| Texto completo | |
| Autor(es): |
Cartarozzi, Luciana Politti
[1, 2]
;
Rieder, Phillip
[1]
;
Bai, Xianshu
[1]
;
Scheller, Anja
[1]
;
Rodrigues de Oliveira, Alexandre Leite
[2]
;
Kirchhoff, Frank
[1]
Número total de Autores: 6
|
| Afiliação do(s) autor(es): | [1] Univ Saarland, CIPMM, Mol Physiol, Bldg 48, D-66421 Homburg - Germany
[2] Univ Estadual Campinas, UNICAMP, Lab Nerve Regenerat, Cidade Univ Zeferino Vaz, Rua Monteiro Lobato 255, BR-13083970 Campinas, SP - Brazil
Número total de Afiliações: 2
|
| Tipo de documento: | Artigo Científico |
| Fonte: | JOURNAL OF NEUROSCIENCE METHODS; v. 299, p. 8-15, APR 1 2018. |
| Citações Web of Science: | 6 |
| Resumo | |
Background: Interactions between motoneurons and glial cells are pivotal to regulate and maintain functional states and synaptic connectivity in the spinal cord. In vivo two-photon imaging of the nervous system provided novel and unexpected knowledge about structural and physiological changes in the grey matter of the forebrain and in the dorsal white matter of the spinal cord. New method: Here, we describe a novel experimental strategy to investigate the spinal grey matter, i.e. the ventral horn motoneurons and their adjacent glial cells by employing in vivo two-photon laser-scanning microscopy (2P-LSM) in anesthetized transgenic mice. Results: After retrograde tracer labelling in transgenic mice with cell-specific expression of fluorescent proteins and surgical exposure of the lumbar intumescence groups of motoneurons could be visualized deeply localized in the ventral horn. In this region, morphological responses of microglial cells to ATP could be recorded for an hour. In addition, using in mice with expression of GCaMP3 in astrocytes, physiological Ca2+ signals could be recorded after local noradrenalin application. Comparison with existing methods: Previous in vivo imaging protocols were restricted to the superficial dorsal white matter or upper layers of the dorsal horn. Here, we modified a multi-step procedure originally established for a root-crush injury. We adapted it to simultaneously visualize motoneurons and adjacent glial cells in living animals. Conclusion: A modified surgery approach is presented to visualize fluorescently labelled motoneurons and glial cells at a depth of more than 200 mu m in the grey matter ventral horn of the mouse spinal cord. (C) 2018 Elsevier B.V. All rights reserved. (AU) | |
| Processo FAPESP: | 15/08297-8 - Análise in vivo da reatividade glial dependente de MHC Classe I após lesão do sistema nervoso |
| Beneficiário: | Luciana Politti Cartarozzi |
| Modalidade de apoio: | Bolsas no Exterior - Estágio de Pesquisa - Doutorado |
| Processo FAPESP: | 13/16134-6 - Estudo dos mecanismos imunomoduladores exercidos pelas células tronco mesenquimais sobre a reatividade das células gliais e correlação com a capacidade regenerativa após axotomia de raízes lombares |
| Beneficiário: | Luciana Politti Cartarozzi |
| Modalidade de apoio: | Bolsas no Brasil - Doutorado |
| Processo FAPESP: | 14/06892-3 - Utilização de células tronco mesenquimais na interface do sistema nervoso central e periférico: reparo de lesões proximais |
| Beneficiário: | Alexandre Leite Rodrigues de Oliveira |
| Modalidade de apoio: | Auxílio à Pesquisa - Temático |