[Show abstract][Hide abstract] ABSTRACT: Delivery of therapeutics to the brain is challenging because many organic molecules have inadequate aqueous solubility and limited bioavailability. We investigated the efficiency of a dendrimer-based formulation of a poorly aqueous soluble drug, haloperidol, in targeting the brain via intranasal and intraperitoneal administration. Aqueous solubility of haloperidol was increased by more than 100-fold in the developed formulation. Formulation was assessed via different routes of administration for behavioural (cataleptic and locomotor) responses, and for haloperidol distribution in plasma and brain tissues. Dendrimer-based formulation showed significantly higher distribution of haloperidol in the brain and plasma compared to a control formulation of haloperidol administered via intraperitoneal injection. Additionally, 6.7 times lower doses of the dendrimer-haloperidol formulation administered via the intranasal route produced behavioural responses that were comparable to those induced by haloperidol formulations administered via intraperitoneal injection. This study demonstrates the potential of dendrimer in improving the delivery of water insoluble drugs to brain.
[Show abstract][Hide abstract] ABSTRACT: The dopamine D2 receptor is considered one of the most important neurotransmitter receptors relevant to behavioral and clinical effects of antipsychotic drugs. Its expression and purification however is met with several challenges. This chapter provides a detailed methodology on the cell-free synthesis of the dopamine D2L receptor, using Escherichia coli (E. coli) lysate in a regenerative dialysis membrane system. This cellfree technique utilizes protein synthesis machinery and exogenous dopamine D2L DNA to synthesize functional protein outside of intact cells. The cell-free system offers various advantages specifically for the expression of transmembrane proteins, like G-protein-coupled receptors, which typically present a significant challenge. Transmembrane protein synthesis via more conventional approaches exhibit a number of innate limitations including protein aggregation, misfolding, and low yield due to cellular toxicity. The cell-free protein synthesis systems allow for the continuous replenishment of depleting precursors and removal of toxic buildup through a size-regulated porous dialysis membrane. As such this system facilitates higher yields of G-protein-coupled receptors when compared to conventional cell-based methods. Furthermore, this method provides the capability to modify the protein product, as it can be designed to incorporate radiolabeled isotopes, unnatural amino acids, solubilizing agents, cofactors, and inhibitors as is relevant for more innovative and specific research questions. Finally, an optimized cell-free system can synthesize high levels of this G-protein-coupled receptor within a few hours of incubation, providing an efficient solution to the challenge of characterizing the dopamine D2 receptor.
[Show abstract][Hide abstract] ABSTRACT: Functional imaging studies in schizophrenic patients have demonstrated metabolic brain abnormalities during cognitive tasks. This study aimed to 1) introduce a novel analysis of brain metabolic function in live animals to characterize the hypo- and hyperfrontality phenomena observed in schizophrenia and following NMDA antagonist exposure, and 2) identify a robust and representative MK-801 treatment regimen that effectively modeled brain metabolic abnormalities as well as a range of established behavioural abnormalities representative of schizophrenia.
The validity of the MK-801 animal model was examined across several established pre-clinical tests, and a novel assessment of brain metabolic function using PET/CT fused imaging. In the present study, MK-801 was administered acutely at 0.1 mg/kg and 0.5 mg/kg, and sub-chronically at 0.5 mg/kg daily for 7 days.
Acute treatment at 0.5 mg/kg-disrupted facets of memory measured through performance in the 8-arm radial maze task and generated abnormalities in sensorimotor gating, social interaction and locomotor activity. Furthermore, this treatment regimen induced hyperfrontality (increased brain metabolic function in the prefrontal area) observed via PET/CT fused imaging in the live rat.
While PET and CT fused imaging in the live rat offers a functional representation of metabolic function, more advanced PET/CT integration is required to analyze more discrete brain regions.
These findings provide insight on the effectiveness of the MK-801 pre-clinical model of schizophrenia and provide an optimal regimen to model schizophrenia. PET/CT fused imaging offers a highly translatable tool to assess hypo- and hyperfrontality in live animals.
No preview · Article · Oct 2013 · Neuropharmacology
[Show abstract][Hide abstract] ABSTRACT: Synapsins are neuronal phosphoproteins crucial to regulating the processes required for normal neurotransmitter release. Synapsin II, in particular, has been implied as a candidate gene for schizophrenia. This study investigated synapsin II mRNA expression, using real-time reverse transcriptase-PCR, in coded dorsolateral prefrontal cortical samples provided by the Stanley Foundation Neuropathology Consortium. Synapsin IIa was decreased in patients with schizophrenia when compared with both healthy subjects and patients with bipolar disorder, whereas synapsin IIb was only significantly reduced in patients with schizophrenia when compared with healthy subjects but not in patients with bipolar disorder. Furthermore, lifetime antipsychotic drug use was positively associated with synapsin IIa expression in patients with schizophrenia. Results suggest that impairment of synaptic transmission by synapsin II reduction may contribute to dysregulated convergent molecular mechanisms, which result in aberrant neural circuits that characterize schizophrenia, while implicating involvement of synapsin II in therapeutic mechanisms of currently prescribed antipsychotic drugs.The Pharmacogenomics Journal advance online publication, 26 March 2013; doi:10.1038/tpj.2013.6.
Full-text · Article · Mar 2013 · The Pharmacogenomics Journal
[Show abstract][Hide abstract] ABSTRACT: Allosteric modulators are emerging as new therapeutics for the treatment of psychiatric illnesses, such as schizophrenia. Conventional antipsychotic drugs are typically dopamine D2 receptor antagonists that compete with endogenous dopamine at the orthosteric site, and block excessive dopamine neurotransmission in the brain. However, they are unable to treat all symptoms of schizophrenia and often cause adverse motor and metabolic side effects. The binding profile of allosteric modulators differs, as they interact with their receptor at a novel binding site and their activity is determined by physiological signaling. In collaboration, our laboratories have synthesized and evaluated over 185 compounds for their allosteric modulatory activity at the dopamine D2 receptor. Of these compounds, PAOPA is among the most potent allosteric modulators, and has been shown to be effective in treating the MK-801 induced preclinical animal model of schizophrenia. The objective of this study was to evaluate PAOPA's ability to prevent and reverse behavioral abnormalities in an amphetamine-sensitized preclinical animal model of schizophrenia. Amphetamine sensitized rats were given PAOPA during sensitization and following sensitization to determine whether PAOPA is able to prevent and reverse behavioral abnormalities. Furthermore, changes in post-mortem dopamine levels were measured by high performance liquid chromatography in various brain regions. The results presented demonstrate that PAOPA is able to prevent and reverse behavioral and biochemical abnormalities in an amphetamine-sensitized animal model of schizophrenia.
No preview · Article · May 2012 · European neuropsychopharmacology: the journal of the European College of Neuropsychopharmacology
[Show abstract][Hide abstract] ABSTRACT: The synthesis of dimethyl derivatives of 5.6.5 spiro bicyclic lactam Pro-Leu-Gly-NH(2) peptidomimetics was carried out to test the hypothesis that by placing methyl groups on the β-methylene carbon of the thiazolidine ring steric bulk would be introduced into the topological space that the β-methylene carbon is believed to occupy in the negative allosteric modulators of the dopamine D(2) receptor. With such a modification, a positive allosteric modulator would be converted into a negative allosteric modulator. This hypothesis was shown to be correct as 3a and 4a where found to be negative allosteric modulators, whereas their unmethylated derivatives were positive allosteric modulators of the dopamine D(2) receptor.
Preview · Article · Apr 2012 · ACS Chemical Neuroscience
[Show abstract][Hide abstract] ABSTRACT: Synapsin II is a synaptic vesicle-associated phosphoprotein that has been implicated in the pathophysiology of schizophrenia. Researchers have demonstrated reductions in synapsin II mRNA and protein in post-mortem prefrontal cortex and hippocampus samples from patients with schizophrenia. Synapsin II protein expression has been shown to be regulated by dopamine D(1) and D(2) receptor activation. Furthermore, behavioral testing of the synapsin II knockout mouse has revealed a schizophrenic-like behavioral phenotype in this mutant strain, suggesting a relationship between dysregulated and/or reduced synapsin II and schizophrenia. However, it remains unknown the specific regions of the brain of which perturbations in synapsin II play a role in the pathophysiology of this disease. The aim of this project was to evaluate animals with a selective knock-down of synapsin II in the medial prefrontal cortex through the use of siRNA technology. Two weeks after continuous infusion of synapsin II siRNAs, animals were examined for the presence of a schizophrenic-like behavioral phenotype. Our results reveal that rats with selective reductions in medial prefrontal cortical synapsin II demonstrate deficits in sensorimotor gating (prepulse inhibition), hyperlocomotion, and reduced social behavior. These results implicate a role for decreased medial prefrontal cortical synapsin II levels in the pathophysiology of schizophrenia and the mechanisms of aberrant prefrontal cortical circuitry, and suggest that increasing synapsin II levels in the medial prefrontal cortex may potentially serve as a novel therapeutic target for this devastating disorder.
No preview · Article · Feb 2012 · Schizophrenia Research
[Show abstract][Hide abstract] ABSTRACT: Curcumin (Curcuma Longa), an extract of the rhizome turmeric, has been used for centuries in the diets and traditional medicines of both Asian and Indian cultures and today remains a popular dietary spice worldwide. Curcumin has a diverse array of favourable properties including its capacity as an antioxidant, anti-inflammatory and anticarcinogen. Furthermore, curcumin has generally been demonstrated to have low toxicity and is capable of crossing the blood brain barrier. However, this hydrophobic molecule displays low bioavailability, which is a particular challenge for treatment of central nervous system disorders. In spite of this limitation, the administration of curcumin has been shown to be effective in the treatment of neurodegenerative disorders such as Alzheimer's disease, Parkinson's disease, stroke and tardive dyskinesia, as well as other brain disorders such as multiple sclerosis, depression, and brain tumours. While curcumin's diverse mechanisms of action in the treatment of disease continue to be an enigma, its therapeutic usefulness and cost effectiveness make it a particularly valuable compound. This chapter summarizes our current understanding of the implications of curcumin in the central nervous system.
[Show abstract][Hide abstract] ABSTRACT: Heat shock proteins are constitutively expressed, but are also induced by heat and other environmental stressors. These proteins play an important role in maintaining correct folding of cellular proteins; protecting the cell from deleterious stresses by inhibiting the aggregation of partially denatured proteins and refolding them to correct confirmations. The Catecholamine Regulated Protein of size 40-kDa (CRP40) is a heat shock-like splice variant of Mortalin-2; its peptide sequence is identical to the carboxyl terminal of Mortalin-2. This protein is found co-localized with tyrosine hydroxylase in the dopaminergic pathways of the midbrain. CRP40 is differentially expressed by dopamine D1 and D2 receptor antagonists; when dopamine concentrations increase, CRP40 is upregulated. Since this increased CRP40 protein expression at high levels of DA is inhibited by treatment of cells with antioxidants, and since the natural oxidation of excess dopamine forms a potent toxic species of oxidant, CRP40 is implicated in the response pathways associated with oxidative stress. Oxidative stress and mitochondrial dysfunction have been implicated in dopaminergic disorders of the brain. CRP40 and Mortalin-2, therefore, are of particularly interest when conducting novel research in the realm of Parkinson's disease and Schizophrenia. Further, as a splice variant of Mortalin-2, CRP40 could play a central role in research involving other Mortalin-related diseases such as stroke and ischemia, carcinogenesis, Alzheimer's disease, and Huntington's disease. With so many unexplored avenues, CRP40 could boast an exciting future in health research.
[Show abstract][Hide abstract] ABSTRACT: Haloperidol (HP) is a widely prescribed antipsychotic drug used for the treatment of mental disorders. However, while providing therapeutic benefits, this drug also causes serious extrapyramidal side effects, such as tardive dyskinesia (TD). Upon chronic administration, HP causes behavioural supersensitivity to dopamine D2 receptor agonists, as well as the development of vacuous chewing movements (VCMs), in an animal model of human TD. Currently, a prevailing hypothesis to account for these behavioural abnormalities implicates oxidative stress. This study was undertaken to examine whether the free radical trapping agent, α-phenyl-N-tert-butylnitrone (PBN), can prevent the development of behavioural supersensitivity to dopamine D2 receptor agonists and the development of VCMs. Additionally, the study examined whether increased synthesis of apoptosis inducing factor (AIF) can result from HP-induced oxidative stress. Male Sprague-Dawley rats were treated with HP in conjunction with PBN, or its vehicle, for 4weeks. After a 24-hour washout period, behavioural observations were recorded along with the estimation of lipid peroxidation and antioxidant enzyme activities. The free radical trapping agent, PBN, prevented the development of behavioural supersensitivity, reduced lipid peroxidation and prevented the reduction of antioxidant enzyme activities. AIF concentrations at the mRNA and protein levels remained unchanged; therefore increased AIF gene expression is unlikely to be involved in HP-induced oxidative stress. The findings of the present study suggest the involvement of striatal free radicals in the development of behavioural supersensitivity, and free radical trapping agents, such as PBN, as possible options for the treatment of extrapyramidal side effects in humans.
[Show abstract][Hide abstract] ABSTRACT: Curcumin (Curcuma Longa Linn), the active component of turmeric, has been shown to be effective in ameliorating several stress and drug-induced disorders in rats and humans. However, it is unclear whether short term curcumin administration can prevent the abnormal oro-facial movements (AOFM) which develop following blockade of dopamine D2 receptors by antagonist such as Haloperidol. The objective of this study is to determine whether short term treatment with curcumin along with Haloperidol can prevent the development of AOFM in rats. Male Sprague Dawley rats were administered curcumin at 200 mg/kg, and Haloperidol at 2 mg/kg daily for 2 weeks, and AOFMs and locomotor activity were assessed at baseline, day 7 and day 14. By day 14, rats receiving concurrent curcumin administration had a significant reduction in the incidence of Haloperidol-induced AOFMs, but no change on the Haloperidol-induced hypolocomotion. There was no spiked increase in locomotor activity in absence of challenge with dopamine D2 receptor agonist. The exact mechanism by which curcumin attenuates AOFMs remains unknown, therefore, we performed a proteomic analysis of the striatal samples obtained from control and curcumin treated groups. A number of proteins were altered by curcumin, among them an antiapoptotic protein, Bcl-XL, was significantly upregulated. These results suggest that curcumin may be a promising treatment to prevent the development of AOFMs and further suggest some therapeutic value in the treatment of movement disorders.