat# A-31573; RRID: AB_2536183 Anti-NeuN-antibody Millipore Cat# ABN91; RRID: AB_11205760 HA-Tag (C29F4) Rabbit mAb Cell Signaling Technology Cat# 3724; RRID: AB_1549585 Goat anti-Chicken IgY (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 Thermo Fisher Scientific Cat# A32931; RRID: AB_2762843 Bacterial and virus strains AAV1-CAG-Flex-GCaMP6s-WPRE-SV40 Gift from Dr. Douglas Kim & GENIE Project Cat# 100835-AAV1; RRID: Addgene_100835 AAV1-CMV-eGFP-Cre-WPRE-SV40 Gift from Dr. James M. Wilson Cat# 105545-AAV1; RRID: Addgene_105545 AAV5-hSyn-hM4D(Gi)-mCherry Gift from Dr. Bryan Roth Cat# 50475-AAV5; RRID: Addgene_50475 AAV1 hSyn FLEx mGFP-2ASynaptophysin-mRuby Gift from Dr. Liqun Luo Cat# 71760-AAV1; RRID: Addgene_71760 AAV2-Ef1a-DIO-ChR2-eYFP Gift from Dr. Karl Deisseroth Cat# 35507-AAV1; RRID: Addgene_35507 AAV5-Ef1a-DIO-eYFP UNC vector core Cat# AAV5-Ef1a-DIO-eYFP AAV2-EF1α-DIO-hM4D(Gi)-mCherry UNC Vector Core Cat# AAV2-EF1α-DIO-hM4D(Gi)-mCherry AAV2-hSyn-DIO-mCherry UNC Vector Core Cat# AAV2-hSyn-DIO-mCherry AAV2-Ef1a-DIO-eArch3.0-eYFP UNC vector core Cat# AAV2-Ef1a-DIO-eArch3.0-eYFP AAV1-DIO-RG Gift from Dr. SeungHee Lee N/A AAV1-DIO-TVA-EGFP Gift from Dr. SeungHee Lee N/A pRV-EnvA-ΔG-tdTomato Gift from Dr. SeungHee Lee N/A AAV2/1-hS-FLEX-TVA-HA-N2cG Kavli Institute for Systems Neuroscience Cat# AAV2/1-hS-FLEX-TVA-HA-N2cG EnvA pseudotyped SADB19 Rb mCherry Kavli Institute for Systems Neuroscience Cat# EnvA pseudotyped SADB19 Rb mCherry EnvA pseudotyped N2cG Rb mCherry Kavli Institute for Systems Neuroscience Cat# EnvA pseudotyped N2cG Rb mCherry Chemicals, peptides, and recombinant proteins D-glucose Sigma Cat# G8270 L-glucose Carbosynth Cat# MG05247 D-fructose Sigma Cat# F0127 Sucrose Sigma Cat# S7903 Sucralose Sigma Cat# 69293 Intralipid Sigma Cat# I141 (Continued on next page) e1 Neuron 113, 1–19.e1–e9, August 20, 2025 .. REAGENT or RESOURCE SOURCE IDENTIFIER Protein Proteinex-18 Proteinex-18 Ensure® Abbott Ensure® original powder D-galactose Sigma Cat# G0750 D-Mannitol Sigma Cat# M4125 Acesulfame K Supelco Cat# 04054 Methyl α-D-glucopyranoside Sigma Cat# 9376 Cholecystokinin octapeptide (sulfated) Bachem Cat#4033010 Peptide Y.Y. (3-36) Tocris Cat# 1618 Liraglutide Tocris Cat# 6517 Amylin Tocris Cat# 3418 Lithium chloride solution Sigma Cat# L7026 Ghrelin(rat) Tocris Cat# 1465 4OH-tamoxifen Sigma Cat# H6278 Clozapine N-oxide (CNO) Sigma Cat# C0832 Dexamethasone sodium phosphate Sigma Cat# PHR1768 TritonTM-X100 Sigma Cat# T8787 DAPI Sigma Cat# D9542 Nissl stain Invitrogen Cat# N21479 Tween® 80 Sigma Cat# P1754 polyglycerol 3-polyglycidyl ether Huntsman Corporation Cat# GE-38 PBS Nalgene Cat# AM9625 sodium carbonate Sigma-Aldrich Cat# S7795 sodium bicarbonate Sigma-Aldrich Cat# S5761 sodium azide Sigma-Aldrich Cat# S2002 PBSN Sigma-Aldrich Cat# P4417 boric acid Sigma-Aldrich Cat# B6768 sodium sulfite Sigma-Aldrich Cat# S0505 sodium dodecyl sulfate Sigma-Aldrich Cat# 75746 sodium hydroxide Sigma-Aldrich Cat# S5881 PES vacuum filter VWR Cat# 10040-440 Triton-X Sigma-Aldrich Cat# T9284 2,2 ′ -Thiodiethanol Sigma-Aldrich Cat# 88561 dimethyl sulfoxide Alfa-Aesar Cat# 36480 Normal donkey serum Abcam Cat# ab7475; RRID: AB_2885042 Fluorescence Mounting Medium Agilent Cat# S3023 Fluoro-Gold Fluorochrome, LLC N/A Silicone Elastomer Sealant Word Precision Instruments Kwik-Cast Ketoprofen Zoetis KETOFEN Enrofloxacin Baytril® N/A Experimental models: Organisms/strains Mouse: Crf-IRES-: B6(Cg)-Crh tm1(cre)Zjh/J The Jackson Laboratory RRID: IMSR_JAX: 012704 Mouse: Crf flox: B Cre 6.129S4(Cg)-Crh tm2.1Maj /J The Jackson Laboratory RRID: IMSR_JAX: 030110 Mouse: ROSA26iDTR: BL/6-Gt (ROSA)26Sor tm1(HBEGF)Awai /J The Jackson Laboratory RRID: IMSR_JAX: 007900 Mouse: Ai14: B6.Cg-Gt(ROSA) 26Sor tm14(CAG-tdTomato)Hze /J The Jackson Laboratory RRID: IMSR_JAX: 007914 (Continued on next page) Neuron 113, 1–19.e1–e9, August 20, 2025 e2
Recombinant:Article Title: Encoding the glucose identity by discrete hypothalamic neurons via the gut-brain axis.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Goat polyclonal anti-Fos Santa Cruz Cat# sc-52-g; RRID: AB_2629503 Rabbit anti-CRF Forver Vale Lab, Salk Institute Cat# CRF; RRID: AB_2314231 Rabbit anti-alpha-CGRP Bachem Cat# T-4032; RRID: AB_518147 Donkey anti-Goat IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 647 Thermo Fisher Scientific Cat# A-21447; RRID: AB_2535864 Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 594 Thermo Fisher Scientific Cat# A-21207; RRID: AB_141637 Donkey anti-rabbit IgG (H+L) highly cross-adsorbed secondary antibody, Alexa Fluor 647 Thermo Fisher Scientific Cat# A-31573; RRID: AB_2536183 Anti-NeuN-antibody Millipore Cat# ABN91; RRID: AB_11205760 HA-Tag (C29F4) Rabbit mAb Cell Signaling Technology Cat# 3724; RRID: AB_1549585 Goat anti-Chicken IgY (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 Thermo Fisher Scientific Cat# A32931; RRID: AB_2762843 Bacterial and virus strains AAV1-CAG-Flex-GCaMP6s-WPRE-SV40 Gift from Dr. Douglas Kim & GENIE Project Cat# 100835-AAV1; RRID: Addgene_100835 AAV1-CMV-eGFP-Cre-WPRE-SV40 Gift from Dr. James M. Wilson Cat# 105545-AAV1; RRID: Addgene_105545 AAV5-hSyn-hM4D(Gi)-mCherry Gift from Dr. Bryan Roth Cat# 50475-AAV5; RRID: Addgene_50475 AAV1 hSyn FLEx mGFP-2ASynaptophysin-mRuby Gift from Dr. Liqun Luo Cat# 71760-AAV1; RRID: Addgene_71760 AAV2-Ef1a-DIO-ChR2-eYFP Gift from Dr. Karl Deisseroth Cat# 35507-AAV1; RRID: Addgene_35507 AAV5-Ef1a-DIO-eYFP UNC vector core Cat# AAV5-Ef1a-DIO-eYFP AAV2-EF1α-DIO-hM4D(Gi)-mCherry UNC Vector Core Cat# AAV2-EF1α-DIO-hM4D(Gi)-mCherry AAV2-hSyn-DIO-mCherry UNC Vector Core Cat# AAV2-hSyn-DIO-mCherry AAV2-Ef1a-DIO-eArch3.0-eYFP UNC vector core Cat# AAV2-Ef1a-DIO-eArch3.0-eYFP AAV1-DIO-RG Gift from Dr. SeungHee Lee N/A AAV1-DIO-TVA-EGFP Gift from Dr. SeungHee Lee N/A pRV-EnvA-ΔG-tdTomato Gift from Dr. SeungHee Lee N/A AAV2/1-hS-FLEX-TVA-HA-N2cG Kavli Institute for Systems Neuroscience Cat# AAV2/1-hS-FLEX-TVA-HA-N2cG EnvA pseudotyped SADB19 Rb mCherry Kavli Institute for Systems Neuroscience Cat# EnvA pseudotyped SADB19 Rb mCherry EnvA pseudotyped N2cG Rb mCherry Kavli Institute for Systems Neuroscience Cat# EnvA pseudotyped N2cG Rb mCherry Chemicals, peptides, and recombinant proteins D-glucose Sigma Cat# G8270 L-glucose Carbosynth Cat# MG05247 D-fructose Sigma Cat# F0127 Sucrose Sigma Cat# S7903 Sucralose Sigma Cat# 69293 Intralipid Sigma Cat# I141 (Continued on next page) e1 Neuron 113, 1–19.e1–e9, August 20, 2025 .. REAGENT or RESOURCE SOURCE IDENTIFIER Protein Proteinex-18 Proteinex-18 Ensure® Abbott Ensure® original powder D-galactose Sigma Cat# G0750 D-Mannitol Sigma Cat# M4125 Acesulfame K Supelco Cat# 04054 Methyl α-D-glucopyranoside Sigma Cat# 9376 Cholecystokinin octapeptide (sulfated) Bachem Cat#4033010 Peptide Y.Y. (3-36) Tocris Cat# 1618 Liraglutide Tocris Cat# 6517 Amylin Tocris Cat# 3418 Lithium chloride solution Sigma Cat# L7026 Ghrelin(rat) Tocris Cat# 1465 4OH-tamoxifen Sigma Cat# H6278 Clozapine N-oxide (CNO) Sigma Cat# C0832 Dexamethasone sodium phosphate Sigma Cat# PHR1768 TritonTM-X100 Sigma Cat# T8787 DAPI Sigma Cat# D9542 Nissl stain Invitrogen Cat# N21479 Tween® 80 Sigma Cat# P1754 polyglycerol 3-polyglycidyl ether Huntsman Corporation Cat# GE-38 PBS Nalgene Cat# AM9625 sodium carbonate Sigma-Aldrich Cat# S7795 sodium bicarbonate Sigma-Aldrich Cat# S5761 sodium azide Sigma-Aldrich Cat# S2002 PBSN Sigma-Aldrich Cat# P4417 boric acid Sigma-Aldrich Cat# B6768 sodium sulfite Sigma-Aldrich Cat# S0505 sodium dodecyl sulfate Sigma-Aldrich Cat# 75746 sodium hydroxide Sigma-Aldrich Cat# S5881 PES vacuum filter VWR Cat# 10040-440 Triton-X Sigma-Aldrich Cat# T9284 2,2 ′ -Thiodiethanol Sigma-Aldrich Cat# 88561 dimethyl sulfoxide Alfa-Aesar Cat# 36480 Normal donkey serum Abcam Cat# ab7475; RRID: AB_2885042 Fluorescence Mounting Medium Agilent Cat# S3023 Fluoro-Gold Fluorochrome, LLC N/A Silicone Elastomer Sealant Word Precision Instruments Kwik-Cast Ketoprofen Zoetis KETOFEN Enrofloxacin Baytril® N/A Experimental models: Organisms/strains Mouse: Crf-IRES-: B6(Cg)-Crh tm1(cre)Zjh/J The Jackson Laboratory RRID: IMSR_JAX: 012704 Mouse: Crf flox: B Cre 6.129S4(Cg)-Crh tm2.1Maj /J The Jackson Laboratory RRID: IMSR_JAX: 030110 Mouse: ROSA26iDTR: BL/6-Gt (ROSA)26Sor tm1(HBEGF)Awai /J The Jackson Laboratory RRID: IMSR_JAX: 007900 Mouse: Ai14: B6.Cg-Gt(ROSA) 26Sor tm14(CAG-tdTomato)Hze /J The Jackson Laboratory RRID: IMSR_JAX: 007914 (Continued on next page) Neuron 113, 1–19.e1–e9, August 20, 2025 e2
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