Developers
Laura Gó
Description of the technology
Although there are multiple methods for analyzing apoptosis in cultured cells (e.g.
Main element of this technology is formulated as a protocol, describing how to detect apoptosis of leukocytes in mouse lymph nodes via the detection of apoptotic caspases. This protocol have previously used to study factors that modulate dendritic cell survival in lymph nodes; however, it can also be used to analyze other leukocytes that migrate to the lymph nodes. Dendritic cells labeled with a fluorescent cell tracker are subcutaneously injected in the posterior footpads of mice. Once the labeled dendritic cells reach the popliteal lymph nodes, the animals are intravenously injected with fluorescent
Practical application
An important advantage protocol according to this technology is its applicability to study the survival of leucocytes and, especially, dendritic cells in vivo. This protocol can also be used to study the survival of other leukocytes that migrate to the lymph nodes. In this regard, the protocol can be used to study any cell type that expresses the chemokine receptor CCR7, which is responsible for driving leukocytes and other cells to the lymph nodes. The cell types that migrate to the lymph nodes include B cells, naive and central memory T cells and some types of monocytes and macrophages.
Notably, some metastatic cells, including cells of melanoma,
The method can also be used to analyze the survival of leukocytes or other cells types, not only in mice but also in other species, provided that the cells to be studied use
Thus, the technology can be promising for application in the study of different diseases (immune diseases, cancer, etc), processes of aging, including cell senescence, drug discovery and others.
Laboratories
- Centro de Investigaciones Biológicas. Consejo Superior de Investigaciones Científicas, Madrid (Spain)
- Centro de Microscopía y Citometría, Universidad Complutense, Madrid (Spain)
- Present address: Department of Microbiology and Immunobiology, Harvard Medical School, Boston (USA)
Links
http://www.nature.com/nprot/journal/v9/n5/full/nprot.2014.078.htmlPublications
- Gó
mez-Caba ñas, L. et al. «Detecting apoptosis of leukocytes in mouse lymph nodes." 9 Nature Protocols (2014): 1102–1112. Delgado-Martin , C. et al. «Chemokine CXCL12 uses CXCR4 and a signaling core formed by bifunctional Akt, extracellularsignal-regulated kinase (ERK)½, and mammalian target of rapamycin complex 1 (mTORC1) proteins to control chemotaxis and survival simultaneously in mature dendritic cells." 286 J. Biol. Chem. (2011): 37222–37236.Rey-Gallardo , A. et al. «Polysialic acid is required forneuropilin-2a /b-mediated control ofCCL21-driven chemotaxis of mature dendritic cells and for their migration in vivo." 21 Glycobiology (2011): 655–662.