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In vivo two-photon imaging of motoneurons and adjacent glia in the ventral spinal cord.
J Neurosci Methods. 2018 04 01; 299:8-15.JN

Abstract

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 μm in the grey matter ventral horn of the mouse spinal cord.

Authors+Show Affiliations

Molecular Physiology, Center for Integrative Physiology and Molecular Medicine (CIPMM), University of Saarland, Building 48, 66421 Homburg, Germany; Laboratory of Nerve Regeneration, State University of Campinas - UNICAMP, Cidade Universitária "Zeferino Vaz", Rua Monteiro Lobato, 255, 13083970 - Campinas-SP, Brazil.Molecular Physiology, Center for Integrative Physiology and Molecular Medicine (CIPMM), University of Saarland, Building 48, 66421 Homburg, Germany.Molecular Physiology, Center for Integrative Physiology and Molecular Medicine (CIPMM), University of Saarland, Building 48, 66421 Homburg, Germany.Molecular Physiology, Center for Integrative Physiology and Molecular Medicine (CIPMM), University of Saarland, Building 48, 66421 Homburg, Germany.Laboratory of Nerve Regeneration, State University of Campinas - UNICAMP, Cidade Universitária "Zeferino Vaz", Rua Monteiro Lobato, 255, 13083970 - Campinas-SP, Brazil. Electronic address: alroliv@unicamp.br.Molecular Physiology, Center for Integrative Physiology and Molecular Medicine (CIPMM), University of Saarland, Building 48, 66421 Homburg, Germany. Electronic address: frank.kirchhoff@uks.eu.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

29408351

Citation

Cartarozzi, Luciana Politti, et al. "In Vivo Two-photon Imaging of Motoneurons and Adjacent Glia in the Ventral Spinal Cord." Journal of Neuroscience Methods, vol. 299, 2018, pp. 8-15.
Cartarozzi LP, Rieder P, Bai X, et al. In vivo two-photon imaging of motoneurons and adjacent glia in the ventral spinal cord. J Neurosci Methods. 2018;299:8-15.
Cartarozzi, L. P., Rieder, P., Bai, X., Scheller, A., Oliveira, A. L. R., & Kirchhoff, F. (2018). In vivo two-photon imaging of motoneurons and adjacent glia in the ventral spinal cord. Journal of Neuroscience Methods, 299, 8-15. https://doi.org/10.1016/j.jneumeth.2018.01.005
Cartarozzi LP, et al. In Vivo Two-photon Imaging of Motoneurons and Adjacent Glia in the Ventral Spinal Cord. J Neurosci Methods. 2018 04 1;299:8-15. PubMed PMID: 29408351.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - In vivo two-photon imaging of motoneurons and adjacent glia in the ventral spinal cord. AU - Cartarozzi,Luciana Politti, AU - Rieder,Phillip, AU - Bai,Xianshu, AU - Scheller,Anja, AU - Oliveira,Alexandre Leite Rodrigues de, AU - Kirchhoff,Frank, Y1 - 2018/02/03/ PY - 2016/11/24/received PY - 2018/01/11/revised PY - 2018/01/28/accepted PY - 2018/2/7/pubmed PY - 2019/9/5/medline PY - 2018/2/7/entrez KW - Glial cells KW - In vivo two-photon imaging KW - Motoneurons KW - Ventral spinal cord SP - 8 EP - 15 JF - Journal of neuroscience methods JO - J. Neurosci. Methods VL - 299 N2 - 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 μm in the grey matter ventral horn of the mouse spinal cord. SN - 1872-678X UR - https://www.unboundmedicine.com/medline/citation/29408351/In_vivo_two-photon_imaging_of_motoneurons_and_adjacent_glia_in_the_ventral_spinal_cord L2 - https://linkinghub.elsevier.com/retrieve/pii/S0165-0270(18)30019-0 DB - PRIME DP - Unbound Medicine ER -