Treatment of addiction

Conditioning of Rats for Place Preference

[0086]Male Sprague-Dawley rats weighing 250-300 g (grams) were maintained under conditions of constant temperature (23° C.) and humidity (50%) in a reverse 12:12 hrs (hours) light / dark cycle, with free access to food and water. In order to determine rats' response to cocaine related cues, a Conditioned Place Preference (CPP) apparatus was used. The CPP apparatus consisted of a 30-cm-(centimeter) high white plastic box, 22×90 cm, divided into two compartments (22×40 cm each) that were separated by two 30-cm-high dividers, creating a narrower, 10-cm-wide section. The two compartments differed; one having black and white striped walls and the other having walls with stripes in different shades of gray. Both compartments had equally spaced perforations on the floor. The middle section was neutral in color, with white walls. A counterbalanced protocol was used for training and testing of rats. For baseline measurements on day 1, each rat was indiv...

Impact on Cocaine Administration and Subsequent Memory Retrieval on PARP-1 Activation in the Brain

[0090]Immediately after completion of the CPP memory retrieval testing, cell nuclei from various memory-related regions such as the CeA, BLA (basolateral amygdala), dentate gyms, striatum, hippocampal cornu ammonis area 3, prefrontal cortex and accumbens nucleus, were isolated for measurement of PARP-1 activation in the cells' nuclei. PARP-1 activation was assayed by immunolabeling of polyADP-ribose polymers formed mainly on activated PARP-1.

[0091]Brain punches were taken from the dentate gyms, striatum, hippocampal cornu-ammonis area, prefrontal cortex, arcuate nucleus, CeA and BLA, from three rats in each group. For extraction of cell nuclei, specific brain tissues were first treated (5-min on ice) with lysis buffer (pH-7.5) containing 0.2% NP-40, 1 mM DTT, 0.1 EDTA, 10 mM KCl, 1.5 mM MgCl2, 1% protease inhibitors (Sigma), and 20 mM Tris-HCl. The lysates were then centrifuged (6,000-r...

Determination of the Effect of CPP without Memory Retrieval Testing on PARP-1 Activation

[0093]In order to determine that the observed increase in PARP-1 activation in Example 2A was attributed to contextual memory retrieval, rather than to consolidation, that is to a gradual stabilization of cocaine-conditioned memory throughout the training process, rats were subjected to 10-day cocaine- or saline-CPP training, but not to the subsequent memory retrieval test. 6 rats were tested in each group. Levels of polyADP-ribosylation were determined as in Example 2A. The levels measured in the CeA of cocaine-CPP rats after training were similar to controls (p>0.05; FIG. 3A).