guinea pig anti dyn antibody  (Neuromics)

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

Journal: Endocrinology

Article Title: Short-Term High-Fat Diet Increases Leptin Activation of CART Neurons and Advances Puberty in Female Mice

doi: 10.1210/en.2017-00452

Figure Lengend Snippet: Representative photomicrographs (magnification ×63) showing immunostaining for (A) NKB in red and (B) DYN in blue and Kiss1-hrGFP neurons in green, and the overlapping images of the rostral arcuate of female mice (PN32) fed an RD or HFD. The images are the maximum intensity projection of a series of 35 individual optical planes. Scale bar = 20 μm.

Article Snippet: The sections were washed, and, after blocking [5% normal horse serum, 0.4% Triton X-100 in tris(hydroxymethyl)aminomethane; pH 7.6], the slices were incubated overnight in different primary antibody combinations: rabbit anti-NKB antibody (1:2500; Bachem, Torrance, CA), guinea pig anti-DYN antibody (1:250; Neuromics, Minneapolis, MN) or rabbit anti–p-STAT3 (1:1000; Cell Signaling Technology, Danvers, MA), and goat anti-CART (1:500; Santa Cruz Biotechnology, Dallas, TX).

Techniques: Immunostaining

Journal: Endocrinology

Article Title: Short-Term High-Fat Diet Increases Leptin Activation of CART Neurons and Advances Puberty in Female Mice

doi: 10.1210/en.2017-00452

Figure Lengend Snippet: Antibody Table

Article Snippet: The sections were washed, and, after blocking [5% normal horse serum, 0.4% Triton X-100 in tris(hydroxymethyl)aminomethane; pH 7.6], the slices were incubated overnight in different primary antibody combinations: rabbit anti-NKB antibody (1:2500; Bachem, Torrance, CA), guinea pig anti-DYN antibody (1:250; Neuromics, Minneapolis, MN) or rabbit anti–p-STAT3 (1:1000; Cell Signaling Technology, Danvers, MA), and goat anti-CART (1:500; Santa Cruz Biotechnology, Dallas, TX).

Techniques: Sequencing

Journal: PLoS ONE

Article Title: Hypothalamic Orexin-A Neurons Are Involved in the Response of the Brain Stress System to Morphine Withdrawal

doi: 10.1371/journal.pone.0036871

Figure Lengend Snippet: Representative photographs showing double-label immunohistochemistry for TH and OXA (NTS, A), CRF and OXA (BNST, B; PVN, D; and CeA, E) and pro-DYN and OXA (NAc, C). Scale bar, 50 µm. Images depict the orexin A innervations (blue-black) of the CRF, pro-DYN and TH neurons (brown) in the extended amygdala, NTS and PVN. Higher magnification images of the boxes areas depicted in each photomicrograph show examples of the areas where OXA axons are opposed to the somata of CRF, TH and pro-DYN cells. Abbreviations: NTS, nucleus of the solitary tract; BNST, bed nucleus of the stria terminalis; NAc, nucleus accumbens; PVN, hypothalamic paraventricular nucleus; CeA, central amygdala; cc, canal central; 3V, third ventricle.

Article Snippet: Following the OXA fibers staining, sections were rinsed in PBS, treated with 3% bovine serum albumin and then incubated with the guinea-pig anti-DYN antibody (1∶2000, Neuromics, Edina, MN, USA), rabbit anti-CRF antibody or anti-TH antibody (1∶6000, Chemicon, Temecula, CA).

Techniques: Immunohistochemistry