ap5  (Alomone Labs)

Journal: The Journal of Cell Biology

Article Title: Translocation of CaMKII to dendritic microtubules supports the plasticity of local synapses

doi: 10.1083/jcb.201202058

Figure Lengend Snippet: Activity-dependent translocation of αCaMKII to dendritic sites near synapses. (A) Hippocampal neuron (12 DIV) expressing mGFP-αCaMKII and mCherry imaged before, during, and after stimulation with 0Mg 2+ /Gly for 5 min. (B, i) Pseudocolor image from dendrite in A. Kymograph (ii–iii) and time-lapse (iv) analyses of the change in the fluorescent intensity ratio (mGFP-αCaMKII/mCherry) over time across the dendrite (i–ii, black line) and across the spine (iii, yellow box, red line). Stimulation period is indicated (Stim). (iv) n = 4 spines and subdendritic regions from the neuron shown in A. Red and white arrows (or brackets) point, respectively, to synaptic and dendritic sites where CaMKII translocated. Bars: (neuron) 10 µm; (dendrite) 5 µm. (C) Normalized ratio (±SEM) of dendritic segment where mGFP-αCaMKII accumulated over total dendritic length after a 5-min 0Mg 2+ /Gly stimulation (Vehicle), in the presence of 50 µM AP5, 1 μM TTX, 10 μM Cd 2+ , or 10 μM CPA. n = 7–24 neurons per condition. *, P

Article Snippet: TTX was from Alomone laboratories; AP5, KN92, KN93, and ionomycin were from EMD Millipore; CPA was from Tocris Bioscience; and bicuculline was from Sigma-Aldrich.

Techniques: Activity Assay, Translocation Assay, Expressing

Journal: Neurobiology of disease

Article Title: HIV and opiates dysregulate K+-Cl− cotransporter 2 (KCC2) to cause GABAergic dysfunction in primary human neurons and Tat-transgenic mice

doi: 10.1016/j.nbd.2020.104878

Figure Lengend Snippet: hNeurons lose KCC2 immunoreactivity after exposure to HIV-Tat ± morphine. ( A ) Representative images of hNeuron cultures immunolabeled for MAP2 and KCC2. Loss of KCC2 immunoreactivity can be noted in cells treated with both 50 nM Tat 1-86 and 50 nM Tat 1-86 + morphine. Rescue is seen in 50 nM Tat 1-86 + AP5 (scale bar = 50 μm). ( B, C ) The percentage of cells immunoreactive for KCC2. Both 6 h ( B ) and 24 h ( C ) exposure to 50 - 100 nM Tat 1-86 ± morphine results in significant decrease in KCC2 immunoreactivity compared to their respective controls ( † p

Article Snippet: hNeuron cultures were treated with HIVsup at 125 - 500 pg/mL [p24] or HIV protein (10 - 100 nM HIV-1 Tat1-86 IIIB (clade B) or 250 pM - 1 nM R5-tropic gp120ADA , X4-tropic gp120IIIB , or dual-tropic gp120 MN; ImmunoDx, Woburn MA) ± 500 nM morphine sulfate for 6 or 24 h. Drug treatments: 10 μM CLP257 (Sigma), 50 μM AP5 (Alomone Labs, Jerusalem IL), 50 nM maraviroc (MVC; BOC Sciences, Shirley, NY) were applied 30 min prior to HIVsup , HIV protein, and morphine for 24 h experiments.

Techniques: Immunolabeling