No / In the November issue (2001) of Neuroscience Letters, Holmin et al. (Neurosci. Lett. 314 (2001) 151) reported that the synthesis of the intermediate filament protein nestin was upregulated by potassium-induced depolarization in the rat cortex. In this letter, we provide supplementary evidence that repeated cortical spreading depression elicited by potassium induces a delayed upregulation of nestin. However, we argue against the authors' conclusion, Nestin expression was N-methyl-D-aspartate (NMDA)-receptor dependent since dizocilpine (MK-801) treatment abolished the response because spreading depression itself is very sensitive to NMDA-receptor block, and the drug treatment was initiated prior to potassium application to the cortex in Holmin et al.'s study.

In this thesis the optical method of Near-Infrared-Spectroscopy (NIRS) is evaluated with regards to its capability of non-invasive detection of Leão´s cortical spreading depression (CSD) and spontaneous peri-infarct-depolarizations (PID). With the NIR-spectrometer NIRO 500 (Hamamatsu, Japan) regional cerebral oxy-genation (rCBO) changes were measured during CSD in 9, and during PID in 10 barbiturate anesthatized rats. The method if NIRS that relies on oxygen-dependent absorption changes of hemo-globin and cytochrome oxydase as well as the high penetrability of biologic tissues for light in the range between 700 and 1000 nm proved suitable to detect and to distinguish both CSD and PID experimentally. This distinction relies on the robust decrease of deoxy- and increase of oxyhemo-globin concentrations (i.e. a relative hyperoxemia) during CSD while PID is cha-racterized by an initial increase of deoxy- and decrease of oxyhemoglobin (relative hypooxemia). Despite the profound anatomical differences between gyrencephalic humans and lyssencephalic rats, the observed patterns of rCBO changes may guide the inter-pretation of future NIRS measurements in patients with migraines with aura (CSD) or stroke (PID). However, for concentration changes of oxydized cytochrome aa3 with its low con-centration compared to the hemoglobins, the pathophysiological interpretation of the data obtained with NIRO 500 is confounded by the limits of attenuation mea-surements at only four wavelengths. A validation of the cytochrome oxydase signal and an improved quantification of all concentration changes is highly desirable and may be achieved by employment of a continuous-wavelength device measuring the full spectral range of the near infrared. It would also allow to measure the mean optical pathlength in the highly scattering tissue and to correct for its physiologically occuring changes e.g. by measurements at the water absorption peak. Similar improvements would enhance the value of the method for further physiolo-gical and pathophysiological studies because NIRS provides the unique opportu-nity to obtain simultaneous data on blood oxygenation as well as the redox state of the mitochondrial cytochrome oxydase.

No description available.

Submitted by (ana.araujo@ufrpe.br) on 2016-08-12T14:04:00Z No. of bitstreams: 1 Rosangela Silveira do Nascimento.pdf: 1012353 bytes, checksum: b5f0d139481ab0bfdd6a1f689175c1f6 (MD5) / Made available in DSpace on 2016-08-12T14:04:00Z (GMT). No. of bitstreams: 1 Rosangela Silveira do Nascimento.pdf: 1012353 bytes, checksum: b5f0d139481ab0bfdd6a1f689175c1f6 (MD5) Previous issue date: 2008-02-29 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / In the present work we analyze the dynamics of electrical cortical activity during the phenomenon of spreading depression (DA) and during the periods before and after this phenomenon. The characteristic of DA is reduced amplitude of spontaneous electrical activity that occurs in neural tissue after the application of stimulus that can be electrical, chemical, mechanical, luminous etc. In order to study properties of time series of electrical cortical activity recorded by ECoG (electrocortiogram) before,during and after DA, we apply Detrended Fluctuation Analysis(DFA). This method is designed to quantify long term correlations (memory) in temporal series such ECoG register. The method was successfully applied in studies of DNA sequences and non-stationary time series as heart rate variability, stride intervals, financial time series etc. The application of DFA results in scaling exponentα that quantifies correlation properties of nonlinear dynamical systems. This experiment indicates if temporal series posses long term correlations. In this work we calculate exponent α for different intervals: control (before the stimulus), after the stimulus, during the avalanche, during DA and after DA for two experimental groups of rats, nourished and malnourished. For both experimental groups the values of exponent α indicates persistent behavior for all intervals except during the avalanche in which correlations degrade. The presence of long term correlations in physiological time series observed in healthy organisms represents complexity that guaranties the organism’s adaptability to stress and disease. The absence of correlations during the avalanche indicates the loss of this complexity. Non-parametric Wilcoxon test was used to compare mean values of exponents α for all intervals of analyzed time series. In cases of nourished rats, the mean values ofα are significantly different for control, stimulus, avalanche, DA and after-DA intervals. Wilcoxon test was also used to compare mean values of α for corresponding intervals for the two experimental groups. The result is significant difference in mean values of α for control, stimulus avalanche, DA and after DA intervals between two experimental groups. The hypothesis that α =0.5 for avalanche intervals was not rejected by test, confirming the loss of correlations in this phase. Comparison of mean values of α for different intervals (control, stimulus, DA and after DA) with avalanche using the Wilcoxon test results in significant difference between two groups. / O presente estudo se propõe a analisar a dinâmica da atividade elétrica cortical durante o fenômeno da depressão alastrante (DA) e nos períodos que antecede e sucede o fenomêno. A DA é caracterizada pela redução da amplitude da atividade elétrica espontânea que ocorre no tecido neural, após a aplicação de um estímulo de natureza elétrica, química, mecânica, luminosa e outros. Visando estudar o comportamento da série temporal da atividade elétrica cortical, registrada no ECoG (eletrocorticograma), durante a DA e nos períodos que precede e sucede o fenômeno, foi aplicado o método do DFA (Detrended Fluctuation Analysis). Este método permite quantificar a existência de correlação de longo alcance (memória) numa série temporal, como é o caso do registro do ECoG. Anteriormente, o método foi aplicado em seqüências de DNA e no estudo de séries temporais não estacionárias, tais como, dinâmica da variabilidade cardíaca, flutuações de eletroencefalograma de humanos, intervalos entre passos sucessivos de humanos, séries econômicas e outros. A aplicação do DFA numa série temporal permite a determinação de um expoente de escalonamento α, que pode contribuir para a compreensão das propriedades dos sistemas dinâmicos não lineares. Este expoenteα revela se a série temporal apresenta correlação de longo alcance ou não. Neste trabalho os expoentes α foram calculados nas fases de controle, estímulo, avalanche, DA e após a DA para o ECoG, em dois grupos experimentais, ratos nutridos e ratos desnutridos. Em ambos os grupos experimentais, os valores obtidos para o expoente de escalonamento α denotam que a série temporal do ECoG apresenta correlação persistente (comportamento da série no presente se mantém no futuro) em todas as fases do processo com exceção da avalanche, período no qual ocorre perda de correlação. A presença de correlação de longo alcance numa série temporal biológica é uma resposta sempre observada em organismos saudáveis cuja complexidade do sinal registrado garante a adaptabilidade do organismo a situações de estresse e/ou distúrbios. Enquanto a ausência de correlação, observada na avalanche, indica a perda de propriedades fractais nos sistemas fisiológicos. O uso do método não-paramétrico de Wilcoxon, para comparar os valores médios dos expoentes α obtidos para o grupo de animais nutrido, durante as fases de controle, estimulação, DA, após DA, revelou que essas diferentes fasesdiferem significativamente. Os valores médios dos expoentes α obtidos para o grupo de animais desnutrido, durante as fases de controle, estimulação, DA, após DA, também não foram significativamente diferentes, quando comparados pelo método de Wilcoxon. Na comparação dos valores médios de α nas fases de controle, estimulação, DA, após DA entre os dois grupos de animais (nutrido e desnutrido) o teste de Wilcoxon revelou que as médias dos expoentes α em cada fase para os animais nutridos diferem significativamente daquelas obtidas para os animais desnutridos. Na avalanche a hipótese de que o expoente α é igual a 0,5, não foi rejeitada pelo teste de Wilcoxon, ou seja, o teste confirmou a perda de correlação nessa fase. Na comparação entre as médias dos expoentes α nos diferentes intervalos (controle, estimulação, DA, após DA) com o valor do expoente α na avalanche, o teste de Wilcoxon acusou diferença significativa tanto no grupo dos nutridos como no grupo dos desnutridos.

Submitted by Fabio Sobreira Campos da Costa (fabio.sobreira@ufpe.br) on 2016-12-01T12:26:58Z No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Tese Mariana Barros.pdf: 1911747 bytes, checksum: f5c0832d347da9436db27522938984b9 (MD5) / Made available in DSpace on 2016-12-01T12:26:59Z (GMT). No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Tese Mariana Barros.pdf: 1911747 bytes, checksum: f5c0832d347da9436db27522938984b9 (MD5) Previous issue date: 2016-02-26 / CAPES / A nutrição no início da vida e o exercício físico podem alterar a excitabilidade cerebral, interferindo em processos de desenvolvimento e em parâmetros eletrofisiológicos. A depressão alastrante cortical (DAC) é um fenômeno relacionado com a excitabilidade cerebral. Em estudos anteriores, demonstramos potenciação da atividade elétrica cortical espontânea (ECoG) após a DAC. Nesta tese investigou-se a influência das condições de lactação e do exercício de natação, no início da vida e na idade adulta, sobre essa potenciação. Ratos Wistar machos foram amamentados em ninhadas com 6 ou 12 filhotes (grupos L6 e L12). A natação, precoce e tardia, foi realizada entre 8-23 e 60-75 dias de vida, respectivamente. O ECoG foi registrado aos 90-120 dias. Com um algoritmo específico (software Matlab™), determinou-se a amplitude do ECoG. Diferenças intergrupos de peso corporal e encefálico (L12<L6) confirmaram a deficiência nutricional do grupo L12. Diferenças, no mesmo animal, entre as amplitudes do ECoG antes e depois da DAC confirmaram a potenciação pós-DAC. Um grupo que não sofreu DAC não apresentou potenciação do ECoG. A natação precoce reduziu, e a tardia aumentou, essa potenciação do ECoG. A deficiência nutricional (condição L12) atenuou esse efeito. Estes dados sugerem que “condição de lactação”, “exercício de natação” e “idade ao exercício” modulam a potenciação do ECoG. A relevância desses resultados na fisiopatologia de doenças neurológicas dependentes da excitabilidade cerebral, como enxaqueca com aura e epilepsia, merecem futura investigação. / Nutrition in early life and physical exercise can alter brain excitability, interfering with development processes and electrophysiological parameters. Cortical spreading depression (CSD) is a phenomenon related to brain excitability. In previous studies, we demonstrated spontaneous brain electrical activity (ECoG) potentiation after CSD. In this thesis investigated the influence of the lactation conditions and swimming exercise in early life and adulthood, on this potentiation. Male Wistar rats were suckled in litters with 6 or 12 pups (L6 and L12 groups). The swimming, early and late, was carried out between 8-23 and 60-75 days of age, respectively. The ECoG was recorded at 90-120 days. With a specific algorithm (software Matlab ™), we determined the amplitude of the ECoG. Intergroup differences in body- and brain- weight (L12 <L6) confirmed the nutritional deficiency of the L12 group. Differences, in the same animal, between ECoG amplitudes before and after the CSD confirmed the post-CSD potentiation. A group that did not suffer CSD displayed no ECoG potentiation. Early swimming reduced, and later increased, this ECoG potentiation. The nutritional deficiency (L12 condition) attenuated this effect. These data suggest that "lactation condition", "swimming exercise" and "age to exercise" modulate ECoG potentiation. The relevance of these findings in the pathophysiology of brain excitability-dependent neurological diseases, such as migraine with aura and epilepsy, deserve further investigation.

No / Cortical spreading depression (CSD) is a transient disruption of local ionic homeostasis that may promote migraine attacks and the progression of stroke lesions. We reported previously that the local inhibition of nitric oxide (NO) synthesis with N¿-nitro-L-arginine methyl ester (L-NAME) delayed markedly the initiation of the recovery of ionic homeostasis from CSD. Here we describe a novel method for selective, controlled generation of exogenous NO in a functioning brain region. It is based on microdialysis perfusion of the NO donor, 2-(N,N-diethylamino)-diazenolate-2-oxide (DEA/NO). As DEA/NO does not generate NO at alkaline pH, and as the brain has a strong acid-base buffering capacity, DEA/NO was perfused in a medium adjusted at alkaline (but unbuffered) pH. Without DEA/NO, such a microdialysis perfusion medium did not alter CSD. DEA/NO (1, 10 and 100 ¿M) had little effect on CSD by itself, but it reversed in a concentration-dependent manner the effects of NOS inhibition by 1 mM L-NAME. These data demonstrate that increased formation of endogenous NO associated with CSD is critical for subsequent, rapid recovery of cellular ionic homeostasis. In this case, the molecular targets for NO may be located either on brain cells to suppress mechanisms directly involved in CSD genesis, or on local blood vessels to couple flow to the increased energy demand associated with CSD

No description available.

In der Pathogenese der Temporallappenepilepsie wird kindlichen hippokampalen Schädigungen eine wesentliche Rolle zugeschrieben. Epileptische Krämpfe und perinatale Asphyxie sind zwei häufige Ursachen dieser Schädigungen. Anhaltende epileptiforme Aktivität im Niedrig-Mg2+-Modell als einer experimentellen Form epileptischer Krämpfe führt in organotypischen hippokampalen Schnittkulturen (OHSK) der Ratte, die als Ersatzsystem des kindlichen Hippokampus verwendet werden, zu Zellschäden. Während dieser Untersuchungen ergab sich der Verdacht auf eine zusätzlich schädigende Wirkung erhöhter Sauerstoffspannungen. In meiner ersten Versuchsreihe konnte ich nachweisen, dass erhöhte Sauerstoffspannungen (60 %, 95 %) verglichen mit 20%-Sauerstoffspannung zu reversiblen und irreversiblen Zellschäden in OHSK führen. Die Zellschäden wurden über Veränderungen reizinduzierter Feldpotentiale, d.h. Abnahme der Amplitude, Zunahme der Latenz und Zunahme des Doppelpulsindex, sowie über die Propidium Jodid (PJ)-Fluoreszenzintensität bestimmt. In der zweiten Versuchsreihe konnte gezeigt werden, dass erhöhte Sauerstoffspannungen auch nach einer Hypoxie im Sinne einer hyperoxischen Reoxygenierung verglichen mit normoxischer Reoxygenierung vermehrt Zellschäden in OHSK zur Folge haben. In der dritten Versuchsreihe konnte ich ausschließen, dass erhöhte Sauerstoffspannungen eine notwendige Bedingung für Zellschäden infolge anhaltender epileptiformer Aktivität sind. Um die zellschädigende Rolle von Spreading Depressions (SDs), die während epileptiformer Aktivität auftreten, zu bestimmen, wurde in der vierten Versuchsreihe eine Methode etabliert, SD-ähnliche Ereignisse isoliert und zuverlässig in normoxischen OHSK auszulösen. Auf diese Weise wiederholt ausgelöste SD-ähnliche Ereignisse führten zu Zellschäden, bestimmt über die Veränderung elektrophysiologischer Eigenschaften von SD-ähnlichen Ereignissen, Abnahme der Feldpotentialamplitude und PJ-Fluoreszenzintensität. / Hippocampal damage during infancy is thought to play an important role in the pathogenesis of temporal lobe epilepsy. Epileptic seizures and perinatal asphyxia are two frequent causes of these damages. Sustained epileptiform activity induced in the low Mg2+-model of epileptic seizures leads to cell damage in organotypic hippocampal slice cultures (OHSC) of the rat, which are used as a surrogate for the infantile hippocampus. During a previous study utilising this model the suspicion arose that increased oxygen tension could have an additional damaging effect. My first series of experiments proved that increased oxygen tension (60 %, 95 %) lead to reversible and irreversible cell damage in OHSC compared to 20%-oxygen tension. Cell damage was determined by alterations of evoked field potentials, i.e. decrement of amplitude, increment of latency and paired pulse index, as well as by propidium iodide fluorescence. The second series of experiments showed that increased oxygen tension applied after an hypoxic period (hyperoxic reoxygenation) result in augmented cell damage compared to normoxic reoxygenation. With the third series of experiments it could be excluded that increased oxygen tension is an essential condition for the occurrence of cell damage due to sustained epileptiform activity. In order to elucidate the damaging role of spreading depressions (SD), which emerge during epileptiform activity, a method was established in the fourth series of experiments that allowed the reliable induction of SD-like events in normoxic OHSC. Repetitive SD-like events induced by this method led to cell damage, assessed by alterations of electrophysiological characteristics of SD-like events, decrement of evoked field potential amplitude and propidium iodide fluorescence.

Traumatic brain injury (TBI) is a leading cause of disability worldwide. Annually, 150 to 200/1,000,000 people become disabled as a result of brain trauma. Axonal degeneration is a critical, early event following TBI of all severities but whether axon degeneration is a driver of TBI remains unclear. Molecular pathways underlying the pathology of TBI have not been defined and there is no efficacious treatment for TBI. Despite this significant societal impact, surprisingly little is known about the molecular mechanisms that actively drive axon degeneration in any context and particularly following TBI. Although severe brain injury may cause immediate disruption of axons (primary axotomy), it is now recognized that the most frequent form of traumatic axonal injury (TAI) is mediated by a cascade of events that ultimately result in secondary axonal disconnection (secondary axotomy) within hours to days. Proposed mechanisms include immediate post-traumatic cytoskeletal destabilization as a direct result of mechanical breakage of microtubules, as well as catastrophic local calcium dysregulation resulting in microtubule depolymerization, impaired axonal transport, unmitigated accumulation of cargoes, local axonal swelling, and finally disconnection. The portion of the axon that is distal to the axotomy site remains initially morphologically intact. However, it undergoes sudden rapid fragmentation along its full distal length ~72 h after the original axotomy, a process termed Wallerian degeneration. Remarkably, mice mutant for the Wallerian degeneration slow (Wlds) protein exhibit ~tenfold (for 2–3 weeks) suppressed Wallerian degeneration. Yet, pharmacological replication of the Wlds mechanism has proven difficult. Further, no one has studied whether Wlds protects from TAI. Lastly, owing to Wlds presumed gain-of-function and its absence in wild-type animals, direct evidence in support of a putative endogenous axon death signaling pathway is lacking, which is critical to identify original treatment targets and the development of viable therapeutic approaches. Novel insight into the pathophysiology of Wallerian degeneration was gained by the discovery that mutant Drosophila flies lacking dSarm (sterile a/Armadillo/Toll-Interleukin receptor homology domain protein) cell-autonomously recapitulated the Wlds phenotype. The pro-degenerative function of the dSarm gene (and its mouse homolog Sarm1) is widespread in mammals as shown by in vitro protection of superior cervical ganglion, dorsal root ganglion, and cortical neuron axons, as well as remarkable in-vivo long-term survival (>2 weeks) of transected sciatic mouse Sarm1 null axons. Although the molecular mechanism of function remains to be clarified, its discovery provides direct evidence that Sarm1 is the first endogenous gene required for Wallerian degeneration, driving a highly conserved genetic axon death program. The central goals of this thesis were to determine (1) whether post-traumatic axonal integrity is preserved in mice lacking Sarm1, and (2) whether loss of Sarm1 is associated with improved functional outcome after TBI. I show that mice lacking the mouse Toll receptor adaptor Sarm1 gene demonstrate multiple improved TBI-associated phenotypes after injury in a closed-head mild TBI model. Sarm1-/- mice developed fewer beta amyloid precursor protein (βAPP) aggregates in axons of the corpus callosum after TBI as compared to Sarm1+/+ mice. Furthermore, mice lacking Sarm1 had reduced plasma concentrations of the phosphorylated axonal neurofilament subunit H, indicating that axonal integrity is maintained after TBI. Strikingly, whereas wild type mice exhibited a number of behavioral deficits after TBI, I observed a strong, early preservation of neurological function in Sarm1-/- animals. Finally, using in vivo proton magnetic resonance spectroscopy, I found tissue signatures consistent with substantially preserved neuronal energy metabolism in Sarm1-/- mice compared to controls immediately following TBI. My results indicate that the Sarm1-mediated prodegenerative pathway promotes pathogenesis in TBI and suggest that anti-Sarm1 therapeutics are a viable approach for preserving neurological function after TBI.