| Autor: |
Gratacap RL; Department of Molecular and Biomedical Sciences, University of Maine., Bergeron AC; Department of Molecular and Biomedical Sciences, University of Maine., Wheeler RT; Department of Molecular and Biomedical Sciences, University of Maine; Graduate School of Biomedical Sciences and Engineering, University of Maine; robert.wheeler@umit.maine.edu. |
| Jazyk: |
angličtina |
| Zdroj: |
Journal of visualized experiments : JoVE [J Vis Exp] 2014 Nov 27 (93), pp. e52182. Date of Electronic Publication: 2014 Nov 27. |
| DOI: |
10.3791/52182 |
| Abstrakt: |
Early defense against mucosal pathogens consists of both an epithelial barrier and innate immune cells. The immunocompetency of both, and their intercommunication, are paramount for the protection against infections. The interactions of epithelial and innate immune cells with a pathogen are best investigated in vivo, where complex behavior unfolds over time and space. However, existing models do not allow for easy spatio-temporal imaging of the battle with pathogens at the mucosal level. The model developed here creates a mucosal infection by direct injection of the fungal pathogen, Candida albicans, into the swimbladder of juvenile zebrafish. The resulting infection enables high-resolution imaging of epithelial and innate immune cell behavior throughout the development of mucosal disease. The versatility of this method allows for interrogation of the host to probe the detailed sequence of immune events leading to phagocyte recruitment and to examine the roles of particular cell types and molecular pathways in protection. In addition, the behavior of the pathogen as a function of immune attack can be imaged simultaneously by using fluorescent protein-expressing C. albicans. Increased spatial resolution of the host-pathogen interaction is also possible using the described rapid swimbladder dissection technique. The mucosal infection model described here is straightforward and highly reproducible, making it a valuable tool for the study of mucosal candidiasis. This system may also be broadly translatable to other mucosal pathogens such as mycobacterial, bacterial or viral microbes that normally infect through epithelial surfaces. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
|
