Several factors contribute to the failure of the central nervous system (CNS) to regenerate after injury. These include inhibition of axonal growth by myelin and glial scar associated molecules, as well as the intrinsic inability of adult CNS neurons to grow long axons in environments that are permissive for younger neurons. Neurons in the peripheral nervous system (PNS) display a much higher capacity to regenerate after injury than CNS neurons, as shown by conditioning lesion experiments and by microtransplantation of dorsal root ganglia neurons into CNS white matter tracts. Our central hypothesis is that neurons of the PNS express specific regeneration associated genes that mediate their enhanced growth response after injury. We have employed a combination of subtractive hybridization, microarray comparison and promoter analysis to probe for genes specific to neurons of the dorsal root ganglia (DRG), using cerebellar granule neurons (CGN) as a reference. We have identified over a thousand different genes, many of whose products form interaction networks and signaling pathways. Moreover, we have identified several dozen transcription factors that may play a role in establishing DRG neuron identity and shape their responses after injury. One of these transcription factors is Signal Transducer and Activator of Transcription 3 (STAT3), previously known to be upregulated in the PNS after a conditioning lesion but not known to be specific to the PNS. Using a real time PCR and immunochemical approaches we have shown that STAT3 is constitutively expressed and selectively active in DRG neurons both in culture and in vivo. We show that the overexpression of wild type STAT3 in cerebellar granule neurons leads to the formation of supernumerary neurites, whereas the overexpression of constitutively active STAT3-C leads to a 20% increase in total neurite outgrowth. It is hoped that the genetic delivery of STAT3-C, potentially combined with co-activators of transcription, will improve functional regeneration of CNS axons in vivo.

Seit den Anfängen empirisch-neurowissenschaftlicher Forschung gilt Sprachkompetenz zuvorderst als eine Leistung der Hirnrinde (Kortex), jedoch wurden v. a. im Zuge sich verbessernder bildgebender Verfahren aphasische Syndrome auch nach Läsionen subkortikaler Hirnregionen, insbesondere der Basalganglien und des Thalamus nachgewiesen. Diese Strukturen liegen in der Tiefe des Gehirns und kommunizieren über weit gefächerte Faserverbindungen mit dem Kortex. In erster Linie werden den Basalganglien senso-motorische Kontrollfunktionen zugewiesen. Dementsprechend werden diverse Erkrankungen, die durch Störungen physiologischer Bewegungsabläufe gekennzeichnet sind (z. B. Morbus Parkinson, Chorea Huntington), auf Funktionsdefekte dieser Strukturen zurückgeführt. Der Thalamus wird häufig als Relaisstation des Informationsaustauschs zwischen anatomisch entfernten Arealen des Nervensystems aufgefasst. Basalganglien und Thalamus werden jedoch auch darüber hinausgehende Funktionen, z. B. zur Bereitstellung, Aufrechterhaltung und Auslenkung von Aufmerksamkeit bei der Bearbeitung kognitiver Aufgaben zugesprochen. In der vorliegenden Arbeit wurde mit elektrophysiologischen Methoden untersucht, ob auf der Ebene von Thalamus und Basalganglien kognitive Sprachleistungen, spezifisch der syntaktischen und semantischen Verarbeitung nachgewiesen werden können und inwieweit sich eventuell subkortikale von kortikaler Sprachverarbeitung unterscheidet. Die Untersuchung spezieller Sprachfunktionen der Basalganglien und des Thalamus ist im Rahmen der operativen Behandlung bewegungsgestörter Patienten mit der sog. Tiefenhirnstimulation (DBS = engl. Deep Brain Stimulation) möglich. Hierbei werden Patienten mit Morbus Parkinson Stimulationselektroden in den Nucleus subthalamicus (STN) implantiert. Bei Patienten mit generalisierten Dystonien erfolgt die Implantation in den Globus pallidus internus (GPI) und bei Patienten mit essentiellem Tremor in den Nucleus ventralis intermedius (VIM). STN und GPI sind Kernareale der Basalganglien, der VIM ist Teil des motorischen Systems. Nach der Implantation besteht die Möglichkeit, direkt von diesen Elektroden elektroenzephalographische (EEG)-Signale abzuleiten und diese mit simultan abgeleiteten Oberflächen-EEG zu vergleichen. In dieser Arbeit wurden DBS-Patienten aus allen genannten Gruppen in Bezug auf Sprachverständnisleistungen untersucht. Neben der Präsentation korrekter Sätze hörten die Patienten Sätze mit syntaktischen oder semantischen Fehlern. In verschiedenen Studien wurden an der Skalp-Oberfläche EKP-Komponenten (EKP = ereigniskorrelierte Potentiale) beschrieben, welche mit der Verarbeitung solcher Fehler in Verbindung gebracht werden. So verursachen syntaktische Phrasenstrukturverletzungen eine frühe links-anteriore Negativierung (ELAN). Dieser Komponente folgt eine späte Positivierung (P600), die mit Reanalyse und Reparaturmechanismen in Verbindung gebracht wird. Semantische Verletzungen evozieren eine breite Negativierung um 400ms (N400). In den thalamischen Ableitungen wurden zwei zusätzliche syntaktische fehlerbezogene Komponenten gefunden, die (i) ~ 80ms nach der Skalp-ELAN und (ii) ~ 70ms vor der Skalp-P600 auftraten. Bei semantischen Verletzungen wurde im Thalamus ein fehlerbezogenes Potential nachgewiesen, welches weitgehend parallel mit dem am Skalp gefundenen Muster verläuft. Aus den Ergebnissen der vorliegenden Studie folgt, dass der Thalamus spezifische Sprachfunktionen erfüllt. Komponenten, die Sprachverarbeitungsprozesse reflektieren, konnten in den Basalganglienstrukturen STN und GPI nicht identifiziert werden. Aufgrund der erhobenen Daten werden zwei getrennte Netzwerke für die Verarbeitung syntaktischer bzw. semantischer Fehler angenommen. In diesen Netzwerken scheint der Thalamus spezifische Aufgaben zu übernehmen. In einem ‚Syntaxnetzwerk’ kommunizieren frontale Hirnstrukturen unter Einbeziehung des Thalamus mit parietalen Hirnstrukturen. Dem Thalamus wurde eine Mediationsfunktion in der syntaktischen Reanalyse zugesprochen. In einem ‚Semantiknetzwerk’ waren keine eindeutig zuordenbaren Prozesse auf thalamischer Ebene nachweisbar. Es wurde eine unscharfe, jedoch aber spezifische Aktivierung des Thalamus über den gesamten Zeitraum der kortikalen semantischen Analyse gezeigt, welche als Integration verschiedener Analysemechanismen gewertet wurde. / Since the beginning of empirical neuroscientific research language competence has been primary localized at the brain cortex. Improved functional neuroimaging techniques were able to localize lesions in structures which caused aphasic syndromes. These syndromes were particularly found after lesions of the basal ganglia and the thalamus These structures are located in the depth of the brain and communicate over widespread fiber connections with the cortex. Sensori-motor control functions are primarily assigned to the basal ganglia. Various diseases, which are characterized by disturbances of physiological courses of motion (e.g. Parkinson’s disease, Chorea Huntington) are attributed to function defects of these structures. The thalamus was originally understood as a simple relay station of information exchange between various cortical areas of the nervous system. However, additional functions were assigned to the basal ganglia and thalamus, e.g. maintenance and deflection of attention while processing of cognitive tasks. Within the present investigation thalamic and basal ganglia achievements in language processing were studied with electro-physiological methods to investigate differences between cortical and subcortical processes. The investigation of special language functions of the basal ganglia and the thalamus is possible in the context of the surgical treatment of movement-disordered patients with "Deep Brain Stimulation (DBS)". Stimulation electrodes have been implanted to patients with Parkinson’s disease into the subthalamic nucleus (STN), patients with generalized dystonia into the globus pallidus internus (GPi), and patients suffering from essential tremor into the ventral intermediate nucleus of the thalamus (VIM). STN and GPi are core areas of the basal ganglia, the VIM is part of the motor system. EEG-signals may be derived directly from these implanted electrodes and be compared to the simultaneously derived signals of a surface EEG. In this thesis DBS patients from all groups mentioned above were examined regarding language understanding achievements. The patients listened to sentences, which were either correct or syntactical or semantical incorrect. Different studies described scalp ERP components (ERP = event related potentials) which occurred after different types of errors in sentences. Thus, syntactic phrase structure violations cause an early left anterior negativity (ELAN). A late positivity (P600) follows this component and was hypothesized as a reflection of syntactical reanalysis and/or repair. Semantic violations evoke a broad negativity around 400ms (N400). In the thalamic EEG two additional syntactic components were identified, which were seen (i) ~ 80ms after the scalp ELAN and (ii) ~ 70ms before the scalp-P600. At thalamic level semantic violations caused a negativity, which parallels largely with the negativity found at the scalp. The results of this study suggest, that the thalamus fulfills specific language functions. In the basal ganglia structures (GPI and STN) no language specific components were found. Due to the collected data two separate networks are suggested for the processing of syntactic and/or semantic errors. In these networks the thalamus seems to fulfill specific tasks. In a "syntax network” frontal brain structures communicate with parietale brain structures via the thalamus. The function of the thalamus in this network is the mediation of the syntactic reanalysis. In a "semantic network” no clearly classified processes were proved at thalamic level, because of a similarity of thalamic and scalp signals. However, during the entire period of the cortical analysis activation of the thalamus was apparent. This activation was rated as integration of different analysis mechanisms.

The structure of afferent neurons is pseudounipolar. Studies suggest that they relay action potentials (APs) to both directions of the T-junctions to reach the cell body and the spinal cord. Moreover, the somata are electrically excitable and shown to be able to transmit the signals to associated satellite cells. Our study demonstrates that this transmission can go further and pass onto passive neighbouring somata, if they are in direct contact with same satellite cells. The neurons activate the satellite cells by releasing ATP. This triggers the satellite cells to exocytose acetylcholine to the neighbouring neurons. In addition, the ATP inhibits the nicotinic receptors of the neurons by activating P2Y receptors and initiating the G-protein-mediated pathway, thus reducing the signals that return to the neurons that initiated the signals. This “sandwich synapse” represents a unique pathway by the ectopic release between the somata and the satellite cells.

The structure of afferent neurons is pseudounipolar. Studies suggest that they relay action potentials (APs) to both directions of the T-junctions to reach the cell body and the spinal cord. Moreover, the somata are electrically excitable and shown to be able to transmit the signals to associated satellite cells. Our study demonstrates that this transmission can go further and pass onto passive neighbouring somata, if they are in direct contact with same satellite cells. The neurons activate the satellite cells by releasing ATP. This triggers the satellite cells to exocytose acetylcholine to the neighbouring neurons. In addition, the ATP inhibits the nicotinic receptors of the neurons by activating P2Y receptors and initiating the G-protein-mediated pathway, thus reducing the signals that return to the neurons that initiated the signals. This “sandwich synapse” represents a unique pathway by the ectopic release between the somata and the satellite cells.

Peripheral nerve injury induces a robust regenerative state in sensory neurons that includes elevated expression of injury/regeneration-associated genes. The molecular signal(s) underlying the transition to the regenerating state are largely unknown. Brain-derived neurotrophic factor (BDNF) is the sole identified neurotrophin that is upregulated in sensory neurons following peripheral nerve injury. As members of the neurotrophin family exert a profound influence on the intact phenotype of sensory neurons, I hypothesize that injury-associated alterations in BDNF expression play a similar role in the injured/regenerating response. Antagonizing endogenous BDNF with a function-blocking antibody prevented increases in injury/regeneration-associated gene expression and decreased the growth capabilities of the injured sensory neurons. However, BDNF was not important for maintaining this cell body response in injured neurons. The elevation of BDNF expression in injured sensory neurons either through intrathecal infusion or electrical stimulation was associated with increased injury/regeneration-associated gene expression in a dose dependent manner and the latter corresponded to increased sensory axonal regeneration. Though BDNF was able to induce and enhance the intrinsic cell body response of injured sensory neurons, exogenous BDNF was not sufficient to induce an injury phenotype in intact sensory neurons. Thus, additional signals are likely induced by the injury response. In conclusion, BDNF plays a critical role in inducing the regenerative state in sensory neurons following injury and strategies aimed at elevating levels of BDNF available to the injured sensory neuron during the inductive phase improve the cell body response.

Peripheral nerve injury induces a robust regenerative state in sensory neurons that includes elevated expression of injury/regeneration-associated genes. The molecular signal(s) underlying the transition to the regenerating state are largely unknown. Brain-derived neurotrophic factor (BDNF) is the sole identified neurotrophin that is upregulated in sensory neurons following peripheral nerve injury. As members of the neurotrophin family exert a profound influence on the intact phenotype of sensory neurons, I hypothesize that injury-associated alterations in BDNF expression play a similar role in the injured/regenerating response. Antagonizing endogenous BDNF with a function-blocking antibody prevented increases in injury/regeneration-associated gene expression and decreased the growth capabilities of the injured sensory neurons. However, BDNF was not important for maintaining this cell body response in injured neurons. The elevation of BDNF expression in injured sensory neurons either through intrathecal infusion or electrical stimulation was associated with increased injury/regeneration-associated gene expression in a dose dependent manner and the latter corresponded to increased sensory axonal regeneration. Though BDNF was able to induce and enhance the intrinsic cell body response of injured sensory neurons, exogenous BDNF was not sufficient to induce an injury phenotype in intact sensory neurons. Thus, additional signals are likely induced by the injury response. In conclusion, BDNF plays a critical role in inducing the regenerative state in sensory neurons following injury and strategies aimed at elevating levels of BDNF available to the injured sensory neuron during the inductive phase improve the cell body response.

In this thesis, the learning of probabilistic relationships between stimulus-action pairs is investigated under the probability learning paradigm. The effect of reward is investigated in the first three experiments. Additionally, the effect of context and uncertainty is investigated in the second and third experiments, respectively. The fourth experiment is the replication of the second experiment with a group of Parkinson patients where the effect of dopamine medication on probability learning is studied. In Experiment 1, we replicate the classical probability learning task by comparing monetary and non-monetary reward feedback. Probability learning behavior is observed in both monetary and non-monetary rewarding feedback conditions. However, no significant difference between the monetary and non-monetary feedback conditions is observed. In Experiment 2, a variation of the probability learning task which includes irrelevant contextual information is applied. Probability learning behavior is observed, and a significant effect is found between monetary and non-monetary feedback conditions. In Experiment 3 / a probability learning task similar to that in Experiment 2 is applied, however, in this experiment, stimulus included relevant contextual information. As expected, due to the utilization of the relevant contextual information from the start of the experiment, no significant effect is found for probability learning behavior. The effect of uncertainty observed in this experiment is a replication of the reports in literature. Experiment 4 is identical to Experiment 2 / except that the subject population is a group of dopamine medicated Parkinson patients and a group of age matched controls. This experiment is introduced to test the suggestions in the literature regarding the enhancement effect of dopamine medication in probability learning based on positive feedback conditions. In Experiment 4, probability learning behavior is observed in both groups, but the difference in learning performance between Parkinson patients and controls was not significant, probably due to the low number of subject recruited in the experiment. In addition to these investigations, learning mechanisms are also examined in Experiments 1 and 4. Our results indicate that subjects initially search for patterns which lead to probability learning. At the end of Experiments 1 and 4, upon learning the winning frequencies, subjects change their behavior and demonstrate maximization behavior, which makes them prefer continuously one option over the other.

Thesis (Ph. D.)--University of Washington, 2002. / Vita. Includes bibliographical references (leaves 115-128).

Autonomic control of cardiac function depends on the coordinated activity generated by neurons within the intracardiac ganglia, and intrinsic feedback loops within the ganglia provide precise control of cardiac function. Both pituitary adenylate cyclase-activating polypeptide (PACAP) and vasoactive intestinal polypeptide (VIP) are important regulators of cell-to-cell signaling within the intracardiac ganglia, and PACAP and VIP action on these ganglia, mediated through associated receptors, play an important role in the regulation of coronary blood flow, cardiac contraction, relaxation, and heart rate. Results reported here using PACAP and VIP provide direct evidence of some of the complex signaling which occurs in neurons of the rat intracardiac ganglia.

Neurological and psychiatric disorders reveal that the basal ganglia subserve diverse functional domains, including movement, reward, and cognitive disorders (e.g., Parkinson's disease, addiction, schizophrenia). Monkey anatomical studies show that the striatum, the input structure of the basal ganglia, receives projections from nearly the entire cerebral cortex with a broad topography of motor, limbic, and association zones. However, until recently, non-invasive methods have not been available to conduct the complete mapping of the cortex to the striatum in humans. The development of functional connectivity magnetic resonance imaging (fcMRI) now allows the identification of functional connections in humans. The present dissertation reports two studies that first create a complete map of corticostriatal connectivity and then more closely examine striatal connectivity with association networks underlying cognition.