Directed Differentiation of Neural Progenitors into Neurons Is Accompanied by Altered Expression of P2X Purinergic Receptors

Neural differentiation has been extensively studied in vitro in a model termed neurospheres, which consists of aggregates of neural progenitor cells. Previous studies suggest that they have a great potential for the treatment of neurological disorders. One of the major challenges for scientists is to control cell fate and develop ideal culture conditions for neurosphere expansion in vitro, without altering their features. Similar to human neural progenitors, rat neurospheres cultured in the absence of epidermal and fibroblast growth factors for a short period increased the levels of beta-3 tubulin and decreased the expression of glial fibrillary acidic protein and nestin, compared to neurospheres cultured in the presence of these factors. In this work, we show that rat neurospheres cultured in suspension under mitogen-free condition presented significant higher expression of P2X2 and P2X6 receptor subunits, when compared to cells cultured in the presence of growth factors, suggesting a direct relationship between P2X2/6 receptor expression and induction of neuronal differentiation in mitogen-free cultured rat neurospheres.

Comparação e validação de vários métodos de mapeamento cortical para o planeamento pré-cirúrgico por Imagem Funcional de Ressonância Magnética

Na análise funcional de imagens do cérebro podem utilizar-se diferentes métodos na identificação de zonas de activação. Tem havido uma evolução desde o método de correlação [19], para outros métodos [9] [14] até o método baseado no modelo linear generalizado que é mais comum ser utilizado hoje e que levou ao pacote de software SPM [15]. Deve-se principalmente à versatilidade que o método tem em realizar testes com diferentes objectivos. Têm sido publicados alguns estudos comparativos. Poucos têm sido quantitativos [20] e quando o são, o número de métodos testados é reduzido[22]. Há muitos estudos comparativos do ponto de vista da estatística envolvida (da matemática) mas que têm em geral apenas ns académicos. Um objectivo deste estudo é comparar os resultados obtidos por diferentes métodos. É de particular interesse averiguar o comportamento de cada método na fronteira do local de activação. As diferenças serão avaliadas numericamente para os seguintes métodos clássicos: t de Student, coeficiente de correlação e o modelo linear generalizado. Três novos métodos são também propostos - o método de picos de Fourier, o método de sobreposição e o método de amplitude. O segundo pode ser aplicado para o melhoramento dos métodos de t de Student, coe ciente de correlação e modelo linear generalizado. Ele pode no entanto, também manter-se como um método de análise independente. A influência exercida em cada método pelos parâmetros pertinentes é também medida. É adoptado um conjunto de dados clínicos que está amplamente estudado e documentado. Desta forma elimina-se a possibilidade dos resultados obtidos serem interpretados como sendo específicos do caso em estudo. Há situações em que a influência do método utilizado na identificação das áreas de activação de imagens funcionais do cérebro é crucial. Tal acontece, por exemplo, quando um tumor desenvolve-se perto de uma zona de activação responsável por uma função importante . Para o cirurgião tornase indispensável avaliar se existe alguma sobreposição. A escolha de um dos métodos disponíveis poderá ter infuência sobre a decisão final. Se o método escolhido for mais conservador, pode verificar-se sobreposição e eliminar-se a possibilidade de cirurgia. Porém, se o método for mais restritivo a decisão final pode ser favorável à cirurgia. Artigos recentes têm suportado a ideia de que a ressonância magnética funcional é de facto muito útil no processo de decisão pré-operatório [12].O segundo objectivo do estudo é então avaliar a sobreposição entre um volume de activação e o volume do tumor. Os programas informáticos de análise funcional disponíveis são variados em vários aspectos: na plataforma em que funcionam (macintosh, linux, windows ou outras), na linguagem em que foram desenvolvidos (e.g. c+motif, c+matlab, matlab, etc.) no tratamento inicial dos dados (antes da aplicação do método de análise), no formato das imagens e no(s) método(s) de análise escolhido(s). Este facto di culta qualquer tentativa de comparação. À partida esta poderá apenas ser qualitativa. Uma comparação quantitativa implicaria a necessidade de ocorrerem três factos: o utilizador tem acesso ao código do programa, sabe programar nas diferentes linguagens e tem licença de utilização de software comercial (e.g. matlab). Sendo assim foi decidido adoptar uma estratégia unificadora. Ou seja, criar um novo programa desenvolvido numa linguagem independente da plataforma, que não utilize software comercial e que permita aplicar (e comparar quantitativamente) diferentes métodos de análise funcional. A linguagem escolhida foi o JAVA. O programa desenvolvido no âmbito desta tese chama-se Cérebro.

Sonic hedgehog signaling regulates mode of cell division of early cerebral cortex progenitors and increases

The morphogen Sonic Hedgehog (SHH) plays a critical role in the development of different tissues. In the central nervous system, SHH is well known to contribute to the patterning of the spinal cord and separation of the brain hemispheres. In addition, it has recently been shown that SHH signaling also contributes to the patterning of the telencephalon and establishment of adult neurogenic niches. In this work, we investigated whether SHH signaling influences the behavior of neural progenitors isolated from the dorsal telencephalon, which generate excitatory neurons and macroglial cells in vitro. We observed that SHH increases proliferation of cortical progenitors and generation of astrocytes, whereas blocking SHH signaling with cyclopamine has opposite effects. In both cases, generation of neurons did not seem to be affected. However, cell survival was broadly affected by blockade of SHH signaling. SHH effects were related to three different cell phenomena: mode of cell division, cell cycle length and cell growth. Together, our data in vitro demonstrate that SHH signaling controls cell behaviors that are important for proliferation of cerebral cortex progenitors, as well as differentiation and survival of neurons and astroglial cells.

Increased hippocampal excitability and impaired spatial memory function in mice lacking VGLUT2 selectively in neurons defined by tyrosine hydroxylase promoter activity

Three populations of neurons expressing the vesicular glutamate transporter 2 (Vglut2) were recently described in the A10 area of the mouse midbrain, of which two populations were shown to express the gene encoding, the rate-limiting enzyme for catecholamine synthesis, tyrosine hydroxylase (TH).One of these populations (‘‘TH– Vglut2 Class1’’) also expressed the dopamine transporter (DAT) gene while one did not ("TH–Vglut2 Class2"), and the remaining population did not express TH at all ("TH-Vglut2-only"). TH is known to be expressed by a promoter which shows two phases of activation, a transient one early during embryonal development, and a later one which gives rise to stable endogenous expression of the TH gene. The transient phase is, however, not specific to catecholaminergic neurons, a feature taken to advantage here as it enabled Vglut2 gene targeting within all three A10 populations expressing this gene, thus creating a new conditional knockout. These knockout mice showed impairment in spatial memory function. Electrophysiological analyses revealed a profound alteration of oscillatory activity in the CA3 region of the hippocampus. In addition to identifying a novel role for Vglut2 in hippocampus function, this study points to the need for improved genetic tools for targeting of the diversity of subpopulations of the A10 area

Effects of Hormonal Supplementation on Nuclear Maturation and Cortical Granules Distribution of Canine Oocytes During Various Reproductive Stages

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Role of Locus coeruleus noradrenergic neurons in cardiorespiratory and thermal control during hypoxia

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Serotonergic neurons in the nucleus raphe obscurus contribute to interaction between central and peripheral ventilatory responses to hypercapnia

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Brain monoaminergic neurons and ventilatory control in vertebrates

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Role of neurokinin-1 expressing neurons in the locus coeruleus on ventilatory and cardiovascular responses to hypercapnia

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Mapping of serotonin-immunoreactive neurons of Anastrepha obliqua Macquart larvae

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

CONTROL of THE CENTRAL CHEMOREFLEX BY A5 NORADRENERGIC NEURONS IN RATS

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Serotonergic neurons in the median raphe nucleus regulate inhibitory avoidance but not escape behavior in the rat elevated T-maze test of anxiety

Rationale: A wealth of evidence supports the involvement of the serotonergic neurons of the median raphe nucleus (MRN) in anxiety. However, it is presently unclear whether serotonergic pathways arising from this nucleus play distinguishing regulatory roles in defensive behaviors that have been associated with specific subtypes of anxiety disorders. Objectives: To evaluate the role of the MRN serotonergic neurons in the regulation of two defensive behaviors, inhibitory avoidance and escape, which have been related, respectively, to generalized anxiety and panic disorders. Methods: Male Wistar rats were submitted to the elevated T-maze test of anxiety after intra-MRN administration of drugs that either non-selectively or selectively change the activity of the serotonergic neurons. Results: Intra-MRN injection of FG 7142 (0.04 and 0.08 nmol) and kainic acid (0.03 and 0.06 nmol), drugs that non-selectively stimulate the MRN serotonergic neurons, facilitated inhibitory avoidance acquisition, but impaired escape performance. Microinjection of muscimol (0.11 and 0.22 nmol), a compound that non-selectively inhibits the activity of the MRN serotonergic neurons, impaired inhibitory avoidance and facilitated escape performance. Both kainic acid and muscimol also changed rat locomotion in the open-field test. Intra-MRN injection of 8-OH-DPAT (0.6-15 nmol) and WAY-100635 (0.18-0.74 nmol), respectively an agonist and an antagonist of somatodendritic 5-HT1A receptors located on serotonergic neurons of the MRN, only affected inhibitory avoidance-while the former inhibited the acquisition of this behavior, the latter facilitated it. Conclusion: MRN serotonergic neurons seem to be selectively involved in the regulation of inhibitory avoidance in the elevated T-maze. This result supports the proposal that 5-HT pathways departing from this nucleus play an important role in anxiety processing, with implications for pathologies such as generalized anxiety disorder.

5-HT1A receptors in the dorsal hippocampus mediate the anxiogenic effect induced by the stimulation of 5-HT neurons in the median raphe nucleus

We evaluated the involvement of dorsal hippocampus (DH) 5-HT1A receptors in the mediation of the behavioral effects caused by the pharmacological manipulation of 5-HT neurons in the median raphe nucleus (MRN). To this end, we used the rat elevated T-maze test of anxiety. The results showed that intra-DH injection of the 5-HT1A/7 agonist 8-OH-DPAT facilitated inhibitory avoidance, an anxiogenic effect, without affecting escape. Microinjection of the 5-HT1A antagonist WAY-100635 was ineffective. In the elevated T-maze, inhibitory avoidance and escape have been related to generalized anxiety and panic disorders, respectively. Intra-MRN administration of the excitatory aminoacid kainic acid, which non-selectively stimulates 5-HT neurons in this brain area facilitated inhibitory avoidance and impaired escape performance, but also affected locomotion. Intra-MRN injection of WAY-100635, which has a disinhibitory effect on the activity of 5-HT neurons in this midbrain area, only facilitated inhibitory avoidance. Preadministration of WAY-100635 into the DH blocked the behavioral effect of intra-MRN injection of WAY-100635, but not of kainic acid. These results indicate that DH 5-HT1A receptors mediate the anxiogenic effect induced by the selective stimulation of 5-HT neurons in the MRN. (c) 2007 Elsevier B.V. and ECNP. All rights reserved.