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Molecular basis of differentiation of stem and neural progenitor cells


The nervous system is formed by network formed by trillions of neuron with different phenotypes and other supporting cell types. The immense phenotypic variety of cells in the CNS is due to the differential development of stem and progenitor cells which give origin to neurons, astrocytes and oligodendrocytes. This process of neurogenesis is directed by the activation of innumerous receptors on cell surfaces of differentiating cells. We have shown in previous studies the participation of kinin-B2, purinergic and cholinergic receptors in neuronal differentiation using P19 embryonic carcinoma cells as an in vitro model. Cholinergic receptor expression was altered when cells were differentiated in the presence of inhibitors of purinergic or kinin-B2 receptors. Based on these results, we are proposing to study the participation of these receptors in the neural differentiation of mouse embryonic and mesenchymaI stem cells as well as in the process of maturation of rat progenitor cells (neurospheres). DNA aptamers will be identified using the SELEX technique from a combinatorial DNA library which is capable to specifically recognize stem cells in order to purify these cells from contaminating ones. Gene expression and receptor activities of kinin-B2, purinergic and nicotinic receptor activity during differentiation of stem and progenitor cells will be determined. The fates of neuronal differentiation will be evaluated in the presence of the specific inhibitor of kinin-B2 receptors. RNA aptamers and RNA interference will be used for subtype-specific inhibition of purinergic P2Y1, P2X2,4,7 receptor activity or gene expression, respectively. This study will contribute to the functional analysis of stem cells differentiated in vitro, and furthermore, verify the participation of kinin-B2 and purinergic receptors in directing neuronal differentiation. Furthermore, the capability of stem and progenitor cells to give origin to regenerative processes in the peripheral nervous system will be evaluated. (AU)

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Scientific publications (16)
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
GLASER, TALITA; CAPPELLARI, ANGELICA REGINA; PILLAT, MICHELI MAINARDI; ISER, ISABELE CRISTIANA; WINK, MARCIA ROSANGELA; OLIVEIRA BATTASTINI, ANA MARIA; ULRICH, HENNING. Perspectives of purinergic signaling in stem cell differentiation and tissue regeneration. PURINERGIC SIGNALLING, v. 8, n. 3, SI, p. 523-537, . (06/61285-9)
OLIVEIRA, SOPHIA L. B.; PILLAT, MICHELI M.; CHEFFER, ARQUIMEDES; LAMEU, CLAUDIANA; SCHWINDT, TELMA T.; ULRICH, HENNING. Functions of Neurotrophins and Growth Factors in Neurogenesis and Brain Repair. Cytometry Part A, v. 83A, n. 1, SI, p. 76-89, . (06/61285-9)
TRUJILLO, CLEBER A.; NEGRAES, PRISCILLA D.; SCHWINDT, TELMA T.; LAMEU, CLAUDIANA; CARROMEU, CASSIANO; MUOTRI, ALYSSON R.; PESQUERO, JOAO B.; CERQUEIRA, DEBORA M.; PILLAT, MICHELI M.; DE SOUZA, HELLIO D. N.; et al. Kinin-B2 Receptor Activity Determines the Differentiation Fate of Neural Stem Cells. Journal of Biological Chemistry, v. 287, n. 53, p. 44046-44061, . (06/61285-9)
MARTINS, ANTONIO H.; ALVES, JANAINA M.; PEREZ, DINELY; CARRASCO, MARIMEE; TORRES-RIVERA, WILMARIE; ETEROVIC, VESNA A.; FERCHMIN, PEDRO A.; ULRICH, HENNING. Kinin-B2 Receptor Mediated Neuroprotection after NMDA Excitotoxicity Is Reversed in the Presence of Kinin-B1 Receptor Agonists. PLoS One, v. 7, n. 2, . (06/61285-9)
FERCHMIN, P. A.; PAGAN, ONE R.; ULRICH, HENNING; SZETO, ADA C.; HANN, RICHARD M.; ETEROVIC, VESNA A.. Actions of octocoral and tobacco cembranoids on nicotinic receptors. Toxicon, v. 54, n. 8, SI, p. 1174-1182, . (06/61285-9)
NERY, ARTHUR A.; RESENDE, RODRIGO R.; MARTINS, ANTONIO H.; TRUJILLO, CLEBER A.; ETEROVIC, VESNA A.; ULRICH, HENNING. Alpha7 Nicotinic Acetylcholine Receptor Expression and Activity During Neuronal Differentiation of PC12 Pheochromocytoma Cells. JOURNAL OF MOLECULAR NEUROSCIENCE, v. 41, n. 3, p. 329-339, . (06/61285-9)
MARTINS, ANTONIO H.; ALVES, JANAÍNA M.; TRUJILLO, CLEBER A.; SCHWINDT, TELMA T.; BARNABÉ, GABRIELA F.; MOTTA, FABIANA L. T.; GUIMARÃES, ALESSANDER O.; CASARINI, DULCE E.; MELLO, LUIZ E.; PESQUERO, JOÃO B.; et al. Kinin-B2 receptor expression and activity during differentiation of embryonic rat neurospheres. Cytometry Part A, v. 73, n. 4, p. 361-368, . (06/61285-9)
NEGRAES, PRISCILLA D.; LAMEU, CLAUDIANA; HAYASHI, MIRIAN A. F.; MELO, ROBSON L.; CAMARGO, ANTONIO C. M.; ULRICH, HENNING. The Snake Venom Peptide Bj-PRO-7a Is a M1 Muscarinic Acetylcholine Receptor Agonist. Cytometry Part A, v. 79A, n. 1, p. 77-83, . (06/61285-9)
SMUC, TINA; AHN, IL-YOUNG; ULRICH, HENNING. Nucleic acid aptamers as high affinity ligands in biotechnology and biosensorics. Journal of Pharmaceutical and Biomedical Analysis, v. 81-82, p. 210-217, . (12/50393-6, 06/61285-9, 12/50880-4)
NERY, ARTHUR A.; MAGDESIAN, MARGARET H.; TRUJILLO, CLEBER A.; SATHLER, LUCIANA B.; JULIANO, MARIA A.; JULIANO, LUIZ; ULRICH, HENNING; FERREIRA, SERGIO T.. Rescue of Amyloid-Beta-Induced Inhibition of Nicotinic Acetylcholine Receptors by a Peptide Homologous to the Nicotine Binding Domain of the Alpha 7 Subtype. PLoS One, v. 8, n. 7, . (06/61285-9, 12/50880-4)
NERY, ARTHUR A.; NASCIMENTO, ISIS C.; GLASER, TALITA; BASSANEZE, VINICIUS; KRIEGER, JOSE E.; ULRICH, HENNING. Human Mesenchymal Stem Cells: From Immunophenotyping by Flow Cytometry to Clinical Applications. Cytometry Part A, v. 83A, n. 1, SI, p. 48-61, . (06/61285-9)
DONA, FLAVIA; CONCEICAO, ISALTINO MARCELO; ULRICH, HENNING; RIBEIRO, ELIANE BERALDI; FREITAS, THALMA ARIANI; ABRAHAO NENCIONI, ANA LEONOR; DA SILVA FERNANDES, MARIA JOSE. Variations of ATP and its metabolites in the hippocampus of rats subjected to pilocarpine-induced temporal lobe epilepsy. PURINERGIC SIGNALLING, v. 12, n. 2, p. 295-302, . (06/06502-4, 06/61285-9)
LAMEU, CLAUDIANA; TRUJILLO, CLEBER A.; SCHWINDT, TELMA T.; NEGRAES, PRISCILLA D.; PILLAT, MICHELI M.; MORAIS, KATIA L. P.; LEBRUN, IVO; ULRICH, HENNING. Interactions between the NO-Citrulline Cycle and Brain-derived Neurotrophic Factor in Differentiation of Neural Stem Cells. Journal of Biological Chemistry, v. 287, n. 35, p. 29690-29701, . (06/61285-9)
GLASER, TALITA; RESENDE, RODRIGO R.; ULRICH, HENNING. Implications of purinergic receptor-mediated intracellular calcium transients in neural differentiation. CELL COMMUNICATION AND SIGNALING, v. 11, . (06/61285-9, 12/50880-4)
MARCONI, MARCELO; SANCHEZ, RAUL; ULRICH, HENNING; ROMERO, FERNANDO. Potassium Current in Mature Bovine Spermatozoa. SYSTEMS BIOLOGY IN REPRODUCTIVE MEDICINE, v. 54, n. 6, p. 231-239, . (06/61285-9)
SARTORE, RAFAELA C.; CAMPOS, PRISCILA B.; TRUJILLO, CLEBER A.; RAMALHO, BIA L.; NEGRAES, PRISCILLA D.; PAULSEN, BRUNA S.; MELETTI, TAMARA; COSTA, ELAINE S.; CHICAYBAM, LEONARDO; BONAMINO, MARTIN H.; et al. Retinoic Acid-Treated Pluripotent Stem Cells Undergoing Neurogenesis Present Increased Aneuploidy and Micronuclei Formation. PLoS One, v. 6, n. 6, p. e20667, . (06/61285-9)

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Filed patent(s) as a result of this research project

POLINUCLEOTIDEOS QUIMICAMENTE MODIFICADOS E PROCESSO DE PRODUÇÃO DE POLINUCLEOTÍDEOS QUIMICAMENTE MODIFICADOS BR1020130217018 - Universidade de São Paulo (USP) . Alexander Henning Ulrich ; Vinicius Bassaneze ; Arthur Andrade Nery ; Jose Eduardo Krieger - August 2013, 26

CHEMICALLY MODIFIED POLYNUCLEOTIDES AND METHOD FOR PRODUCING CHEMICALLY MODIFIED POLYNUCLEOTIDES US14/915,170 - Universidade de São Paulo (USP) . José Eduardo Krieger ; Alexander Henning Ulrich ; Arthur Andrade Nery ; Vinícius Bassaneze - February 2016, 26

CHEMICALLY MODIFIED POLYNUCLEOTIDES AND METHOD FOR PRODUCING CHEMICALLY MODIFIED POLYNUCLEOTIDES EP20140839532 - Universidade de São Paulo (USP) . José Eduardo Krieger ; Alexander Henning Ulrich ; Arthur Andrade Nery ; Vinícius Bassaneze - August 2014, 25

CHEMICALLY MODIFIED POLYNUCLEOTIDES AND METHOD FOR PRODUCING CHEMICALLY MODIFIED POLYNUCLEOTIDES PCT/BR2014/000294 - Universidade de São Paulo (USP) . Alexander Henning Ulrich ; Arthur Andrade Nery ; José Eduardo Krieger ; Vinícius Bassaneze - August 2014, 25