tetrodotoxin  (Alomone Labs)

Journal: Cells

Article Title: Long-Term Dynamic Changes of NMDA Receptors Following an Excitotoxic Challenge

doi: 10.3390/cells11050911

Figure Lengend Snippet: nNOS (+) neurons show a decreased expression of the NMDA receptor subunit GluN1. ( A ) The pictogram illustrates the “patch-like” procedure employed to isolate mRNA obtained from single nNOS (+) neurons. The same approach was employed to isolate nNOS (−) neurons employed as control. ( B ) Bar graphs illustrate changes in mRNA levels of the indicated genes measured by real- time PCR in nNOS (+) neurons when compared to the general population of nNOS (−) cells (relative Nos1 expression in nNOS (−): 1.07 ± 0.22 vs. 2.92 ± 0.25 in nNOS (+) neurons, p

Article Snippet: Cells were incubated for 1 h at RT with anti-GluN1 antibody (1:200, Alomone, Israel) and anti-NOS1 antibody (1:50, Santa Cruz Biotechnology, Dallas, TX, USA) in the blocking solution.

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: bioRxiv

Article Title: Long-term dynamic changes of NMDA receptors following an excitotoxic challenge

doi: 10.1101/2022.01.03.474836

Figure Lengend Snippet: nNOS (+) neurons show a decreased expression of the NMDA receptor subunit GluN1. (A) The pictogram illustrates the “patch-like” procedure employed to isolate mRNA obtained from single nNOS (+) neurons. The same approach was employed to isolate nNOS (-) neurons employed as control. (B) Bar graphs illustrate changes in mRNA levels of the indicated genes measured by real-time PCR in nNOS (+) neurons when compared to the general population of nNOS (-) cells (relative Nos1 expression in nNOS (-): 1.07 ± 0.22 vs. 2.92± 0.25 in nNOS (+) neurons, p

Article Snippet: Cells were incubated for 1 h at RT with anti-GluN1 antibody (1:200, Alomone) and anti-NOS1 antibody (1:50, Santa Cruz Biotechnology) in the blocking solution.

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: Translational Psychiatry

Article Title: Alteration of NMDA receptor trafficking as a cellular hallmark of psychosis

doi: 10.1038/s41398-021-01549-7

Figure Lengend Snippet: Tuning GluN2B-NMDAR NMDAR surface dynamics alters sensorimotor gating in adults. a Schematic description of the experimental workflow: control of MAM-exposed pups received intracerebral injections during the second postnatal week of NMDAR surface diffusion modulators. b Representative trajectories (50 ms acquisition) of surface GluN2B-NMDAR on hippocampal neurons exposed to TAT-non sense (TAT-NS, 10 µM), TAT-2B (10 µM), control IgG (10 µg/ml), or IgG direct against extracellular epitope of the GluN2B-NMDAR (10 µg/ml). Scale bars = 500 nm (left/right). c The startle response, i.e., pre-pulse inhibition (PPI), was measured at P60 and compared between conditions ( n = 10–16 rats per group; * P

Article Snippet: Endogenous surface GluN2B-NMDAR or transfected GluN2A-SEP, GluN3A-SEP, GluA1-SEP, GABA-gamma2-GFP, or Dopamine D1-YFP were immune stained in live neurons using a rabbit polyclonal anti-GluN2B (1:200; Alomone, Israel) or mouse monoclonal anti-GFP (1:500, Roche, Switzerland), respectively.

Techniques: Diffusion-based Assay, Inhibition

Journal: Translational Psychiatry

Article Title: Alteration of NMDA receptor trafficking as a cellular hallmark of psychosis

doi: 10.1038/s41398-021-01549-7

Figure Lengend Snippet: Cerebrospinal fluid (CSF) from patients with SCZSD alters the surface dynamics of synaptic NMDAR. a Schematic description of the experimental workflow. CSF from patients with schizophrenic spectrum disorders (SCZSD, n = 12), affective spectrum disorders (AffectSD, n = 9) subarachnoid hemorrhage (SAH, n = 9 patients), brain polytrauma ( n = 3), hemophagocytic lymphohistiocytosis (HLH, n = 1), were collected. Then, single nanoparticle (QDot) tracking of GluN2B-NMDAR was performed onto cultured hippocampal neurons exposed to CSF. b Representative trajectories (50 ms acquisition) of surface GluN2B-NMDAR-QD complexes in dendritic spines of neurons exposed to various CSFs. Scale bar = 300 nm. c Diffusion coefficient (µm 2 /s) of synaptic GluN2B-NMDAR values after exposure for 15–30 min to patients’ CSF. Each plotted dot corresponds to the median diffusion coefficient value of one patient CSF. The mean and standard deviation (SD) are represented for each condition. Right panel , Comparison of GluN2B-NMDAR synaptic diffusion coefficient values after exposure for 15–30 min to patients’ CSF (Control, n = 2757 trajectories; SCZSD, n = 5849; AffectSD, n = 2410; Polytrauma, n = 825; SAB, n = 3957; *** p

Article Snippet: Endogenous surface GluN2B-NMDAR or transfected GluN2A-SEP, GluN3A-SEP, GluA1-SEP, GABA-gamma2-GFP, or Dopamine D1-YFP were immune stained in live neurons using a rabbit polyclonal anti-GluN2B (1:200; Alomone, Israel) or mouse monoclonal anti-GFP (1:500, Roche, Switzerland), respectively.

Techniques: Cell Culture, Diffusion-based Assay, Standard Deviation

Journal: Translational Psychiatry

Article Title: Alteration of NMDA receptor trafficking as a cellular hallmark of psychosis

doi: 10.1038/s41398-021-01549-7

Figure Lengend Snippet: NMDAR surface dynamics and synaptic plasticity are developmentally impaired in the MAM model. a Schematic description of the experimental workflow. Cultured hippocampal networks were made from rat pups. Examples of immuncytochemical staining of neurons (NeuN, green), nucleus (DAPI, blue), and glial cells (GFAP, red) in cultured network at 10 days in vitro from control or MAM-exposed pups. b Representative trajectories (50 ms acquisition) of surface GluN2B-NMDAR-QD complexes on neurons from control or MAM-exposed pups at three developmental stages: days in vitro 6–10 (D8), days in vitro 10–15 (D12), days in vitro 15–22 (D20). Scale bars = 5 µm/300 nm (left/right). Right, Comparison of GluN2B-NMDAR diffusion coefficient control or MAM-exposed conditions (D8 cont, n = 1916 trajectories; D8 MAM, n = 1220; D12 cont, n = 2055; D12 MAM, n = 946; D20 cont, n = 1288; D20 MAM, n = 1317; ** p

Article Snippet: Endogenous surface GluN2B-NMDAR or transfected GluN2A-SEP, GluN3A-SEP, GluA1-SEP, GABA-gamma2-GFP, or Dopamine D1-YFP were immune stained in live neurons using a rabbit polyclonal anti-GluN2B (1:200; Alomone, Israel) or mouse monoclonal anti-GFP (1:500, Roche, Switzerland), respectively.

Techniques: Cell Culture, Staining, In Vitro, Diffusion-based Assay

Journal: Translational Psychiatry

Article Title: Alteration of NMDA receptor trafficking as a cellular hallmark of psychosis

doi: 10.1038/s41398-021-01549-7

Figure Lengend Snippet: DISC1 downregulation alters NMDAR surface dynamics. a Schematic description of the experimental workflow: we compared the GluN2B-NMDAR surface dynamics onto neurons in which we downregulated either IL1RAPL1 or DISC1. b Representative trajectories (50 ms acquisition) of surface GluN2B-NMDAR-QD complexes on neurons transfected with either a scrambled siRNA (DISC1 Scr) or DISC1 siRNA (DISC1 Kd). Scale bar = 300 nm. Below , Cumulative distributions of synaptic GluN2B-NMDAR instantaneous diffusion coefficient (µm 2 /s) in DISC1 Scr and DISC1 Kd (Scr, n = 976 trajectories; Kd, n = 662; *** p

Article Snippet: Endogenous surface GluN2B-NMDAR or transfected GluN2A-SEP, GluN3A-SEP, GluA1-SEP, GABA-gamma2-GFP, or Dopamine D1-YFP were immune stained in live neurons using a rabbit polyclonal anti-GluN2B (1:200; Alomone, Israel) or mouse monoclonal anti-GFP (1:500, Roche, Switzerland), respectively.

Techniques: Transfection, Diffusion-based Assay