John J. Wagner

D1/5 receptor-mediated enhancement of LTP requires PKA, Src family kinases, and NR2B-containing NMDARs

The efficacy of the D1/5 agonist SKF38393 (100nM-60microM) to increase long-term potentiation (LTP) in the CA1 region was investigated in the rat hippocampal slice preparation. The receptor specificity of this enhancing effect was confirmed using the D1/5 antagonist SKF83566 (2microM). Although the ability of D1/5 receptors to increase both the persistence and the early magnitude of LTP has previously been linked to activation of the cAMP/PKA pathway, the subsequent molecular events leading to the enhancement of LTP have not been characterized. In experiments using SKF38393 (20microM), a requirement for the activation of both protein kinase A (PKA) and Src family tyrosine kinase pathways was demonstrated, as pretreatment with either H89 (10microM) or PP2 (10microM) kinase inhibitors prevented the D1/5-mediated enhancement of LTP. In addition, NMDA receptors containing the NR2B subunit were identified as a potential downstream target for this signaling pathway, as pretreatment with the selective antagonist Ro 25-6981 (1microM) also prevented the D1/5-mediated enhancement of LTP. The results identify a crucial role for NR2B-containing NMDA receptors in the modulation of LTP by D1/5-receptors in the CA1, suggesting that endogenously released dopamine may act through this mechanism as a modulator of hippocampal-dependent learning and memory tasks.

Cocaine-induced modulation of long-term potentiation in the CA1 region of rat hippocampus

In order to further characterize the actions of cocaine on synaptic activity in the hippocampus, recordings of field excitatory postsynaptic potentials in the CA1 region of the rat hippocampal slice preparation were used to monitor drug effects on long-term potentiation (LTP) evoked in response to stimulation of the Schaffer collateral pathway. Cocaine had dose-dependent, biphasic effects on the magnitude of LTP at these excitatory synapses in the stratum radiatum ranging from a significant enhancement of LTP at intermediate drug concentrations (5-10 microM), to an inhibition of LTP at a relatively high drug concentration (30 microM). The local anesthetic lidocaine had only inhibitory effects on the induction of LTP at all concentrations examined (10-75 microM), whereas the monoamine transporter antagonists, WIN 35348 (1 microM) or GBR 12935 (5 microM) significantly enhanced the magnitude of LTP. The D(2)-like dopamine receptor antagonist, eticlopride was effective in preventing this action of cocaine, whereas pretreatment with the D(1/5) antagonist, SCH 23390 was ineffective. These results suggest that endogenously released dopamine, in the presence of cocaine (5-10 microM), can act via D(2)-like receptors to significantly increase the magnitude of LTP in the CA1 region of the hippocampus.

Neurochemical and electrophysiological deficits in the ventral hippocampus and selective behavioral alterations caused by high-fat diet in female C57BL/6 mice.

Mounting experimental evidence, predominantly from male rodents, demonstrates that high-fat diet (HFD) consumption and ensuing obesity are detrimental to the brain. To shed additional light on the neurological consequences of HFD consumption in female rodents and to determine the relatively early impact of HFD in the likely continuum of neurological dysfunction in the context of chronic HFD intake, this study investigated effects of HFD feeding for up to 12weeks on selected behavioral, neurochemical, and electrophysiological parameters in adult female C57BL/6 mice; particular focus was placed on the ventral hippocampus (vHIP). Selected locomotor, emotional and cognitive functions were evaluated using behavioral tests after 5weeks on HFD or control (low-fat diet) diets. One week later, mice were sacrificed and brain regional neurochemical (monoamine) analysis was performed. Behaviorally naïve mice were maintained on their respective diets for an additional 5-6weeks at which time synaptic plasticity was determined in ex vivo slices from the vHIP. HFD-fed female mice exhibited increased: (i) locomotor activity in the open field testing, (ii) mean turn time on the pole test, (iii) swimming time in the forced swim test, and (iv) number of marbles buried in the marble burying test. In contrast, the novel object recognition memory was unaffected. Mice on HFD also had decreased norepinephrine and dopamine turnover, respectively, in the prefrontal cortex and the vHIP. HFD consumption for a total of 11-12weeks altered vHIP synaptic plasticity, evidenced by significant reductions in the paired-pulse ratio and long-term potentiation (LTP) magnitude. In summary, in female mice, HFD intake for several weeks induced multiple behavioral alterations of mainly anxiety-like nature and impaired monoamine pathways in a brain region-specific manner, suggesting that in the female, certain behavioral domains (anxiety) and associated brain regions, i.e., the vHIP, are preferentially targeted by HFD.

The effects of extinction training in reducing the reinstatement of drug-seeking behavior : Involvement of NMDA receptors

Although the process of extinction has been well documented for various forms of behavioral responses, the effects of extinction on the reinstatement of drug-seeking behavior are relatively understudied. In this report, the effectiveness of an extinction training protocol to reduce primed reinstatement responses was compared with the effectiveness of an equivalent period of enforced abstinence. We found that extinction training performed in the drug taking environment significantly reduced reinstatement behavior subsequently primed by either contextual cues, conditioned cues, or cocaine infusion. The ability of extinction to reduce cocaine primed reinstatement was blocked by the systemic administration of the competitive NMDAR antagonist ((+/-)CPP, 5mg/kg i.p.) administered prior to each extinction training session. Interestingly, this pharmacological intervention had no impact on the effectiveness of extinction to reduce drug-seeking behavior primed by either contextual cues or conditioned cues. These results suggest that an extinction training experience involves multiple mechanisms that can be dissociated into nonNMDAR and NMDAR dependent components with respect to the type of reinstatement (i.e. context-, conditioned stimuli (CS)-, or drug-induced) being assessed.

D-serine facilitates the effects of extinction to reduce cocaine-primed reinstatement of drug-seeking behavior.

Male Sprague Dawley rats were allowed to self-administer cocaine (0.5 mg/kg) during 90 min sessions for a period of 15 days. On day 16, rats were either held abstinent in their home cage environment or experienced an extinction session in which the active lever had no programmed consequences. Facilitating N-methyl-D-aspartate (NMDA) receptor activity with the coagonist D-serine (100 mg/kg i.p.) before or following the extinction session significantly reduced the subsequent cocaine-primed reinstatement of drug-seeking behavior tested on day 17. D-serine significantly reduced drug-primed reinstatement only when combined with extinction, and its effectiveness when administered following the training session suggested that an enhancement of consolidation of extinction learning had occurred. In contrast, D-serine treatment did not reduce sucrose-primed reinstatement, indicating that the beneficial effects of this adjunct pharmacotherapy with extinction training were specific to an addictive substance (cocaine) and did not generalize to a natural reward (sucrose).

Postsynaptic dopamine D3 receptor modulation of evoked IPSCs via GABAA receptor endocytosis in rat hippocampus

Dopamine is known to be an important modulator of learning and memory processes, but its mechanisms of action at the cellular level are diverse and are not fully characterized. In the hippocampus, pharmacologically isolated monosynaptic IPSCs were measured using the whole‐cell voltage‐clamp recording technique. Both electrically evoked and spontaneous miniature GABAA receptor currents were recorded from CA1 pyramidal neurons in slices obtained from mature rats in the presence of the D3‐selective agonist PD128907. The activation of D3 receptors inhibited synaptic GABAergic input without affecting presynaptic function or passive membrane properties. Inhibition of IPSCs evoked from stratum radiatum occurred via regulation of dynamin‐dependent trafficking of the GABAA receptor, as inclusion of dynamin inhibitory peptide (50 μM) in the recording solution prevented the inhibitory effects of PD128907 (1 μM). This effect of D3 receptor activation could be prevented by intracellular application of either an inhibitor of protein kinase A (PKI, 20 μM) or an activator of protein kinase A (8‐OH‐cAMP, 50 μM). Neither synchronous IPSCs evoked from the stratum oriens nor asynchronous miniature IPSCs recorded from the stratum radiatum were affected by D3 agonist. The induction of long‐term potentiation (LTP) of the extracellular field response in both the stratum radiatum and stratum oriens demonstrated that only potentiation in the stratum radiatum was significantly enhanced by PD128907 (1 μM). Our results suggest that the activation of D3 receptors can modulate GABAA receptor endocytosis in the hippocampus in a lamina specific manner, and thereby alter the efficacy of GABAergic transmission in the stratum radiatum of the CA1 region through a postsynaptic mechanism of action.

Simultaneous expression of cocaine-induced behavioral sensitization and conditioned place preference in individual rats

Conditioned place preference and locomotor sensitization are rodent behavioral models commonly used to investigate the actions of drugs of abuse. However, few studies have examined both paradigms in the same group of animals. We were interested in developing a combined protocol which successfully induced both conditioned place preference and sensitization simultaneously in cocaine-treated Sprague-Dawley rats in order to test the hypothesis that the magnitude of these two phenomena would be positively correlated. We used an open-field with a removable place preference insert to assess these measures independently. Cocaine-conditioned animals demonstrated a significant shift in preference for the drug-paired compartment and a sensitized locomotor response which was not observed in saline-conditioned animals challenged with cocaine. There was no significant relationship between locomotor sensitization and conditioned place preference in individual animals. We further examined these results with respect to each rats initial response to cocaine, response to a novel environment and central zone entries in an open-field. Locomotor sensitization demonstrated an inverse correlation with the initial cocaine response. In contrast, conditioned place preference demonstrated an inverse correlation with the centre response. These results demonstrate that the combination of the acute cocaine response and the centre response in a novel open-field environment can be used to indicate the propensity of a given rat to exhibit either behavioral sensitization or conditioned place preference; however, it seems that sensitization and place preference are not necessarily co-expressed to a similar extent in the same individual animal.

D-serine treatment reduces cocaine-primed reinstatement in rats following extended access to cocaine self-administration.

The most intractable feature of drug addiction is the high rate of relapse, even following extended periods of abstinence from drug-taking. Evidence suggests that allowing rats extended access to cocaine self-administration leads to behavioral characteristics in these animals that are consistent with the development of addiction in humans. In the current study, rats were allowed to self-administer cocaine over a total of 22 daily sessions, the final seven of which were long-access (LgA) sessions of 6 h duration. Assessments of reinstatement of drug-seeking behavior were made following reintroduction to the drug-taking environment and noncontingent priming with either conditioned stimulus (CS) or cocaine in both extinguished and abstinent subject groups. Three separate groups of rats were treated with either saline or D-serine (100 mg/kg i.p.) administered 2 h prior to, or immediately following, each extinction training session. Saline-treated LgA rats were resistant to the effects of extinction training to reduce noncontingent priming of reinstatement of drug-seeking behavior with either CS or cocaine. In contrast, treatment with D-serine either before or immediately following the sessions resulted in a significant enhancement in the ability of extinction training to reduce cocaine-primed reinstatement of drug-seeking behavior. These results suggest that D-serine can act to enhance the consolidation of extinction learning in LgA rats, and is therefore a promising adjunctive agent along with behavioral therapy for the treatment of cocaine addiction.

Cocaine Enhancement of Long-Term Potentiation in the CA1 Region of Rat Hippocampus: Lamina-Specific Mechanisms of Action

There is an expanding body of work characterizing dopaminergic modulation of synaptic plasticity in the hippocampus CA1 region, an area known to be involved in learning and memory. However, in vitro studies to date have focused almost exclusively on the proximal and distal apical dendritic layers (strata radiatum and lacunosum moleculare, respectively). In this report, we establish that dopaminergic activity can enhance long‐term potentiation (LTP) in the basal dendritic layer (stratum oriens) of CA1 in the rat hippocampal slice preparation. Application of the D1/5 agonist SKF38393 (20 μM) significantly increased the magnitude of basal LTP of the fEPSP response following high‐frequency stimulation of the Schaffer collateral/commissural inputs in the stratum oriens layer. In addition, endogenous dopamine (DA) activity facilitated by the presence of cocaine (6 μM) was also capable of enhancing the magnitude of basal LTP. Prior application of the D1/5 antagonist SKF83566 (2 μM) prevented this effect of cocaine, indicating that endogenously released dopamine was exerting its LTP‐enhancing effect in stratum oriens via activation of D1/5 receptors. This final result stands in contrast with the previously characterized effects of cocaine on apical LTP in the stratum radiatum, which instead have been shown to require D3 receptor activation. These observations demonstrate that dopaminergic mechanisms resulting in the enhancement of hippocampal LTP are lamina specific at Schaffer collateral/commissural synapses in the CA1 region. Synapse 2010.

Alterations in synaptic plasticity coincide with deficits in spatial working memory in presymptomatic 3xTg-AD mice

Alzheimers disease is a neurodegenerative condition believed to be initiated by production of amyloid-beta peptide, which leads to synaptic dysfunction and progressive memory loss. Using a mouse model of Alzheimers disease (3xTg-AD), an 8-arm radial maze was employed to assess spatial working memory. Unexpectedly, the younger (3month old) 3xTg-AD mice were as impaired in the spatial working memory task as the older (8month old) 3xTg-AD mice when compared with age-matched NonTg control animals. Field potential recordings from the CA1 region of slices prepared from the ventral hippocampus were obtained to assess synaptic transmission and capability for synaptic plasticity. At 3months of age, the NMDA receptor-dependent component of LTP was reduced in 3xTg-AD mice. However, the magnitude of the non-NMDA receptor-dependent component of LTP was concomitantly increased, resulting in a similar amount of total LTP in 3xTg-AD and NonTg mice. At 8months of age, the NMDA receptor-dependent LTP was again reduced in 3xTg-AD mice, but now the non-NMDA receptor-dependent component was decreased as well, resulting in a significantly reduced total amount of LTP in 3xTg-AD compared with NonTg mice. Both 3 and 8month old 3xTg-AD mice exhibited reductions in paired-pulse facilitation and NMDA receptor-dependent LTP that coincided with the deficit in spatial working memory. The early presence of this cognitive impairment and the associated alterations in synaptic plasticity demonstrate that the onset of some behavioral and neurophysiological consequences can occur before the detectable presence of plaques and tangles in the 3xTg-AD mouse model of Alzheimers disease.