Stephanie C. Licata

Abuse and dependence liability of benzodiazepine-type drugs : GABAA receptor modulation and beyond

Over the past several decades, benzodiazepines and the newer non-benzodiazepines have become the anxiolytic/hypnotics of choice over the more readily abused barbiturates. While all drugs from this class act at the GABA(A) receptor, benzodiazepine-type drugs offer the clear advantage of being safer and better tolerated. However, there is still potential for these drugs to be abused, and significant evidence exists to suggest that this is a growing problem. This review examines the behavioral determinants of the abuse and dependence liability of benzodiazepine-type drugs. Moreover, the pharmacological and putative biochemical basis of the abuse-related behavior is discussed.

Neurochemistry of Drug Action: Insights from Proton Magnetic Resonance Spectroscopic Imaging And Their Relevance to Addiction

Proton magnetic resonance spectroscopy (1H MRS) is a noninvasive imaging technique that permits measurement of particular compounds or metabolites within the tissue of interest. In the brain, 1H MRS provides a snapshot of the neurochemical environment within a defined volume of interest. A search of the literature demonstrates the widespread utility of this technique for characterizing tumors, tracking the progress of neurodegenerative disease, and for understanding the neurobiological basis of psychiatric disorders. As of relatively recently, 1H MRS has found its way into substance abuse research, and it is beginning to become recognized as a valuable complement in the brain imaging toolbox that also contains positron emission tomography, single‐photon‐emission computed tomography, and functional magnetic resonance imaging. Drug abuse studies using 1H MRS have identified several biochemical changes in the brain. The most consistent alterations across drug class were reductions in N‐acetylaspartate and elevations in myo‐inositol, whereas changes in choline, creatine, and amino acid transmitters also were abundant. Together, the studies discussed herein provide evidence that drugs of abuse may have a profound effect on neuronal health, energy metabolism and maintenance, inflammatory processes, cell membrane turnover, and neurotransmission, and these biochemical changes may underlie the neuropathology within brain tissue that subsequently gives rise to the cognitive and behavioral impairments associated with drug addiction.

Quantification of J-resolved proton spectra in two-dimensions with LCModel using GAMMA-simulated basis sets at 4 Tesla

A two‐dimensional, J‐resolved magnetic resonance spectroscopic extraction approach was developed employing GAMMA‐simulated, LCModel basis‐sets. In this approach, a two‐dimensional J‐resolved (2D‐JPRESS) dataset was resolved into a series of one‐dimensional spectra where each spectrum was modeled and fitted with its theoretically customized LCModel template. Metabolite levels were derived from the total integral across the J‐series of spectra for each metabolite. Phantoms containing physiologic concentrations of the major brain chemicals were used for validation. Varying concentrations of glutamate and glutamine were evaluated at and around their accepted in vivo concentrations in order to compare the accuracy and precision of our method with 30 ms PRESS. We also assessed 2D‐JPRESS and 30 ms PRESS in vivo, in a single voxel within the parieto‐occipital cortex by scanning ten healthy volunteers once and a single healthy volunteer over nine repeated measures. Phantom studies demonstrated that serial fitting of 2D‐JPRESS spectra with simulated LCModel basis sets provided accurate concentration estimates for common metabolites including glutamate and glutamine. Our in vivo results using 2D‐JPRESS suggested superior reproducibility in measuring glutamine and glutamate relative to 30 ms PRESS. These novel methods have clear implications for clinical and research studies seeking to understand neurochemical dysfunction. Copyright

Low-frequency oscillations measured in the periphery with near-infrared spectroscopy are strongly correlated with blood oxygen level-dependent functional magnetic resonance imaging signals

Abstract. Low-frequency oscillations (LFOs) in the range of 0.01–0.15 Hz are commonly observed in functional imaging studies, such as blood oxygen level-dependent functional magnetic resonance imaging (BOLD fMRI) and functional near-infrared spectroscopy (fNIRS). Some of these LFOs are nonneuronal and are closely related to autonomic physiological processes. In the current study, we conducted a concurrent resting-state fMRI and NIRS experiment with healthy volunteers. LFO data was collected simultaneously at peripheral sites (middle fingertip and big toes) by NIRS, and centrally in the brain by BOLD fMRI. The cross-correlations of the LFOs collected from the finger, toes, and brain were calculated. Our data show that the LFOs measured in the periphery (NIRS signals) and in the brain (BOLD fMRI) were strongly correlated with varying time delays. This demonstrates that some portion of the LFOs actually reflect systemic physiological circulatory effects. Furthermore, we demonstrated that NIRS is effective for measuring the peripheral LFOs, and that these LFOs and the temporal shifts between them are consistent in healthy participants and may serve as useful biomarkers for detecting and monitoring circulatory dysfunction.

Evaluating the effects of systemic low frequency oscillations measured in the periphery on the independent component analysis results of resting state networks

Independent component analysis (ICA) is widely used in resting state functional connectivity studies. ICA is a data-driven method, which uses no a priori anatomical or functional assumptions. However, as a result, it still relies on the user to distinguish the independent components (ICs) corresponding to neuronal activation, peripherally originating signals (without directly attributable neuronal origin, such as respiration, cardiac pulsation and Mayer wave), and acquisition artifacts. In this concurrent near infrared spectroscopy (NIRS)/functional MRI (fMRI) resting state study, we developed a method to systematically and quantitatively identify the ICs that show strong contributions from signals originating in the periphery. We applied group ICA (MELODIC from FSL) to the resting state data of 10 healthy participants. The systemic low frequency oscillation (LFO) detected simultaneously at each participants fingertip by NIRS was used as a regressor to correlate with every subject-specific IC time course. The ICs that had high correlation with the systemic LFO were those closely associated with previously described sensorimotor, visual, and auditory networks. The ICs associated with the default mode and frontoparietal networks were less affected by the peripheral signals. The consistency and reproducibility of the results were evaluated using bootstrapping. This result demonstrates that systemic, low frequency oscillations in hemodynamic properties overlay the time courses of many spatial patterns identified in ICA analyses, which complicates the detection and interpretation of connectivity in these regions of the brain.

Anti-conflict effects of benzodiazepines in rhesus monkeys: relationship with therapeutic doses in humans and role of GABAA receptors

Rationale and ObjectivesConflict procedures are used to study mechanisms underlying the anxiolytic effects of benzodiazepines (BZs). We established a conflict procedure with rhesus monkeys in order to examine the role of GABAA receptors in the anxiolytic-like effects of BZs.MethodsFour rhesus monkeys responded under a two-component multiple schedule in which responding was maintained under a fixed-ratio schedule of food delivery in the absence (non-suppressed responding) and presence (suppressed responding) of response-contingent electric shock.ResultsConventional BZs (alprazolam, flunitrazepam, clonazepam, nitrazepam, lorazepam, bromazepam, diazepam, flurazepam, clorazepate, chlordiazepoxide) engendered increases in the average rates of suppressed responding at low to intermediate doses and decreased the average rates of non-suppressed responding at higher doses. Positive correlations were observed when the therapeutic potencies of BZs in humans were compared with potencies to increase the rates of suppressed responding (R2=0.83) or decrease the rates of non-suppressed responding (R2=0.60). The 5-HT1A agonist buspirone increased the rates of suppressed responding, although the effects were modest, whereas the opioid morphine lacked anti-conflict effects. The BZ antagonist flumazenil also modestly increased the rates of suppressed responding. A relatively low dose of flumazenil enhanced, while a high dose blocked, alprazolam’s anti-conflict effects. Compounds selective for α1 subunit-containing GABAA receptors (zolpidem, zaleplon, CL218,872) engendered relatively weak increases in the rates of suppressed responding.ConclusionsA rhesus monkey conflict procedure was established with predictive validity for therapeutic doses in people and provided evidence that anxiolytic-like effects of BZs can occur with relatively low intrinsic efficacy at GABAA receptors and are reduced by α1GABAA receptor selectivity.

Alterations in brain-derived neurotrophic factor in the mouse hippocampus following acute but not repeated benzodiazepine treatment.

Benzodiazepines (BZs) are safe drugs for treating anxiety, sleep, and seizure disorders, but their use also results in unwanted effects including memory impairment, abuse, and dependence. The present study aimed to reveal the molecular mechanisms that may contribute to the effects of BZs in the hippocampus (HIP), an area involved in drug-related plasticity, by investigating the regulation of immediate early genes following BZ administration. Previous studies have demonstrated that both brain derived neurotrophic factor (BDNF) and c-Fos contribute to memory- and abuse-related processes that occur within the HIP, and their expression is altered in response to BZ exposure. In the current study, mice received acute or repeated administration of BZs and HIP tissue was analyzed for alterations in BDNF and c-Fos expression. Although no significant changes in BDNF or c-Fos were observed in response to twice-daily intraperitoneal (i.p.) injections of diazepam (10 mg/kg + 5 mg/kg) or zolpidem (ZP; 2.5 mg/kg + 2.5 mg/kg), acute i.p. administration of both triazolam (0.03 mg/kg) and ZP (1.0 mg/kg) decreased BDNF protein levels within the HIP relative to vehicle, without any effect on c-Fos. ZP specifically reduced exon IV-containing BDNF transcripts with a concomitant increase in the association of methyl-CpG binding protein 2 (MeCP2) with BDNF promoter IV, suggesting that MeCP2 activity at this promoter may represent a ZP-specific mechanism for reducing BDNF expression. ZP also increased the association of phosphorylated cAMP response element binding protein (pCREB) with BDNF promoter I. Future work should examine the interaction between ZP and DNA as the cause for altered gene expression in the HIP, given that BZs can enter the nucleus and intercalate into DNA directly.

Contribution of α1 subunit-containing γ-aminobutyric acidA (GABAA) receptors to motor-impairing effects of benzodiazepines in squirrel monkeys

RationaleBenzodiazepines (BZs) are effective anxiolytics and hypnotics, but their use is limited by unwanted side effects, such as motor impairment.ObjectivesTo assess the contribution of α1 subunit-containing γ-aminobutyric acidA (GABAA) receptor subtypes to the motor-impairing effects of BZs, the present study evaluated two observable measures of motor coordination (balance on a pole, resistance to hind-limb flexion) engendered by nonselective and selective BZ-site agonists in squirrel monkeys.Materials and methodsMultiple doses of nonselective BZs (triazolam, alprazolam, diazepam, and chlordiazepoxide) and α1 subunit-preferring agonists (zolpidem and zaleplon) were administered to adult male squirrel monkeys (N = 4–6), and experimenters rated the monkey’s ability to balance on a horizontal pole (“ataxic-like effects”), as well as the degree of resistance to hind-limb flexion (“myorelaxant-like effects”).ResultsAdministration of all BZ-type drugs resulted in ataxic-like and myorelaxant-like effects. Pretreatment with the α1 subunit-preferring antagonist β-carboline-3-carboxylate-t-butyl ester (βCCT) attenuated the ataxic-like effects engendered by both types of drugs. However, βCCT was largely ineffective at blocking the ability of both BZs and non-BZs to induce myorelaxant-like effects.ConclusionsThese experiments demonstrate dose-dependent motor impairment in squirrel monkeys using quantitative behavioral observation techniques. Altogether, these findings suggest a lack of a prominent role for α1 subunit-containing receptors in the alteration of hind-limb flexion, a putative measure of myorelaxation, induced by BZ-type drugs in monkeys.

Zolpidem reduces the blood oxygen level-dependent signal during visual system stimulation.

Zolpidem is a short-acting imidazopyridine hypnotic that binds at the benzodiazepine binding site on specific GABA(A) receptors to enhance fast inhibitory neurotransmission. The behavioral and receptor pharmacology of zolpidem has been studied extensively, but little is known about its neuronal substrates in vivo. In the present within-subject, double-blind, and placebo-controlled study, blood oxygen level-dependent functional magnetic resonance imaging (BOLD fMRI) at 3 Tesla was used to assess the effects of zolpidem within the brain. Healthy participants (n=12) were scanned 60 min after acute oral administration of zolpidem (0, 5, 10, or 20mg), and changes in BOLD signal were measured in the visual cortex during presentation of a flashing checkerboard. Heart rate and oxygen saturation were monitored continuously throughout the session. Zolpidem (10 and 20mg) reduced the robust visual system activation produced by presentation of this stimulus, but had no effects on physiological activity during the fMRI scan. Zolpidems modulation of the BOLD signal within the visual cortex is consistent with the abundant distribution of GABA(A) receptors localized in this region, as well as previous studies showing a relationship between increased GABA-mediated neuronal inhibition and a reduction in BOLD activation.

Modest abuse-related subjective effects of zolpidem in drug- naïve volunteers

Recent case reports suggest that the short-acting benzodiazepine-like hypnotic, zolpidem, may have abuse potential among individuals who have no personal history of abusing drugs or alcohol, particularly at doses higher than those recommended for treating insomnia. This study recruited drug-naive volunteers to assess the subjective effects of multiple doses of zolpidem (0, 5, 10, or 20 mg) administered in a within-subject double-blind design. Participants (n=11) answered computerized questionnaires (Addiction Research Center Inventory, visual analog scales, and a hypothetical Drug versus Money Choice) to address the hypothesis that a supratherapeutic dose (20 mg) would increase ratings of abuse-related subjective effects, while lower therapeutic doses (5 and 10 mg) would not. Although participants rated some effects as negative at 10 and 20 mg, the highest dose engendered predominantly positive abuse-like effects such as ‘High’, ‘Like’, and ‘Good Effects’. However, no dose of zolpidem was chosen over money (