guinea pig anti ppdyn antibody  (Neuromics)

Journal: bioRxiv

Article Title: Parabrachial neuron types categorically encode thermoregulation variables during heat defense

doi: 10.1101/2020.06.11.138370

Figure Lengend Snippet: a-c , Changes in basal T core ( a ), basal physical activity ( b ), and basal energy expenditure ( c ) after blocking of POA-projected LPB Pdyn neurons (n = 6 each; 4-6 weeks post viral injection). d-g , Changes of energy expenditure ( d-e ), and physical activity ( f-g ) during warm challenge ( d, f ) or cold challenge ( e, g ), respectively, after blocking of POA-projected LPB Pdyn neurons (n = 6 each; 6-8 weeks post viral injection). (– 360 – 0 min) and (0 – 360 min) represent the averaged EE between t = (– 360 – 0 min) and t = (0 – 360 min), respectively. All data are shown as mean ± s.e.m. All the p-values are calculated based on ordinary two-way ANOVA with Bonferroni’s corrections (right panels in a-g ). *p ≤ 0.05; ns, not significant.

Article Snippet: Primary antibodies include chicken anti-GFP (Abcam, #ab13970, 1: 1000), rabbit anti-Neuropeptide S (Abcam, #ab18252, 1: 500), rabbit anti-cfos (Synaptic systems, #226003, 1: 10000), guinea pig anti-cfos (Synaptic systems, #226004, 1: 500), rat anti-RFP (Chromotek, #5F8, 1: 1000), rabbit anti-Glutamate (Sigma, #G6642-.2ML, 1: 1000; see ( - ) for validation.), rabbit anti-Dynorphin A (Phoenix Pharmaceuticals, #H-021-03, 1:500) and Guinea pig anti-ProDynorphin (GeneTex, GTX10280, 1: 500).

Techniques: Activity Assay, Blocking Assay, Injection

Journal: Cell reports

Article Title: Dissecting the Roles of GABA and Neuropeptides from Rat Central Amygdala CRF Neurons in Anxiety and Fear Learning

doi: 10.1016/j.celrep.2019.08.083

Figure Lengend Snippet: (A) Injection schematic and representative image of bilateral AAV infection in CeA CRF neurons (DAPI counterstain). Scale bar, 500 μm. (B) Knockdown of VGAT in CeA CRF neurons reduced time spent in the open arms (t(22) = 3.158, **p = 0.0046; n = 12 for both groups) and entries into the open arms (t(22) = 7.858, ****p < 0.0001; n = 12 for both groups) on the elevated plus maze (EPM). (C) Knockdown of VGAT reduced time spent in the center(t(22) = 2.156, *p = 0.0423; n = 12 for both groups) and entries into the center(t(22) = 2.407, *p = 0.0249; n = 12 for both groups) of the open field (OF). (D) Knockdown of CRF, DYN, or NTS did not change time spent in the open arms of the EPM (F(3,39) = 0.5965, p = 0.6211, one-way ANOVA; n = 13 sh Con , 11 sh Crh , 10, sh Dyn , and 9 sh Nts ) but did increase the number of entries into the open arms (F(3,39) = 2.139, p = 0.0092, one-way ANOVA; n = 13 sh Con , 11 sh Crh , 10, sh Dyn , and 9 sh Nts ; *p < 0.05 compared with sh Con by Dunnett’s test). (E) Knockdown of CRF, DYN, or NTS did not change baseline anxiety-like behavior in the open field (time in the center: F(3,39) = 1.610, p = 0.2026; entries into the center: F(3,39) = 1.477, p = 0.2356; n = 13 sh Con , 11 sh Crh , 10, sh Dyn , and 9 sh Nts ). Data are represented as mean ± SEM.

Article Snippet: The blot was probed with 1:200 dilution of goat anti-CRF antibody (Santa Cruz Biotechnology, Dallas, TX, sc-1761) or 1:1000 dilution of guinea pig anti-DYN antibody (Neuromics, GP10110) in 5% milk overnight at 4°C with shaking.

Techniques: Injection, Infection, Knockdown

Journal: Cell reports

Article Title: Dissecting the Roles of GABA and Neuropeptides from Rat Central Amygdala CRF Neurons in Anxiety and Fear Learning

doi: 10.1016/j.celrep.2019.08.083

Figure Lengend Snippet: (A)Top, example image of dual infection of CeA CRF neurons with a cocktail of AAVs carrying shRNA and hM3Dq. Scale bar, 200 μm. Bottom, experimental protocol. (B) Knockdown of CRF led to more time spent on the open arms (t(17) = 3.613, **p = 0.0021; n = 9 sh Con and 10 sh Crh ) and more entries into the open arms (t(17) = 6.468, ****p < 0.0001; n = 9 sh Con and 10 sh Crh ) of the elevated plus maze after activation of CeA CRF neurons with hM3Dq and CNO (2 mg/kg i.p.). (C) Knockdown of CRF did not alter anxiety-like behavior in the open field (center time: t(17) = 0.854, p = 0.5669; center entries: t(17) = 0.208, p = 0.8376; n = 9 sh Con and 10 sh Crh ) after activation of CeA CRF neurons with hM3Dq and CNO (2 mg/kg i.p.). (D) Knockdown of DYN led to more time spent on the open arms (t(18) = 5.151, ****p < 0.0001; n = 9 sh Con and 11 sh Dyn ) and more entries into the open arms (t(18) = 5.589, ****p < 0.0001; n = 9 sh Con and 10 sh Dyn ) of the elevated plus maze after activation of CeA CRF neurons. (E) Knockdown of DYN led to more time spent in the center (U = 15, **p = 0.0074; n = 9 sh Con and 11 sh Dyn ) and more entries into the center (U = 17.5, *p = 0.013;n = 9 sh Con and 11 sh Dyn ) of the open field after activation of CeA CRF neurons. (F) Knockdown of NTS did not change the time spent on the open arms (t(17) = 0.4315, p = 0.6716; n = 10 sh Con and 9 sh Nts ) or entries into the open arms (t(17) = 0.5536, p = 0.5871; n = 10 sh Con and 9 sh Nts ) of the elevated plus maze after activation of CeA CRF neurons. (G) Knockdown of NTS did not change the time spent in the center (U = 30, p = 0.2428; n = 10 sh Con and 9 sh Nts ) and more entries into the center (t(17) = 1.319, p = 0.2046; n = 10 sh Con and 9 sh Nts ) of the open field after activation of CeA CRF neurons. Data are represented as mean ± SEM.

Article Snippet: The blot was probed with 1:200 dilution of goat anti-CRF antibody (Santa Cruz Biotechnology, Dallas, TX, sc-1761) or 1:1000 dilution of guinea pig anti-DYN antibody (Neuromics, GP10110) in 5% milk overnight at 4°C with shaking.

Techniques: Infection, shRNA, Knockdown, Activation Assay

Journal: Cell reports

Article Title: Dissecting the Roles of GABA and Neuropeptides from Rat Central Amygdala CRF Neurons in Anxiety and Fear Learning

doi: 10.1016/j.celrep.2019.08.083

Figure Lengend Snippet: (A) Injection schematics and example of viral expression in CeA CRF neurons infected with hM4Di-mCherry DREADD (left) and shRNA with EGFP (right). Scale bar, 200 μm. (B)Top, experimental protocol for fear conditioning. Bottom, chemogenetic inhibition of CeA CRF neurons with hM4Di and CNO(2 mg/kg i.p.) did not affect freezing during fear conditioning but disrupted contextual fear retrieval during the first minute (U = 1, ***p = 0.0003; n = 8 for both groups), as well as cued fear retrieval (t(14) = 4.846, ***p = 0.0003; n = 8 for both groups). (C) Top, experimental protocol. Bottom, shRNA-mediated knockdown of CRF and DYN, but not NTS, in CeA CRF neurons blunted contextual fear retrieval during the first minute (F(3,38) = 12.53, p < 0.0001, one-way ANOVA; n = 13 sh Con , 11 sh Crh , 9 sh Dyn , and 9 sh Nts ; ***p<0.0001 sh Crh compared with sh Con and **p = 0.0054 sh Dyn compared with sh Con by Dunnett’s test). Knockdown of CRF and DYN also reduced cued freezing, yet knockdown of NTS enhanced cued freezing (F(3,39) = 34.13, p < 0.0001; n = 13 sh Con , 11 sh Crh , 10 sh Dyn , and 9 sh Nts ; ****p < 0.0001 sh Crh compared with sh Con , ****p < 0.0001 sh Dyn compared with sh Con , and *p = 0.0115 sh Nts compared with sh Con by Dunnett’s test). (D) Knockdown of Vgat in CeA CRF neurons did not affect contextual fear learning (t(13) = 0.6684, p = 0.5156; n = 8 sh Con and 7 sh Vgat ) or cued fear learning (t(13) = 0.0125, p = 0.9902; n = 8 sh Con and 7 sh Vgat ). Data are represented as mean ± SEM.

Article Snippet: The blot was probed with 1:200 dilution of goat anti-CRF antibody (Santa Cruz Biotechnology, Dallas, TX, sc-1761) or 1:1000 dilution of guinea pig anti-DYN antibody (Neuromics, GP10110) in 5% milk overnight at 4°C with shaking.

Techniques: Injection, Expressing, Infection, shRNA, Inhibition, Knockdown

Journal: Cell reports

Article Title: Dissecting the Roles of GABA and Neuropeptides from Rat Central Amygdala CRF Neurons in Anxiety and Fear Learning

doi: 10.1016/j.celrep.2019.08.083

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: The blot was probed with 1:200 dilution of goat anti-CRF antibody (Santa Cruz Biotechnology, Dallas, TX, sc-1761) or 1:1000 dilution of guinea pig anti-DYN antibody (Neuromics, GP10110) in 5% milk overnight at 4°C with shaking.

Techniques: Virus, Recombinant, RNAscope, Multiplex Assay, cDNA Synthesis, Gene Expression, Bicinchoninic Acid Protein Assay, Software