Category: Essays

Materials and Methods – Animals

Male B6129S adult mice (8-10 weeks old; weighing 25-35g) were supplied from breeding colonies in our facilities at the University of Miami, Miller School of Medicine, Miami FL as previously described in detail (Balda et al., 2006). Following weaning (postnatal day 21), mice were housed in single-sex groups; males were used for the current study. […]

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The effect of phosphodiesterase inhibitors on the extinction of cocaine-induced conditioned place preference in mice

everal phosphodiesterase inhibitors (PDEi) improve cognition, suggesting that an increase in brain cAMP and cGMP facilitates learning and memory. Since extinction of drug seeking behavior requires associative learning, consolidation and formation of new memory, the present study investigated the efficacy of three different PDEi in extinction of cocaine-induced conditioned place preference (CPP) in B6129S mice. […]

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Previous studies have indicated the significance of variations in the daily schedule of cocaine within the conditioning phase, rather than the dose of cocaine, on the development of drug-associated memory (Itzhak & Anderson, 2012; Conrad et al., 2013). While conditioning by fixed daily doses of cocaine resulted in relatively low magnitude of place preference and […]

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Experiment 3: Reconsolidation of Fix-C and Esc-C memory

Administration of MK-801 before the CPP test (pre memory retrieval) prevented a reduction in place preference for Fix-C [F(1,48)=8.904; p=0.004; group effect] but it had no effect on Esc-C (Fig 2.3B,C). For Fix-C, there was a significant time effect [F(2,48)=48.437; p<0.001] and a significant group x time interaction [F(2,48)=3.246; p=0.048]. For Esc-C, there was a […]

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Experiment 2: Acquisition of Esc-C and Fix-C memory

The effect of the NR2B antagonist ifenprodil on acquisition of place preference was investigated because of the observed increase in NR2B subunit expression in the hippocampus. Additionally, because inhibition of nNOS was shown to block Fix-C memory acquisition (Itzhak et al., 1998) and since nNOS activity is regulated by calcium entry through NMDAR (Christopherson et […]

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Immunoblot Analysis

Twenty-four hours after conditioning, mice were tested for the expression of place preference (20min) then sacrificed. Bi-lateral hippocampus was dissected and flash frozen on dry ice. Tissues were homogenized on ice in 200μl RIPA buffer (4.5mM Tris- HCl pH7.4, 150mM NaCl, 1% NP-40, 0.5% sodium deoxycholate, 0.1% SDS (Boston Bioproducts, Ashland, MA) supplemented with Complete […]

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Experiment 1: Contribution of NMDAR subunits in formation of Fix-C and Esc-C memory

To determine whether NMDAR subunits were differentially regulated in mice conditioned by Fix-C and Esc-C (Itzhak & Anderson, 2012) we performed quantitative real-time polymerase chain reaction (qPCR) and immunoblot analyses (n=3-4 mice/group). Bilateral hippocampus was dissected 24h after conditioning and subsequently analyzed. In the hippocampus NR2A and NR2B are the predominant NR2 subunits that comprise […]

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Materials and Methods (Conditioning procedure)

Subjects Male C57BL/6J mice (8 weeks old) were purchased from Jackson Laboratories (Bar Harbor, Maine). Mice were housed in groups of 5/cage with food and water adlibitum and were acclimatized to the vivarium for one week before experiments began. Animal care was in accordance with the Guide for the Care and Use of Laboratory Animals […]

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Variations in the stimulus salience of cocaine reward influences drug-associated contextual memory

Drugs of abuse act as reinforcers because they influence learning and memory processes resulting in long-term memory of drug reward. We have previously shown that mice conditioned by fixed daily dose of cocaine (Fix-C) or daily escalating doses of cocaine (Esc-C) resulted in short- and long-term persistence of drug memory, respectively, suggesting different mechanisms in […]

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Research objectives and hypothesis

An important target for combating drug addiction is to understand the neurobiological mechanisms that sub-serve relapse to drug use (Mantsch et al., 2010). Drug addiction is thought to usurp the neural mechanisms of learning and memory (Hyman, 2005) and affect long term plasticity as a result of changes in gene expression (McClung & Nestler, 2008). […]

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