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After 60 sec, mice were guided to the platform location before retrieval from the pool. Mice were placed on one end of the catwalk and allowed free exploration for 3 min or until they have 5 compliant trials, whichever comes first. For probe trials, the platform was removed and each mouse was allowed to swim for 60 sec. 2 h following the 16 h probe trial, all mice underwent the visible platform training, where a local cue (pole built using legos) was placed above the hidden platform. Potency values were generated using wells treated with a broad concentration range of LNA oligomer, from which the resulting normalized Tau/Tuj-1 and Tuj-1 values were analyzed compared to normalized values from saline control samples. Calcium oscillation reduction in the neuronal cells was measured for the oligomers of the invention and compared with that of the control cells (i.e., the calcium oscillations in the neuronal cells that are not treated with the oligomers).



Therapeutic proteins that reduce calcium oscillations to levels below 70% of control will be expected to negatively impact CNS network activity in vivo and electrophysiologic spontaneous neuronal activity in vitro. To measure primary cortical neuron spontaneous calcium oscillation, rat primary cortical neurons were prepared from Sprague-Dawley rat embryos (E19). The effect of small molecules on calcium oscillations will be measured using substantially the same method provided in Example 2. To measure primary cortical neuron spontaneous calcium oscillation, rat primary cortical neurons will be prepared. Effect of oligomers on primary neuronal spontaneous calcium oscillations was measured under two conditions, in the presence and absence of 1 mM MgCl2 as described previously (Murphy et. Cells will be loaded with a cell permanent fluorescent calcium dye. This is also something that you will not resolve within yourself overnight. Raw data will be exported, and best webcam show spike amplitude and frequency will be calculated. Data were exported and classified using the Catwalk software. An average of classified trials was used for data analysis. Each session consisted of 2 trials with a 10-min inter-trial interval. She tells her friends: 'I don't mind that. This site was designed keeping in mind iPhones and smartphones. So, don't pick a username that could give someone a clue about your real world identity or don't post profile pictures to a public site that you wouldn't be comfortable with anyone you know seeing.



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