Navegando por Palavras-chave "GFP"

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    Cloning and expression of transgenes using linear vectors in Trypanosoma cruzi
    (Elsevier B.V., 2014-06-01) los Angeles Curto, Maria de; Lorenzi, Hernan A.; Moraes Barros, Roberto R. [UNIFESP]; Souza, Renata T. [UNIFESP]; Levin, Mariano J.; Franco da Silveira, José [UNIFESP]; Schijman, Alejandro G.; Inst Invest Ingn Genet & Biol Mol INGEBI CONICET; Universidade Federal de São Paulo (UNIFESP)
    The identification of new targets for vaccine and drug development for the treatment of Chagas' disease is dependent on deepening our understanding of the parasite genome. Vectors for genetic manipulation in Trypanosoma cruzi basically include those that remain as circular episomes and those that integrate into the parasite's genome. Artificial chromosomes are alternative vectors to overcome problematic transgene expression often occurring with conventional vectors in this parasite. We have constructed a series of vectors named pTACs (Trypanosome Artificial Chromosomes), all of them carrying telomeric and subtelomeric sequences and genes conferring resistance to different selection drugs. in addition, one pTAC harbours a modified GFP gene (pTAC-gfp), and another one carries the ornithine decarboxilase gene from Crithidia fasciculata (pTAC-odc). We have encountered artificial chromosomes generated from pTACs in transformed T. cruzi epimastigotes for every version of the designed vectors. These extragenomic elements, in approximately 6-8 copies per cell, remained as linear episomes, contained telomeres and persisted after 150 and 60 generations with or without selection drugs, respectively. the linear molecules remained stable through the different T. cruzi developmental forms. Furthermore, derived artificial chromosomes from pTAC-odc could complement the auxotrophy of T. cruzi for polyamines. Our results show that pTACs constitute useful tools for reverse functional genetics in T. cruzi that will contribute to a better understanding of T. cruzi biology. (C) 2014 Published by Elsevier B.V. on behalf of Australian Society for Parasitology Inc.
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    Distribution and proliferation of bone marrow cells in the brain after pilocarpine-induced status epilepticus in mice
    (Wiley-Blackwell, 2010-08-01) Longo, Beatriz [UNIFESP]; Romariz, Simone [UNIFESP]; Blanco, Miriam Marcela [UNIFESP]; Vasconcelos, Juliana Fraga; Bahia, Luciana; Pereira Soares, Milena Botelho; Mello, Luiz E. [UNIFESP]; Ribeiro-dos-Santos, Ricardo; Universidade Federal de São Paulo (UNIFESP); Hosp Sao Rafael
    P>The distribution of bone marrow cells in brain areas during the acute period after pilocarpine-induced status epilepticus (SE) was investigated here. To achieve this, we generated chimeric mice by engrafting bone marrow cells from enhanced green fluorescent protein (eGFP) transgenic mice. GFP+ bone marrow-derived cells were found throughout the brain, predominantly in the hippocampus. As expected, these cells exhibited the characteristics of microglia. the pattern of distribution, proliferation, and differentiation of GFP+ cells changes as a function of intensity and time following SE. This pattern is also a consequence of the inflammatory response, which is followed by the progressive neuronal damage that is characteristic of the pilocarpine model.
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    Participation of bone marrow-derived cells in hippocampal vascularization after status epilepticus
    (W B Saunders Co Ltd, 2014-05-01) Romariz, Simone A. [UNIFESP]; Garcia, Karina de O. [UNIFESP]; Paiva, Daisylea de Souza [UNIFESP]; Bittencourt, Simone [UNIFESP]; Covolan, Luciene [UNIFESP]; Mello, Luis Eugenio [UNIFESP]; Longo, Beatriz Monteiro [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)
    Purpose: Diseases such as temporal lobe epilepsy, brain trauma and stroke can induce endothelial cell proliferation and angiogenesis in specific brain areas. During status epilepticus (SE), bone marrow-derived cells are able to infiltrate and proliferate, dramatically increasing at the site of injury. However, it is still unclear whether these cells directly participate in vascular changes induced by SE.Method: To investigate the possible role of bone marrow-derived cells in angiogenesis after seizures, we induced SE by pilocarpine injection in previously prepared chimeric mice. Mice were euthanized at 8 h, 7 d or 15 d after SE onset.Results: Our results indicated that SE modified hippocampal vascularization and induced angiogenesis. Further, bone marrow-derived GFP(+) cells penetrated through the parenchyma and participated in the formation of new vessels after SE. We detected bone marrow-derived cells closely associated with vessels in the hippocampus, increasing the density of blood vessels that had decreased immediately after pilocarpine-induced SE.Conclusion: We conclude that epileptic seizures directly affect vascularization in the hippocampus mediated by bone marrow-derived cells in a time-dependent manner. (C) 2014 British Epilepsy Association. Published by Elsevier B.V. All rights reserved.