receptor guinea pig polyclonal antibody  (Neuromics)

Journal: Nature Communications

Article Title: Altered surface mGluR5 dynamics provoke synaptic NMDAR dysfunction and cognitive defects in Fmr1 knockout mice

doi: 10.1038/s41467-017-01191-2

Figure Lengend Snippet: mGluR5 and GluN1 are more confined within the synaptic compartment in Fmr1 KO neurons. ( a ) Representative surface distribution of mGluR5-QD (upper panel) and GluN1-QD (lower panel) in a 500-frame stack (each dot represents the detection of a single receptor during 50-ms acquisition time), revealing the synaptic site as a trapping zone (green). ( b ) Relative fractions of synaptic mGluR5-QD (left panel) and GluN1-QD (right panel) particles. These values are increased in Fmr1 KO neurons (mGluR5-QD: WT, 8.053 ± 0.504 %, n = 16 dendritic fields from 3 cultures; Fmr1 KO, 15.95 ± 0.685 %, n = 14 dendritic fields from 3 cultures; ***P < 0.001, t = 9.44, df = 28, unpaired Student’s t -test; GluN1-QD: WT, 8.318 ± 0.382 %, n = 63 dendritic fields from 6 cultures; Fmr1 KO, 10.02 ± 0.446 %, n = 51 dendritic fields from 6 cultures; **P < 0.01, t = 2.91, df = 112, unpaired Student’s t -test)

Article Snippet: For single-molecule tracking experiments, neurons were first exposed for 10 min to either mouse monoclonal anti-NH 2 mGluR5 antibody (1:20) , mouse monoclonal anti-GluA2 AMPA receptor (AMPAR) subunit antibody (Millipore, #MAB397, 1:200), or rabbit polyclonal anti-GluN1 NMDA receptor (NMDAR) subunit antibody (Alomone Laboratories, #AGC-001, 1 : 200 and Supplementary refs. – ) at 37 °C.

Techniques:

Journal: Nature Communications

Article Title: Altered surface mGluR5 dynamics provoke synaptic NMDAR dysfunction and cognitive defects in Fmr1 knockout mice

doi: 10.1038/s41467-017-01191-2

Figure Lengend Snippet: Disruption of the mGluR5/Homer scaffold increases surface mGluR5/NMDAR co-clustering. ( a ) Cultured WT and Fmr1 KO hippocampal neurons were triple-labeled for mGluR5, GluN1 and Homer1. ( b , c ) Left: Representative image of Homer/mGluR5 and Homer/GluN1 co-localization; middle: Distribution of co-localized mGluR5/Homer1 or GluN-/Homer1 clusters; right: mGluR5/Homer1 and GluN1/Homer1 clusters as percentage of total mGluR5 or GluN1 signal (mGluR5: WT, 9.53 ± 0.959 %, n = 27; Fmr1 KO, 34.14 ± 4.598 %, n = 20; ***P < 0.001, t = 6, df = 45, unpaired Student’s t -test; GluN1: WT 15.75 ± 1.841 %, n = 29; Fmr1 KO 37.15 ± 5.324 %, n = 18; ***P < 0.001, t = 4.47, df = 45, unpaired Student’s t -test). ( d ) Left: Representative image showing mGluR5/GluN1/Homer1 colocalization; middle: Distribution of co-localized mGluR5/GluN1/Homer1 labeling; right: Co-localized mGluR5/GluN1/Homer1 clusters as percentage of synaptic GluN1 signal (WT, 63.97 ± 2.414 %, n = 26; Fmr1 KO, 72.14 ± 2.081 %, n = 23; *P < 0.05, t = 2.53, df = 47, unpaired Student’s t -test). ( e ) TAT-mGluR5ct peptide increased mGluR5/GluN1 co-clustering at synapses in WT neurons, whereas TAT-mGluR5mu or TAT-mGluR5ct (both 5 µM, 1 h) had no effect in Fmr1 KO neurons; Left: Representative images and distribution of mGluR5/GluN1/Homer1-co-labeling signal in control and TAT-mGluR5mu or TAT-mGluR5ct treated WT and Fmr1 KO neurons. Right: Co-localized mGluR5/GluN1/Homer1-positive signals as percentage of synaptic GluN1 signal (WT: 63.97 ± 2.414 %, n = 26; WT TAT-mGluR5mu: 61.19 ± 3.489 %, n = 14; WT TAT-mGluR5ct: 71.79 ± 1.528 %, n = 22; WT vs. WT TAT-mGluR5ct, *P = 0.043; WT TAT-mGluR5mu vs. WT TAT-mGluR5ct * P = 0.017, F (2, 59) = 4.87; Fmr1 KO: 72.14 ± 2.081 %, n = 23; Fmr1 KO TAT-mGluR5mu: 67.32 ± 2.832 %, n = 18; Fmr1 KO TAT-mGluR5ct: 67.58 ± 2.69 %, n = 26; Fmr1 KO vs. Fmr1 KO TAT-mGluR5ct P = 0.391; Fmr1 KO TAT-mGluR5mu vs. Fmr1 KO TAT-mGluR5ct P = 0.997, F (2, 64) = 1.13). P values by one-way ANOVA test with Tukey’s Multiple Comparison test. n = dendritic fields from 3 cultures. Scale bar = 10 μm ( a ) 2 μm ( b , c , d , e )

Article Snippet: For single-molecule tracking experiments, neurons were first exposed for 10 min to either mouse monoclonal anti-NH 2 mGluR5 antibody (1:20) , mouse monoclonal anti-GluA2 AMPA receptor (AMPAR) subunit antibody (Millipore, #MAB397, 1:200), or rabbit polyclonal anti-GluN1 NMDA receptor (NMDAR) subunit antibody (Alomone Laboratories, #AGC-001, 1 : 200 and Supplementary refs. – ) at 37 °C.

Techniques: Cell Culture, Labeling

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Mast cell expression of the serotonin 1A receptor in guinea pig and human intestine

doi: 10.1152/ajpgi.00421.2012

Figure Lengend Snippet: A purified polyclonal 5-HT1A receptor antibody recognized a 45-kDa band on Western blots, indicative of 5-HT1A receptor expression, in 50-μg protein extracts from guinea pig and human mucosa with attached submucosal plexus and from guinea pig and human myenteric plexus-longitudinal muscle preparations. The same antibody recognized 5-HT1A receptor protein on Western blots. A: absence of protein recognition by the primary antibody for the lysis buffer used for guinea pig mucosal-submucosal plexus preparations. B: antibody recognition of a 45-kDa band in extract obtained from mucosal-submucosal plexus preparations from guinea pig small intestine. C: antibody recognition of a 45-kDa band in extract obtained from myenteric plexus-longitudinal (Long) muscle preparations from guinea pig small intestine. D: absence of protein recognition by the primary antibody for the lysis buffer used for guinea pig myenteric plexus-longitudinal muscle preparations. E: recognition of 5-HT1A receptor protein by the antibody used for guinea pig preparations. F: absence of protein recognition by the primary antibody for the lysis buffer used for human mucosal-submucosal plexus preparations. G: antibody recognition of a 45-kDa band in extract obtained from mucosal-submucosal plexus preparations from human small intestine. H: antibody recognition of a 45-kDa band in extract obtained from myenteric plexus-longitudinal muscle preparations from human small intestine. I: absence of protein recognition by the primary antibody for the lysis buffer used for human myenteric plexus-longitudinal muscle preparations. J: recognition of 5-HT1A receptor protein by the antibody used for human preparations. Primary antibody (5 ml, 1:200 dilution) was obtained from ImmunoStar (catalog no. 24504); secondary antibody was donkey horseradish peroxidase (10 ml, 1:5,000 dilution) 5-HT1A receptor protein (catalog no. ab152462, Abcam).

Article Snippet: A purified rabbit polyclonal 5-HT 1A receptor antibody (catalog no. 20079, ImmunoStar) and a guinea pig polyclonal 5-HT 1A receptor antibody (catalog no. 550469, BD Bioscience, Franklin Lakes, NJ), which labeled 5-HT 1A receptor protein on Western blots, recognized a corresponding 45-kDa band in protein extracts from whole-mount myenteric and submucosal plexuses and from intact intestinal segments obtained from three guinea pigs and three human Roux-en-Y gastric bypass surgeries ( ).

Techniques: Purification, Western Blot, Expressing, Lysis

Journal: Neural Regeneration Research

Article Title: Similar effects of substance P on learning and memory function between hippocampus and striatal marginal division

doi: 10.4103/1673-5374.131603

Figure Lengend Snippet: Neurokinin 1 receptor-positive neurons in the hippocampus as shown by immunofluorescence staining. (A) The structure of the hippocampus. (B, C) Neurokinin 1 receptor-positive immunofluorescence-labeled neurons expressed in the CA1 field of the hippocampus. The scale bar is 250 μm in A, and 50 μm in B and C. Fluorescent stain is Cy3; positive expression is shown red. Arrows identify neurons. Hip: Hippocampus.

Article Snippet: The sections were then incubated in guinea pig anti-neurokinin 1 receptor polyclonal antibodies (1:1,000; Chemicon) diluted in 0.01 mol/L PBS (pH 7.4) with 1% bovine serum albumin, 0.3% TritonX-100 and 0.05% sodium azide (NaN 3 ) for 36–48 hours at 4°C.

Techniques: Immunofluorescence, Staining, Labeling, Expressing

Journal: Neural Regeneration Research

Article Title: Similar effects of substance P on learning and memory function between hippocampus and striatal marginal division

doi: 10.4103/1673-5374.131603

Figure Lengend Snippet: Neurokinin 1 receptor-positive neurons in the neostriatum (CPU, GP, MrD) as shown by immunofluorescence staining. (A) A stippled pattern of neurokinin 1 receptor-positive neurons in both the medial striatum and more caudally in the dorsolateral part of the striatum. (B) Neurokinin 1 receptor-positive fibers were dense in the MrD. (C) The neurokinin 1 receptor-positive fusiform neurons in the MrD were moderate in size and had two spiny primary dendrites emerging dorsoventrally from the two poles of the cell bodies. The scale bar is 250 μm in A, and 50 μm in B and C. Fluorescent stain is Cy3; positive expression is shown red. Arrows identify neurokinin 1 receptor-positive neurons. CPU: Caudate putamen; GP: Globus pallidus; MrD: marginal division.

Article Snippet: The sections were then incubated in guinea pig anti-neurokinin 1 receptor polyclonal antibodies (1:1,000; Chemicon) diluted in 0.01 mol/L PBS (pH 7.4) with 1% bovine serum albumin, 0.3% TritonX-100 and 0.05% sodium azide (NaN 3 ) for 36–48 hours at 4°C.

Techniques: Immunofluorescence, Staining, Expressing

Journal: Neural Regeneration Research

Article Title: Similar effects of substance P on learning and memory function between hippocampus and striatal marginal division

doi: 10.4103/1673-5374.131603

Figure Lengend Snippet: Comparison of learning and memory in a Y-maze after blockade of neurokinin 1 receptor mRNA. The data are expressed as mean ± SD, with eight rats in each group. The distribution of the values was checked for normality. The comparisons between the groups were conducted by one-way analysis of variance followed by Student's t -test. a P < 0.01, b P < 0.05, vs . pre-operation. Hipb: Bilateral hippocampus injection group; MrDb: bilateral MrD injection group; Hipi: unilateral hippocampus injection group; MrDi: unilateral MrD injection group; NSb: normal control group received 2 μL normal saline; MrD: marginal division.

Article Snippet: The sections were then incubated in guinea pig anti-neurokinin 1 receptor polyclonal antibodies (1:1,000; Chemicon) diluted in 0.01 mol/L PBS (pH 7.4) with 1% bovine serum albumin, 0.3% TritonX-100 and 0.05% sodium azide (NaN 3 ) for 36–48 hours at 4°C.

Techniques: Injection