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All mice were allowed to sit on the platform for 10 sec after each training trial. Mice were anesthetized using isoflurane anesthetic (1.5-4%). The mouse to be injected, weighing 20-30 g, was held by the loose skin at the back of the neck with the thumb and first fingers of one hand. Hacked social media pix and vids, public sex and nudity, clothed men using naked guys, showers and locker rooms, nude dancers, fraternity boys hazing, dates and hookups, sex parties, lesbian sex gif pranks, muscle worship. Using retractors or forceps, the muscle was peeled caudally to expose the cisternal membrane at the base of the skull. For intrathecal injections in rats, intrathecal catheters were surgically implanted using methods described by Yaksh and Rudy, Physiol. Administration Routes-Intra-Cerebroventricular or Intrathecal Injections. Intracerebroventricular injections were performed using a Hamilton micro syringe fitted with a 27 or 30-gauge needle, according to the method of Haley and McCormick. 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.



As discussed in Example 2, the effect of small molecules on primary neuronal spontaneous calcium oscillations will be measured under two conditions, in the presence and absence of 1 mM MgCl2 as described previously (Murphy et. Tau antisense oligonucleotide impact on spontaneous calcium oscillations in primary neurons is summarized in FIG. 4. See Murphy et al., J. Neurosci. 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. Small molecules 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. 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. 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).



Addition of small molecule will inhibit AMPA receptor mediated oscillations. Raw data will be exported, and spike amplitude and frequency will be calculated. I scrutinize at the road trucker and toward the raw smooch became severely very mammoth. Cells were plated 25,000 cells/well onto 384 well poly-D-lysine coated FLIPR plates (Greiner Bio-One) in 25 .mu.l/well Neurobasal media containing B27 supplement and 2 mM glutamine (day 1 in vitro, DIV1). Objects were cleaned thoroughly with 25% ethanol between training and testing sessions and between subjects, and were cleaned again at the end of the day with mild detergent. Mice received 2 sessions per day for 4 consecutive days. 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. Exploration was recorded using ObjectScan tracking software (Cleversys, Reston, Va.). All behavior was recorded with a video tracking system (Cleversys Inc). Note how warmly these sexy beasts welcome visitors our erotic video chat. Think carefully about who you choose to chat to and why they might want to video chat with you.



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