Nitric Oxide (NO) is a powerful endogenous free radical that has numerous biological functions including vasodilation and serves as a post synaptic second messenger in the central nervous system (CNS). Numerous studies implicate NO• involvement in CNS disorders such as schizophrenia and drug abuse. These studies address the direct in vivo determination of an FDA-approved NO• donor (nitroglycerin) on extracellular levels of NO• in the frontal cortex and core of the nucleus accumbens in a lightly anesthetized rat. State-of-the-art in vivo amperometric recording techniques coupled with a novel 4-channel low noise pre-amplifier system and new generation microelectrode arrays (MEAs) will be used to record extracellular levels of NO• at 100Hz before and during nasal administration of either placebo (1) or nitroglycerin. This studies will determine the feasibility of measuring NO• in the CNS while administering the NO• donor nasally and determine the amplitude and kinetic time course effects of a nasally delivered NO• donor in two rat brain areas, the frontal cortex and core of the nucleus accumbens.

Empathy plays a crucial role in our relationships with others and enhances personal well-being. The brain areas that are critical for the experience of on-line empathy and empathic behavior are not known. The current study investigated the neural substrates of empathy through the examination of whether the ventromedial prefrontal cortex (VMPC) is critical for empathy. For the first time, on-line empathic experience and behavior were measured in patients with brain damage to the VMPC. Six patients with bilateral damage to the VMPC were case-matched on specific demographic and neuropsychological criteria to two comparison groups: a brain damage group and a healthy adult group. On-line empathy was induced in an ecologically-valid manner in which the participant experienced live the sorrow of another person. The participant thought they would be playing an economic game against two opponents. However, during the study the participant overheard their game opponent experience deep sadness, revealing that it was the anniversary of their son's death (empathic induction.) A comparison neutral induction involved the participant overhearing their opponent converse with the research assistant about a neutral topic. On-line empathic experience was measured by a questionnaire completed before and after the inductions. Empathic behavior was measured implicitly through an economic game. It was defined as the degree of behavioral change on the game as a result of the empathic induction (after accounting for baseline behavior.) The economic game used to measure empathic behavior was the Repeated Fixed Opponent variant of the well-validated Ultimatum Game. This particular variant had not been studied in participants of a similar age range to the patient sample (younger and older adults). Furthermore, there is evidence for some aging-related differences in behavior on economic games, providing additional rationale to examine the behavior of healthy younger and older adults on the game. Consequently, game behavior of younger and older adults was measured and then used to implement a model of healthy game behavior in the experiment that investigated empathy in patients with damage to the VMPC. Patients with damage to the VMPC experienced poor on-line empathy and showed poor empathic behavior. Patients with brain damage to the VMPC reported significantly less on-line empathy than patients with brain damage to other regions. Empathic behavior was not shown by patients with damage to the VMPC as a result of the empathic induction and their behavior was significantly different from both the healthy and the brain damage comparison groups which showed increased empathic behavior due to the empathic induction. A specific role for the VMPC region in empathy was demonstrated by the finding that patients with damage to this region had less on-line empathy and empathic behavior than patients with brain damage to other regions. This study showed that the VMPC region of the brain is critical for empathy. Further research is needed to elucidate whether patients with brain damage to the VMPC show decreased empathic behavior in all domains or whether it is specific to monetary decision-making.

Thesis (M.Sc. (Pharmacology))--North-West University, Potchefstroom Campus, 2008.

Schizophrenia may result from altered gene expression leading to abnormal neurodevelopment. In a search for genes with altered expression in schizophrenia, cDNA library subtractive hybridization experiments using post-mortem human frontal cerebral cortices from schizophrenia individuals and neurological controls were performed. I found the mRNA of two neurodevelopmentally important genes, Nogo (RTN4) and calcium/calmodulin-dependent protein kinase II beta (CaMKIIβ), to be overexpressed in post-mortem frontal cortex tissues from patients who suffered with schizophrenia. I used the quantitative real-time polymerase chain reaction method to determined the mRNA levels of these genes in tissues from age- and sex-matched individuals. Nogo is a myelin-associated protein which inhibits the outgrowth of neurites and nerve terminals. The gene produces three splice variants, Nogo-A, B and C. I found Nogo-C mRNA to be overexpressed by 26% in schizophrenia. I also found a 17% reduction of Nogo-B mRNA in samples from individuals who had been diagnosed with severe depression. Furthermore, I showed that there is a direct correlation between the expression of both Nogo-A and -C and the presence of a CAA insert in the 3’UTR of the Nogo gene. CaMKII is a kinase localized at the postsynaptic density. The holoenzyme is primarily composed of the subunits α and β, encoded by two separate genes. It influences the expression of many neuroreceptors, in particular receptors of the glutamatergic pathway. CaMKII also mediates neural maturation during puberty, a time of onset of schizophrenia. The expression of CaMKIIα was elevated 29% in frontal cortex tissues of patients who suffered from depression. The expression of CaMKIIβ was elevated 27% in tissues of schizophrenia patients and 36% in tissues of patients diagnosed with depression. Upregulation of CaMKIIβ was associated with the presence of the CAA insert in at least one copy of the Nogo gene in a group containing both healthy subjects and patients with mental illness, possibly linking the CaMKII and Nogo pathways. The values for the expression of Nogo, CaMKIIα and CaMKIIβ were normalized to β-glucuronidase expression to minimize the effects of mRNA degradation. These results confirm that upregulation of Nogo-C and CaMKIIβ is likely associated with schizophrenia.

Schizophrenia may result from altered gene expression leading to abnormal neurodevelopment. In a search for genes with altered expression in schizophrenia, cDNA library subtractive hybridization experiments using post-mortem human frontal cerebral cortices from schizophrenia individuals and neurological controls were performed. I found the mRNA of two neurodevelopmentally important genes, Nogo (RTN4) and calcium/calmodulin-dependent protein kinase II beta (CaMKIIβ), to be overexpressed in post-mortem frontal cortex tissues from patients who suffered with schizophrenia. I used the quantitative real-time polymerase chain reaction method to determined the mRNA levels of these genes in tissues from age- and sex-matched individuals. Nogo is a myelin-associated protein which inhibits the outgrowth of neurites and nerve terminals. The gene produces three splice variants, Nogo-A, B and C. I found Nogo-C mRNA to be overexpressed by 26% in schizophrenia. I also found a 17% reduction of Nogo-B mRNA in samples from individuals who had been diagnosed with severe depression. Furthermore, I showed that there is a direct correlation between the expression of both Nogo-A and -C and the presence of a CAA insert in the 3’UTR of the Nogo gene. CaMKII is a kinase localized at the postsynaptic density. The holoenzyme is primarily composed of the subunits α and β, encoded by two separate genes. It influences the expression of many neuroreceptors, in particular receptors of the glutamatergic pathway. CaMKII also mediates neural maturation during puberty, a time of onset of schizophrenia. The expression of CaMKIIα was elevated 29% in frontal cortex tissues of patients who suffered from depression. The expression of CaMKIIβ was elevated 27% in tissues of schizophrenia patients and 36% in tissues of patients diagnosed with depression. Upregulation of CaMKIIβ was associated with the presence of the CAA insert in at least one copy of the Nogo gene in a group containing both healthy subjects and patients with mental illness, possibly linking the CaMKII and Nogo pathways. The values for the expression of Nogo, CaMKIIα and CaMKIIβ were normalized to β-glucuronidase expression to minimize the effects of mRNA degradation. These results confirm that upregulation of Nogo-C and CaMKIIβ is likely associated with schizophrenia.

Working memory (WM) capacity – the capacity to maintain and manipulate information in mind – plays an essential role in high-level cognitive functions. An important determinant of WM capacity is the ability to resolve interference of previously encoded but no longer relevant information (proactive interference: PI). Four different mechanisms of PI resolution involving binding and inhibition have been proposed in the literature, although debate continues regarding their role. Braver et al. (2007) introduced an important distinction in the PI resolution literature, proposing two general types of PI control mechanisms that occur at different time points: proactive control (involves preparation in advance of the interference) and reactive control (occurs after interference occurs). This thesis proposed that among these four functions involving binding and inhibition, item inhibition and binding could be involved in proactive control, while familiarity inhibition and episodic inhibition could be involved in reactive control. The question is which mechanism in each pair is indeed involved in proactive control and reactive control respectively, and how these proactive control and reactive control mechanisms work together to resolve PI. In addition, do these mechanisms play a role in the relationship between PI resolution and WM? In an individual differences study, individuals’ ability to resolve PI was assessed in memory tasks, with two versions of each that encouraged the use of either proactive or reactive control. In addition, measures were obtained of individuals’ ability of binding and inhibition in tasks that had minimal memory demands. Regression analyses showed contributions of binding and inhibition to PI resolution and WM. Moreover, these functions are responsible for the correlation between PI resolution and WM. In a neuroimaging study, the neural basis of proactive control was examined by comparing two memory tasks that differed in their demand on binding and inhibition. In addition, the brain regions engaged in reactive control was examined by contrasting trials involving interference or not. The thesis showed that item inhibition carried out by the left inferior frontal cortex (IFC) is involved in proactive control while episodic inhibition carried out by the left IFC and the posterior parietal cortex is involved in reactive control.

Aging is typically associated with declining mental abilities, most prominent for some forms of memory. There are, however, large inter-individual differences within the older population. Some people experience rapid decline whereas others seem almost spared from any adverse effects of aging. This thesis examined the neural underpinnings of such individual differences by using longitudinal observations of episodic memory change across 15-20 years, combined with structural and functional magnetic resonance imaging of the brain. Study I found significant correlations between volume and activity of the hippocampus (HC), and memory change over a 6-year period. That is, individuals with decline in HC function also had declining memory. In contrast, Study II showed that successfully aged individuals, who maintained high memory scores over 15-20 years, had preserved HC function compared to age-matched elderly with average memory change. The successful agers had HC activity levels comparable to those of young individuals, as well as higher frontal activity. Study III revealed that individual differences in memory ability and brain activity of elderly reflect both differential age-related changes, and individual differences in memory ability that are present already in midlife, when age effects are minimal. Specifically, memory scores obtained 15-20 years earlier reliably predicted brain activity in memory-relevant regions such as the frontal cortex and HC. This observation challenges results from previous cross-sectional aging studies that did not consider individual differences in cognitive ability from youth. Collectively the three studies implicate HC and frontal cortex function behind heterogeneity in cognitive aging, both substantiating and qualifying previous results from cross-sectional studies. More generally, the findings highlight the importance of longitudinal estimates of cognitive change for fully understanding the mechanisms of neurocognitive aging. / <p>At the time of the doctoral defense, the following paper was unpublished and had a status as follows: Paper 3: Manuscript.</p>

<p>Schizophrenia is a complex and debilitating psychiatric illness characterized by positive symptoms like hallucinations and delusions and negative symptoms like blunting of affect, avolition, and poverty of thought. This constellation of symptoms is hypothesized to result from dopaminergic dysfunction, glutamatergic dysfunction, and dysfunctional stress-reactivity. Prior to the onset of schizophrenia there is a prodromal period when individuals begin to experience sub-clinical symptoms and decreased functioning. This period is important to study not only to help elucidate biologic mechanisms of the illness but also to potentially alter the course of the illness through early treatment. The difficulty of studying this period lies in its recognizing it prospectively. To address this researchers have begun to study the at-risk mental state, a state that is associated with a high but not inevitable risk of conversion to psychosis. The studies described in this dissertation are aimed at a neurofunctional characterization of the at-risk mental state in three primary domains: reward-anticipation, hippocampus-dependent learning, and stress-reactivity. Individuals at-risk for psychosis and age-matched healthy volunteers underwent functional magnetic resonance imaging while performing tasks targeting these domains. In the reward-anticipation task, at-risk individuals showed decreased ventral tegmental area (VTA) and dorsolateral prefrontal cortex (DLPFC) responses to reward anticipation. In the hippocampus-dependent learning task, at-risk individuals showed deficits in hippocampus-dependent memory, decreased VTA engagement, and increased DLPFC activation during learning of associations between items. In the stress-reactivity task, at-risk individuals showed increased activation in the bed nucleus of the stria terminalis/basal forebrain (BNST), anterior cingulate cortex (ACC), and medial prefrontal cortex (mPFC) in response to neutral faces. Collectively, these experiments show that neurofunctional deficits in reward-anticipation, hippocampus-dependent learning, and stress-reactivity are present in the putative prodrome, prior to the onset of psychosis. Regions implicated are those that would be expected based on current models of schizophrenia and neurofunctional studies in those with frank psychosis.</p> / Dissertation

Attention-deficit/hyperactivity disorder (ADHD) is one of the most common disorders of childhood. It is theorized to be caused by catecholamine dysfunction in the striatum (Str) and frontal cortex (FC). The spontaneously hypertensive rat (SHR) has been used as a model for ADHD because of its attention deficits, impulsiveness, and hyperactivity. Prior studies of dopamine (DA) in the Str and FC have revealed conflicting results in the SHR compared to control, indicative of a need for a better understanding of DA dynamics in this model. In addition to the DA hypothesis, studies have begun implicating glutamate in the etiology of ADHD. Previous evaluations of the SHR model of ADHD found that the SHR have increased α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor activity and elevated calcium levels in the FC, suggesting that altered glutamatergic neurotransmission exists in the SHR. The first set of studies presented here suggest that increased surface expression of DA transporters may exist in the SHR model of ADHD, lowering basal DA levels. Second, we discovered that the glutamate system in the FC of the SHR model of ADHD is hyperfunctional, thus raising the possibility that targeting glutamate dysfunction in the FC could lead to the development of novel therapeutics for the treatment of ADHD. The third and fourth set of studies explored glutamate signaling in the awake rodent to fully understand glutamate neurotransmission as well as the effects of methylphenidate (MPH) on glutamate signaling in the prelimbic cortex, a region heavily implicated in ADHD. The SHR displayed similar phasic glutamate signaling compared to control; however, in the SHR but not the WKY control, chronic treatment with MPH lowered phasic glutamate amplitude. Additionally, intermediate treatment with MPH increased tonic glutamate in the SHR only, whereas chronic MPH treatment increased tonic levels in both the SHR and WKY compared to saline. Taken together, this body of work characterizes DA and glutamate signaling in the anesthetized SHR model of ADHD. Additionally, glutamate dynamics and the effects of the stimulant medication MPH were explored in the awake animal, providing evidence that glutamate is a likely target for future neuropharmacology for the treatment of ADHD.

Thesis (M.Sc. (Pharmacology))--North-West University, Potchefstroom Campus, 2008.