The positive symptoms of schizophrenia primarily result from an excess of high affinity D2-like receptors (i.e. D2High receptors). First-generation antipsychotics, such as haloperidol, are D2-like antagonists that can cause severe extrapyramidal effects. Aripiprazole, a dopamine and serotonin partial agonist, has fewer side effects, making it tolerable for adults and children. Extrapyramidal effects (e.g. Parkinsonism, dystonia, and akathisia) are among the most problematic side effects produced by antipsychotic compounds, which likely result from an excess of D2-like receptors in the dorsal striatum. In order to examine the effects of repeated antipsychotic treatment on dopamine system functioning, this thesis compared the molecular effects of repeated haloperidol and aripiprazole administration on D2High receptors, as well as various indices of dopamine second messenger system functioning. Preadolescent and adult rats were pretreated with haloperidol or aripiprazole for 11 consecutive days. After either a 4- or 8-day drug abstinence period dorsal striatal tissue was extracted. [35S]GTPγS binding assays were conducted to assess the effects of repeated haloperidol and aripiprazole treatment on the efficacy and potency of D2-like receptors. PKA subunits and components of the Akt pathway were measured using Western Blots. Results showed that repeated treatment with haloperidol or aripiprazole did not significantly affect D2-like receptor efficacy or potency in young or adult rats. In both age groups, haloperidol significantly increased the expression of PKA-Cα, PKA-Cβ, and PKA-RII, but not p-PKA. Haloperidol also significantly increased PKA-Cβ and PKA-RII levels relative to aripiprazole. Repeated administration of haloperidol significantly increased p-GSK-3β levels in young and adult rats, but neither haloperidol nor aripiprazole significantly affected GSK-3β, Akt, or p-Akt levels. Overall, the results of this thesis indicate that repeated aripiprazole and haloperidol treatment differentially affects D2 signaling pathways in the dorsal striatum. Aripiprazole has less extreme or prolonged effects on D2 receptor signaling pathways than haloperidol, as evidenced by the lack of post-treatment upregulation in the cAMP and Akt pathways. Upregulation of D2-like receptors and, in turn, upregulation of proteins in the cAMP and Akt pathways may be partially responsible for the side effects induced by long-term antipsychotic treatment.

Background: Multiple sclerosis (MS) is a chronic, neurological disease characterized by targeted destruction on central nervous system (CNS) myelin. The autoimmune theory is the most widely accepted explanation of disease pathology. Circulating Th-1 cells become activated by exposure to CNS-specific antigens such as myelin basic protein. The activated Th-1 cells secrete inflammatory cytokines, which are pivotal for inflammatory responses. We hypothesize that enhanced production of inflammatory cytokines triggers cellular events within the dorsal root ganglia (DRG) and/or spinal cord, facilitating the development of neuropathic pain (NPP) in MS. NPP, the second worst disease-induced symptom suffered by patients with MS, is normally regulated by DRG and/or spinal cord. Objective: To determine gene and protein expression levels of tumor necrosis factor-alpha (TNF ) within DRG and/or spinal cord in an animal model of MS. Methods: Experimental autoimmune encephalomyelitis (EAE) was induced in adolescent female Lewis rats. Animals were sacrificed every 3 days post-disease induction. DRG and spinal cords were harvested for protein and gene expression analysis. Results: We show significant increases in TNF expression in the DRG and of EAE animals at peak disease stage, as assessed by clinical symptoms. Conclusion: Antigen-induced production of inflammatory cytokines such as TNF within the DRG identifies a potential noel mechanism for MS-induced NPP.

Background: Multiple sclerosis (MS) is a chronic, neurological disease characterized by targeted destruction on central nervous system (CNS) myelin. The autoimmune theory is the most widely accepted explanation of disease pathology. Circulating Th-1 cells become activated by exposure to CNS-specific antigens such as myelin basic protein. The activated Th-1 cells secrete inflammatory cytokines, which are pivotal for inflammatory responses. We hypothesize that enhanced production of inflammatory cytokines triggers cellular events within the dorsal root ganglia (DRG) and/or spinal cord, facilitating the development of neuropathic pain (NPP) in MS. NPP, the second worst disease-induced symptom suffered by patients with MS, is normally regulated by DRG and/or spinal cord. Objective: To determine gene and protein expression levels of tumor necrosis factor-alpha (TNF ) within DRG and/or spinal cord in an animal model of MS. Methods: Experimental autoimmune encephalomyelitis (EAE) was induced in adolescent female Lewis rats. Animals were sacrificed every 3 days post-disease induction. DRG and spinal cords were harvested for protein and gene expression analysis. Results: We show significant increases in TNF expression in the DRG and of EAE animals at peak disease stage, as assessed by clinical symptoms. Conclusion: Antigen-induced production of inflammatory cytokines such as TNF within the DRG identifies a potential noel mechanism for MS-induced NPP.

Age-related hearing loss, presbycusis, is a complex disorder involving the interaction of both peripheral and central neurological deficits. Central auditory dysfunction may contribute to poor temporal discrimination of complex sounds such as speech. This research is timely since our population over 60 years old is increasing rapidly due to advances in medicine and nutrition as well as the advancing age of baby boomers. This study was designed to provide a better understanding of age-related changes in dorsal cochlear nucleus (DCN) physiology. DCN was chosen because it receives direct input from the auditory nerve and much is known about its neuronal morphology, physiology and circuitry. In young animals, DCN output neurons, fusiform cells, receive excitatory inputs from the acoustic nerve, which is modulated and shaped by inhibitory glycinergic inputs from nearby vertical cells. A number of studies in rodents suggested an age-related impairment of glycinergic neurotransmission. To access the functional impact of reduced putative glycinergic input in the central auditory system, this study compared the physiological responses of DCN neurons from young adult and aged rats in response set of simple and more complex acoustic stimuli. Single-unit extracellular recordings were made from two groups of DCN neurons: fusiform cells and cartwheel cells. Fusiform cells reflect the culmination of DCN processing, therefore were good candidates for studying the effect of aging on one ascending auditory stream. Two specific aims were directed at fusiform cell: SA1) Examine the effects of aging on fusiform cell response properties to simple tone burst stimuli; SA2) Examine the effect of aging on DCN output neuron response to complex temporal stimuli. A third aim, SA3) Examine the impact of aging on the response properties of cartwheel cells, a DCN inhibitory interneuron. Fusiform cells recorded from aged rats displayed significantly higher maximum discharge rates to characteristic frequency (CF) tones, fewer nonmonotonic CF rate-level functions and more wide-chopper type post-stimulus time histograms (PSTHs) when compared to neurons from young adult rats. These findings were consistent with an age-related loss of inhibitory glycinergic input. To elucidate how loss of inhibition could lead to functional deficits in temporal processing, fusiform cells were challenged to encode sinusoidally amplitude modulated (SAM) tones. DCN output neurons were presented with SAM tones at three modulation depths at 30 dB above hearing level/response threshold with the carrier frequency set to each unit’s CF. Temporal synchronicity to the SAM envelope was measured using vector strength from temporal modulation transfer functions (tMTFs). Firing rate to SAM tones was also assessed in rate modulation transfer functions (rMTFs). DCN output neurons from aged rats showed no loss of rate response (rMTF) but displayed a selective loss of temporal precision to SAM tones with significant age-related changes in peak vector strength (best modulation frequency), and the shape and category of tMTF. Wide-chopper PSTH types had significantly lower vector strength values than buildup and pauser PSTHs in both young and aged fusiform cells. Since a significantly greater proportion of aged neurons exhibited wide-chopper responses, this could explain, in part, the loss of temporal processing. The age-related response changes in the present study mimicked results from earlier studies were glycine inhibition onto young adult fusiform cells was pharmacologically blocked. Cartwheel cells receive excitatory inputs from granule cell parallel fibers as well as somatosensory dorsal column nucleus and project glycinergic inputs onto DCN output neurons. They appear to play a role in the integration of auditory and somatosensory inputs such as sensing head position. Aged cartwheel neurons exhibited signs of disinhibition showing increased spontaneous activity, increased maximum discharge rates and altered rate-level functions. The observed age-related changes in cartwheel cells are consistent with deafferentation studies using acoustic trauma. Collectively, the changes in DCN output neurons and cartwheel cells reflect a potentially maladaptive age-related neuroplasticity in response to a loss of excitatory acoustic nerve input. These in vivo extracellular findings were consistent with a global downregulation of glycinergic input within the DCN of aged rats. This reduced inhibition may contribute to functional deficits, particularly in activities that require precise timing of events such as response to speech-like stimuli.

WINKELER, Georgia Freire Paiva. Efeitos agudos da administração de pressão positiva contínua em vias aéreas de modo não invasivo sobre o parênquima pulmonar de voluntários sadios nas posições supina e prona : alterações na tomografia computadorizada de alta resolução. 2006. 133 f. Dissertação (Mestrado em Medicina) - Universidade Federal do Ceará, Fortaleza, 2006. / Submitted by denise santos (denise.santos@ufc.br) on 2011-10-27T11:45:56Z No. of bitstreams: 1 2006_dis_gfpwinkeler.pdf: 6533137 bytes, checksum: 7461eccbce139fb3d37c13df801fe7a5 (MD5) / Approved for entry into archive by Eliene Nascimento(elienegvn@hotmail.com) on 2011-11-01T13:21:03Z (GMT) No. of bitstreams: 1 2006_dis_gfpwinkeler.pdf: 6533137 bytes, checksum: 7461eccbce139fb3d37c13df801fe7a5 (MD5) / Made available in DSpace on 2011-11-01T13:21:03Z (GMT). No. of bitstreams: 1 2006_dis_gfpwinkeler.pdf: 6533137 bytes, checksum: 7461eccbce139fb3d37c13df801fe7a5 (MD5) Previous issue date: 2006 / Introduction: Noninvasive positive-pressure ventilation (NIPPV) is an effective means of treating patients with acute respiratory failure and its use has been well established in cardiogenic pulmonary edema and in exacerbations of chronic obstructive pulmonary disease (COPD), reducing the need for endotracheal intubation and improving survival. Furthermore the continuous positive airway pressure (CPAP) – a mode of NIPPV – is the recommended treatment for obstructive sleep apnea syndrome (OSAS), where frequently there is no abnormality in pulmonary parenchyma. Also in the acute respiratory distress syndrome (ARDS), the application of positive end-expiratory pressure (PEEP) may result in alveolar recruitment of nonaerated units as well as in overinflation of the aerated lung areas. Alveolar overinflation is considered an important factor related to ventilator-induced lung injury causing higher mortality. The prone position has beneficial effects on oxygenation in these patients and the additive effect of PEEP with this maneuver is debatable. High resolution computed tomography (HRCT) is an excellent imaging method to evaluate the effects of positive pressure and prone position on pulmonary parenchyma. Objectives: To evaluate the effects of CPAP applied by a nasal mask on pulmonary inflation in normal subjects in supine and prone positions. Patients and methods: This is an interventionist study that evaluated eight healthy volunteers. A protocol of HRCT of the lung was performed in three regions: at the apex (2 cm above the aortic level), hilum (1 cm below the carina) and basis (2 cm above the right diaphragm) in the supine position, without and with CPAP of 5, 10 and 15 cmH2O. Also HRCT slices were performed in the prone position at the lung basis, without and with CPAP of 10 cmH2O. All HRCT slices were obtained at the functional residual capacity. Each CPAP level was maintened at least five minutes and the period between the different levels of CPAP was similar. For analysis the results were divided into regions ventral, medial and dorsal and with slices of apex, hilum and basis together. The mean lung densities (MLD) and the percentual of units with densities lower than -950 UH (overinflated) were calculated for each region. Results: There was no difference between the MLD of apex, hilum and basis for the same level pressure. In the supine position, there were a MLD reduction and an increase of the number of pixels on hyperinflated areas according to CPAP levels: without CPAP -761 HU e 7,25%; CPAP 5: -780 HU e 8,57%; CPAP 10: -810 HU e 11,62%; CPAP 15: -828 UH e 14,65% (p< 0,05). The same occurred in the prone position without CPAP: -759 UH e 6,30% and with CPAP 10: -803 UH e 9,94% (p < 0,05). It was observed a crescent ventro-dorsal density gradient in supine position that was inverse in prone position. At CPAP of 10 cmH2O there was lower percentage of pixels on hyperinflated areas in the prone position than in supine. In the non dependent lung regions (ventral in supine and dorsal in prone) there were lower percentage of pixels on hyperinflated areas and higher on normoaerated areas in the prone position than in supine with little differences in the dependent regions. Conclusions: Non invasive CPAP in normal subjects induces progressive overdistension with increase of pressure levels in supine and prone positions. CPAP of 10 cmH2O causes less overdistension of the non dependent regions than the same level of CPAP in supine position, without inducing significant overinflation of the dependent regions. So that the prone position causes a more homogeneous air distribution through the lungs. / Introdução: A ventilação não invasiva com pressão positiva (VNI) vem tendo uma crescente utilidade na prática clínica e o seu uso está bem estabelecido em casos de edema agudo de pulmão e nas exacerbações da doença pulmonar obstrutiva crônica (DPOC), diminuindo a necessidade de intubação orotraqueal e melhorando a sobrevida. Além disso, a pressão positiva contínua em vias aéreas (CPAP) – modo de VNI – constitui o tratamento de escolha para pacientes portadores da síndrome de apnéia obstrutiva do sono (SAOS), onde geralmente não há alteração no parênquima pulmonar. Ainda a aplicação de níveis elevados de pressão positiva expiratória final (PEEP) no manejo da síndrome do desconforto respiratório agudo (SDRA) está associada tanto ao recrutamento alveolar como à hiperdistensão de áreas previamente normoaeradas, com resultados ainda indefinidos quanto ao impacto na sobrevida. Um dos recursos para melhora da oxigenação nestes pacientes é a posição prona e os efeitos da associação desta manobra com pressão positiva permanecem controversos. A tomografia computadorizada de alta resolução (TCAR) constitui um excelente método de imagem para avaliação qualitativa e quantitativa do parênquima pulmonar. O emprego da TCAR pode auxiliar na investigação dos efeitos da CPAP de modo não invasivo sobre o parênquima pulmonar, contribuindo para a elucidação dos efeitos fisiológicos da pressão positiva e da posição prona. Objetivos: Avaliar e comparar os efeitos de diferentes níveis de CPAP de modo não invasivo sobre o parênquima pulmonar em indivíduos sadios nas posições supina e prona. Casuística e métodos: Estudo intervencionista com oito voluntários sadios, sem doença cardiopulmonar. Foram realizados cortes tomográficos de alta resolução em três regiões: ápice (2 cm acima do arco aórtico), hilo (1 cm abaixo da carina) e base (2 cm acima do diafragma) na posição supina, sem CPAP (basal) e com CPAP de 5, 10 e 15 cmH2O; e na posição prona, corte em base, sem CPAP e com CPAP de 10 cmH2O. A seqüência das posições e da ordem das pressões aplicadas foi randomizada. Aguardava-se um período de no mínimo 5 minutos após completa adaptação da máscara para realização do exame e o mesmo período de tempo entre um nível de pressão e outro. Os dados foram analisados agrupando-se os cortes tomográficos das três regiões e por subdivisões em regiões ventral, medial e dorsal, sendo calculadas as médias das densidades pulmonares e o percentual do número de unidades com densidade menor que -950 UH (hiperaeradas) para cada uma das regiões. Resultados: Não houve diferença das médias das densidades pulmonares entre ápice, hilo e base para o mesmo nível de pressão. Na posição supina, houve redução da densidade pulmonar e aumento do percentual de pixels nas áreas hiperaeradas com níveis crescentes de pressão: basal -761 UH e 7,25%; CPAP 5: -780 UH e 8,57%; CPAP 10: -810 UH e 11,62%; CPAP 15: -828 UH e 14,65% (p < 0,05). O mesmo foi observado na posição prona: basal -759 UH e 6,30%; CPAP 10: -803 UH e 9,94% (p < 0,05). Este aumento da aeração também foi observado nas regiões ventral, medial e dorsal. Foi encontrado um gradiente crescente no sentido ventro-dorsal de densidades pulmonares na posição supina e o inverso na posição prona. A CPAP de 10 cmH2O, na posição prona, ocasionou menor aumento do percentual de pixels nas áreas hiperaeradas em relação à supina. Nas regiões não dependentes do pulmão (ventral em supina e dorsal em prona), observou-se um menor percentual de pixels nas áreas hiperaeradas e aumento nas normoaeradas na posição prona em relação à supina, praticamente sem diferença nas regiões dependentes. Conclusões: A aplicação de diferentes níveis de CPAP, de modo não invasivo, em voluntários sadios, resultou em maior aeração com níveis crescentes de pressão e maior homogeneização da aeração pulmonar, tanto na posição supina como na prona. Houve menor hiperaeração nas regiões não dependentes na posição prona, em relação à supina, sem CPAP e com CPAP de 10 cmH2O, com melhor distribuição da aeração pulmonar naquela posição.

Stroke is the third leading cause of death and the primary cause of adult long-term disability in Canada. Despite advances in rehabilitation research, stroke survivors experience an unusually high incidence of depressive symptoms which undermine recovery outcomes by reducing patient motivation levels. Human and animal studies have linked the incidence of post-stroke depression and the extent of prefrontal cortex (PFC) damage. The PFC and the lateral habenula (LHb) are limbic structures that are strongly connected to the serotonergic dorsal raphe nucleus (DRN), a key neuronal hub for mood regulation. We hypothesized that PFC stroke produces a depressive phenotype by triggering maladaptive reorganization in mood-related networks. We used viral and optogenetic strategies to functionally characterize PFC and LHb projections to DRN. Moreover, we found that PFC stroke causes a time-dependent remodeling of LHb inputs to DRN 5-HT neurons which results in altered postsynaptic glutamate receptor number and subunit composition. This remodeling likely reflects a homeostatic upregulation of LHb-DRN synapses in response to stroke-induced challenge to network activity. Since these synapses encode stress and aversion, potentiation of this pathway could contribute to depressive symptoms following stroke. However, more work will be needed to identify the behavioral and network-level consequences of altered LHb-DRN dynamics. Thus, a deeper understanding of circuit mechanisms implicated in post-stroke depression will provide insights into this disease and open new treatment avenues to improve recovery.

Injury-induced sensitization of nociceptors contributes to pain states and the development of chronic pain. Inhibiting activity-dependent mRNA translation through mechanistic target of rapamycin and mitogen-activated protein kinase (MAPK) pathways blocks the development of nociceptor sensitization. These pathways convergently signal to the eukaryotic translation initiation factor (eIF) 4F complex to regulate the sensitization of nociceptors, but the details of this process are ill defined. Here we investigated the hypothesis that phosphorylation of the 5' cap-binding protein eIF4E by its specific kinase MAPK interacting kinases (MNKs) 1/2 is a key factor in nociceptor sensitization and the development of chronic pain. Phosphorylation of ser209 on eIF4E regulates the translation of a subset of mRNAs. We show that pronociceptive and inflammatory factors, such as nerve growth factor (NGF), interleukin-6 (IL-6), and carrageenan, produce decreased mechanical and thermal hypersensitivity, decreased affective pain behaviors, and strongly reduced hyperalgesic priming in mice lacking eIF4E phosphorylation (eIF4E(S209A)). Tests were done in both sexes, and no sex differences were found. Moreover, in patch-clamp electrophysiology and Ca2+ imaging experiments on dorsal root ganglion neurons, NGF-and IL-6-induced increases in excitability were attenuated in neurons from eIF4ES209A mice. These effects were recapitulated in Mnk1/2(-/-) mice and with the MNK1/2 inhibitor cercosporamide. We also find that cold hypersensitivity induced by peripheral nerve injury is reduced in eIF4ES209A and Mnk1/2 (-/-) mice and following cercosporamide treatment. Our findings demonstrate that the MNK1/2-eIF4E signaling axis is an important contributing factor to mechanisms of nociceptor plasticity and the development of chronic pain.

No description available.

Endomorphin 1 and 2 are two tetrapeptides recently isolated from bovine as well as human brains and proposed to be the endogenous ligand for the μ- opiate receptor Opioid compounds expressing μ-receptor preference are generally potent analgesics. The spinal cord dorsal horn is considered to be an important site for the processing of sensory information including pain. The discovery that endomorphins produced greater analgesia in mice upon intrathecal as compared to intracerebroventricular injections raises the possibility that dorsal horn neurons may represent the anatomic site upon which endomorphins exert their analgesic effects. We report here the detection of endomorphin 2-immunoreactive fiber-like elements in superficial layers of the rat dorsal horn by immunohistochemical techniques. Whole-cell patch recordings from substantia gelatinosa neurons of cervical spinal cord slices revealed two conspicuous effects of exogenously applied endomorphin 1 and 2: (i) depression of excitatory postsynaptic potentials evoked by stimulation of dorsal root entry zone, and (ii) hyperpolarization of substantia gelatinosa neurons. These effects were reversed by the selective μ-opiate receptor antagonist β-funaltrexamine. Collectively, the detection of endomorphin-like immunoreactivity in nerve fibers of the superficial layers and the inhibitory action of endomorphins on substantia gelatinosa neurons provide further support for a potential role of these two peptides in spinal nociception.

Nociceptin, also referred to as orphanin FQ, is believed to be the endogenous ligand for the ORL1. Nociceptin, when injected intracerebroventricularly to mice, produced hyperalgesia in behavioral tests. Recent studies have demonstrated the presence of ORL1 transcript in the spinal cord, and ORL1-like immunoreactivity has been localized to nerve fibers and somata throughout the spinal cord. Here, we report the localization of nociceptin-like immunoreactivity to fiber-like elements of the superficial layers of the rat dorsal horn by immunohistochemical techniques. Whole-cell recordings from substantia gelatinosa neurons in transverse lumbar spinal cord slices of 22-26-day-old rats showed that exogenous nociceptin at low concentrations (100-300 nM) depressed excitatory postsynaptic potentials evoked by stimulation of dorsal rootlets without causing an appreciable change of resting membrane potentials and glutamate- evoked depolarizations. At a concentration of 1 μM, nociceptin hyperpolarized substantia gelatinosa neurons and suppressed spike discharges. The hyperpolarizing and synaptic depressant action of nociceptin was not reversed by the known opioid receptor antagonist naloxone (1 μM). Our result provides evidence that nociceptin-like peptide is concentrated in nerve fibers of the rat dorsal horn and that it may serve as an inhibitory transmitter within the substantia gelatinosa.