Thesis (M.Sc. (Pharmacology))--North-West University, Potchefstroom Campus, 2007.

Posttraumatic stress disorder (PTSD) is an anxiety disorder that may result from an exposure to a severely traumatic life-event. It is characterised by a delayed onset of psychological and physical symptoms including re-experiencing the event, avoidance of reminders associated with the trauma, increased autonomic arousal and distinct memory deficits. This disorder is also characterised by a maladaptive hypothalamic-pituitary-adrenal (HPA)-axis response and altered monoamine concentrations in the hippocampus and pre-frontal cortex. The Time Dependent Sensitization (TDS) model is a putative animal model of PTSD that is based on the concept of repeated trauma, using three acute stressors (TS) of intense severity followed by a mild situational reminder (RS) on day 7 subsequent to the acute stressors. The aims of this study were to determine if the Triple Stressor (TS) induces stress and if the situational reminder (RS) is necessary for the maintenance of the stress response over time and whether these two stress responses are qualitatively and quantitively different. This was done to further validate the TDS model and to characterize the development and progression of the stress-related pathology of PTSD. Methods used were High Performance Liquid Chromatography (HPLC) with electrochemical detection (biochemical correlates) for quantifying the monoamines dopamine (DA), noradrenaline (NA) and serotonin (5-HT) concentrations in the hippocampus and pre-frontal cortex (PFC); radio immuno assay (RIA) for the determination of plasma corticosterone concentrations (neuroendocrine parameter) and the use of the Elevated Plus Maze (EPM) to detect anxiety-like behaviour (behavioural analyses). The study was subdivided into an Acute and Re-Stress study (n = 10). In the Acute Study rats were exposed to TS as the only stressor. Group 1 was sacrificed immediately after TS, Group 2 was sacrificed 3 days post TS and Group 3 on day 7 post TS. In the Re-Stress Study both TS and RS were used as stressors. Group 4 was sacrificed immediately after the situational reminder, Group 5 was sacrificed 3 days post RS and Group 6 on day 7 post RS. A group of unstressed rats were used as Control. The results of this study found corticosterone concentrations elevated immediately after the TS (p<0.05). Exposure to the RS resulted in a profound hypocortisolism (p<0.05). These results indicate a possible disturbance in the regulation of the HPA-axis, which manifests as an enhanced negative feed-back upon re-introduction of the stressful situation. Changes in MA concentrations were evident. Although no definite fixed trend is apparent in this study, it is evident that the TDS model does induce monoamine dysregulation. Hippocampal NA. DA and 5-HT concentrations were noted to be elevated on day 7 post TS (p<0.05). On day 7 post RS only hippocampal 5HT was decreased significantly (p<0.05). Behavioural analyses indicate that stress related anxiety was not sustained after the TS but 7 days after the exposure to the RS rats were most anxious (p<0.05). The results confirm that the TDS model does induce PTSD-like symptoms in rats and that the situational reminder (RS) is necessary for the maintenance of the stress response. This model may be useful in the investigation of future experimental pharmacological interventions in the management of PTSD. / Thesis (M.Sc. (Pharmacology))--North-West University, Potchefstroom Campus, 2005.

Posttraumatic stress disorder (PTSD) is an anxiety disorder that may result from an exposure to a severely traumatic life-event. It is characterised by a delayed onset of psychological and physical symptoms including re-experiencing the event, avoidance of reminders associated with the trauma, increased autonomic arousal and distinct memory deficits. This disorder is also characterised by a maladaptive hypothalamic-pituitary-adrenal (HPA)-axis response and altered monoamine concentrations in the hippocampus and pre-frontal cortex. The Time Dependent Sensitization (TDS) model is a putative animal model of PTSD that is based on the concept of repeated trauma, using three acute stressors (TS) of intense severity followed by a mild situational reminder (RS) on day 7 subsequent to the acute stressors. The aims of this study were to determine if the Triple Stressor (TS) induces stress and if the situational reminder (RS) is necessary for the maintenance of the stress response over time and whether these two stress responses are qualitatively and quantitively different. This was done to further validate the TDS model and to characterize the development and progression of the stress-related pathology of PTSD. Methods used were High Performance Liquid Chromatography (HPLC) with electrochemical detection (biochemical correlates) for quantifying the monoamines dopamine (DA), noradrenaline (NA) and serotonin (5-HT) concentrations in the hippocampus and pre-frontal cortex (PFC); radio immuno assay (RIA) for the determination of plasma corticosterone concentrations (neuroendocrine parameter) and the use of the Elevated Plus Maze (EPM) to detect anxiety-like behaviour (behavioural analyses). The study was subdivided into an Acute and Re-Stress study (n = 10). In the Acute Study rats were exposed to TS as the only stressor. Group 1 was sacrificed immediately after TS, Group 2 was sacrificed 3 days post TS and Group 3 on day 7 post TS. In the Re-Stress Study both TS and RS were used as stressors. Group 4 was sacrificed immediately after the situational reminder, Group 5 was sacrificed 3 days post RS and Group 6 on day 7 post RS. A group of unstressed rats were used as Control. The results of this study found corticosterone concentrations elevated immediately after the TS (p<0.05). Exposure to the RS resulted in a profound hypocortisolism (p<0.05). These results indicate a possible disturbance in the regulation of the HPA-axis, which manifests as an enhanced negative feed-back upon re-introduction of the stressful situation. Changes in MA concentrations were evident. Although no definite fixed trend is apparent in this study, it is evident that the TDS model does induce monoamine dysregulation. Hippocampal NA. DA and 5-HT concentrations were noted to be elevated on day 7 post TS (p<0.05). On day 7 post RS only hippocampal 5HT was decreased significantly (p<0.05). Behavioural analyses indicate that stress related anxiety was not sustained after the TS but 7 days after the exposure to the RS rats were most anxious (p<0.05). The results confirm that the TDS model does induce PTSD-like symptoms in rats and that the situational reminder (RS) is necessary for the maintenance of the stress response. This model may be useful in the investigation of future experimental pharmacological interventions in the management of PTSD. / Thesis (M.Sc. (Pharmacology))--North-West University, Potchefstroom Campus, 2005.

Posttraumatic stress disorder (PTSD) is classified as an anxiety disorder and the characteristic symptoms (re-experiencing, avoidance as well as numbing of general responsiveness and hyperarousal) of this disorder develop in response to a traumatic event. The disorder is characterised by hypothalamic-pituitary-adrenal (HPA) axis abnormalities linked with changes in cortisol moreover, the hippocampus and cortex also play a role in the neurobiology. With regard to the neurochemistry of this disorder it is known that gamma amino butyric acid (GABA) is involved however, the precise role of GABA in PTSD and how stress changes GABA concentrations in the brain are still not fully understood. Another aspect regarding PTSD that has not been clearly defined is the treatment of PTSD. Classic anxiolytics such as diazepam is expected to relieve the anxiety linked with PTSD. Studies with this group of drugs have however not produced the concrete evidence needed to establish it as a treatment of choice for PTSD and subsequently other classes of drugs have been investigated as possible treatment options for PTSD. Among these is lamotrigine, which in a clinical study was found to be effective in alleviating symptoms of PTSD. Moreover, a possible pharmacokinetic-pharmacodynamic relationship for each of these drugs has also not been elucidated. In order to elude on some of these uncertainties, an animal model of PTSD, time dependent sensitisation (TDS), was used. GABA levels in the rat hippocampus and frontal cortex were determined at two different time intervals following the TDS procedure (1 day and 7 days post re-stress). High performance liquid chromatography (HPLC) with electrochemical (EC) detection was used to determine gamma amino butyric acid (GABA) concentrations. To investigate the possible anxiolytic effects of diazepam and lamotrigine in this model, as well as a possible pharmacokinetic-pharmacodynamic relationship for each drug, pharmacokinetic profiles for both drugs were established in order to find the times of peak and trough levels of each drug. Blood samples were collected at different time intervals after drug administration either from the tail vein of rats (lamotrigine) or directly from the heart (diazepam). Subsequently, drug concentrations at each time interval were determined by means of HPLC with ultraviolet (UV) detection. The behaviour of rats was analysed using the elevated plus-maze (EPM) at peak or trough concentrations of the drugs and this was performed after either acute administration of the drug, or after a 14 day chronic treatment regime. GABA levels in the hippocampus were not found to change statistically significantly in response to stress at either 1 day or 7 days post re-stress. In the frontal cortex, however, GABA levels increased in response to stress at 1 day post re-stress, with a statistically insignificant, but strong trend towards an increase, at 7 days post re-stress. With regard to the pharmacokinetic profiles, the peak concentration of diazepam was found to occur at 60 minutes, with lamotrigine's peak at 120 minutes. The behavioural studies indicated that acute treatment with diazepam 3 mg/kg resulted in a statistically significant increase in both ratio open arm entries and ratio time spent in the open arms at peak level of the drug. After acute treatment with diazepam 3 mg/kg a statistically significant decrease in ratio time spent in open arms was also found when the ratio time spent in open arms at peak level of the drug and the ratio time spent in open arms at trough level of the drug was compared. In response to chronic treatment with diazepam 3 mg/kg for 14 days, test animals exhibited an increase in the ratio open arm entries at trough level of the drug, with a statistically insignificant yet definite trend towards an increase at peak level. Acute treatment with lamotrigine 10 mg/kg resulted in no statistically significant change in EPM parameters. In response to chronic treatment, however, a statistically significant increase was found in ratio time spent in open arms at peak level of the drug, with a statistically insignificant trend towards an increase at trough level. From the results of this study, we may therefore conclude that GABA-levels in the brain are definitely affected, but in different ways, following TDS-stress. A pharmacokinetic-pharmacodynamic relationship between the drugs' levels and aversive behaviour could also be established. Furthermore it appears that more sustained anxiolytic effects are evident following chronic treatment with both drugs than with acute administration of these drugs. / Thesis (M.Sc. (Pharmacology))--North-West University, Potchefstroom Campus, 2006

Posttraumatic stress disorder (PTSD) is an anxiety disorder that may follow exposure to severe emotional trauma and presents with various symptoms of anxiety, hyperarousal and cognitive anomalies. Interestingly, only 10-30% of an exposed population will go on to develop full-blown PTSD. Cholinergic neurotransmission is implicated in anxiety as well as other typical manifestations of PTSD, particularly cognitive changes. The frontal cortex and hippocampus regulate and in turn are affected by stress, and have also been implicated in the underlying neuropathology of PTSD. These areas are densely innervated by cholinergic neurons originating from the basal forebrain. In this study, the time dependent sensitization (TDS) model was used to induce symptoms of PTSD in animals. The study was designed to determine the long-term effects of an intense, prolonged aversive procedure on central muscarinic acetylcholine receptor (mAChR) characteristics and the correlation if any of those findings to cognitive aspects and general arousal as characteristics associated with PTSD. In order to achieve this goal, male Sprague-Dawley rats were exposed to the TDS stress paradigm with behavioral/neuro-receptor assessments performed on day 7 post re-stress (duration of each experiment in whole is 14 days). Acoustic startle reflex (ASR) was used to determine emotional state (hyperarousal), while the conditioned taste aversion (CTA) paradigm was implemented in order to assess aversive memory. Muscarinic receptor binding studies were performed in the frontal cortex and hippocampus. Moreover, both the stress-exposed and control animals were pre-tested in the acoustic startle chamber in order to attempt to separate stress sensitive from stress-resilient animals based on predetermined ASR criteria. The ASR niodel was previously validated in our laboratory, while the CTA model was validated in this project before application. In the CTA model, an i.p. injection with lithium chloride (LiCl) (associated with digestive malaise), was used as unconditioned stimulus (US) and was paired with a saccharinlcyclamate drinking solution as conditioned stimulus (CS) to induce aversion to the novel taste (CS) when presented in the absence of the US. Population data of animals tested in the ASR experiment indicated no statistical significant difference between stressed and control animals. However, when each animal was assessed individually, 22.5 % of the exposed population displayed all increase above the predetermined criteria of 35 % in startle response, indicating a state of heightened arousal. In contrast, only 4.2 O h of control animals (no stress) displayed an increase in arousal based on the above mentioned criteria. Muscarinic receptor densities (Bm,) in the total population of animals exposed to stress showed a statistical significant increase in both the hippocampus and frontal cortex when compared to controls, with no changes in & values observed in either one of the areas. In the CTA experiment, TDS stress was implemented as US paired with a saccharinlcyclamate drinking solution as CS. An acute session of prolonged stress (as used in the TDS model) effectively induced aversion to a novel taste and a subsequent reminder of the stress (restress) paired with the CS sustained the acquire adversive memory. Furthermore, LiCl was reintroduced as US in order to assess the effect of prior exposure to two types of stress (acute and TDS) on subsequently acquired CTA memory. Prior exposure to acute stress had no significant effect on subsequently acquired aversive memory when measured either 3- or 7 days post-conditioning (CS-US). Stress-restress (TDS) exposure, however, indicated a significant decrease in aversive memory from 3- to 7 days post-conditioning (CS-US) as well as a significant decrease in aversive memory between the control- and the TDS group 7 days post-conditioning. The mAChR density (B,,) in the frontal cortex; but not in the hippocampus, was elevated at the same point in time (7 days post CS-US pairing) that CTA memory was impaired following TDS stress (stress-restress). Ultimately, these data support an association between altered cholinergic receptors and hyperarousallanxiety in an animal model of PTSD. The data also support the phenomenon of individual susceptibility to stress in animals that parallels that observed in humans exposed to severe trauma. Impaired aversive memory (CTA) is a consequence of prior exposure to TDS stress, but not acute stress, and is likewise mediated by an altered central cholinergic transmission displayed as an increase in mAChRs in the frontal cortex. The lack of studies regarding the influence of the cholinergic system in PTSD related behavior earns ,this project value as inimitable PTSD research. / Thesis (M.Sc. (Pharmacology))--North-West University, Potchefstroom Campus, 2007.

O córtex pré-frontal medial (PFCm) é caracterizado por entradas glutamatérgicas e dopaminérgicas que convergem sobre os mesmos neurônios alvos. Devido à escassa informação sobre as bases anatômicas das interações entre a dopamina (DA) e o glutamato (Glu), mapeamos a distribuição de subunidades (Su) de receptores (Rs) de Glu do tipo AMPA, NMDA e kainato no PFCm e investigamos a sua expressão em neurônios contendo a fosfoproteína DARPP-32 e em interneurônios. Os resultados mostram que as Su GluR2/3 dos Rs do tipo AMPA são as mais amplamente distribuídas no PFCm e expressas em todos os neurônios DARPP-32+. GluR2/3 é também amplamente co-localizado com as Su NMDAR1 dos Rs de Glu do tipo NMDA e GluR5/6/7 dos Rs do tipo kainato. Em contraste, as Su GluR1 e GluR4 são somente fracamente expressos no PFCm e não são co-localizados com DARPP-32, porém com GABA ou parvalbumina. Os resultados indicam que as Su GluR2/3, NMDAR1 e GluR5/6/7 são amplamente expressos em neurônios piramidais DARPP-32+ enquanto GluR1 e GluR4 são predominantemente expressos em interneurônios do PFCm. / The medial prefrontal cortex (PFCm) is characterized by glutamatergic and dopaminergic afferents that converge on the same target neurons. Since there is only limited information about the anatomical bases for interactions between dopamine (DA) and glutamate (Glu), we mapped the distribution of AMPA, NMDA and kainate Glu receptor (Rs) subunits (Su) in the PFcm and investigated their expression in neurons containing the phosphprotein DARPP-32 and in interneurons. Results show that the Su GluR2/3 of AMPA type Rs are the most prominently distributed in the PFCm and expressed in all neurons DARPP-32+. GluR2/3 is also widely co-localized with the NMDA type Su NMDAR1 and the Kainate Su GluR5/6/7. In contrast, the Su GluR1 and GluR4 are only weakly expressed in the PFCm and are not colocalized with DARPP-32 but with GABA or parvalbumin. Results indicate that the Su GluR2/3, NMDAR1, and GluR5/6/7 are prominently expressed in DARPP-32+ pyramidal neurons, whereas GluR1 and GluR4 are predominantly expressed by interneurons in the PFC.

La capacité à évaluer les résultats nos actions est fondamentale pour adapter et optimiser notre comportement et dépend d’un système superviseur chargé d’évaluer l’action, détecter les erreurs, déclencher des corrections.Le réseau neuronal sous-jacent la supervision de l’action n’a pas été complètement caractérisé chez l’homme.Dans une première étude nous avons enregistré dans l’Aire Motrice Supplémentaire (AMS) des LFP évoqués par les réponses et modulés par la performance. Des LFP évoqués exclusivement par les erreurs ont été enregistrés plus tardivement dans le cortex préfrontal médian.Dans la deuxième étude, nous avons observé que les activités de hautes-fréquences gamma sont, elles aussi, modulées par la performance des sujets, mais dans un vaste réseau frontal et extra-frontal.Dans une troisième étude, utilisant des enregistrements simultanés électroencéphalographiques (EEG) et magnétoencéphalographiques (MEG), nous observé une activité évoquée par un feedback interne sur l’EEG (mais pas en MEG), alors qu'une activité évoquée par le feedback externe était bien visible sur les enregistrements MEG, indiquant que les générateurs de ces deux activités cérébrales, sont différents. Nos résultats montrent une implication de l’AMSp dans la supervision de l’action chez l’homme, bien plus importante que ce que l’on soupçonnait auparavant. L’AMS évalue précocement, et de façon continue, l’action en cours et elle engage vraisemblablement des structures préfrontales en cas d’erreur seulement. Le traitement de l’erreur d’action, selon qu'il se fonde sur des informations internes ou externes est certainement sous-tendu par des réseaux corticaux différents. / The capacity to evaluate the outcome of our actions is fundamental for adapting and optimizing behaviour. This capability depends on an action monitoring system in charge of assessing ongoing actions, detecting errors, and evaluating outcomes.Electrical brain activity evoked by negative outcomes is thought to originate within the medial part of the frontal cortex. Nonetheless, the underlying neuronal network is incompletely characterised in humans.In the two first studies, we investigated the anatomical substrates of action monitoring in humans using intracerebral local field potential (LFP) recordings of cerebral cortex from epileptic patients. Response evoked LFPs sensitive to outcome were recorded from the Supplementary Motor Area proper (SMA), while LFPs evoked exclusively by errors were recorded later in the medial prefrontal cortex. High-gamma-frequency activity (60-180 Hz) was modulated as a function of action outcome in a vast frontal and extra-frontal network.In a third study using simultaneous recording of electroencephalography (EEG) and magnetoencephalography (MEG), we found that error related activity was detected by EEG (but not by MEG), while feedback-related activity was detected by MEG, indicating that the sources of these two forms of outcome-modulated brain activity are different.To conclude the SMA is much more involved in action monitoring than previously thought. SMA rapidly and continuously assesses ongoing actions and likely engages more rostral prefrontal structures in the case of error. Processing of action errors and of negative externally delivered feedback therefore appears to be supported by distinct cortical networks.

Background: Although several neurophysiological models have been proposed for panic disorder with agoraphobia (PD/AG), there is limited evidence from functional magnetic resonance imaging (fMRI) studies on key neural networks in PD/AG. Fear conditioning has been proposed to represent a central pathway for the development and maintenance of this disorder; however, its neural substrates remain elusive. The present study aimed to investigate the neural correlates of fear conditioning in PD/AG patients. Method: The blood oxygen level-dependent (BOLD) response was measured using fMRI during a fear conditioning task. Indicators of differential conditioning, simple conditioning and safety signal processing were investigated in 60 PD/AG patients and 60 matched healthy controls. Results: Differential conditioning was associated with enhanced activation of the bilateral dorsal inferior frontal gyrus (IFG) whereas simple conditioning and safety signal processing were related to increased midbrain activation in PD/AG patients versus controls. Anxiety sensitivity was associated positively with the magnitude of midbrain activation. Conclusions: The results suggest changes in top-down and bottom-up processes during fear conditioning in PD/AG that can be interpreted within a neural framework of defensive reactions mediating threat through distal (forebrain) versus proximal (midbrain) brain structures. Evidence is accumulating that this network plays a key role in the aetiopathogenesis of panic disorder.

Rats were administered nicotine (0.3 mg/kg) for 11 consecutive days before and after an electrolytic medial frontal cortex lesion. Behavioral testing was arranged so that the rats were tested on the RAM 1 day after drug administration followed by behavioral testing on the MWT 19 days after drug treatment, or tested on the MWT 1 day after drug administration followed by testing on the RAM4 days after drug treatment. Results of MWT testing showed that regardless of the drug/behavioral testing interval, lesioned rats given nicotine demonstrated enhancement relative to saline-treated animals. Results of RAM testing showed that nicotine improved performance in non-lesioned rats compared to non-lesioned rats given saline. Four days after drug administration, nicotine improved performance in lesioned rats to levels of non-lesioned rats regardless of drug treatment. A second experiment was implemented to determine if the previous training on the MWT affected performance on the RAM.

Die Suche nach einem Zielobjekt in einer komplexen visuellen Szene ist ein alltäglicher Wahrnehmungsvorgang und ein etabliertes experimentelles Paradigma für die Untersuchung selektiver Aufmerksamkeit. Einem klassischen Modell zufolge ist der Suchprozeß seriell: Die Objekte werden nacheinander vom Aufmerksamkeitsfokus selektiert und so für die Identifikation bereitgestellt. Ein Alternativmodell postuliert einen parallelen Suchprozeß, bei dem alle Objekte in der Szene gleichzeitig vom Sehsystem verarbeitet werden. Beide Modelltypen sind gleich gut mit den Resultaten bisheriger Verhaltensexperimente kompatibel. In der vorliegenden Arbeit wurden die neuronalen Grundlagen des Suchprozesses mit Hilfe der funktionellen Magnetresonanztomographie (fMRT) im menschlichen Gehirn untersucht. Es ist bekannt, dass das frontale Augenfeld (FEF) und drei Subregionen (AIPS, PIPS und IPTO) des posterioren parietalen Cortex (PPC) den im seriellen Suchmodell postulierten Teilprozeß der Verschiebung des Aufmerksamkeitsfokus (ohne Augenbewegungen) kontrollieren. In Experiment 1 wurde geprüft, ob diese Regionen auch am Suchprozeß beteiligt sind. Dazu wurde das fMRT-Signal zwischen einer schwierigen Suche nach einer Verknüpfung zweier visueller Merkmale und einer einfachen Suche nach einem einzelnen Merkmal verglichen. Motorische Anforderungen und Reizmuster waren in beiden Bedingungen so ähnlich wie möglich und in Kontrollexperimenten wurde sichergestellt, dass Aktivierungsunterschiede zwischen beiden Bedingungen keine motorischen oder sensorischen Prozesse reflektieren, sondern spezifisch den Prozeß der Verknüpfungssuche. FEF, AIPS, PIPS und IPTO wurden differentiell aktiviert. In Experiment 2 wurde getestet, ob die Beteiligung dieser Areale an der visuellen Suche von der Notwendigkeit einer Merkmalsverknüpfung abhängt. Dazu wurde eine schwierige Merkmalssuche mit der einfachen Merkmalssuche verglichen und kontrolliert, dass auch dieser Vergleich sensorische und motorische Faktoren eliminierte. Differentielle Aktivierungen in diesem Experiment reflektierten nun nicht mehr den Merkmalsverknüpfungsprozeß, sondern allein die höhere Schwierigkeit der Suche. Auch hier fand sich eine differentielle Aktivierung des FEF, AIPS, PIPS und IPTO. Dabei unterschieden sich die Schwierigkeit auf der Verhaltensebene wie auch die differentielle Aktivierung von PIPS auf der neuronalen Ebene nicht zwischen Verknüpfungs- und schwieriger Merkmalssuche. Die Ergebnisse demonstrieren, dass das FEF und drei Subregionen des PPC an der schwierigen visuellen Suche beteiligt sind. Dies ist gut mit der Annahme eines seriellen und nur schwer mit der eines parallelen Suchprozesses vereinbar. Darüber hinaus suggerieren die Befunde, dass der Beitrag des PPC und FEF zur visuellen Suche nicht auf den Prozeß der Merkmalsverknüpfung beschränkt ist, sondern allgemeiner die Anforderung an den Suchprozeß reflektiert. / The search for a target object in a complex visual scene is an all-day process of visual perception and an established experimental paradigm for the study of selective attention. A classical model postulates a serial search process. That is, objects are selected sequentially by the focus of attention and are thereby routed to the identification stage. An alternative model postulates a parallel search process, in which all objects within the scene are processed simultaneously. Both models are equally consistent with the current behavioural data. In this thesis, the neural basis of the search process in the human brain was investigated with functional magnetic resonance imaging (fMRI). The frontal eye field (FEF) and three sub-regions (AIPS, PIPS und IPTO) of the posterior parietal cortex (PPC) are known to control the shifting of the focus of attention in space (without eye movements), which is postulated by the serial model to be an essential sub-process of visual search. Experiment 1 tested whether the same areas are also engaged in the search process. The fMRI signal was compared between a difficult search for a feature conjunction and an easy search for a single feature. Motor requirements and stimuli were as similar as possible across conditions and control experiments demonstrated that activation differences between conditions do not reflect sensory or motor factors, but rather the process of conjunction search. The FEF, AIPS, PIPS, and IPTO were differentially activated. Experiment 2 tested whether the involvement of these areas in visual search depends on the necessity for conjoining features. A difficult feature search was compared with the easy feature search. This comparison also eliminated sensory and motor factors according to control experiments. Differential activations in this experiment did not reflect the feature conjunction process, but only the higher search difficulty. Again, a differential activation of the FEF, AIPS, PIPS, and IPTO was found. The conjunction and the difficult feature search did not differ in their difficulty at the behavioral level as well as in PIPS activation strength at the neural level. The results show that the FEF and three PPC sub-regions contribute to difficult visual search. This is consistent with the assumption of a serial, but much less consistent with the assumption of a parallel, search process. Furthermore the results suggest that the contribution of the PPC and FEF to visual search is not restricted to the feature conjunction process, but more generally reflects the demands on the search process.