Journal Article

PUBDB-2024-05617

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png Resolving exit strategies of mycobacteria in Dictyostelium discoideum by combining high‐pressure freezing with 3D ‐correlative light and electron microscopy

Franzkoch, R. ; Anand, A. ; Breitsprecher, L. ; Psathaki, O. E. (Corresponding author) ; Barisch, C. (Corresponding author)CSSB*

2024
Wiley-Blackwell Oxford [u.a.]

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Please use a persistent id in citations: doi:10.1111/mmi.15205  doi:10.3204/PUBDB-2024-05617

Abstract: The infection course of Mycobacterium tuberculosis is highly dynamic and comprises sequential stages that require damaging and crossing of several membranes to enable the translocation of the bacteria into the cytosol or their escape from the host. Many important breakthroughs such as the restriction of mycobacteria by the autophagy pathway and the recruitment of sophisticated host repair machineries to the Mycobacterium-containing vacuole have been gained in the Dictyostelium discoideum/M. marinum system. Despite the availability of well-established light and advanced electron microscopy techniques in this system, a correlative approach integrating both methods with near-native ultrastructural preservation is currently lacking. This is most likely due to the low ability of D. discoideum to adhere to surfaces, which results in cell loss even after fixation. To address this problem, we improved the adhesion of cells and developed a straightforward and convenient workflow for 3D-correlative light and electron microscopy. This approach includes high-pressure freezing, which is an excellent technique for preserving membranes. Thus, our method allows to monitor the ultrastructural aspects of vacuole escape which is of central importance for the survival and dissemination of bacterial pathogens.

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Contributing Institute(s):

1. CSSB-Forschungszentrum Borstel (CSSB-FZB-CB)

Research Program(s):

1. 899 - ohne Topic (POF4-899) (POF4-899)
2. DFG project G:(GEPRIS)531703706 - Entschlüsselung der molekularen Mechanismen des Austritts von Orientia tsutsugamushi aus der Wirtszelle (531703706) (531703706)
3. SFB 1557 P01 - Umbau und Nutzung der Lipid-Trafficking-Maschinerie der Wirtszelle durch pathogene Mykobakterien (P01) (516902060) (516902060)
4. SFB 944 P25 - Funktionale Bedeutung des Lipidtransports für die mykobakterielle Infektion in Dictyostelium (P25) (442222172) (442222172)

Experiment(s):

1. No specific instrument

Appears in the scientific report 2024

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Database coverage:
; ; ; BIOSIS Previews ; Biological Abstracts ; Clarivate Analytics Master Journal List ; Current Contents - Life Sciences ; DEAL Wiley ; Ebsco Academic Search ; Essential Science Indicators ; IF < 5 ; JCR ; Nationallizenz ; SCOPUS ; Science Citation Index Expanded ; Web of Science Core Collection

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